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BIOENERGY CROPS AND BIOREMEDIATION BIOENERGY CROPS AND BIOREMEDIATION - A REVIEW A Contract Report by ADAS for the Department for Food, Environment and Rural Affairs Final Report AUGUST 2002 BIOENERGY CROPS AND BIOREMEDIATION CONTENTS AUTHOR(S) PAGE EXECUTIVE SUMMARY 1. INTRODUCTION 1 1.1 BACKGROUND 1 1.2 THE ENERGY CROPS IN QUESTION 2 1.2.1 1.2.2 2 3 1.3 1.4 SRC willow and poplar Miscanthus 4 4 4 5 THE ISSUES 1.3.1 1.3.2 1.3.3 2. M Bullard & P Nixon Land availability Waste availability The role of energy crops 6 REFERENCES WASTE UTILISATION: ENVIRONMENTAL & CROP EFFECTS 2.1 NITROGEN & PHOSPHORUS – LOSSES TO WATER 2.1.1 2.1.2 2.1.3 2.1.4 2.1.5 2.2 2.2.3 P Johnson Introduction Nutrient requirements of biomass crops Use of organic manures or wastes Landfill leachate & other urban aqueous wastes References GASEOUS LOSSES 2.2.1 2.2.2 7 Gaseous losses of nutrients Emissions of radiatively active gases from soils & organic waste materials References 7 7 7 11 14 16 J King 19 19 21 37 BIOENERGY CROPS AND BIOREMEDIATION 2.3 HEAVY METALS & OTHER TOXINS 2.3.1 2.3.2 2.3.3 2.3.4 2.3.5 2.3.6 2.4 2.4.3 2.4.4 3. F Nicholson 42 42 43 47 51 52 52 C Britt Introduction Agricultural and municipal wastes Industrial wastes References BIOREMEDIATION OF CONTAMINATED SITES 56 56 60 66 68 P Nixon 73 3.1 INTRODUCTION 73 3.2 PHYTOREMEDIATION 73 3.3 POTENTIAL ENVIRONMENTAL PROBLEMS 78 3.3.1 3.3.2 78 3.4 4. PAGE Introduction Concentrations of contaminants in livestock manures & other wastes Contaminant uptake/removal by biomass crops Other pathways of contaminant movement and environmental effects Conclusions References IMPACTS ON BIODIVERSITY 2.4.1 2.4.2 AUTHOR(S) Bioaccumulation of pollutants Release of metals during combustion 78 79 REFERENCES WASTE UTILISATION AND BIOREMEDIATION: THE ROLE OF GMOs 4.1 INTRODUCTION 4.2 PHYTOREMEDIATION OF INORGANIC POLLUTANTS 4.2.1 4.2.2 4.2.3 4.2.4 4.2.5 4.2.6 Root uptake Transport within plants Hyperaccumulation Metallothioneins Phytochelatins Transformation of toxic elements N Smith 81 81 82 82 83 83 84 84 85 BIOENERGY CROPS AND BIOREMEDIATION AUTHOR(S) 4.3 5. PHYTOREMEDIATION OF ORGANIC POLLUTANTS N Smith PAGE 86 4.4 PLANT TRANSFORMATION 88 4.5 RISKS 89 4.6 DISCUSSION 90 4.7 REFERENCES 91 LEGISLATION & CODES OF PRACTICE AFFECTING RECYCLING & LAND APPLICATION OF ORGANIC WASTES 5.1 INTRODUCTION 5.2 UK WASTE REGULATIONS & DIRECTIVES 5.2.1 5.2.2 5.2.3 5.2.4 Codes of practice Odours & gaseous emissions Water pollution Land application, including bioremediation, of organic materials 5.2.5 Draft Soil Strategy - England 5.2.6 Nitrate 5.2.7 Phosphate 5.2.8 Sewage sludge 5.2.9 Safe Sludge Matrix 5.2.10 Sewage sludge use in forestry and on restored land 5.2.11 Waste Management Licensing Regulations 5.3 G Hickman 98 98 98 100 101 102 103 103 104 105 106 106 108 109 UK LEGISLATION, NOT PRIMARILY AIMED AT WASTE DISPOSAL, WHICH MAY AFFECT ORGANIC WASTES 110 5.3.1 5.3.2 5.3.3 5.3.4 110 111 111 111 Animal By-Products Order Plant Health (GB) Order, 1993 Planning controls Contaminated land BIOENERGY CROPS AND BIOREMEDIATION 5.4 EC LEGISLATION: PROPOSALS THAT MAY AFFECT ORGANIC WASTE 5.4.1 5.4.2 5.4.3 5.4.4 5.5 6. PAGE G Hickman 112 112 Introduction EC Sludge Directive – Working Document 3rd Draft EC Biological Treatment of Waste Directive – Working Document 2nd Draft EC Landfill Directive 112 113 114 115 REFERENCES CONCLUSIONS & RESEARCH RECOMMENDATIONS 6.1 6.2 AUTHOR(S) CONCLUSIONS RESEARCH RECOMMENDATIONS C Britt 118 118 119 BIOENERGY CROPS AND BIOREMEDIATION EXECUTIVE SUMMARY Introduction 1. The Government's aim is for the UK to produce 10% of all electricity from renewable sources by 2010. A viable biomass industry is central to this target. If energy crops and forest residues are to provide fuel for an additional electricity capacity of 1,500 MW, around 125,000 ha of energy crops will be needed. Current plantings are of willow (Salix) and poplar (Populus) short rotation coppice (SRC) and Miscanthus but, although planting grants are available, uptake has so far been slow. 2. Using energy crops for the disposal of agricultural, municipal or even industrial wastes, or growing them on low-value ‘brown-field’ sites (e.g. capped landfill sites, mining spoils and contaminated ex-industrial land), may provide additional revenue and improve their appeal to growers. On brown-field sites energy crops also offer real opportunities for site stabilisation and bioremediation of contaminated soils. 3. This report summarises the findings of a desk study, which investigated the opportunities and potential problems associated with systems that utilise energy crops as disposal sites for waste materials or for the phytoremediation of contaminated land. The study included an extensive review of the literature and expert analysis of the issues. The main objectives of the study were: a) To review opportunities for bioremediation with energy crops. b) To evaluate the likely magnitude of available land and waste application rates. c) To quantify possible nutrient and metal losses, via leaching in soils and water. d) To quantify nutrient and metal uptake by energy crops. e) To evaluate the potential for use of genetically modified organisms to improve the bioremediation/waste utilisation capabilities of energy crops. f) To evaluate the likely magnitude of atmospheric emissions of CH4, N2O and CO2 from wastes applied to energy crops. g) To evaluate the impact of waste application, and/or siting on contaminated soils, on biodiversity associated with energy crops. h) To review and summarise the legal framework affecting disposal of farm and non-farm wastes, and consider how relevant UK and EU legislation helps or hinders the use of energy crops for bioremediation. i) To identify research requirements. Nitrogen and Phosphorus – Losses to Water 4. Energy crops require little additional nutrients in the planting year. In particular, extra nitrogen is not required, as mineralisation from soil organic matter will supply sufficient quantities of this nutrient and applying more will increase nitrate losses by leaching. Once established, atmospheric deposition and nutrient return in leaf fall are likely to be significant factors in energy crop nutrient cycles. 5. There is conflicting evidence on the N requirements of energy crops from the second year onwards, possibly arising from the dynamics of nutrients within the crop. Dense SRC crops have a higher proportion of bark in the harvested wood, and bark contains a BIOENERGY CROPS AND BIOREMEDIATION much higher concentration of nutrients. In SRC and Miscanthus, Danish work suggests leaching does not occur, even when organic manures are applied at rates well above standard recommendations. However, these could be mineralised, and leached from the soil at harvest, or after restoration of the site to agriculture. There is also the likelihood of large CO2 losses from the soil following site restoration. 6. To avoid nutrients leaching into surface or ground-water, only manures or slurries with a low available N content should be applied prior to planting energy crops, or in the establishment year. Organic wastes with higher available N contents might be applied from the second year onwards. Applications should be guided by the DEFRA/NAWAD Code of Good Agricultural Practice for the Protection of Water (the ‘Water Code’), and take into account crop off-take. 7. The Water Code specifies that a maximum of 250 kg ha-1 of organic nitrogen can be applied in any twelve month period. This figure is used throughout this review, for illustrative purposes. It should be noted that this figure is for total organic nitrogen and that available nitrogen levels will depend on the type of organic manure/waste being applied. For low available nitrogen materials, in non-sensitive catchments, 500 kg N ha-1 is allowed every two years. With the expansion of nitrate vulnerable zones (NVZs) in 2002 at least 55% of England will fall within sensitive catchments. 8. Regular applications of sewage sludge or manures are likely to rapidly increase soil phosphate levels and the potential for phosphate losses by leaching. Energy crops have a relatively low demand for phosphate and should not be used for the continued disposal of high phosphate organic wastes. 9. Irrigating crops with the leachate from buried waste requires further research. Gaseous Losses 10. Increased gaseous flux of nitrogen and sulphur may occur as a result of the application of organic wastes to energy crops. Ammonia volatilisation from manures and slurries inevitably occurs and, typically, 40-50% of the plant available N can be lost within six hours of application. Whilst the most effective way of reducing ammonia volatilisation from slurries is to inject the slurry into the soil, this is not an option in energy crops, due to the mass of roots or rhizomes. Application methods and rates of manure use that greatly reduce soil air-space or gaseous exchange will increase emissions of nitrous oxide (N2O) and methane (CH4) ‘greenhouse gases’. 11. Manure applications are likely to closely follow crop harvests, exacerbating soil compaction caused by harvesting machinery. These factors suggest that losses of N2O from energy crops following organic waste application are likely to be high but losses have not yet been quantified. Emissions of N as N2O from sewage sludge, over a full year, are likely to be of the order of 1-2% of applied N – similar to those from livestock wastes. 12. Production of energy crops will result in net increases in soil organic matter (through microbial decomposition of the leaf litter and turnover of fine roots) and below-ground carbon, stored in a semi-permanent root system. This may be enhanced by regular applications of organic wastes. BIOENERGY CROPS AND BIOREMEDIATION 13. Using existing figures, the net effects of applying organic wastes to energy crops on the contribution to radiatively active gases in the atmosphere and carbon sequestration were estimated. If livestock wastes are applied to SRC every three years, after harvest, at the maximum rates allowed by the DEFRA Water Code, the estimated benefits of carbon sequestration may be reduced by the effects of CH4 and N2O emissions by between 27 and 36%. The corresponding figures estimated for Miscanthus, which sequesters slightly more carbon, suggest a 24-31% reduction in C sequestration. If wastes were applied annually to Miscanthus nearly all of the carbon sequestration benefits would be negated by N2O (mainly) and CH4 emissions, by approximately 6875%. These estimates are based on sequestration/emissions over a full economic rotation of 25 years. 14. With the above provisos, the applications of moderate quantities of organic wastes to energy crops is likely to be beneficial to soil sustainability and to make modest contributions of carbon sequestration. As well as providing an important source of plant nutrients and valuable quantities of organic matter, livestock manures and other organic wastes applied to fuel crops may contain compounds that can be harmful when applied to plant ecosystems in the human food chain. Heavy Metals and Other Contaminants 15. Heavy metals [including arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), lead (Pb), mercury (Hg), nickel (Ni) and zinc (Zn)] cannot be broken down into less harmful by-products, so phytoremediation strategies focus on their accumulation in above-ground plant parts and removal from the contaminated site. Uptake is affected by the type of waste and the soil type. 16. Organic pollutants can often be completely broken down by plants into less harmful metabolites. Research on hybrid poplars has demonstrated their ability to take up and effectively degrade or deactivate a number of other contaminants, including atrazine, 1,4-dioxane, TNT and trichloroethylene. 17. Biosolids, rather than livestock manures, will be a more important source of heavy metals and organic contaminants to biomass crops. Although research evidence is limited, the current consensus is that organic contaminants in biosolids applied to agricultural land are unlikely to cause significant environmental or human health problems. However, whilst the risk of soil metal contamination from biosolids is recognised and regulated for, there is no current control over inputs of heavy metals from livestock manures, and care should be taken to ensure that soil concentrations do not become excessively high, especially where pig and poultry manures are applied. 18. High yielding biomass crops offer good potential for the phytoremediation of sites contaminated with heavy metals. Willows have been shown to take up large amounts of Cd and Zn. Different biomass crops, species and clones may show large differences in efficiency of heavy metal uptake. There can also be large differences in the concentration of metals in different plant parts. For phytoremediation purposes, it is desirable for metals to be concentrated in the harvested parts of the plant e.g. SRC stems/wood. 19. The maximum annual applications of high-metal content organic wastes, such as sewage sludge or poultry manure, to willow SRC crops are likely to lead to a net BIOENERGY CROPS AND BIOREMEDIATION accumulation of metals (e.g. Zn, Cu, Pb, Ni and Cr) in the soil, as application rates would exceed crop uptake. However, on average soils the maximum soil -1 concentration of 200 mg Zn kg would be reached in around 110 years for sludge and 160 years for poultry manure. 20. Metals removed from sites in energy crops will primarily be concentrated in the ash after combustion or gasification, except mercury which may volatilise and be emitted from the stack to be deposited onto soils in the vicinity of the combustion plant. Other metals may be discharged into the atmosphere, via particulates in the flue gases, and subsequently re-deposited onto land. 21. Recovery of metals from ash may be technically possible, but very expensive. The disposal of ash with high heavy metal concentrations may pose problems. Impacts on Biodiversity 22. Research has shown the value of willow and poplar SRC as a habitat for certain animal groups, including phytophagous insects, songbirds (especially warblers) and pheasants. There has been very little research on the ecology of Miscanthus or other energy grasses. 23. The extent and composition of the ground flora will, to a large degree, determine the biodiversity of animal species within the crop. In most SRC plantations the ground flora is either sparse, due to effective weed control measures, or of limited diversity and dominated by species of low conservation value, typical of agricultural weed communities or disturbed land. Species such as bramble may invade coppice later in the cycle. One extensive survey of SRC plantations showed common nettle to be the most frequent species (81% of all sites). 24. Animal groups known to be well represented in SRC plantations include herbivorous invertebrates that feed on the crop foliage, like willow beetles, and their predators and parasites; insectivorous woodland birds (e.g. thrushes, tits and warblers); and mammals such as rabbits, roe deer and wood mice. Populations of ground-dwelling species will be heavily dependent upon the extent and species composition of the ground flora. 25. The application of waste materials might be expected to have significant effects on the flora and fauna of energy crops. However, no evidence was found in the literature of research that directly addressed this issue. There was only a limited amount of published research on the ecological effects of applying relevant agricultural, urban or industrial wastes to other types of vegetation. 26. Thick applications of waste materials, particularly those that are slow to decompose, may have a ‘mulching’ effect and suppress the ground flora, whilst organic wastes may provide a valuable additional food source for soil and ground-dwelling microorganisms and invertebrates. Mulching would have a net detrimental effect, whilst provision of a supplementary food source would be generally positive. Both would have ‘knock-on’ effects up through the food chain. 27. The addition of nutrients through organic waste applications will also have indirect effects on the growth and composition of the ground flora, and provide crop nutrients. BIOENERGY CROPS AND BIOREMEDIATION However the potential ecological effects of nutrient inputs from manures, slurries and sludges can probably be regarded as minimal, as similar quantities of nutrients would otherwise be applied in inorganic fertilisers. What may be more significant are the effects of additional water supplied in slurries or dirty water. 28. The bioaccumulation of heavy metals or organic toxins in animal tissues, following the application of contaminated wastes to energy crops, is an important concern. However, although high levels of heavy metals (e.g. Cd) in the livers and kidneys of insectivorous shrews and deer species have been observed in US studies, generally the heavy metal contents of these mammalian organs were not considered to have important effects on the health of the animals concerned. It is not known what the effects of low concentrations of compounds like polycyclic aromatic hydrocarbons (PAHs) in animal manures and slurries might have on soil-inhabiting invertebrate groups in biomass crops. 29. At the top of the food chain, predators such as foxes, stoats and birds of prey, might appear to be at greatest risk, but the relatively large ranges of most predator species lessens the degree of exposure to contaminated prey. The risks would, however, greatly increase in the event of a significant area of waste-treated biomass crop being situated in a single location. Sewage contaminated with pathogens pose another risk to wildlife, and although, in most cases, risks may be low, research has shown that in some situations deer may preferentially graze areas that have been treated with sewage sludge, thus increasing their exposure risk. Bioremediation of Contaminated Sites 29. The necessity to decontaminate polluted sites is recognised, both socially and politically, because of the increasing importance placed on environmental protection and human health. As the number of sites and levels of contamination rise, so does the need to develop effective and affordable methods for decontamination. 30. Phytoremediation is the term used to describe the use of plants to mitigate the effects of contamination. There are four fundamental processes that make up phytoremediation: phyto-immobilisatio, phyto-stabilisation, phyto-extraction and phyto-volatilisation. Phytoremediation is a low-cost option, particularly suited to large sites that have relatively low levels of contamination. 31. Many species of Salix, Populus and Miscanthus have characteristics of ‘pioneer’ species – with adaptations for growth on poor sites, under harsh conditions. Willow varieties differ significantly in their levels of tolerance and rates of uptake of heavy metals (see Exececutive Summary 15). 32. Results from pot studies indicate that Miscanthus crops could be successfully grown on contaminated land, although high levels of heavy metals may reduce crop productivity. Most heavy metals accumulate in the roots and rhizomes, rather than in the harvested aerial parts. BIOENERGY CROPS AND BIOREMEDIATION Waste Utilisation and Bioremediation: the Role of Genetically Modified Organisms 33. The genetic modification of biomass crops to improve pollutant uptake, transport, accumulation and tolerance, offers the potential to dramatically increase the effectiveness of phytoremediation of organic compounds and metals from contaminated sites. A single GM energy crop might be produced to efficiently take up several different pollutants. 34. Poplar species have been the subject of a considerable amount of research on the introduction and over-expression of foreign genes. The first reports of poplar transformation were published in the late 1980s. The potential genetic transformation of willows and Miscanthus has received relatively little attention. However, for Miscanthus, research on embryogenic suspension culture, and other in vitro propagation systems, and the transformation of callus tissue via microprojectile bombardment, has progressed to the point that the generation of transgenic Miscanthus clones can be expected in the relatively near future. 35. Genetically modified biomass crops offer the potential for dramatically improved phytoremediation capabilities, but careful consideration must be given to the environmental consequences before such crops are exploited. Primary considerations should include the potential for bioaccumulation of toxic pollutants in animal populations (a problem that is likely to be exacerbated by hyperaccumulation of pollutants in the stems and leaves of GM plants) and the prevention of any possible gene flow into ‘conventional’ crop plants/native plant populations. Legislation and Codes of Practice Affecting the Recycling and Land Application of Organic Wastes 36. The use of wastes, sludges and manures on land generally, and on biomass crops is covered by UK legislation and codes of practice, and EC directives. The regulations are continually changing as old legislation is updated and new controls and recommendations are issued, that must be considered before organic waste materials are applied to land. 37. The DEFRA/NAWAD Codes of Good Agricultural Practice for the Protection of Water, Air and Soil (1998) are statutory codes under Section 97 of the Water Resources Act 1991 and give guidance on best practice for avoiding pollution. Most farmers and other operators applying organic wastes to land follow the guidance of the Codes and all Water Operators have signed up to compliance with them. Compliance is also a basic requirement for participation in most Farm Assurance Schemes. As with all Codes of Practice failure to comply with the Codes is not an offence, but would be taken into account in any legal action taken as a result of a pollution incident. 38. Under the Pollution Prevention and Control (England & Wales) Regulations 2000, industrial installations are regulated by the Environment Agency or local authorities. The impacts of these relatively new regulations are still hard to assess, but they are not expected to hinder the use of wastes on energy crops. The recycling of nutrients in livestock manures is excluded from EC and UK regulations on the use of ‘waste materials’ spread to land. BIOENERGY CROPS AND BIOREMEDIATION 39. Legislation covering water catchments, and the earlier compliance measures to meet the requirements of EC Nitrate Directive (91/676/EEC) are reviewed and the effects of compliance commented on. The recently announced expansion of the areas covered by Nitrate Vulnerable Zones make the restrictions far more widely applied, but the ability of growers of biomass crops to comply remains much the same; the Action Programme limits to nitrogen use are those in the Codes of Practice. 40. The issue of phosphate enrichment of water is becoming increasingly important. The Soil and Water Codes provide guidance on limiting phosphate losses and recommend that farmers should avoid total P inputs in excess of crop off-take. The apparently low P requirements of bioenergy crops could limit the total amount of organic wastes that can be applied. 41. Sewage sludge applications to land, and biomass crops, are regulated by EC Directive 86/278/EEC (1986), and should comply with the Water Code, with applications supplying no more than 250 kg total N ha-1 yr-1. For farmyard manure, the Water Code allows an option for low N sewage products, such as sludge cake, to be applied at 500 kg total N ha-1 in every other year (in non-sensitive catchments only – see Executive Summary 6). Revised regulations will adopt the provisions of the Safe Sludge Matrix, and introduce standards for pathogen content in sludges. They will also ban the use of untreated sewage sludge on all crops from 31 December 2005 (They have been banned on all food crops since December 1999). 42. The use of sewage sludge on non-agricultural land, and for land restoration, is controlled by the Waste Management Licensing Regulations 1994. These regulations do not, however, cover wastes arising from agriculture. Also excluded are numerous specified materials for use in agriculture. Disposal of exempt materials must, however, result in agricultural or ecological benefits, and are generally subject to a maximum annual application rate of 250 t ha-1. 43. The Town and Country Planning Act 1990 (as amended by the Planning and Compensation Act 1991) may limit the use of certain wastes on energy crops that might be planted after mineral extraction has ceased, whilst requirements of the Environmental Protection Act 1990 and Environment Act 1995 may act as drivers for the planting of biomass crops for the bioremediation of contaminated land identified by local authorities as potential risks to human health or the environment. 44. A new proposed EC Sludge Directive may severely restrict the recycling of sewage sludge, and certain other organic materials to agricultural land. It is likely to significantly reduce the maximum levels for heavy metals in soils, introduce limits on levels of heavy metals and organic compounds in sludges, and include treatment standards based on pathogen reduction. It is also likely to confirm the principle that mixing any sludge with other non-regulated wastes makes the entire combined waste stream subject to the conditions of the Directive. 45. Similarly the proposed EC Biological Treatment of Biowaste Directive (or ‘Compost Directive’) would severely restrict the treatment and application to land of many mixed wastes and industrial wastes. The proposed maximum application rates for organic wastes are generally much lower than are currently applied in the UK. Both the Sludge and Compost Directives are unlikely to be enacted in UK legislation before 2005. BIOENERGY CROPS AND BIOREMEDIATION 46. The EC Landfill Directive 1993/31 requires significant reductions in the quantities of putrescible materials disposed of in landfill sites. Targets have been set for 2006, 2009 and 2016. An estimated five million tonnes of organic material is disposed of annually within the UK household waste stream. The Directive is driving waste management companies and waste producers to look at land spreading and bioremediation, as a possible route to divert this material away from landfill. BIOENERGY CROPS AND BIOREMEDIATION CHAPTER 1 INTRODUCTION MIKE BULLARD & PETER NIXON 1.1 BACKGROUND In response to now incontrovertible evidence of human-induced global warming due to the emission of atmospheric warming gases (IPCC, 2000), regional, national and European targets for both emissions reduction and renewable energy generation have been set (DTI, 1999; EC, 1998). For the UK, a target of 10% of electricity generation from renewables has been set for 2010. Central to the government’s renewable energy targets is the stimulation of viable biomass crops (DTI, 1999). It is anticipated that 1,500 MW of new electrical capacity might come from energy crops and forestry residue combustion, and it has been proposed that 125,000 ha of energy crop planting might be needed in order to meet this target. In the UK, the energy crops most likely to be grown are short rotation coppice (SRC) willow and poplar and Miscanthus grass. These high-yielding species are eligible for establishment grants of up to £1,600 ha-1 on IACS-registered arable or grassland, or other grazed grassland (see MAFF, 2000 for full details). Sufficient grants are currently available to enable 23,000 ha of crop to be established; yet less that 2,000 ha have been planted in England and Wales (At the time of writing all of this is willow coppice, mainly established by ‘Project Arbre’ in Yorkshire, Nottinghamshire and Lincolnshire). Major disincentives for uptake of the crops are: 1. Energy generation schemes (the ‘end user’) are coming on-stream at a much slower rate than anticipated, thus demand for the crops is lower than anticipated. 2. Establishment grants only cover 40% or 50% of establishment costs (for Miscanthus and SRC, respectively), leaving a deficit of at least £1,000 ha-1. 3. Energy crops require a long-term commitment for relatively low financial reward. Other less risky cropping options are, therefore, more attractive. 4. Yields from the first plantings have been disappointing and this has suppressed interest. For these reasons, crop uptake has been low. In order to encourage energy crop planting it is, therefore, necessary to consider other initiatives where additional economic benefits can accrue from growing the energy crop. Increasing interest is being given to the concept of disposal of agricultural and municipal wastes on energy crops. This potentially provides organic matter and nutrients needed for crop growth at a low cost, whilst enabling controlled disposal of wastes on a non-food crop. Indeed, some grower organisations recommend the application of wastes to provide the necessary nutrients for SRC production. 1 BIOENERGY CROPS AND BIOREMEDIATION Alternatively, it is pertinent to consider biomass crop opportunities on nonagricultural land, where growing the crop may provide an additional benefit to the land owner. For example, the biomass crop may provide essential site stabilisation (and, possibly, decontamination) - with the sale of biomass an additional, but nonessential, benefit. Many are now proposing that energy crops should be grown on brown-field sites (e.g. capped landfill sites, quarries, mine spoils or contaminated ex-industrial land), in order to facilitate bioremediation of heavy metals and toxins. Another advantage here is that land prices are much lower than for agricultural land, and energy crops are a true added benefit to a situation where the major priority is decontamination of the land. Whilst both routes offer advantages, the true extent of the advantages and the magnitude of the land availability for such cropping are poorly understood. Equally, there are also a number of potential hazards from using these disposal routes and/or sites. Whilst there is information in the public domain about specific examples of waste disposal or bioremediation, there is no consolidation of the subject in a way that would allow policy makers to make clear judgements on the needs and opportunities posed. For DEFRA and Government to support such energy cropping initiatives, there must be clear information on the magnitude of the risks and opportunities that such disposal routes present. This review brings together detailed information on these issues and also identifies areas requiring further research. 1.2 THE ENERGY CROPS IN QUESTION Any crop or agricultural residue that can be presented in a relatively dry form (i.e. at approximately 25% moisture content, or less) is suitable for thermo-chemical conversion. Material with high moisture content can be efficiently used in combined heat and power (CHP) plants. The purpose of growing energy crops is to maximise the yield of ligno-cellulose that can be harvested and presented to a power station in a useable form. The two crops that offer this yield advantage are now described in more detail: 1.2.1 SRC willow and poplar Short rotation coppice (SRC) consists of densely planted, high-yielding varieties of willow, and occasionally poplar, harvested on a two to five year cycle, although most commonly every three years. SRC is a woody, perennial crop, the rootstock or stools remaining in the ground after harvest with new shoots emerging the following spring. An SRC plantation should be viable for up to 30 years before replanting becomes necessary. The osier (Salix viminalis), a native shrub or small tree, is the parent species of the majority of willow varieties grown as energy crops. Willow SRC is mechanically planted in the spring. Unrooted hardwood cuttings, produced by specialist breeders, are inserted into cultivated soil, using equipment 2 BIOENERGY CROPS AND BIOREMEDIATION specifically designed for fast and efficient planting. The planted material grows rapidly in the first year, reaching up to 4 m in height - depending on soil conditions. During the first winter after planting, stems are cut back to ground level to encourage the growth of multiple stems, that is the true coppice. Generally three years after cutback, and again during the winter, the crop is harvested. The equipment used for harvesting will depend on the requirements of the customer/end-user, that is their fuel specification, but in all cases the harvesting equipment will have been specifically developed or have involved modification of existing harvesting machinery. The majority of other operations - such as land preparation, spraying and fertilising - can be completed utilising conventional farm machinery. In the UK, yields achievable from willow SRC at first harvest are normally expected to be between 7 and 12 oven dry tonnes (odt) per hectare per year (odt ha-1 yr-1), depending on ground conditions and efficiency of establishment. Yields should increase at second and third harvests, as the stools mature. Plant breeding programmes are continuing to identify further new willow varieties, that will produce higher yields and demonstrate improved resistance to pests and diseases. 1.2.2 Miscanthus Miscanthus species are woody, perennial, rhizomatous grasses, originating from Asia, which have the potential for very high rates of growth. Miscanthus may be familiar to many as a flowering garden ornamental, but it is the non-flowering forms that are of interest agriculturally. Miscanthus is spring-planted, and canes produced during the summer are harvested in winter. This growth pattern is repeated every year for the lifetime of the crop, which will be at least 15 years. Miscanthus differs from short rotation coppice willow in that it gives an annual harvest, and thus an annual income to the farmer. Miscanthus spreads naturally by means of underground storage organs (rhizomes). However, their spread is slow and the risk of uncontrolled invasions of hedgerows or fields is, consequently, considered to be very low. Rhizomes can be split and the pieces re-planted to produce new plants. All propagation, maintenance and harvest operations can be done with conventional farm machinery. In the UK, long-term average harvestable yields from a mature crop have exceeded 13 odt ha-1 yr-1 at the most productive experimental sites. These high yields suggest that the crop has the potential to make an important contribution to the UK’s commitments to energy generation from renewable sources. 3 BIOENERGY CROPS AND BIOREMEDIATION 1.3 THE ISSUES The disposal of agricultural and municipal wastes to land and the use of contaminated sites for non-food cropping are two areas of increased concern for policy-makers. 1.3.1 Land availability The Environment Agency has estimated that there are between 50,000 and 300,000 ha of contaminated land across the country – distributed between some 100,000 sites. Of these sites, between 5,000 and 20,000 require remedial action, as specified under legislation. In the absence of any formal surveys it is difficult to provide accurate figures on the extent of contaminated land in each region. However, ‘derelict land’ - defined as 'land so damaged by industrial or other development as to be incapable of beneficial use without treatment', occupies an estimated 39,000 ha of land across England (DoE, 1993). Mines and quarries represent a significant area. A 1994 survey of land for mineral workings in England indicated that just under 14,500 ha of land had planning permission for surface disposal of mineral working deposits. Nearly 9,900 ha were actually affected by tipping (although some sites were in the process of being reclaimed), and over 6,000 ha had planning conditions for reclamation. Thus, as much as 1-1.5% of the UK land area may be considered derelict or in need of remediation, and much of this may be potentially available for energy cropping. 1.3.2 Waste availability As can be seen from Table 1.1, significant quantities of wastes are produced each year from various land-based sectors in the UK (DEFRA, 2001). Table 1.1. Estimated total annual waste production in the UK, by sector Sector Annual raisings (Mt) % of total 86 116 16 43 30 26 51 73 428 20 27 <1 6 10 7 6 12 17 100 Agriculture 3 Mining and quarrying 1 Sewage sludge 2 Dredged spoils 1 Municipal 4 Commercial 2 Industrial 2 Demolition and construction 5 TOTAL 1 2 Estimates for 1997 Estimates for 1998/99 3 4 Estimates for 1999 Estimates for 1999/2000 4 5 6 Estimates for 2000 Figures based on estimated dry wt (26.5 Mt total wet wt.) BIOENERGY CROPS AND BIOREMEDIATION 1.3.3 The role of energy crops Energy crops have the potential to utilise agricultural and municipal wastes, and to stabilise or clean up contaminated land. From the perspective of waste disposal/utilisation, energy crops offer the following potential benefits: they are not going to enter the human food chain; they are perennial crops, thus allowing long-term breakdown of organic matter in soils prior to renovation to food cropping; they produce large quantities of biomass that, theoretically, requires large quantities of nutrients, and thus are a sink for the nutrients in waste. From the perspective of bioremediation of contaminated sites, they offer the following potential solutions: they utilise land that would otherwise have no agricultural value; they are non-food crops that will not enter the human food chain; they are perennial crops which may act as ‘excluders’ of contaminants in the soil; alternatively, they may act as ‘tolerators’ of the contaminants, actively taking up the elements which, in some instances, can then be recovered during biomass combustion; the crops can also act as bioremediators of liquid leachates produced from rainfall onto landfill and other contaminated sites; in these situations, they may also act as recipients of agricultural and municipal wastes. There are attendant risks with such systems. With respect to the application of agricultural and municipal wastes, these include: risks of leaching nutrients applied in sludges into groundwater; risks of increased atmospheric emissions of ‘greenhouse’ gases, associated with global warming. For example, sewage sludge can increase the emission of nitrous oxide (Scott et al., 1998) and maybe methane, which although emitted in trace amounts are more effective greenhouse gases (CH4 = 32 CO2 equivalents; N2O = 150 CO2 equivalents; Bouwman, 1990); risks of contaminant accumulation in the production system, which are then emitted from power station stacks upon combustion of the biomass; negative impacts on the biodiversity associated with energy crops. With respect to bioremediation, the risks include: risks of contaminant accumulation in the production system, which are then emitted from power station stacks upon combustion of the biomass; negative impacts on the biodiversity associated with energy crops. The practices, therefore, have the potential of negating some benefits derived from saving fossil fuels by growing energy crops. There is a current lack of evidence on the thresholds of application that are acceptable in different situations and this is, in part, due to a lack of primary research in many areas. 5 BIOENERGY CROPS AND BIOREMEDIATION In order to assist DEFRA and other stakeholders in the production of such research priorities the following review condenses the research that has been carried out to date on these issues, and draws parallels with other cropping systems. 1.4 REFERENCES Bouwman, A F (Ed.) (1990). Soils and the Greenhouse Effect. John Wiley and Sons, Chichester. 575 pp. DEFRA (2001). The Environment in Your Pocket 2001. http://www.defra.gov.uk/ environment/statistics/eiyp. Department for Environment, Food and Rural Affairs. DoE (1993). Survey of Derelict Land in England and Wales. Department of Environment. DTI (1999). New and Renewable Energy: Prospects in the UK for the 21st Century. HMSO, London. DTI (1999). Waste Strategy 2000 for England and Wales: Parts 1 & 2 - ISBN 0 10 146932 2 / 0 10 146933 0. HMSO, London. EC (1998). Energy for the Future: Renewable Sources of Energy. European Commission White Paper. IPCC (2001). Third Assessment Report – Climate Change 2001:The Scientific Basis. Special report on emissions scenarios (Eds. J Houghton, Y Ding, D J Griggs, M Noguer, P J van der Linden, X Dai, K Maskell & CA Johnson). Intergovernmental Panel on Climate Change, United Nations. MAFF (2000). England Rural Development Plan: Energy Crop Scheme Explanatory Booklet. HMSO, London. 24pp. Scott, A; Ball, B C; Crichton, I J & Aitken, M N (1998). Nitrous oxide emissions from grassland amended with sewage sludge. Short communication in Soil use and Management, 14. 55. 6 BIOENERGY CROPS AND BIOREMEDIATION CHAPTER 2 WASTE UTILISATION: ENVIRONMENTAL AND CROP EFFECTS 2.1 NITROGEN AND PHOSPHORUS – LOSSES TO WATER PADDY JOHNSON 2.1.1 Introduction Consideration of the potential benefits or disadvantages of applying novel waste materials to short rotation coppice or graminaceous biomass crops should be preceded by a philosophical assessment. Are the materials being applied to improve crop growth, as a method of waste disposal or in order to reduce adverse environmental effects? Indeed will the use of biomass crops for cleaning up wastes, for example exploitation of the ability of Salix to remove cadmium, increase other adverse environmental effects? Possibly this is beyond the scope of this study. This section of the report, therefore, starts with a review of the nutrient requirements of biomass crops, so that the potential for increasing potential losses of nutrients to the environment can be assessed. It is unfortunate that there appears to be little published information on the beneficial (or otherwise) effects of the application of many of the wastes which it is being suggested could be applied to biomass crops, and none on the consequences of removing the biomass crop once its useful life has ended. It is important that long term effects are discovered, for example if biomass crops are used for the disposal of sewage sludge, will there be a large loss of nitrogen and carbon dioxide to the environment when the plantation is removed and the land returned to conventional agriculture? 2.1.2 Nutrient requirements of biomass crops The nitrogen cycle in the soil is complex. Mineralisation of N from soil organic matter plus crop residues, together with inputs from the atmosphere, can supply large quantities of available N. Mineralisation is likely to be particularly high in the year of planting following disturbance of the soil (Shepherd et al., 1996), and decrease thereafter - unless weed control is by mechanical means. Applied or mineralised nitrogen can be lost from the soil by leaching or by denitrification in wet warm conditions. A balance sheet approach to N fertilisation has some attractions, but many of the factors to be taken into account are not well understood. Several authors have indicated that no nitrogen is required in the planting year for short rotation coppice (Ford-Robertson et al., 1991; Kopp et al., 1993; Ledin & Alriksson, 1996; McLaughin et al., 1985). Indeed, application of soluble fertiliser at planting could have a deleterious effect on growth, as it raises the osmotic potential of the soil solution and may thus slow root development. Few countries give a recommendation for some nitrogen at this stage (Table 2.1); in subsequent years recommendations vary. 7 BIOENERGY CROPS AND BIOREMEDIATION Table 2.1. Nitrogen recommendations from various countries Source Recommendation in establishment year Subsequent years 45 100-150 0 ? 0 0 ? 60 122-217 50? 100-135 112-168 Ledin & Alriksson (1996), Sweden Chambers & Mitchell (1995), UK van Veen (1981), Netherlands Frison (1995), Italy Canada USA Note: Data from Ledin & Willebrand (1996) unless otherwise indicated. Similarly there are suggestions that Miscanthus does not respond to the application of nitrogen (SORGHAL, 1997). Nitrogen requirements post-planting Experience in the United States, summarised by Christopherson (1996), suggests that, in poplar plantations on sites with moderate or good levels of fertility and good weed control, N is unlikely to be required for at least two years. Furthermore, Christopherson (1996) suggests that, due to the recycling of nitrogen in leaf fall, applications to poplar plantations after canopy closure can be greatly reduced or even omitted on some sites. On sandy loam and loamy sand soils in Denmark, Mortensen et al., (1998) obtained a significant increase in yield where N was applied at 75 kg ha-1 in the second year on one site, but there was a non-significant reduction in yield in the third year (Table 2.2). Despite low mineral nitrogen content of the soils, responses at these sites may have been restricted by low water availability. McLaughlin (1985) needed to wait until the third year before recording any increase in yield of poplar wood related to the application of nitrogen. In Northern Ireland, Dawson (1999) has suggested that there was no response to the application of N for at least two rotations when the coppice was grown on reasonably fertile exarable land. Table 2.2. Response of SRC to nitrogen applications Site Soil Type Foulam Jyndevad * loamy sand sandy Year 1 Year 2 Year 3 0N 75 kg ha-1 N 0N 75 kg ha-1 N 0N 75 kg ha-1 N 1.25 1.32 2.68* 3.71* 8.71 6.93 1.62 1.64 3.52 4.94 3.67 3.61 Statistically significant response (P < 0.05) 8 BIOENERGY CROPS AND BIOREMEDIATION Christopherson (1996) suggested that a leaf analysis level of 3% N in the dry matter in mid-July, indicates a sufficiency of nitrogen in the system. This is a leaf concentration slightly above that used for top fruit in the UK (MAFF, 2000). Leaf analysis could be the best way of assessing nitrogen need and should be investigated. Hall et al. (1996) recommended that growers of short rotation coppice should be discouraged from applying nitrogen fertiliser. It is possible that Miscanthus does not respond to nitrogen applications during post establishment years (SORGHAL, 1997). Other nutrients There is a paucity of reports on the response of biomass crops to nutrients other than nitrogen. Therefore a balance sheet approach for the maintenance of soil reserves may be the best approach for deriving crop requirements. An exception is a report by Frison (1995), which differentiates between tree populations, based on the fact that the majority of the nutrients are in the bark and thus the percentage of bark will be much higher at the higher populations (Table 2.3). In the early years of growth uptake and potential removal of phosphate and potash will be small. Of these two nutrients only phosphate carries an environmental threat. Except at very low soil levels applications of these nutrients based on a balance sheet approach would seem appropriate. Table 2.3. Nutrient off-takes in timber of Populus x canadensis clone I-214, kg per oven-dry tonne (Frison, 1995) Nitrogen Phosphate Potash 75,000 trees ha-1 6.9 3.1 5.2 10,000 trees ha-1 5.6 2.1 4.2 -1 1.6 0.7 2.4 277 trees ha Losses of nitrogen by leaching Nitrogen applied at planting or early in the life of the crop is readily lost from the soil by leaching. Mortensen et al., (1998) reported that where 75 kg ha-1 N was applied at planting, leaching losses of N were 32 kg ha-1 higher than where no N was applied. Christian & Riche (1998) showed large leaching losses of N under establishing Miscanthus, which were further increased by the application of N fertiliser on a site already rich in mineralised N. On a moisture retentive soil in west Norfolk, poplars were grown at 3.5 m spacings and fertilised with ammonium nitrate (60, 180 and 300 kg ha-1) or pig slurry applied in spring to supply similar amounts of nitrogen. Where the higher rates of N were 9 BIOENERGY CROPS AND BIOREMEDIATION applied, both tree height and incremental diameter improved dramatically. However, there was a downside, which could be partially controlled by other management techniques. Nitrate leaching was significantly increased (Johnson, 1995) at the higher nitrogen rates (Table 2.4). The experiment showed that where manures are applied, environmental effects needed to be taken into account. If manures were applied for several years and nitrate losses controlled by husbandry (grass would not be the preferred option as it will reduce the amount of moisture available for tree growth), there was a potential for a large build-up of soil N to occur. This might be mineralised when the soil is disturbed at harvest or the plantation removed. Table 2.4. Nitrate-N losses (kg ha-1) by leaching at ADAS Terrington, Norfolk over two winters N applied in slurry (kg ha-1) Weed control Ammonium nitrate 0 60 180 300 300 Cultivation 86 83 133 265 212 Herbicide 48 17 32 110 200 Grass strip 3 5 2 24 39 Weed control 0.002 17.4 4 P SE df N rate < 0.0001 36.6 24 Interaction 0.002 36.6 24 Further monitoring of this site (Johnson, 2000) during winter 1999/2000, after the original treatments had been abandoned some five years earlier, showed much reduced levels of nitrogen loss (Table 2.5), although drainage was very low. Table 2.5. N loss and mean N concentration in winter 1999/2000, estimated from porous ceramic cups installed at 90 cm depth Cult Nil N Grass NH4NO3 Cult Grass Pig slurry Cult Grass Fertiliser SEM Prob. Cover SEM Prob. N loss (kg ha-1) 0.27 0.08 0.38 0.05 0.99 0.16 0.08 0.01 ** 0.03 0.02 ** Mean N conc. (mg l-1) 1.31 0.39 1.81 0.24 4.74 0.79 0.37 0.01 ** 0.13 0.02 ** ** Statistically significant response (P < 0.01) Note: Based on drainage of 21 mm. 10 BIOENERGY CROPS AND BIOREMEDIATION Jorgenson (1999a & 1999b) has published results for both willow and Miscanthus that show that, even if organic manures are applied at rates well above standards for Denmark, then in established plantations nitrate leaching is not enhanced. What we do not know is what will happen when the plantations are returned to agriculture or whether in this instance how much of the applied nitrogen was lost by volatilisation as ammonia. Hall et al. (1996) suggested that the build-up of soil organic matter, and thus soil nitrogen, would be beneficial in terms of soil structure and moisture holding capacity when the land is returned to cropping. They state that the nitrogen will be only slowly mineralised. A comparison may be made with the situation where old grassland is ploughed up, though organic matter and soil nitrogen levels may not be as high unless large quantities of organic material has been ‘disposed’ of on the site. Using the changes in organic matter contents described by Johnson and Prince (1991), Whitmore et al. (1992) calculated that very large losses of nitrate occur in the first five years or so after ploughing. If soil nitrogen levels increase during the growth of biomass crops there is likely to be a large loss of nitrogen by leaching following the return of the land to normal cropping. There may also be an increased loss following any harvesting operation as, unless soil conditions are perfect, the trafficking will act like a cultivation - and cultivation has been shown to increase nitrogen mineralisation. There will also be a large loss of carbon, as carbon dioxide, from the soil. 2.1.3 Use of organic manures or wastes Organic manures have two forms of benefit when added to soil. Firstly, they add nutrients and secondly, they act as soil conditioners and help improve the moisture holding capacity of the soil. Nutrients in organic manures from human, industrial and farmyard animal origin contain nutrients in ‘available’ and ‘slowly available’ forms. Standard tables for many of these products are presented in ‘Fertiliser Recommendations for Agricultural and Horticultural Crops’ (MAFF, 2000). If manures are applied prior to SRC planting, the value of phosphate and potash in the manures can be assessed using total contents. This is true unless soil values are low, when the available nutrient levels should be used, as is standard practice for agricultural crops. Available nitrogen contents of organic manures can be assessed using the ammonium nitrogen contents (and uric acid contents for avian manures). The amounts which will be available for plant uptake can be predicted by using the decision support system MANNER (Chambers et al., 1999). As indicated earlier in this report, losses of nitrogen by leaching in the winter following planting of SRC are elevated. Also, no nitrogen is required in the establishment year. Logically, it follows that if organic manures are to be applied prior to planting, they should contain the minimal quantity of ‘available’ nitrogen. Analysis would show this as ammoniacal nitrogen (poultry manures should be analysed for uric acid N as well). Therefore, it is concluded that poultry manures, animal slurries and liquid digested sewage sludge, which contain a high proportion of available nitrogen, should not be applied prior to planting or in the establishment year because it will increase the risk of N loss by leaching. Van den Berg (taken from a translation with no details of the journal) reported that annual applications of pig slurry after planting and then annually, in an uncontrolled 11 BIOENERGY CROPS AND BIOREMEDIATION experiment in the Netherlands, increased the growth of poplars above that from applications of inorganic fertiliser over a five-year period. Spring and summer applications were marginally better than autumn applications, indicating that at least some of the nutrients from the pig slurry were lost, possibly by leaching. In a second set of experiments using calf slurry, there was also an indication from the growth parameters measured, that slurry applications in early summer, improved growth more than mineral fertiliser. This occurred on all sites except one, where rooting was shallow and may have been damaged by traffic during spreading. Some of the improvement in growth may have been as a result of the extra water from the slurry. Water is probably the greatest limitation to growth of biomass crops (Samson, 1993). In the USA, Sidle & Kardos (1979) reported elevated levels of nitrate loss following very high applications of nitrogen in bio-solids (1400 and 3000 kg ha-1). Aschmann et al., (1992) also reported increased losses in the year following application of waste water sludge, with greatly reduced losses in the second year, except where high levels of nitrogen had been applied. Prior to planting and during the preparation of a site for SRC, an application of a bulky organic manure, such as well-rotted farmyard manure, sludge cake, or other material with a low available nitrogen content, can be beneficial. Similar yields of biomass to those achieved with artificial fertiliser have been recorded in Sweden, where biosolids have been applied (Ledin et al., 1998). Post-planting applications of materials with higher available nitrogen contents, particularly in the second year of growth, would be possible, but care should be taken not to exceed the N requirements of the crop. There will also be an increased risk of losses of nitrogen as ammonia (see Section 2.2). These views are in general agreement with those of Riddle-Black (1998) for biosolids, though she does not distinguish between materials with high available nitrogen contents used pre-planting and those with low levels, which is important for environmental reasons, as discussed earlier. Bulky organic materials tend to improve soil structure and moisture holding capacity, particularly of light soils, and this can be advantageous. The nitrogen supplied by these materials will be released slowly by mineralisation and this has advantages, both from environmental and nutritional points of view. Applications have in the past, been shown to improve the quality of unrooted rose cuttings (Johnson, 1977). It would be expected that a similar effect would be seen with SRC, a view also held by Dawson (1999). If organic manures are applied, their nutrient content should be taken into account when deciding whether any manufactured fertiliser is to be applied. Standard nutrient contents for animal manures are contained in standard texts (MAFF, 1994). The Code of Good Agricultural Practice for the Protection of Water (MAFF, 1998) specifies that a maximum of 250 kg ha-1 of organic nitrogen can be applied in any twelve month period. This figure is used throughout this review, for illustrative purposes. It should be noted that this figure is for total organic nitrogen and that available nitrogen levels will depend on the type of organic manure/waste being applied. For low available nitrogen materials, in non-sensitive catchments, 500 kg N ha-1 is allowed every two years. With the expansion of nitrate vulnerable zones (NVZs) in 2002 at least 55% of England will fall within sensitive catchments. 12 BIOENERGY CROPS AND BIOREMEDIATION Phosphate loss to the environment Phosphate losses are most likely to occur when water erosion occurs. Water-driven erosion occurs when there is a lack of vegetation preventing the impact of rainfall on the soil surface leading to surface flow. Therefore, the danger period with biomass crops is during the establishment period. Malik et al. (2000) have shown reduced erosion by growing cover vegetation during this period, but all other vegetation will compete with the biomass crop for water, which is the major restraint to yield. Thornton et al. (1998) found variable results when comparing coppice with conventional agricultural crops, with higher losses than from cotton but less than from maize. Tolbert & Wright (1998) indicated no differences in erosive soil loss between coppice and conventional crops in the establishment year, but give no indication as to what happens once the biomass crop is fully established. A reduction in erosion following establishment might, however, be expected as raindrop impact would be reduced by interception by leaves and branches, particularly so in Miscanthus, and soil surface stability would increase as organic matter contents increase. If biomass crops are to be used for bioremediation, growers will need to decide whether or not to use competing cover crops to reduce early erosion risks, considering factors like rainfall, soil type and topography. Many organic waste materials contain large amounts of phosphate. Typical figures for total phosphate and other nutrients in animal manures and sewage sludge are given in Tables 2.6 and 2.7 (MAFF, 2000). Table 2.6. Typical total nutrient content of livestock manures (fresh weight basis) Manure Type Solid manures Cattle farmyard manure Pig farmyard manure Sheep farmyard manure Duck manure Layer manure Poultry litter Slurries/liquids Dairy Beef Pig Dirty water Separated cattle slurries (liquid portion) Strainer box Weeping wall Mechanical separator Dry matter (%) 25 25 25 25 30 60 6.0 6.0 4.0 < 1.0 Nitrogen (N) Phosphate (P2O5) 6.0 7.0 6.0 6.5 16 30 kg t-1 3.5 7.0 2.0 5.5 13 25 3.0 2.3 4.0 0.3 kg m-3 1.2 1.2 2.0 trace Phosphate applied at 250 kg ha-1 N 146 250 83 211 203 208 100 130 125 kg m-3 1.5 3.0 4.0 1.5 2.0 3.0 ND = No data 13 0.3 0.5 1.2 50 62 85 BIOENERGY CROPS AND BIOREMEDIATION Table 2.7. Typical total nutrient content and available phosphorus of sludges (kg t-1 or kg m-3 fresh weight) Digested liquid (kg m-3) Digested cake (kg t-1) Thermally dried (kg t-1) Lime stabilised (kg t-1) Dry matter Total nitrogen Total phosphate (%) (N) (P2O5) 4 25 95 40 2.0 7.5 35 6.0 1.5 9.0 45 8.0 Phosphate applied at 250 kg ha-1 N 187 299 321 333 Where applications of these materials are made at the maximum nitrogen rate (250 kg ha-1) recommended in the Code of Good Agricultural Practice for the Protection of Water (MAFF, 1998), large quantities of phosphate will also be added. This is particularly the case with sewage sludges. Many other industrial organic wastes may also have large contents of phosphate. The Water Code suggests that, because of the increased risk of phosphate leaching, applications of phosphate should not exceed crop removal once soil indices reach 4/5. If regular applications of organic wastes are made, phosphate applications will certainly exceed eventual removal in the biomass crop and result in the elevation of soil available phosphate levels. Alriksson (1998) suggests yields of Salix spp. can range from 16-54 t DM ha-1 over a five-year period. Using the highest phosphate content given by Frison (1992) (Table 2.3), this gives a phosphate off-take ranging from 50 to 167 kg ha-1 over 5 years, far less than would be applied by one application of many organic manures. This confirms that there is the potential for rapid increases in soil phosphate levels, and thus potential for phosphate loss by leaching, if regular applications are made. SORGHAL (1997) indicates that experience in Belgium with Miscanthus is that phosphate removal in a 20t DM ha-1 crop is less than 30 kg ha-1 per year. Therefore, if UK codes of practice are to be followed, it can be concluded that biomass crops cannot be used continually for the disposal of high phosphate content organic wastes. 2.1.4 Landfill leachate and other urban aqueous wastes Dilute materials have the advantage of supplying large quantities of water, which can commonly be the limiting factor to the growth of a biomass crop (Samson, 1993), particularly so when the crop is being grown on poor soils, such as those found on restored landfill sites. Application can be made using overhead irrigators, but with some materials this may cause leaf scorch, so trickle irrigation may be the preferred method. Early results from an experiment where landfill leachate is being applied to a SRC crop using trickle irrigation are extremely promising (Farrow, pers. comm.). 14 BIOENERGY CROPS AND BIOREMEDIATION Apart from the possibility of toxic chemicals arising from material disposed of in the landfill, the expected main potentially polluting compounds are ammonia, which is toxic to fish, and salt (NaCl) which is phytotoxic at only moderately high levels (recent work in Sweden is suggesting that it is elevated sodium levels and not chloride which are reducing willow growth). These would alter the ecosystem of fresh water streams, if allowed to contaminate them. Land-based disposal of leachate is common in practice. In greenhouse experiments Wong & Leung (1989), in Hong Kong, found that yields of Acacia confusa were reduced when irrigated with landfill leachate; even when the leachate was diluted with fresh water. However, Cureton et al. (1991) in Canada found they could increase the growth of Salix babylonica and hybrid Populus nigra x maximowiczii by irrigating with landfill leachate in a lysimeter experiment. The leachate used had an electrical conductivity of 10,000 ms cm-1. However, chlorosis of leaves was observed and leaf senescence occurred 5-6 weeks earlier than normal. In what appears to be follow-up work, Shrive et al., 1994 reported on field experiments with the same two species on a clay soil in Ontario. Further investigations by the same group from University of Guelph concentrated on the effect of leachate sprays on the foliage of established trees. It was found that leachate spraying caused a decline in transpiration, but after four years no significant growth effects had been observed. The abstract of this paper concludes that “Treatment and disposal of MSW leachates in tree plantation may offer a low technology, low cost option to municipalities.” Menser et al. 1979 reported that a number of tree species were leachate-tolerant, but in 1983 there was a significant mortality rate in all species tested. Hasselgren (1998) has reported enhanced growth of willow by using landfill leachate in Sweden, that he attributed to the nutrients contained in the leachate. A general conclusion (DoE, 1997) is that waste waters with conductivities greater than 2000-4000 ms cm-1 should not be used for irrigation of trees. Many undiluted leachates will have higher levels. Many leachates contain high levels of nitrogen, and willows have been shown to be effective scavengers for this nutrient (Edwards et al., 1998). It has also been shown (Wong et al., 1990) that allowing landfill leachate to percolate through soil removes many of the potentially harmful substances within the leachate. Thus, any water draining from a site where leachate has been irrigated onto the land is likely to be potentially less harmful to the environment. In Sweden, Johansson & Elowson (1997) have shown that drainage water containing high concentrations of nitrate can be ‘cleaned’ by irrigating to excess on willow plantations. 15 BIOENERGY CROPS AND BIOREMEDIATION 2.1.5 References Alriksson, B (1987). Influence of site factors on Salix growth with emphasis on nitrogen response under different conditions. BioBase European Energy Crops Internetwork. Aschmann, S G; McIntosh, M S; Angle, J S & Hill, R L (1992). Nitrogen movement under a hardwood forest amended with liquid waste water sludge. Agriculture, Ecosystems and Environment 38. 249-263. Boerjesson, P (1999). Environmental effects of energy crop cultivation in Sweden. 1: identification and quantification. Biomass and Bioenergy 16. 137-154. Chambers, B J; Lord, E I; Nicholson, F A & Smith, K A (1999). Predicting nitrogen availability and losses following applications of manures to arable land: MANNER. Soil Use and Management 15(3). 137-143. Chambers, B J & Mitchell, R (1995). Crop Nutrition and Water Relations. In: Arable Energy Coppice (Eds C Britt, M Heath & M Buckland). ADAS, Oxford. 170 pp. Christian, D G & Riche, A B (1998). Nitrate leaching losses under Miscanthus grass planted on a silty clay loam soil. Soil Use and Management 14(3). 131135. Christopherson, N (1996). Tending of short rotation forests – USA. In: Handbook on How to Grow Short Rotation Forests (Eds S Ledin & E Willebrand). Swedish University of Agricultural Sciences/ International Energy Agency (IEA), Uppsala, Sweden. 3.9.1-3.9.2. Cureton, P M; Groenvelt, P H & McBride, R A (1991). Landfill leachate recirculation: Effects on vegetation vigor and clay surface cover infiltration. Journal of Environmental Quality 20. 17-24. DoE (1997). The Potential for Woodland Establishment on Landfill Sites. Edwards, R R; Greaves, M P & Jackson, M B (1998). The potential for the use of willows as components of practical buffer zones. Proceedings of Long Ashton Willow Research Open Day October 1998. Ford-Robertson, J B; Walters, M P & Mitchell, C P (1991). Production and economics of wood fuel crops for energy forestry. Proceedings Wood - Fuel for Thought. ETSU, Harwell, Oxon.. 265-282. Frison, G (1992). Choice of site and establishing short rotation forestry. In: Handbook on How to Grow Short Rotation Forests (Eds S Ledin & A Alriksson). Swedish University of Agricultural Sciences/International Energy Agency (IEA), Uppsala, Sweden. Hall, R L; Allen, S J; Rosier, P T W; Smith, D M; Hodnett, M G; Roberts, J M; Hopkins, R; Davies, H N; Kinniburgh, D G & Goody, D C (1996). Hydrological effects of short rotation energy coppice. ETSU B/W5/00275/REP. Hasselgren, K (1998). Use of municipal waste products in energy forestry: highlights from 15 years of experience. Biomass and Bioenergy 15(1). 71-74. 16 BIOENERGY CROPS AND BIOREMEDIATION Johansson, U & Elowson, S (1997). Cleaning of drainage water from agricultural land using a willow plantation. Proceedings of NJF Seminar No 270, Alternative Use of Agricultural Land; Foulum, Denmark. Johnson, P A & Prince, J M (1991). Changes in organic matter in fen silt soils. In: Advances in Soil Organic Matter Research (Ed. W S Wilson), Royal Society of Chemistry, Cambridge, 1991. Johnson, P A (1995). Utilisation of pig slurry on poplars grown for wood: Effects of husbandry on nitrogen losses by leaching. Proceedings of the ADAS Terrington poplar seminar. July 1995. Johnson, P A (2000). Residual effects of nitrogen applications and weed control techniques on nitrate leaching in a poplar plantation. Report to the Fourth PAMUCEAF Management Committee Meeting, Torun, Poland. Jorgensen, U (1999a). Nitrate leaching from Miscanthus and willow after application of municipal sludge. BioBase European Energy Crops Internetwork Jorgensen, U (1999b). Nitrate leaching from Miscanthus and willow after application of animal manures. BioBase European Energy Crops Internetwork Ledin, S & Willebrand, E (1996) (Eds). Handbook on How to Grow Short Rotation Forests. Swedish University of Agricultural Sciences/International Energy Agency (IEA), Uppsala, Sweden. Ledin, S & Alriksson, A (1996). Choice of site and establishing short rotation forests -Sweden. In: Handbook on How to Grow Short Rotation Forests (Eds S Ledin & E Willebrand). Swedish University of Agricultural Sciences/ International Energy Agency (IEA), Uppsala, Sweden. 2.6.1-2.6.12. MAFF (1998). Code of Good Agricultural Practice for the Protection of Water (The Water Code). Ministry of Agriculture, Fisheries and Food, London. MacLoughlin, M S; Pope, P E & Hansen, E A (1985). Nitrogen fertilisation and ground cover in a hybrid poplar plantation: effects on nitrate leaching. Journal of Environmental Quality 14 (2). 241-245. Malik, R K; Green, T H; Brown, G F & Mays, D (2000). Use of cover crops in short rotation hardwood plantations to control erosion. Biomass and Bioenergy 18. 479-487. Menser, H A; Winant, W M & Bennett, O (1979). Spray irrigation - a land disposal practice for decontaminating leachate from sanitary landfills. Mortensen, J; Nielsen, K H & Jorgensen, U (1998). Nitrate leaching during establishment of willow (Salix viminalis) on two soil types and two fertilization levels. Biomass and Bioenergy 15 (6). 457-466. Riddle-Black, D (1998). Development of a water industry manual for biosolids use in short rotation coppice. Biomass and Bioenergy 15 (1). 101-107. Samson, R; Girouard, P; Omielan, J & Henning, J (1993). Integrated production of warm season grasses and agroforestry for low cost biomass production. In: Proceedings First Biomass Conference of the Americas: Energy, environment, agriculture and industry. Burlington, Vermont. National Renewable Energy Laboratory, Golden, Connecticut. 17 BIOENERGY CROPS AND BIOREMEDIATION Shepherd, M A; Stockdale, E A; Powlson, D S & Jarvis, S C (1996). The influence of organic nitrogen mineralization on the management of agricultural systems in the UK. Soil Use and Management 12 (2). 76-85. Shrive, S C; McBride, R A & Gordon, A M (1994). Photosynthetic and growth responses of two broad leaf tree species to irrigation with municipal landfill leachate. Journal of Environmental Quality 23 534-542. Sidle, R C & Kardos, L T (1979). Nitrate leaching in a sludge treated forest soil. Soil Science Society of America Journal 43. 278-82. SORGHAL (1997). Agronomic aspects of the Miscanthus crop in Belgium. BioBase European Energy Crops Internetwork. Thornton, F C; Joslin, J D; Bock, B R; Houston, A; Green, T H; Schoenholtz, S; Pettry, D & Tyler, D D (1998). Environmental effects of growing woody crops on agricultural land: First year effects on erosion and water quality. Biomass and Bioenergy 15 (1). 57-69. Tolbert, V R & Wright, L L (1998). Environmental enhancement of US biomass crop technologies: research results to date. Biomass and Bioenergy 15 (1). 93100. van Veen, J A; Bretelerm, H; Olie, J J & Frissel, M J (1991). Nitrogen and energy balance of a short rotation poplar forest system. Netherlands Journal of Agricultural Science 29. 163-172. Whitmore, A P; Bradbury, N J & Johnson, P A (1992). Potential contribution of ploughed grassland to nitrate leaching. Agriculture, Ecosystems and Environment 39. 221-233. Wong, M H & Leung, C K (1989). Landfill leachate as irrigation water for trees and vegetable crops. Waste Management and Research 7 (4). 311-324. Wong, M H; Leung, C K & Lan, C Y (1990). Decontamination of landfill leachate by soils with different textures. Biomed Environment Science 3 (4). 429-442. 18 BIOENERGY CROPS AND BIOREMEDIATION 2.2 GASEOUS LOSSES JOHN KING 2.2.1 Gaseous losses of nutrients Of the six major plant nutrient elements (C, H, O, N, P and S) all except phosphorus (P) have a significant gaseous flux element in their normal cycling through the soilplant-air system. Indeed, for C, H and O this is the main method of exchange, in the forms of CO2, H2O and O2, during photosynthesis and respiration. No additional burden on these processes is envisaged due to the addition of organic waste materials to energy crops, with a proviso on water relations, which is considered elsewhere in this review. This leaves the elements of nitrogen and sulphur as possibly having their biogeochemical cycles altered by the addition of organic wastes, such that increased gaseous flux may occur following their application to energy crops. Nitrogen Nitrogen resources may be lost from the soil/organic waste surface in several forms; NH3, N2O, NOx and N2, depending on edaphic conditions and the nature of the organic material applied. The amounts lost as N2O , N2 and NOx are minor and insignificant in nutrient terms, and the N2O flux will be discussed in the following section. Of major concern is the potential loss of nitrogen nutrition to the crop from the efflux of ammonia gas, which is a characteristic of manure applications in agriculture. From the earliest proposals that the valuable nutrient potential of manures should be exploited fully, the problem of post-application losses has also been recognised (Smith & Chambers, 1993). Leaching losses of nitrogen were shown to be controlled more by soil conditions and time of application, whereas gaseous losses occurred within hours and were more a feature of the manure type and its mode of application. Ammonia volatilisation from manures is both a local pollution, problem of objectionable odours, and a contribution to the diffuse pollution of atmospheric nitrogen deposition. It is also responsible for a major loss of nutrients to the crop, being anything from 7 to 84% of slurry ammonium-N applied (Frost et al., 1990). Of this loss 40-50% occurs within 6 hrs, 70% within 24 hours, and over 90% within 5 days of application (Smith & Chambers, 1993). Besides manure factors, soil moisture and environmental conditions (windspeed, temperature and rainfall) also play a part in determining overall losses (Jarvis & Pain, 1990). The most effective abatement practice on this loss for slurries, is to inject the slurry into the soil which reduced losses to 2% of ammonium-N compared with surface applied losses of 74 & 48% (autumn and spring respectively) (Thompson et al., 1987). This is now common practice for applications to grassland, but is not an option open to applications on short rotation coppice sites, or even Miscanthus sites, where woody and perennial root systems are present. Ammonia volatilises from manures, and in particular slurries, because free ammonia (dissolved) is in equilibrium with ammonium ions, and outgasses when the liquid infiltrates into the soil. The equilibrium is such that NH3 is favoured at 19 BIOENERGY CROPS AND BIOREMEDIATION the high pHs found in slurry (pH 7-8) (Harrison & Webb, 2001), and volatilisation will also be aided by higher temperatures and windspeeds (Stevens & Laughlin, 1997). Paradoxically, solid manure does not necessarily emit much lower ammonia concentrations than slurry, especially if the manure is rich in urine. Menzi et al. (1997) measured 60% of the total ammonium-N lost as NH3 from solid manures (compared to 52% for slurry), which constituted 10% of total applied N compared with the 25% for slurry. Chambers et al. (1997) also found a mean emission factor of 65% of applied ammonium-N for FYM (35% for poultry manure). It is the lower proportion of total manure nitrogen which is in the ammonium form in solid manures, which leads to similar, even slightly lower, NH3 emission rates than slurries, even though solid manures tend to contain more total nitrogen (Chambers et al., 1999a). The fact that emissions are largely governed by such a well defined and easily measured fraction of manures, over only a brief period, has allowed ammonia volatilisation to be modeled fairly successfully to determine the nutrient values of manures for following crops. The ADAS model ‘MANNER’ has been used here to obtain typical losses of ammonia-N for various manures applied and also that potentially available to a crop (Chambers et al., 1999a) (Table 2.8). The table assumes a recommended maximum application rate of 250 kg ha-1 N for each manure type (as for Tables 2.10-2.17), on the 1st April at the end of soil drainage, which means that no leaching loss was accounted for. The manures were not incorporated at all and the model made no adjustment for soil type (only in the leaching component). Table 2.8. The calculated ammonia emission and plant available N after surface broadcast applications of various manures to give 250 kg ha-1 N as soon as drainage has ceased in the spring after harvest, according to ‘MANNER’. No Cattle addition FYM Application rate t ha-1 DM Application rate t ha-1 C Application rate kg ha-1 N Pig FYM Cattle slurry Pig slurry Sewage sludge 0.00 0.00 0.00 10.50 4.73 250.00 9.00 4.05 250.00 4.90 2.21 250.00 2.40 1.08 250.00 8.30 3.74 250.00 Potential plant N kg ha-1 0 81 83 138 160 58 Volatilised ammonia N kg ha-1 Available plant N kg ha-1 0 0 40 41 41 42 46 92 39 121 24 34 As can be seen from Table 2.8 the volatilised ammonia loss of potentially available nitrogen to the crop is considerable - between 24 and 46 kg ha-1 for all cases; which constitutes between 32 and 98% of that which is eventually available to the current crop. The figures in this table are also derived from rather benign conditions. If manure is applied earlier in the season, at the end of February or early March, then losses will also be exacerbated by leaching. 20 BIOENERGY CROPS AND BIOREMEDIATION 2.2.2 Emissions of radiatively active gases from soils and organic waste materials The ‘greenhouse effect’ - whereby increased concentrations in the troposphere of certain gases from anthropogenic sources cause the retention of infra-red radiation and thereby global warming of the atmosphere (Dickinson & Cicerone, 1986) - is now a well documented and accepted phenomenon (IPCC, 1997). Concern over this is high enough for international agreements to have been sought limiting emissions of the major gas concerned, CO2 (The ‘Kyoto protocols’). This concern is a major reason for the perceived viability of bioenergy crops as an increasing component of the UK’s energy generation programme - as they are believed to be ‘carbon neutral’ (DTI, 1999). This means that although their combustion in power stations will generate CO2, this will merely re-cycle that which they previously fixed from the atmosphere during photosynthesis, and will once again be fixed by succeeding crops. When considered on a ‘per unit area of land’ basis, this expected neutral balance of carbon input and output will be radically altered by the addition of organic wastes as fertilisers to the crops, during early stages in the rotation. The use of such materials also complicates the picture in that they are known to emit other ‘greenhouse gases’ (depending upon application and edaphic conditions), which are more effective than CO2 at trapping infra-red radiation. The gases in question are chiefly methane (CH4) and nitrous oxide (N2O), which have approximately 32 and 150 times the potential to absorb infra-red radiation respectively, as CO2 (Bouwman, 1990). Houghton et al. (1996) however, put the effect due to N2O as high as 280 times that for CO2, and more recently Smith et al. (2001) quoted the same value for N2O and a CH4 forcing value of 56 CO2 equivalents. All of the gases are associated with organic wastes, though the actual flux emitted will vary according to the nature of the material, edaphic conditions and the way in which it is applied to the soil. Both N2O and CH4 are produced by soil processes that take place under anaerobic conditions, so methods of application that minimise the soil air-space or gaseous exchange will exacerbate their emission. Similarly, the application of very large amounts can lead to anaerobic zones being created. The factors that govern the emission of the two main gases and their possible emission rates will be considered here in separate sections for each gas. Their likely emissions following the application of the major organic waste materials will be considered in turn, and a final section will discuss the likely perturbation to the CO2 flux cycle during the crop rotation, and overall carbon dioxide equivalent balance per unit area of land. Evolution of nitrous oxide (N2O) The production of N2O in soils is a result of natural soil processes and occurs in all soil types at a low level. It is both a minor by-product of the ubiquitous process of nitrification, and also a major product of the process of denitrification which only occurs under anaerobic conditions (Bouwman, 1990). 21 BIOENERGY CROPS AND BIOREMEDIATION During nitrification NO2 is produced in the first reaction step mediated by Nitrosomonas species, according to Equation 1 - before further oxidation by Nitrobacter species to the nitrate ion. Equation 1 NH4+ + 3/2 O2 NO2- + 2H + H2O + E Some of the NO2-, however, is reduced to N2O by nitrifying bacteria, when oxygen is partially limiting or soil acidity is high (Firestone & Davidson, 1989). Therefore some flux of N2O from soil under energy crops should be expected following the ammonification of organic N in plant litter. The magnitude, however, is unknown as emission rates under willow, poplar or Miscanthus plantations have not been measured - but would probably be of the order of 5-25 g N ha-1 d-1, similar to that found under beech (Fagus sylvatica) and alder (Alnus) forests by Mogge et al. (1996) (although denitrification will also contribute to this). Any activity that increases the ammonium concentration in the soil will tend to increase N2O flux during its subsequent nitrification. Therefore, the application of urea and ammonium salt fertiliser will exacerbate this pathway for N2O loss if the edaphic conditions are favourable. The application of nitrogenous organic waste materials to bioenergy plantations may also constitute a net increase in N2O loss by nitrification, but this will be spread over a long period of time, as the material decomposes, and may not be significantly above background rates of emission. Several factors linked to the application of inorganic waste material to soil favour the more productive formative pathway for N2O in the soil, that of denitrification. In this process anaerobic conditions lead to NO3- ions in soil being used as an electron donor for respiration of certain facultative anaerobic micro-organisms. The general reduction pathway for this process is that in Equation. 2, in which N2O is an intermediate result, before the full result of the pathway to molecular nitrogen (Bouwman, 1990). As such it may be emitted from the soil surface, alongside nitrogen gas. Equation 2 NO3- NO2- NO N2O N2 + H2O The microbial requirements for this process are the lack of molecular oxygen, available NO3- ions as a substrate, and an accessible carbon source for energy (Dendooven et al., 1997). Naturally denitrifying organisms must also be present, and other controlling factors are the temperature and pH of the soil medium. The full denitrification process is not a problem for climate change issues, as N2 is radiatively neutral; but factors which increase the ratio of N2O:N2 emission, such as lowered pH, increased access to simple sugars as a C source and periods of anaerobicity, all act to repress NO2- reduction and so lead to a greater efflux of N2O relative to N2 (Dendooven et al., 1997). The emission rate of N2O was reduced by a decline in pH from 6 to 4, because of an increased lag time before NO2- reduction progressed (Ellis et al., 1997). However, below pH 4 denitrification effectively ceases and N2O production in acid forest soils at these low pHs is due mainly to nitrification processes. Although SRC species will tolerate a wide range of soil pH, their yield is reduced on acid soils, and liming to a range of 5.5-8.0 is advised for planting (Britt et al., 1995). Denitrification is therefore unlikely to be inhibited by pH on SRC sites. 22 BIOENERGY CROPS AND BIOREMEDIATION The addition of an organic waste to soil will not only increase the nitrate nitrogen available for denitrification (after mineralisation and ammonification from the organic material), but also the available carbon substrate required. Furthermore, the presence of large quantities of water, such as slurry wastes provide, and the active consumption of oxygen by mineralisation processes, will increase the likelihood and extent of anaerobic sites for denitrification within the soil (Dobbie et al., 1999). This last effect will be exacerbated by the soil compacting activity of manure and/or slurry transporting and spreading machinery (Smith & Chambers, 1993), and be most damaging on already compacted soils such as derelict land or clay soils susceptible to the formation of plough pans. The very act of harvesting the crop in the winter period will lead to compaction on clay sites (Britt et al., 1995), and therefore the conditions during the next rotation will be such that losses of N2O after organic waste application are likely to be high. Just how high is again open to speculation though, as no studies quantifying this loss have been found in the literature, and the following sections extrapolate emission rates from other land use situations. Livestock slurries The application of cattle and pig slurry wastes to AEC land can be seen as a sensible re-cycling of waste material from intensive livestock units with considerable nutrient value to the crop. Slurries can contain of the order of 0.330.44% N, 0.15-0.22% phosphorus and 0.41-0.78% potash on a fresh weight basis (Smith & Chambers, 1993). Approximately half of the nitrogen is in the ammonium form, and considerable quantities can be lost as ammonia gas. Nitrogen lost as N2O from clayey sand and sandy loam soils in Denmark was increased twofold by the application of pig slurry compared with mineral nitrogen fertiliser (Maag et al., 1996). The increase amounted to 1-3 kg ha-1 a-1 N for additions of 25-36 kg ha-1 N as organic N, when 60-90 kg ha-1 N was also applied in mineral form. In earlier studies in Denmark by Christensen (1983), the proportion of applied N lost as N2O, rose from 8.2% when applied as NH4NO3 (200 kg ha-1 N), to 37% when applied as cattle slurry (492 kg ha-1 N) (in Fowler et al., 1997). Over a year this amounted to 182 kg ha-1 N lost as N2O. Emissions of N2O from either mineral or slurry additions to soil are highest immediately after application. Peak values of up to 1.6 kg ha-1 d-1 N were found on a Scottish clay loam by McTaggart et al. (1997), though this fell within 10 days to background levels. In their studies, McTaggart et al. (1997) showed that, when compared with slurry, mineral fertiliser nitrogen was actually the greater source of N2O and that soil compaction was the biggest factor in increasing emissions. However, although slurry in addition to NH4NO3 meant no increase in N2O flux on two occasions, an additional 1.5-2.6 kg ha-1 N flux was apparent in the 60 days after application on the other two occasions (spring and summer), when 75 kg ha -1 N was applied as slurry. In general, however, the N2O loss after the application of slurry constitutes a higher proportion of applied ammonium-N (0.1-4.0%) than that from mineral-N fertiliser (0.1-0.9%) (Eichner, 1990; Harrison & Webb, 2001). The timing of slurry applications has proved a most influential factor in controlling the scale of N2O emissions. Generally emissions (N2O + N2) in the autumn or 23 BIOENERGY CROPS AND BIOREMEDIATION winter ranged from 23-29% of applied ammonium-N compared with 4-5.5% for spring or summer applications (Stevens & Laughlin, 1997). In the autumn and winter, nitrate-N formed by the nitrification of ammonium-N in slurry is not removed by crop uptake and is thereby open to denitrification. In the spring and summer, crop uptake removes nitrate from potential denitrification, and soil moisture deficits also mean anaerobic conditions are less likely. However, after heavy rain or in compacted soil conditions, the higher soil temperatures of summer can lead to high emission rates (McTaggart et al., 1997). Emission rates of N2O (and N2) from slurry have been shown to demonstrate a trend of flux proportional to application rate (Paul et al., 1993), similar to the relationship for mineral N application (Eichner, 1990). However, evidence from field trials is less clear, but supports the presumed trend (Velthof & Onoema, 1993). Measured emission rates from soil amended with slurry are difficult to summarise, due to variable application rates and methods, additional treatments and fertiliser additions, and a range of alternative measurement techniques used. However, Table 2.9 is an attempt to quantify the emissions found relative to the amount of slurry and ammonium-N applied. The figures vary widely, but for typical agricultural total nitrogen applications of 25-125 kg ha-1 in one application, or split into two, an estimated 0.5-15 kg N is lost as N2O in the year after application, broadly in proportion to the amount applied. These figures are generally in agreement, though tending to be higher than the 2% “rule of thumb” value Eichner derived for the proportion of applied manure N lost as N2O. However, in their UK inventory of N2O emissions from farmed livestock, Chadwick et al. (1999) quoted emission factors of 0.4% for pig slurry and 0.3% for cattle slurries, which is only a quarter of those quoted previously. Solid livestock manures Fewer studies have been made of emission rates from solid manures. Although ammonia losses from such manures can be very high, the drier nature of the material means that denitrification, and therefore high N2O losses, are less of a problem. Nevertheless, if heavy rain follows manure application then N2O fluxes can be significant. Coyne et al., 1995 report such a scenario for poultry manure applied to tilled soil at the high rate of 448 kg ha-1 N and measured fluxes of 1.3-3.2 kg ha-1 d-1 N2O-N loss, whereas Cates & Keeney (1987) measured annual losses of 3.6-5.2 kg ha-1 N2O-N. Both of these values appear high compared to those in Table 2.9, but are for very high application rates. Lessard et al. (1996), on the other hand, measured N2O fluxes that amounted to only 1% of applied dairy manure nitrogen under maize. They found efflux rates of 4.1 x 10-2 to 1.2 x 10-1 kg ha-1 d-1 N2O-N from applications of 170 and 339 kg ha-1 N respectively. These were only significantly above background rates in the first seven days after application and 67% of total emissions occurred during the first seven weeks. Set against the general picture from short-term studies, which measure the highest fluxes in the initial stages after application, is the study of Chang et al. (1998), which measured emissions over one year from plots which had received manure 24 BIOENERGY CROPS AND BIOREMEDIATION over 21 years previously. These barley plots had had cattle feedlot manure applied and incorporated every year at rates from 60-180 t ha-1 (as well as 0), and the annual N2O emissions ranged from 11 to 56 kg ha-1 a-1 N2O-N (0.7 kg ha-1 a-1 in the 0 plots). At dry matters of 58% and total N contents of 1.6% dry matter, these amounts again accounted for approximately 2-3% of the total N applied to the plots. A warning is given by this study, that prolonged application of manures can raise emissions, and that predictions based on short-term studies may underestimate efflux in such situations. Chadwick et al. (1999) quoted a seemingly high figure of 5.9% N loss from solid manures when expressed as a percentage of ammonium-N, but as a proportion of total N (typically 10% for solid manures, Menzi et al., 1997) is only 0.6%, which is more in line with their conservative estimates of flux quoted for slurries. Sewage sludge Despite the huge amount of work that has been done on the impact that sewage sludge has on soil microbial processes when applied to land (Smith, 1991), very little is reported in the literature about emissions of greenhouse gases after application. Digested sludge cake has a broadly similar nitrogen content to farmyard manure (Aitken, 1997), and so can be expected to behave similarly in stimulating N2O emission from land by providing both nitrogen and carbon resources to soil nitrification and denitrification processes. In a study where large amounts of sludge cake (16.7 and 83.5 t ha-1 DM) were applied to a sandy loam soil in an arable rotation the mean daily N2O efflux was measured over a five month period from a sown barley crop by Mosier et al. (1982) in Colorado, USA, and compared to similar amounts of N applied as ammonium nitrate fertiliser. At the lower rate (corresponding to 71 kg ha-1 N) rates were of the order of 4 g ha-1 d-1 N2O-N above the control, slightly higher than that for 56-112 kg ha-1 N applied as ammonium nitrate (about 2.5-3.5 g ha-1 d-1). At the higher rate (356 kg ha-1 N), however, 2.2 g ha-1 d-1 N2O-N was emitted above the background, much in excess of the 6 g ha-1 d-1 emitted after 224 kg ha-1 of ammonium nitrate N. Over the full period this amounted to emissions of 0.8 and 1.0% of the applied N evolved as N2O. As for livestock wastes, emissions were highest in the initial post-application period and 69% of those over the 155 day measurement period evolved during the first six weeks. Peaks after this followed fluctuations in soil water content. The effect of rainfall events in leading to peaks in N2O emission from grassland after sewage sludge application, was shown in the only reported UK study by Scott et al. (1998). The full report of this experiment (Scott et al., 2000), details the emission pattern after rates of 185 t DM ha-1 sludge (1.37% N) had been applied for three years (2,535 kg ha-1 a-1 N) to grassland, on an imperfectly drained sandy clay loam soil in southern Scotland. Initial peak diurnal fluxes of 48 to 120 g ha -1 d-1 N2O-N were experienced, which declined to about 10 g ha-1 d-1 above background after about 30 days. This was punctuated though by similar order peaks, following rainfall events, to those initially experienced, illustrating the controlling influence of the interaction between soil permeability and incident rainfall. The cumulative total efflux over a period of six months from August to March, was 10-20 kg ha-1 N2O-N above that from plots fertilised at a similar rate with ammonium nitrate (480 kg ha-1 a-1 N) and represents 0.4-0.8% of the N applied as sludge. 25 BIOENERGY CROPS AND BIOREMEDIATION Table 2.9 Nitrous oxide emission rates from livestock slurry manures cited in the literature Soil type Additional features Slurry type Sandy loam Grassland Dairy Amount of total N applied (kg ha-1 d-1 N) 492 Sandy loam Grassland Cattle 200 Sandy Grassland Dairy 1.4-4.2 Sedge Peat (38% OM) Clayey sand Grassland Cattle 0.7-0.9 Arable Cattle 36-45 Sandy loam Clay loam Arable Imperfectly drained grassland Free draining grassland Arable unknown 25-54 44-75 Dairy 44-124 Sandy loam Sandy loam Sandy loam over Loamy sand Silty clay loam Sandy Amount applied (t ha-1 DM) 45.1-50.1 Pig Amount of NH4+-N applied (kg ha-1 d-1 N) 27.7-72.9 9.6 24.3-25.6 11-380 Pig 100-162 Arable Pig 225 Arable Cattle & Pig 80-100 Long term flux rate (g ha-1 d-1 N) Estimated flux over one year (kg ha-1 a-1 N) 182 0.23-530 0.4-5.3 11-200 18.2 1-5 1-85 1-2 0.4-31 0.56-2.2 0.2-0.8 25-42 9.1-15.3 2.5-12 0.52-1.82 30 10 0.3-1.3 5-25 3 1.1-1.8 0.2-1.6 32-56 66-90 Arable Short term N2O flux rate (g ha-1 d-1 N) 12-180 83-96 52-65 26 Reference Christensen, 1983 Egginton & Smith, 1986 Velthof & Onoema 1993 Jars et al., 1994 Maag et al., 1996 “ McTaggart et al., 1997 Chadwick, 1997 Ferm et al., 1999 Weslien et al., 1998 Arcara et al., 1999 Petersen 1999 BIOENERGY CROPS AND BIOREMEDIATION The above evidence suggests that over a full year the emission of nitrogen as N2O is likely to be of the order of 1-2% of that applied, similar to livestock wastes. Evolution of methane Methane (CH4) is also produced in soils under anaerobic conditions, when the inorganic hydrogen acceptor ions such as the nitrates have been used up (section 2.1) and the fermentation of organic molecules commences (Bouwman, 1990). Nitrates repress this formation by both delaying the onset until they have been reduced, but also by direct toxicity to methanogenesis. Sulphates will also act similarly. Methane production after the application of large quantities of organic wastes to soil, will therefore only be likely under prolonged anaerobic conditions. Methanogenesis can occur mainly by two processes, either using acetates as a substrate according to Equation 3, or by the reduction of CO2 as in Equation 4 (Batjes & Bridges, 1992): Equation 3 Equation 4 CH3COOH > CH4 + CO2 CO2 + 4H2 > CH4 + 2H2O. Methane is also widely consumed by methanotrophic organisms in aerobic soil (Batjes & Bridges, 1992) and by some ammonium oxidising organisms. Aerobic soil can therefore act as a sink rather than a source of methane (Schütz et al., 1990), its presence inducing greater growth and activity of methanotrophic organisms (Bender & Conrad, 1995). For these reasons methane emissions are usually only associated with natural wetlands or rice paddy soils where anaerobic conditions are maintained as an equilibrium, and their effects outweigh that of any aerobic topsoil. Certain situations such as landfill sites where large quantities of organic material are buried under anaerobic conditions also become net methane emitters. Methane from livestock and sewage wastes The huge production of animal and human waste also contributes to the global budget of atmospheric methane, both directly from ruminant animals in the field, but also the storage and processing of waste materials. Generally livestock wastes are highly conducive to methane generation, containing both the organic substrates required and suitable active microorganisms. The actual amounts produced are however highly dependent on the diet of the producer animals and the way in which the manure is treated. Grain-fed housed animals have been found to produce more methane than those kept extensively on low grade forage (Hogan, 1993), and Jarvis et al. (1995) found an inverse relationship between production and the C:N ratio of cattle dung. The main controlling factor in the handling of livestock wastes for methane production is undoubtedly the way in which it is stored, and liquid based storage systems (slurry, etc.) produce the most. Contact with oxygen will suppress methane production for the reasons given above, 27 BIOENERGY CROPS AND BIOREMEDIATION as will extremes of pH, because methanogens require conditions around neutrality (pH 6.6-8.0; Hogan, 1993). Hardly any citations of methane emissions from field-spread manures were found (Sommer et al. (1996) (and none for sewage sludge) though some workers had measured the efflux from cattle and sheep dung (Lodman et al., 1993; Williams, 1993; Jarvis et al., 1995). These studies produced broadly comparable results, that emissions were highest immediately after deposition and lasted over a period of about 10 days, declining with increasing aerobicity of the dung. Values ranged from about 300-2000 mg CH4 m-2 in the field, varying according to type of animal and feeding regime; but amounting roughly to a mean of 74 mg kg-1 wet weight of application. By contrast Willison et al. (1996) found that additions of FYM to soil increased the methanotrophic activity by ammonium fertilisation from mineralising organic nitrogen! Any emissions from applied manure or sewage sludge are therefore likely to be small - highly dependent on both the amount and type of material applied and the soil hydrology immediately after application, and limited to the immediate post application period (1014 days). Indeed, Sommer et al. (1996) only measured 30 g of CH4-C ha-1 emitted from pig slurry spread on bare soil, and all of that within the first day after application. Neither Sommer et al. (1996), nor Jarvis et al. (1995) found that the soil hydrology altered emissions, which suggests that it is only a temporary event dependant entirely on the conditions of the manure, supporting the view that some at least is dissolved CH4 volatilising out of the manure, rather than true methanogenesis (Sommer et al. (1996). A predictive emission rate is impossible to give and only estimates for livestock wastes can be attempted. National and international inventories for greenhouse gas emissions (Hogan, 1993; IPCC, 1997) tend to estimate emissions across the entire manure-handling chain. FYM drylot storage and slurry/lagoon systems with higher emission factors produce most methane during the storage phase. Although it should be noted that the use of manures entails this necessary step, and thereby CH4 emission, this would happen regardless of their use on energy crop sites. Consequently, it should not affect the argument about carbon flux mitigation by energy cropping being reduced by on-site greenhouse gas emissions. However, to produce an estimate for this report we have used generic figures for the amount of volatile solids (VS) in all types of fresh manure (10-12% dry matter) and a methane conversion factor (MCF) for the daily spread or pasture manure handling systems (0.1-2%, over 10-300C range), taken from Hogan (1993) (from the work of Hashimoto, also used by the IPCC, 1997). Soil carbon balance The amount of carbon added to the soil in various organic wastes will, of course, depend upon their composition and the amounts applied. However, the mineralisation of these organic resources follows a fairly predictable pattern of the mineralisation of any vegetation derived material, described roughly by an exponential relationship (Jenkinson, 1990). As a rule of thumb, 66% of added vegetable material is lost as CO2 within a year of 28 BIOENERGY CROPS AND BIOREMEDIATION incorporation. During this year a higher proportion will be lost immediately after incorporation and a smaller proportion will be cycled through the soil microbiota and into the soil organic matter (SOM). In agricultural soils whose organic matter content is at equilibrium, this amount added replaces that lost from mineralisation of soil organic matter for annual additions. Models for carbon turnover exist, such as that of Jenkinson (1990), which can be used to predict the decline in added carbon (and the new soil organic matter equilibrium) over a crop rotation, but an approximation of 10% has been used here. This represents the amount remaining after two years at the initial rate of loss, but it is recognised that this rate declines exponentially as easily decomposed material is respired and only recalcitrant matter remains, and 10% is seen as a median value for recalcitrant material which may remain for several years. In addition to any increase in equilibrium soil organic matter level due to organic waste application, there will be a small increment from litter deposition and turnover. Litter deposition from willows grown on a Swedish peat bog showed slightly retarded decomposition dynamics compared to the “rule of thumb” mentioned above, losing about half their mass over three years and only declining at a very low rate after that (Slapokas and Granhall, 1991). Given that temperatures will be lower and the peat surface wetter in their study than most UK mineral soil sites, we can still assume that up to half the deposited litter carbon will still comprise a deep litter mulch on the soil surface after three years, at the end of the harvest rotation. If leaf litter constitutes 30-40% of the above ground woody biomass, as is the case for alder plantations (Rytter et al., 1989), then willow crops producing a typical annual yield increment of 12 t ha -1 DM will deposit about 11-14 t ha-1 DM as litter over a three year rotation of which half may constitute a standing litter layer from the first harvest onwards (about 5-7 t ha-1 DM). This layer, and the turnover of fine roots, will also contribute an estimated 4-10 t ha-1 DM annual turnover, leading to a small increment in soil organic matter of the order of 0.5 t ha-1 a-1 carbon (Börjesson, 1999). There will also be a net increase in below ground carbon during the first energy crop rotation, caused by the development of a semi-permanent woody root system. In most trees the root system can be the sink for a considerable proportion of the annual carbon increment; for example 11% for fine roots and 31% for coarse roots were recorded in a grey alder plantation in Sweden (Elowson & Rytter, 1993), but the coarse root standing biomass tends to stabilise around 20% over all species (Ericsson et al., 1996). Using this mean figure, and typical UK willow annual biomass production of 12 t ha-1 a-1 DM (Bullard, pers. comm.), then 2.4 t ha-1 DM will be deposited after one year and 9.6 t ha-1 DM at the first harvest of willow after four years. This latter figure is likely to be the standing mass throughout the remainder of the 25 years of the full economic rotation of an energy crop plantation. The energy grass Miscanthus apportions an even greater component of carbon allocation to its root system in the initial building phase of growth, putting about 1 t ha-1 DM in the first year and 810 t ha-1 DM after two years, until it develops a standing root biomass of 29 BIOENERGY CROPS AND BIOREMEDIATION about 15 t ha-1 DM in the third year. The root mass remains at this level for the remainder of the 25 year plantation, for a crop delivering 15 t ha-1 a-1 DM of shoot material (Bullard, pers. comm.). The carbon-equivalence balance of a site Using guide figures from the literature detailed above, the net effects of typical energy crops and organic waste fertiliser practice, on the contribution to radiatively active gases in the atmosphere, and carbon sequestration, can be estimated on a site basis. To do this, one mole of methane has been taken as being equivalent to 56 moles of CO2 and one mole of N2O as being equivalent to 280 moles of CO2 (Smith et al., 2001). Increments of carbon into a site are considered a positive gain to carbon sequestration, whilst losses of greenhouse gases are considered a loss to carbon sequestration value, and for all organic waste and vegetation materials the carbon concentration has been taken as a universal mean of 45% DM (Palm & Rowland, 1997). Livestock wastes and sewage sludge have been assumed to be applied at the maximum loading allowed for arable land which gives an equivalent of 250 kg ha-1 N (Chambers et al., 1999b) using typical nitrogen contents (Anon, 2000). For cattle and pig FYM and digested sewage sludge cake, this means application rates of 10.5, 9.0 and 8.3 t ha-1 DM respectively, and for cattle and pig slurry, 4.9 and 2.4 t ha-1 DM. The suggested rotation for willow coppice is that they are cut after one year (the cuttings left on site), when they can be fertilised by an application of organic waste material, and thereafter every three years until the full economic rotation of 25 years has elapsed. Organic waste materials would be applied after every three-year harvest. The carbon sequestration potential for such a site is calculated in Table 2.10 for the first year, Table 2.11 for the first harvest period, and in Table 2.12 for the full 25 year period. In the first year with no waste application, all plots register a small positive contribution to carbon sequestration on the site (Table 2.10). In the fourth year after one application of waste, there has been a net contribution of carbon to the site by all treatments, but the effect of potential greenhouse gas emissions has been to reduce the effectiveness of this to a negative impact, relative to a site with no addition (Table 2.11). Over the full economic rotation this effect becomes even more marked, reducing the benefits of carbon sequestration by between 27 and 36%. A similar scenario for Miscanthus, gives the values in Table 2.13, 2.14 & 2.15 respectively, again only applying organic wastes in every third year, even though Miscanthus is harvested every year. The amount of carbon sequestered is less than willow in the early stages (due mainly to no standing litter being accounted for, and a smaller root system), but the same effect of greenhouse gas emissions reducing the effectiveness of this sequestration is also apparent (Tables 2.12 & 2.13). After 25 years the 30 BIOENERGY CROPS AND BIOREMEDIATION carbon sequestered is slightly greater than willow coppice, and again its effectiveness is reduced by 24-31% (Table 2.13). If wastes are applied at every opportunity then the carbon sequestration potential increases considerably, even after only four years (Table 2.16). However, the attenuation of this by greenhouse gas emission also increases, as can be seen from the 30-39% reduction in effectiveness displayed in Table 2.16. Over the full 25 years, this amounts to large quantities of extra carbon in the topsoil (Table 2.17), but a 68-75% negation of the benefits of this with respect to greenhouse gas production. This is because, all the losses of N2O and CH4 occur in the initial post application period and, in the annual application scenario of Table 2.17, the annual emission of these gases overwhelmingly outweighs the minor accrual of soil carbon. What is not taken into account in the balances given in Tables 2.10-2.17 is the CO2 emissions from the 90% of applied organic waste that mineralises, nor the carbon in the harvestable biomass of crop. The crop biomass is considered to be entirely neutral in that the same amount of carbon taken out of the atmosphere in its formation is returned on incineration (and through plant respiration). The CO2 respired during organic waste respiration is a greenhouse gas emitted from the site, but it was considered that this would have been returned to the atmosphere whatever the treatment of the material either by mineralisation or incineration. The accrual of soil organic matter from its application is a genuine, though temporary, sequestration of carbon to the site which would not otherwise occur, and the contribution from N2O and CH4 to greenhouse gas emissions, are considered to be different to CO2, as their production is more dependant upon handling and management issues, and technically need not occur if perfectly aerobic conditions could be maintained. The estimates in Tables 2.10-2.12, of the impact of various waste additions on the balance of carbon sequestration and greenhouse gas emissions from an SRC site, are based upon very rough figures from work in normal agricultural contexts. As such, they may not necessarily reflect the situation when large amounts of waste are disposed of to coppice sites on reclaimed land, where soil compaction by reclamation plant or large-scale harvesting equipment may create a soil hydrology that exacerbates the likelihood of greenhouse gas production. Nor do they take any account of other possible waste applications that involve irrigation (by waste water or landfill leachate, etc.) on a regular basis, which again will create conditions conducive to greenhouse gas emissions. Of particular concern would be the annual application of waste to Miscanthus, which is likely to produce even worse conditions than suggested in Tables 2.16 and 2.17 - as the accrual of organic matter from the waste is probably under-estimated in these tables, and this would act to retain moisture and make anaerobiosis more frequent and extensive. With these provisos however, the practice of applying moderate amounts of organic wastes to energy crops is likely to be both beneficial to the soil 31 BIOENERGY CROPS AND BIOREMEDIATION sustainability of the site and make a modest contribution of carbon sequestration to the overall balance of factors which contribute to the forcing of climate change. 32 BIOENERGY CROPS AND BIOREMEDIATION Table 2.10. Carbon equivalent fluxes (C t ha-1) on a willow coppice site one year after planting. 1st year - willow No addition Cattle FYM Pig FYM Cattle slurry Pig slurry Sewage sludge Application rate t ha-1 DM Application rate t ha-1 C Application rate kg ha-1 N 0.00 0.00 0.00 10.50 4.73 250.00 9.00 4.05 250.00 4.90 2.21 250.00 2.40 1.08 250.00 8.30 3.74 250.00 C in litter (6 t ha-1 DM) C in roots (2.4 t ha-1 DM) C in SOM (0.5 t ha-1 yr-1) 2.70 1.08 0.50 2.70 1.08 0.50 2.70 1.08 0.50 2.70 1.08 0.50 2.70 1.08 0.50 2.70 1.08 0.50 N20 efflux (t ha-1 CO2-C equivalent) CH4 efflux (t ha-1 CO2-C equivalent) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Total C sequestered Site balance of C sequestration 4.28 4.28 4.28 4.28 4.28 4.28 4.28 4.28 4.28 4.28 4.28 4.28 Table 2.11. Carbon equivalent fluxes (C t ha-1) on a willow coppice site after one harvest, four years after planting, with organic wastes applied after one year. Application rates the same as Table 2.10. 4th year - willow No addition Cattle FYM Pig FYM Cattle slurry C in litter (6 t ha-1 DM) C in roots (9.6 t ha-1 DM) C in SOM (0.5 t ha-1 yr-1 ) C from waste to SOM (10%) 2.70 4.32 2.00 0.00 2.70 4.32 2.00 0.47 2.70 4.32 2.00 0.41 2.70 4.32 2.00 0.22 2.70 4.32 2.00 0.11 2.70 4.32 2.00 0.37 N20 efflux (t ha-1 CO2-C equivalent) CH4 efflux (t ha-1 CO2-C equivalent) 0.00 0.00 0.60 0.44 0.60 0.37 0.60 0.20 0.60 0.10 0.60 0.35 Total C sequestered Site balance of C sequestration 9.02 9.02 9.49 8.46 9.43 8.45 9.24 8.44 9.13 8.43 9.39 8.45 33 Pig slurry Sewage sludge BIOENERGY CROPS AND BIOREMEDIATION Table 2.12. Carbon equivalent fluxes (C t ha-1) on a willow coppice site after the full economic rotation of 25 years, with organic wastes applied after one year, and again after every three year harvest. Application rates the same as Table 2.10. 25th year - willow C in litter (6 t ha-1 DM) C in roots (2.4 t ha-1 DM) C in SOM (0.5 t ha-1 yr-1) C from waste to SOM (10%) N20 efflux (t ha-1 CO2-C equivalent) CH4 efflux (t ha-1 CO2-C equivalent) Total C sequestered Site balance of C sequestration No addition Cattle FYM Pig FYM Cattle slurry Pig slurry Sewage sludge 2.70 4.32 12.50 0.00 2.70 4.32 12.50 3.78 2.70 4.32 12.50 3.24 2.70 4.32 12.50 1.76 2.70 4.32 12.50 0.86 2.70 4.32 12.50 2.99 0.00 0.00 4.80 3.49 4.80 2.99 4.80 1.63 4.80 0.80 4.80 2.76 19.52 19.52 23.30 15.01 22.76 14.97 21.28 14.85 20.38 14.79 22.51 14.95 Table 2.13. Carbon equivalent fluxes (C t ha-1) on a Miscanthus plantation one year after planting. 1st year - Miscanthus No addition Cattle FYM Pig FYM Cattle slurry Application rate t ha-1 DM Application rate t ha-1 C Application rate kg ha-1 N 0.00 0.00 0.00 10.50 4.73 250.00 9.00 4.05 250.00 4.90 2.21 250.00 2.40 1.08 250.00 8.30 3.74 250.00 C in litter (0 t ha-1 DM) C in roots (0.8 t ha-1 DM) C in SOM (0.5 t ha-1 yr-1) 0.00 0.36 0.50 0.00 0.36 0.50 0.00 0.36 0.50 0.00 0.36 0.50 0.00 0.36 0.50 0.00 0.36 0.50 N20 efflux (t ha-1 CO2-C equivalent) CH4 efflux (t ha-1 CO2-C equivalent) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Total C sequestered Site balance of C sequestration 0.86 0.86 0.86 0.86 0.86 0.86 0.86 0.86 0.86 0.86 0.86 0.86 34 Pig slurry Sewage sludge BIOENERGY CROPS AND BIOREMEDIATION Table 2.14. Carbon equivalent fluxes (C t ha-1) on a Miscanthus plantation four years after planting, with organic wastes applied after one year. Application rates the same as Table 2.13. 4th year - Miscanthus No addition Cattle FYM Pig FYM Cattle slurry Pig slurry Sewage sludge C in litter (6 t ha-1 DM) C in roots (9 t ha-1 DM) C in SOM (0.5 t ha-1 yr-1) C from waste to SOM (10%) 2.70 4.05 2.00 0.00 2.70 4.05 2.00 0.47 2.70 4.05 2.00 0.41 2.70 4.05 2.00 0.22 2.70 4.05 2.00 0.11 2.70 4.05 2.00 0.37 N20 efflux (t ha-1 CO2-C equivalent) CH4 efflux (t ha-1 CO2-C equivalent) 0.00 0.00 0.60 0.44 0.60 0.37 0.60 0.20 0.60 0.10 0.60 0.35 Total C sequestered Site balance of C sequestration 8.75 8.75 9.22 8.19 9.16 8.18 8.97 8.17 8.86 8.16 9.12 8.18 Table 2.15. Carbon equivalent fluxes (C t ha-1) on a Miscanthus plantation after the full economic rotation of 25 years, with organic wastes applied after one year, and again every three years. Application rates the same as Table 2.13. 25th year - Miscanthus C in litter (6 t ha-1 DM) C in roots (15 t ha-1 DM) C in SOM (0.5 t ha-1 yr-1) C from waste to SOM (10%) N20 efflux (t ha-1 CO2-C equivalent) CH4 efflux (t ha-1 CO2-C equivalent) Total C sequestered Site balance of C sequestration No addition Cattle FYM Pig FYM Cattle Slurry Pig Slurry Sewage Sludge 2.70 6.75 12.50 0.00 2.70 6.75 12.50 3.78 2.70 6.75 12.50 3.24 2.70 6.75 12.50 1.76 2.70 6.75 12.50 0.86 2.70 6.75 12.50 2.99 0.00 0.00 4.80 3.49 4.80 2.99 4.80 1.63 4.80 0.80 4.80 2.76 21.95 21.95 25.73 17.44 25.19 17.40 23.71 17.28 22.81 17.22 24.94 17.38 35 BIOENERGY CROPS AND BIOREMEDIATION Table 2.16. Carbon equivalent fluxes (C t ha-1) on a Miscanthus plantation four years after planting, with organic wastes applied every year, after year one. Application rates the same as Table 2.13. 4th year - Miscanthus No addition Cattle FYM Pig FYM Cattle slurry Pig slurry Sewage sludge C in litter (6 t ha-1 DM) C in roots (9 t ha-1 DM) C in SOM (0.5 t ha-1 yr-1) C from waste to SOM (10%) 2.70 4.05 2.00 0.00 2.70 4.05 2.00 1.89 2.70 4.05 2.00 1.62 2.70 4.05 2.00 0.88 2.70 4.05 2.00 0.43 2.70 4.05 2.00 1.49 N20 efflux (t ha-1 CO2-C equivalent) CH4 efflux (t ha-1 CO2-C equivalent) 0.00 0.00 2.40 1.75 2.40 1.50 2.40 0.81 2.40 0.40 2.40 1.38 Total C sequestered Site balance of C sequestration 8.75 8.75 10.64 6.49 10.37 6.47 9.63 6.42 9.18 6.38 10.24 6.46 Table 2.17. Carbon equivalent fluxes (C t ha-1) on a Miscanthus plantation after the full economic rotation of 25 years, with organic wastes applied every year, after year one. Application rates the same as Table 2.13. 25th year - Miscanthus C in litter (6 t ha-1 DM) C in roots (15 t ha-1 DM) C in SOM (0.5 t ha-1 yr-1) C from waste to SOM (10%) N20 efflux (t ha-1 CO2-C equivalent) CH4 efflux (t ha-1 CO2-C equivalent) Total C sequestered Site balance of C sequestration No addition Cattle FYM Pig FYM Cattle slurry 2.70 6.75 12.50 0.00 2.70 6.75 12.50 11.34 2.70 6.75 12.50 9.72 2.70 6.75 12.50 5.29 2.70 6.75 12.50 2.59 2.70 6.75 12.50 8.96 0.00 0.00 14.40 10.48 14.40 8.98 14.40 4.89 14.40 2.40 14.40 8.28 21.95 21.95 33.29 8.41 31.67 8.29 27.24 7.95 24.54 7.75 30.91 8.23 36 Pig slurry Sewage sludge BIOENERGY CROPS AND BIOREMEDIATION 2.2.3 References Aitken, M N (1997). Use of sewage sludge on agricultural land. SAC Technical Note T450. Scottish Agricultural College, Edinburgh. Anon. (2000). Fertiliser Recommendations for Agricultural and Horticultural Crops (RB209). The Stationary Office, London. 177 pp. Arcara, P G; Gamba, C; Bidini, D & Marchetti, R (1999). 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Nitrous oxide emissions from barley plots treated with ammonium nitrate or sewage sludge. Journal of Environmental Quality 11. 78-81. Paul, J W; Beauchamp, E G & Zhang, X (1993). Nitrous and nitric oxide emissions during nitrification and denitrification from manureamended soil in the laboratory. Canadian Journal of Soil Science 73. 539-553. Palm, C A & Rowland, A P (1997). A minimum dataset for characterization of plant quality for decomposition. In: Driven by Nature: Plant litter quality and decomposition (Eds. G Cadisch & K Giller). CABI, Wallingford, Oxon.. 379-392. Petersen, S O (1999). Nitrous oxide emissions from manure and inorganic fertilisers applied to spring barley. Journal of Environmental Quality, 28. 1610-1618. Rytter, L; Slapokas’ T & Granhall, U (1989). Woody biomass and litter production of a fertilised grey alder plantations on a low-humified peat bog. Forest Ecology and Management, 28. 161-176. Schütz, H; Seiler, W & Renneberg, H (1990). Soil and land use related sources and sinks of methane (CH4) in the context of the global methane budget. In, Soils and the Greenhouse Effect (Ed. A Bouwman). John Wiley and Sons, Chichester, UK. 269-288. Scott, A; Ball, B C; Crichton, I J & Aitken, M N (1998). Short communication - Nitrous oxide emissions from grassland amended with sewage sludge. Soil Use and Management 14. p 55. Scott, A; Ball, B C; Crichton I J & Aitken M N (2000). Nitrous oxide and carbon dioxide emissions from grassland amended with sewage sludge. Soil Use and Management 16. 36-41. Slapokas, T & Granhall, U (1991). Decomposition of litter in fertilised short-rotation forests on a low-humified peat bog. Forest Ecology & Management 41. 143-165. 40 BIOENERGY CROPS AND BIOREMEDIATION Smith, K A & Chambers, B J (1993). Utilising the nitrogen content of organic manures on farms - problems and practical solutions. Soil Use and Management 9. 105-112. Smith, S R (1991). Effects of sewage sludge application on soil microbial processes and soil fertility. Advances in Soil Science 16. 191-212. Smith, P; Goulding, K W T; Smith, K A; Powlson, D S; Smith, J U; Falloon, P & Coleman, K (2001). Including trace gas fluxes in estimates of the carbon mitigation potential of UK agricultural land. Soil Use and Management 16. 251-259. Sommer, S G; Sherlock, R R & Khan, R Z (1996). Nitrous oxide and methane emissions from pig slurry amended soils. Soil Biology and Biochemistry 28. 1541-1544. Stevens, R J & Laughlin, R J (1997). The impact of cattle slurries and their management on ammonia and nitrous oxide emissions from grassland. In: Gaseous Emissions from Grasslands (Eds. S C Jarvis. & B F Pain). CABI, Wallingford, Oxon. 233-256. Thompson, R B; Ryden, J C & Lockyer, D R (1987). Fate of nitrogen in cattle slurry following surface application or injection to grassland. Journal of Soil Science 38. 689-700. Velthof, G L & Onoema, O (1993). Nitrous oxide flux from nitric-acidtreated cattle slurry applied to grassland and semi-controlled conditions. Netherlands Journal of Agricultural Science 41. 81-93. Weslien, P; Klemedtsson, L; Svensson, L; Galle, B; KasimirKlemedtsson, A & Gustafsson, A (1998). Nitrogen losses following application of pig slurry to arable land. Soil Use and Management 14. 200-208. Williams, D J (1993). Methane emissions from manure of free-range dairy cows. Chemosphere 26. 179-187. Willison, T W; Cook, R; Müller, A; Powlson, D S (1996). CH4 oxidation in soils fertilised with organic and inorganic-N; differential effects. Soil Biology and Biochemistry 28. 135-136. 41 BIOENERGY CROPS AND BIOREMEDIATION 2.3 HEAVY METALS AND OTHER CONTAMINANTS FIONA NICHOLSON 2.3.1 Introduction Livestock manures and other organic wastes applied to energy crops are an important source of plant nutrients such as nitrogen (N) and phosphorus (P), and also supply valuable quantities of organic matter to the soil. However, they may also contain a range of contaminants that can be harmful to plant, ecosystem or human health. Heavy metals Among the most important contaminants are the heavy metals. Zinc (Zn), copper (Cu) and nickel (Ni) are essential trace elements for plant growth, although they can be phytotoxic if present at excessive levels in the soil (Alloway, 1990). However, chromium (Cr), cadmium (Cd), lead (Pb), arsenic (As) and mercury (Hg) have no known biological function and can be harmful if they enter the wider environment. Because heavy metals cannot be broken down into less harmful by-products, phytoremediation strategies focus on their accumulation in above-ground plant parts and subsequent removal from the contaminated site. Heavy metals may be present in different forms in manures and organic wastes, which could affect the amount and rate available for plant uptake. For example, metals in animal manures will be present mainly in soluble forms or bound to organic matter. In contrast, biosolids or industrial wastes may contain a large proportion of metals strongly bound to mineral particles (in particular Fe, Al and Mn oxides), that are much less available to plants. Once incorporated into the soil, a number of soil properties will affect plant metal uptake including pH, cation exchange capacity, redox conditions, organic matter content and clay content. Organic contaminants Other contaminants often present in waste materials include a wide range of organic compounds with the potential to exert a health or environmental hazard. These include polynuclear aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), phthalate acid esters, phenols, polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs), herbicides and organochlorine pesticides, which all have different toxicities and environmental effects. In contrast to heavy metals, organic pollutants can often be completely broken down by plants into less harmful metabolites. There is no evidence to suggest that organic contaminants from different wastes will behave differently once in the soil, however, soil properties may affect plant uptake and degradation rates. 42 BIOENERGY CROPS AND BIOREMEDIATION Other contaminants Manures and wastes may also contain pathogenic micro-organisms which have the potential to cause plant, animal or human disease. This form of contamination is not considered further in this review, although it may be an important issue when considering the application of livestock and other organic manures to biomass crops. 2.3.2 Concentrations of contaminants in livestock manures and other wastes Livestock manures There is good recent information available on heavy metal concentrations in livestock manures collected from farms in England and Wales (Nicholson et al., 1999). This study of 85 manure samples found that the Zn and Cu concentrations of pig and poultry manures were higher than for cattle manures due to supplementation of these metals in pig and poultry diets (Table 2.18). Heavy metal loading rates where manures are applied at Code of Practice (MAFF, 1998) rates of 250 kg ha-1 total N were calculated and are also given in Table 2.18. There is much less comprehensive information available on the levels of organic contaminants in manures, and very little from the UK. Raszyk et al., 1998 looked at concentrations of 16 PAHs in feedstuffs, drinking water, stable dust, pig slurry, road dust and soil on three pig farms and two cattle farms in the Czech Republic in 1995 and 1996. The average sum of 16 PAHs for pig slurry was 543.2 g kg-1 dry matter, the average sum of seven carcinogenic PAHs was 47.0 g kg-1 dry matter and the average BaP concentration was 2.3 mu g kg-1 of dry matter. Chrysene, benzo(a)anthracene and benzo(b)fluoranthene were the dominant PAH carcinogens on pig and cattle farms, whilst among the other PAHs, phenanthrene, fluoranthene and pyrene were dominant. In Switzerland, Berset & Holzer, 1995 reviewed the present state of contamination of agricultural soils and manures (sewage sludge, liquid manure, and compost) with PAHs and PCBs. Overall PAH concentrations in cattle slurries were 87-309 g kg-1 (mean 165 g kg-1), pig slurries 66339 g kg-1 (mean 143 g kg-1), sewage sludge 1.7-15 mg kg-1 (mean 6.3 mg kg-1) and compost 0.8-2.7 mg kg-1 (mean 2 mg kg-1). PCB concentrations were 20 g kg-1 in cattle slurries, 37 g kg-1 in pig slurries and 32 g kg-1 in compost. A qualitative analysis of the environmental samples showed that besides the 16 PAHs frequently used for quantification, mainly alkylated derivatives as well as N-S- and-O-PAHs were detected. Some work has also been undertaken looking at the inputs and outputs of various organic contaminants in dairy cattle. A three month study in the UK found that the total PCB content (53 congeners) in fresh faeces from lactating dairy cows averaged 1.3 ( 0.42) g kg-1 (Thomas et al., 1999). 43 BIOENERGY CROPS AND BIOREMEDIATION Preliminary, unpublished data from Lancaster University (G. Thomas, pers. comm.) suggest faeces concentrations of 31.2 ng kg-1 furans and 62.5 ng kg-1 dioxins. Other studies have quantified the percentage of organic contaminant intake which was measured in manure, although manure concentrations were not specifically reported (Welsch-Pausch & McLachlan, 1998; McLachlan & Richter, 1998; Stephens et al., 1995). From the limited information available, it appears that levels of PCDD/Fs, PAHs and PCBs are much lower in animal manures than in biosolids. In addition, animal manures are often stored for relatively long periods prior to land spreading (6 -12 months) and during this time it would be expected that some organic contaminants would degrade, and the concentrations reduced compared to fresh excreta. Biosolids Heavy metals may often be present at relatively high levels in biosolids both from industrial and domestic sources, although in recent years concentrations have decreased, due mainly to improved trade effluent controls and the adoption of cleaner manufacturing technologies (Smith, 1996). Inputs of heavy metals to agricultural soils are controlled under the Sludge (Use in Agriculture) Regulations (SI, 1989), which enforces provisions of EC Directive 86/278/EEC, by means of sludge concentration limits, soil limits and maximum permitted application rates. A survey of biosolids production, treatment, recycling and disposal in the UK was undertaken for the financial year 1996/7 (Gendebien et al., 1999). The report includes information on weighted average heavy metal concentrations in biosolids used in agriculture and from this heavy metal loading rates can be derived, assuming sludges are applied at rates equivalent to 250 kg ha-1 total N (Table 2.18). The range of organic contaminants known to be present in biosolids is extensive and diverse. For example, 332 different compounds were identified in German biosolids by Drescher-Kaden et al. (1992). Concentrations of organic pollutants in biosolids were not measured as part of the UK sewage sludge survey (Gendebien et al., 1999), as there are currently no limit values set for biosolids or soils in the UK. However, Smith (1996) summarised the literature published to date and reported concentration ranges for some of the principal groups of organic contaminants. The highest concentrations were for linear alkylbenzene sulphonates (LAS) at 50-15000 mg kg-1 ds, alkylphenols (100-3000 mg kg-1 ds) and phthalates (1-100 mg kg-1 ds). 44 BIOENERGY CROPS AND BIOREMEDIATION Table 2.18. Heavy metal concentrations and loading rates from organic manures and other wastes (application rates equivalent to 250 kg ha-1 total nitrogen, except for paper sludge where the industry maximum of 15 t ha -1 was used). Manure type Dry matter (%) Total N (kg t-1 or m-3) Concentration Zn Cu Ni Livestock manures1 Cattle FYM Pig FYM Dairy slurry Beef slurry Pig slurry Broiler/turkey litter Layer manure 25 25 10 10 10 60 30 6.0 7.0 4.5 3.5 7.0 29 15 17 60 17 17 65 130 175 4.0 42 4.5 4.5 47 19 27 0.7 1.3 0.6 0.6 1.4 2.4 3.0 Biosolids2 15 6.5 119 85 8.6 Paper sludge3 22 14 13 1.0 1 2 3 0.3 Pb Cr (g t-1 or m-3) 0.6 0.8 0.7 0.7 0.8 2.0 2.7 33 0.5 0.5 0.6 0.6 0.6 0.7 1.2 1.7 24 1.5 Source : Nicholson et al., 1999 Source : Gendebien et al., 1999 Source : Davis & Rudd, 1998 45 Loading rate As Cd Zn Cu Ni Pb Cr (kg ha-1) As Cd 0.3 0.2 0.2 0.2 0.2 0.3 0.1 0.06 0.06 0.03 0.03 0.04 0.33 0.39 0.7 2.1 0.9 1.2 2.3 1.1 2.9 0.2 1.5 0.3 0.3 1.7 0.2 0.5 0.03 0.05 0.03 0.04 0.05 0.02 0.05 0.03 0.03 0.04 0.05 0.03 0.02 0.05 0.02 0.02 0.03 0.04 0.02 0.01 0.03 0.01 0.01 0.01 0.02 0.01 <0.01 <0.01 0.003 0.002 0.002 0.002 0.001 0.003 0.007 0.9 0.50 4.6 3.3 0.33 1.27 0.92 0.03 0.019 - <0.25 4.7 0.8 0.06 0.03 0.17 - 0.005 BIOENERGY CROPS AND BIOREMEDIATION Industrial wastes Approximately 6.8 million tonnes (fresh weight) of industrial wastes are recycled to land in England and Wales each year (Gendebien et al., 2001). Various types of wastes are applied including a range of vegetable and animal wastes (5.05 Kt), food industry waste (1,409 Kt), paper sludge (322 Kt), rocks, subsoils and contaminated soil/subsoil (15 Kt), cement waste (12.2 Kt), other biological treatment plant sludges (4.3 Kt), textile waste (3.5 Kt) and small quantities of leather and tannery wastes, fly ash and other mineral wastes (Gendebien et al., 2001). Data on the composition of selected industrial wastes are reported by Gendebien et al. (2001). These were derived from a limited number of sources in the UK over a period of ten years and may not be entirely representative, although they do provide a broad indication of the composition of the wastes. As an example, heavy metal application rates for paper sludge (Table 2.18) were calculated using the median heavy metal analysis of 25 samples reported by Davis & Rudd (1998) and assuming the industry maximum application rate of 15 t ha-1. Note that this data should be treated with some caution, due to the variable nature of paper sludge from different mills and production processes. There is little information on other contaminants in industrial wastes. Paper sludges are known to contain dioxins and organohalogens, especially where chlorine bleaching was used, and wastes from the textile and tanning industries may contain organic dyes and pesticide residues (Anon, 1998). Summary Assuming an application rate equivalent to 250 kg ha-1 total N, biosolids will be a more important source of heavy metals and organic contaminants to biomass crops than livestock manures. Whilst the risks of soil metal contamination from biosolids applications are recognised and regulated for, there is no current control over inputs of heavy metals from livestock manures and care should be taken to ensure that soil concentrations do not become excessively high, especially where pig and poultry manures are applied. Industrial wastes may supply large amounts of certain metals (e.g. Zn from de-inked paper sludge and Cr from tanning industry wastes). The scientific consensus is that, based on the limited information available, organic contaminants in biosolids applied to agricultural land are unlikely to cause significant environmental or human health problems (Smith, 1996). 46 BIOENERGY CROPS AND BIOREMEDIATION 2.3.3 Contaminant uptake/removal by biomass crops Heavy metals Phytoremediation strategies for heavy metal contaminated sites have tended to concentrate on the use of metal tolerant or hyper-accumulator species (e.g. Brassica spp., Thlaspi caerulescens) which usually produce relatively small quantities of biomass. However, high biomass producers such as willow (Salix spp.) have been increasingly used as potential phytoremediator crops. In a Swedish study, high Cd concentrations were found in willow shoots and the authors concluded that Salix viminalis could potentially remove significant amounts of Cd from the soil, although it would take 12-20 years to return a soil with 600 g ha-1 (c. 0.15 mg kg-1) Cd to half its original value (Eriksson & Ledin, 1999). Nielsen (1994), also found that willow takes up large amounts of Cd and Zn, but Labrecque et al., 1995) reported that Salix discolor and Salix viminalis were less able to take up Ni, Hg, Cu and Pb. There are large differences in the efficiency of metal uptake between different biomass crops, plant species and clones. The EU-funded BIORENEW project is investigating bioremediation of contaminated land using biomass crops and is currently assessing the metal tolerance of different willow varieties (Watson et al., 1999). Other European work has looked at acclimation of Salix species to metal (Zn, Cu and Cd) stress in an attempt to induce increased resistance (Punshon & Dickinson, 1997), and at differences in tolerance and metal accumulation between different Salix clones (Landberg et al., 1996). Similarly, work in the USA has looked at differences in metal uptake between Salix clones grown hydroponically (Punshon & Dickinson, 1999), whilst Rugh et al., (1998) reported initial work on the potential of transgenic yellow poplar trees to remediate Hg polluted soils. Reported concentrations of metals in biomass crops are given in Table 2.19, although often these are from plants grown on contaminated or heavily sludged soils. There can be large differences in concentrations between different plant parts. For example, Punshon & Dickinson (1997) reported the magnitude of Cu accumulation in the order roots > wood > new stem > leaves; whereas for Cd the order was leaves > stem > wood > roots. Roots, wood and stems can immobilise metals for a number of years, as opposed to leaves which are shed annually returning any metals they contain to the soil. It is most useful in terms of remediation potential for metals to accumulate in the harvested plant parts (i.e. wood, stems and leaves). Note that the time of sampling can also influence the heavy metal concentration in the leaves (Riddel-Black, 1994) as can interactions with other soil heavy metals (Punshon & Dickinson, 1997). 47 BIOENERGY CROPS AND BIOREMEDIATION Table 2.19. Heavy metal concentrations in biomass crops and calculated metal uptake rates. Metal Crop Concentration Comments Source Metal contaminated soils Metal contaminated soils 8-30 year old plantations 8-30 year old plantations Field contaminated with Cd Heavy sludge applications Plants grown in culture solutions Sludge applications made Borjesson (1999) Felix et al. (1999) Dickinson (1997) Dickinson (1997) Eriksson & Ledin (1999) Eriksson & Ledin (1999) Felix (1997) Riddell-Black (1994) Punshon & Dickinson (1997) Labrecque et al. (1995) (mg kg-1 DM) Cd Salix - shoots Salix viminalis - aerial tissues Salix -foliage Salix - wood Salix viminalis - stem-wood Salix viminalis - leaves Salix viminalis - aerial tissues Salix (4 spp) - stems Salix (4 spp) -aerial tissues Salix (2 spp) - leaves 0.4 - 3.9 22 43.9 76.4 0.35 - 2.43 0.31 - 1.96 8.3-22.2 3.3-7.7 <100 <1.8 Cu Salix - foliage Salix - wood Salix caprea - foliage and woody branches Salix (7 spp) - foliage and woody branches Salix (4 spp)-stems Salix (4 spp) -aerial tissues Salix (2 spp) - leaves 4.2 4.6 681 183-509 5.9- 8.2 25-100 <20.7 Metal contaminated soils Metal contaminated soils 3-year old trees on highly contaminated soil 1-year old trees on highly contaminated soil Heavy sludge applications Plants grown in culture solutions Sludge applications made Dickinson (1997) Dickinson (1997) Dickinson (2000) Dickinson (2000) Riddell-Black (1994) Punshon & Dickinson (1997) Labrecque et al. (1995) Zn Salix - foliage Salix - wood Salix (3 spp) -roots Salix (3 spp) - stems Salix (4 spp)-stems Salix (2 spp) - leaves 87.1 77.3 <50 <8 95-156 <560 Metal contaminated soils Metal contaminated soils Plants grown on mining spoil Plants grown on mining spoil Heavy sludge applications Sludge applications made Dickinson (1997) Dickinson (1997) Dickinson et al. (1994) Dickinson et al. (1994) Riddell-Black (1994) Labrecque et al. (1995) Pb Salix - foliage Salix - wood Salix (2 spp) - leaves 17.3 157.4 <6.5 Metal contaminated soils Metal contaminated soils Sludge applications made Dickinson (1997) Dickinson (1997) Labrecque et al. (1995) Ni Salix (4 spp)-stems Salix (2 spp) - leaves 0.9-1.4 <20 Heavy sludge applications Sludge applications made Riddell-Black (1994) Labrecque et al. (1995) Hg Salix (2 spp) - leaves <20 Sludge applications made Labrecque et al. (1995) 48 BIOENERGY CROPS AND BIOREMEDIATION Soil factors such as soil pH and clay content can strongly influence plant heavy metal uptake. Some researchers have reported manipulating soil conditions or using soil amendments (e.g. chelating agents) to increase metal uptake. For example, Kayser et al. (2000) investigated the efficiency of several crops (including Salix viminalis) at phyto-extracting Zn, Cd and Cu from calcareous soils and found that plant accumulation of these metals increased by a factor of 2-3 where nitrilotriacetate (NTA) and elemental sulphur were applied to the soil. However, the use of soil amendments to increase metal solubility can also increase the risks of leaching or downward migration of the metals. It is also important to note that metals may be present in wastes and soils in different forms, and that some metal fractions are so strongly bound to mineral particles that they may never be taken up by plants and removed from the soil. There has been relatively little work published specifically looking at metal uptake following organic manure applications to biomass crops. However, metal uptake rates from a study where sewage sludge was applied to Salix crops at 12.5 t ds ha-1 yr-1 for 6 years (Hasselgren, 1999) are reported in Table 2.20. Typical UK sludge and poultry manure metal application rates can be compared with the stem uptake rates reported by Hasselgren (1999) and indicate that all metals would be added at higher rates than they could be removed by the crop, leading to a net accumulation in the soils (Table 2.20). Table 2.20. Salix stem uptake of sludge heavy metals, amounts of metals supplied by UK sludges and poultry manure applied at 250 kg ha -1 total N yr-1, and calculated metal accumulation rates. Heavy metal Zn Cu Pb Cd Ni Cr 1 2 3 Average stem Amount of metal metal uptake supplied by UK rate (g ha-1 yr-1)1 sludge (g ha-1 yr-1) 600 59 30 11 8.2 4.8 Net metal accumulation (g ha-1 yr-1) 4562 328 1273 19 328 904 3962 269 1243 8 320 899 Amount of metal supplied by poultry manure2 (g ha-1 yr-1) Net metal accumulation (g ha-1 yr-1)3 2900 500 50 7 50 30 2300 441 20 4 42 25 Source : Hasselgren (1999). Laying hen manure (see Table 2.18). Assumes metals applied in poultry manure are taken up at the same rate as those in sludge. Labrecque et al. (1995) calculated the transfer coefficients (TC) of metals from soils to two Salix species where dried, pelleted sludge was applied. Cd and Zn were the most readily transferred metals, with 15-25% of added sludge metals taken up into the biomass, followed by Ni, Hg, Cu and Pb. Sludge applications increased levels of metals in the soil proportionately with the amounts applied in the sludge. However, this did not necessarily translate into an increased plant concentration. The authors found there was a good relationship between soil and plant concentrations of Cd and Zn, while plant concentrations of Cu, Hg, Ni and Pb were less dependent on the soil concentrations. 49 BIOENERGY CROPS AND BIOREMEDIATION In a later study, Labrecque et al. (1998) reported the differences between the amounts of metal introduced into the soil with dried, pelleted sludge applied at different rates (100, 200 and 300 kg ha-1 of available N) and those removed following biomass harvest. The net accumulation of Zn in the soil after two years of sludge applications ranged from c. 2.5 to c. 8.4 kg ha-1 and for Cu from c. 1.4 to c. 4.4 kg ha-1 (Table 2.21). The amount of Zn removed by the crop increased slightly with increased sludge application rate, whilst the amount of Cu removed was similar at all application rates. Analysis of soil (0-20 cm depth) confirmed that soil Zn and Cu levels had generally increased compared with the pre-planting levels; however levels of soil Ni, Cr and Cd were largely unaffected by the two sludge applications. Table 2.21. Net soil accumulation of metals after 2 years of sludge applications. Metal Sludge content (mg kg-1 ds) Amount of metal applied (kg ha-1) Amount of metal removed in biomass (kg ha-1)1 Net soil metal accumulation (kg ha-1)2 Zinc 404 3.03 6.06 9.09 0.57 0.65 0.72 2.46 5.41 8.37 Copper 201 1.51 3.02 4.53 0.07 0.11 0.09 1.44 2.91 4.44 Source: Labrecque et al., 1998 1 Calculated by difference 2 Mean results of 2 sites, 2 species and 2 planting densities These results indicate that Salix crops can remove heavy metals from soils, although the removal rate depends on the metal concerned, soil properties, the plant species/variety grown and the amount of biomass produced. However, where organic manure applications are made, metals will be added at greater rates than they can be removed by a biomass crop such as Salix, leading to an accumulation in the soil. Assuming a typical UK biosolids application to Salix at a rate equivalent to 250 kg N yr-1 would lead to a net soil accumulation of c. 4 kg Zn ha-1 yr-1 (Table 2.20), and assuming that the soil currently has the UK median Zn concentration of 82 mg kg-1 (McGrath & Loveland, 1992), then it would take around 115 years for the soil to reach the limit concentration of 200 mg kg -1 (DoE, 1996). Similarly, for poultry manure applications it would take around 160 years to reach the soil limit concentration for Zn. 50 BIOENERGY CROPS AND BIOREMEDIATION Other contaminants Hybrid poplar trees have been used to uptake, hydrolyse and dealkylate the pesticide atrazine to less toxic metabolites (Burken & JL, 1997; Burken & Schnoor, 1996). Results indicated that poplar cuttings were able to take up the majority of the applied atrazine that was not strongly sorbed to the soil organic fraction. Hybrid poplar cuttings have also been used to uptake and translocate 1,4dioxane (a suspected carcinogen) to leaf surfaces (Aitchison et al., 2000). Around 80% of the dioxane taken up by poplars was transpired to the atmosphere where it can be readily dispersed and photo-degraded. Other researchers in the USA (Orchard et al., 2000) have used poplars to take up the high explosive 2,4,6trinitrotoluene (TNT) and the carcinogenic degreaser trichloroethylene (TCE). This work has looked exclusively at the remediation of contaminated sites and has not addressed the use of these plants to take up contaminants from organic manures or other wastes. Ultimate fate of metals and other contaminants in biomass crops At harvest, the above-ground parts of biomass crops will be removed from the growing site and transported to the power station. During combustion all the heavy metals, except Hg, will be concentrated in the resultant ash. Because Hg is a volatile metal, during combustion it will be released in the gaseous state, and eventually be oxidised and scavenged by wet or dry deposition processes at various distances from the source (Steinnes, 1990). Some researchers have found elevated levels of Hg in soils near crematoria caused by the volatilisation of Hg in dental fillings during cremation, which was subsequently re-deposited to the nearby soil (Nieschmidt & Kim, 1997). A similar situation could occur near power stations where biomass crops grown on Hg contaminated soils are burnt. Metals on particulates in the flue gases may be discharged to the atmosphere, via the stack, and subsequently re-deposited onto land. However, appropriate technology to improve the quality of stack emissions should minimise this problem, although there may be considerable expense involved. Metals in the bottom and fly ash could potentially be recovered for re-use, however this is likely to be very expensive and the quantities of ash generated are unlikely to make recovery an economic proposition. It is more likely that the ash will be buried in landfill sites where conditions may be such that metal containing leachates could enter the environment. 2.3.4 Other pathways of contaminant movement and environmental effects Hasselgren (1999) reported that most of the metals applied to Salix crops in sewage sludge remained in the upper 10 cm soil layer, with total soil Zn and Cu concentrations increasing by 17 and 4 times, respectively, where sludge was applied at 12.5 t ds ha-1 yr-1 for 6 years. Hasselgren (1998) also found that where sewage sludge was applied to Salix at rates up to 20 t ha-1 ds, the groundwater (at 1-2.5 m) had double the concentration of Cu, Pb and Hg, and 3-5 times the 51 BIOENERGY CROPS AND BIOREMEDIATION concentration of Zn and Ni after sludge applications, although Cd concentrations were below detection limits. Most previous research indicates that heavy metals applied in biosolids would accumulate in the upper soil layers, with relatively little lost via leaching (Smith, 1996). Nevertheless, there could be a greater risk of leaching if soil amendments are used to increase plant metal uptake. Salix leaves have been shown to accumulate high concentrations of some metals and a further risk to the environment could come where animals consume these leaves and accumulate high levels of metals in their body tissues. 2.3.5 Conclusions 1. Biomass crops can remove heavy metals and organic contaminants from soils, although the removal rate depends on contaminant and soil properties, the plant species/variety grown and the amount of biomass produced. 2. Where organic manure (livestock manure, biosolids, industrial waste) applications are made, heavy metals will generally be added at greater rates than they can be removed by a biomass crop, leading to an accumulation in the soil. 3. Very little is known about the types and quantities of organic contaminants in organic manures or how quickly they could be removed or broken down by biomass crops. 4. After combustion, if the ash is buried in landfill sites, conditions may be such that heavy metal leachates could enter the environment. 2.3.6 References Aitchison, E; Kelley, S; Alvarez, P & Schnoor, J (2000). Phytoremediation of 1,4,-dioxane by hybrid poplar trees. Water Environment Research 72 (3). 313-321. Alloway, B J (1990). Heavy Metals in Soils. Blackie, London. Anon. 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Investigation of the criteria for and guidance on the landspreading of industrial wastes. Final report to the Environment Agency. WRc Report 4088/7. Environment Agency R&D Technical Report P193. Dickinson, N (2000). Strategies for sustainable woodland on contaminated soils. Chemosphere 41. 259-263. Dickinson, N M (1997). Rehabilitation and remediation of metal-contaminated soils using trees. 4th International Conference on the Biogeochemistry of Trace Elements, Berkeley, California. 437-8. Dickinson, N M; Punshon, T; Hodkinson, R B & Lepp, N W (1994). Metal tolerance and accumulation in willow. Willow Vegetation Filters for Municipal Wastewaters and Solids, Uppsala, Sweden. 121-127. DoE (1996). Code of Practice for Agriculture Use of Sewage Sludge. Department of the Environment. Drescher-Kaden, U; Bruggeman, R; Matthes, B & Matthies, M (1992). Contents of organic pollutants in German sewage sludges. 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Environmental Pollution 102, 129-137. 55 BIOENERGY CROPS AND BIOREMEDIATION 2.4 IMPACTS ON BIODIVERSITY CHRIS BRITT 2.4.1 Introduction Although there has been a vast amount of research on the environmental impacts of applying sewage sludge, farmyard manures and slurries to agricultural land and forestry, this has been heavily biased towards studies of impacts on water, soil and air quality – with relatively little work on the effects on biodiversity. There has been almost no research on the ecological impacts of applying various waste materials to short rotation coppice or energy grasses. Because of this paucity of directly relevant information, provisional assumptions must be based on an interpretation of indirectly relevant experience (e.g. biodiversity impacts of waste applications to agricultural or forest crops), within the context of conditions prevailing within ‘typical’ energy crops. Such an interpretation firstly requires an overview of energy crops as habitats for wildlife. Ecology of bioenergy crops The ecology of short rotation willow and poplar crops in the UK, Sweden and elsewhere has been quite extensively studied. Although biodiversity in any single plantation is heavily influenced by factors such as intensity of management, adjacent land uses and proximity to semi-natural woodland, previous research has demonstrated the potential value of SRC as a habitat for certain groups of invertebrates and birds. These include numerous phytophagous insect species, including pest species such as the blue willow beetle (Phratora vulgatissima) (Sage & Tucker, 1997 & 1998a), and songbirds (Sage & Robertson, 1996; Coates & Say, 1999). In contrast, there has been very little research on the ecology of Miscanthus or other grasses grown as bioenergy crops. Vegetation Surveys in 1993 and 1996 have provided quite a comprehensive picture of the ground flora composition in SRC plantations (Sage & Tucker, 1998b). Over the two surveys, the most frequently recorded National Vegetation Classification (NVC) types were various weed and tall herb communities – with an increase in woodland and scrub communities in 1996. The most frequently recorded species was the common nettle (Urtica dioica), which occurred in 81% of plots in both years. Rosebay willowherb (Chamaenerion angustifolium) declined sharply between the two surveys. In total, 151 plant species were recorded, with 19 of these present in at least 25% of all plots in one or both years. The mean numbers of plant species recorded per plot, in five 10 m2 quadrats, were 13.5 in 1993 and 13.8 in 1996. The total number of species recorded per plot ranged from 3-24 in 1993 and 4-31 in 1996. A detailed study of five SRC sites in southern England gave similar results (Coates & Say, 1999). Although the total numbers of plant species recorded were generally quite high (up to 120 at a single site), possibly due to inadequate weed 56 BIOENERGY CROPS AND BIOREMEDIATION control at some sites and the close proximity of semi-natural habitats at all, certain general trends were recognised in vegetation succession. For example: Small annual ruderal species, such as scarlet pimpernel (Anagallis arvensis), groundsel (Senecio vulgaris) and common chickweed (Stellaria media), were present at low levels during the early stages of the cropping cycle, and then tended to disappear. Larger ruderals, such as common orache (Atriplex patula), cleavers (Galium aparine), knotgrass (Polygonum aviculare), rosebay willowherb (Chamaenerion angustifolium), thistles (Cirsium spp.) and docks (Rumex spp.), occurred at high levels early in the cropping cycle and persisted at reduced levels. Semi-ruderal grasses with a degree of shade tolerance, such as creeping bent (Agrostis stolonifera), cocksfoot (Dactylis glomerata) and Yorkshire fog (Holcus lanatus), persisted into the cropping cycle. Bramble (Rubus fruticosus agg.) invaded coppice crops later in the cycle. In SRC where weed control was quite good, and the crop canopy was rapidly closed, ground vegetation from the third year after planting onwards provided approximately 0-15% cover. Coates & Say (1999) described the vegetation type dominating most plots at this time as “mixed semi-ruderal.” The overall picture, therefore, is of a ground flora that is either very sparse – because of effective weed control measures and/or heavy shading from the SRC crop – or of limited diversity and dominated by species of low conservation value, typical of agricultural weed communities or disturbed land. There are occasional exceptions of course, particularly where herbicide inputs have been negligible or nil, and where there are nearby seed sources of woodland species – but the requirements of good SRC husbandry and the relative scarcity of semi-natural woodland will always ensure that such sites are atypical. Soil biodiversity There is some evidence that willow short rotation coppice plantations can support large earthworm populations, and that earthworms play a significant role in the decomposition of leaf litter (Šlapokas & Granhall, 1991). Research in Sweden has also demonstrated differences between willow species in the rates of leaf decomposition, with Salix daphnoides seemingly more attractive than S. fragilis or S. viminalis to all soil organisms – including earthworms (Šlapokas & Granhall, 1991). Ground-dwelling invertebrates In an upland (almost 300 m altitude), willow SRC plantation in mid-Wales, ground-dwelling invertebrates, captured in pitfall traps during the first and second growing seasons, were fairly typical of the previous land-use (Slater et al., 1997). The study site was weedy, however, and the authors suggest that in “well managed” SRC plantations pitfall traps are likely to capture few herbivorous species and more scavengers and carnivores. Canopy invertebrates The large leaf area within a fast-growing willow or poplar SRC crop provides an important food resource for a wide range of herbivorous invertebrates, which can 57 BIOENERGY CROPS AND BIOREMEDIATION sometimes occur at very high densities. These herbivorous, or phytophagous, species can, in turn, support a diversity of predatory and parasitic invertebrate species; as well as numerous insectivorous birds and mammals. Reddersen (2001) conducted a survey of commercial willow (Salix viminalis) SRC plantations in Denmark, to quantify catkin abundance. He concluded that the abundance of flowers in years 2-4 after harvest (willow crops did not flower in the year after harvest), although variable, was sufficiently high to make willow SRC a potentially important source of nectar for bees and other nectar-feeding insects. Birds Short rotation coppice provides a good habitat for many insectivorous, woodland bird species – including thrushes (Turdidae), tits (Paridae) and warblers (Sylviidae). Warblers particularly favour SRC, probably because of the abundant invertebrate prey that this habitat supports and because it is structurally similar to early stages of regenerating traditional coppice and to willow carr – two habitats for which these species are known to have a preference. For example, Fuller & Green (1998), studying bird populations in adjacent stands of recent, old and thinned/singled small-leaved lime (Tilia cordata) coppice, found that willow warbler (Phylloscopus trochilus), garden warbler (Silvia borin) and chiffchaff (Phylloscopus collybita) were all far more abundant in the recent coppice. Thrush (Turdidae) densities also tended to be higher in recent coppice. Chaffinches, however, were least frequently recorded in the recent coppice. Willow SRC has also been shown to provide a good habitat for pheasants (Phasianus colchicus). Mammals There is little published information on mammal populations in SRC or energy grass plantations. Among the mammal species that are known to use SRC crops, rabbits (Oryctolagus cuniculus) and roe deer (Capreolus capreolus) are frequently common (Sage & Tucker, 1998b). A survey of five SRC sites in southern England, by Coates & Say (1999), recorded 13 mammal species. Four species were thought to be common – rabbit, brown hare (Lepus capensis), mole (Talpa europaeus) and wood mouse (Sylvaemus sylvaticus), although this assumption is based purely on casual observations for the first three (i.e. only small mammals, including wood mice, were formally recorded). The other nine species recorded in SRC were hedgehog (Erinaceus europaeus), common shrew (Sorex araneus), serotine (Vespertilio serotinus) and pipistrelle (Pipistrellus pipistrellus – both subspecies) bats, bank vole (Clethrionomys glareolus), harvest mouse (Micromys minutus), brown rat (Rattus norvegicus), badger (Meles meles) and roe deer. Other species likely to utilise SRC plantations would include predatory species such as the fox (Vulpes vulpes), stoat (Mustela erminea), weasel (Mustela nivalis) and polecat (Putorius putorius). Slater et al. (1997) compared small mammal populations in newly established willow SRC (during the first year only), and in a nearby rough grassland pasture, at an upland site (almost 300 m) in mid-Wales. They recorded higher numbers of wood mice in SRC and higher numbers of field voles (Microtus agrestis) in the 58 BIOENERGY CROPS AND BIOREMEDIATION pasture. Destruction of the ground flora, during SRC crop establishment, resulted in the initial loss of field voles. Consequently no field voles were trapped in the first spring after planting, but small numbers were recorded in the autumn. It is unlikely that SRC plantations with high standards of weed control will support large vole populations. Almost weed-free SRC crops will also provide relatively poor habitats for other small mammal groups, such as mice and shrews – due to a lack of low ground cover (absence of nesting sites and exposure to predators) and inadequate food resources (e.g. invertebrates, seeds and fruits). Indeed, an unpublished PhD project by Bodnor (cited by Sage & Tucker, 1998b) found that both abundance and diversity of small mammals, such as wood mice, were greater in weedy SRC plots. Bats make disproportionate use of rivers and lakes for hunting (Vaughan et al., 1997), but the frequently large populations of arboreal invertebrates in SRC may make this a suitable habitat for foraging woodland bat species such as the 45 kHz pipistrelle (Pipistrellus pipistrellus), Myotis spp. and, possibly, horseshoe bats (Rhinolophus spp.) and Plecotus spp. Further research is needed to confirm the extent of usage of energy crops by bat species, and to evaluate the potential role of energy crops in bat conservation. Effects of waste applications The application of waste materials to energy crops might be expected to have significant effects on the flora and fauna within bioenergy crops, such as SRC and Miscanthus. However, an extensive review of the literature revealed no evidence of research that looked directly at the ecological effects of applying farm, urban or industrial waste products to energy crops. There has also been only a limited amount of research into the ecological effects of applying such materials to other types of vegetation. Consequently, although relevant research will be referred to when possible, much of this section must inevitably rely largely upon ‘educated supposition’. For example, it can safely be assumed that thick applications (or patchy distribution) of some materials, particularly wastes that are relatively slow to decompose, will suppress ground flora development. Although this ‘mulching’ of weeds will be beneficial to the crop, it will have generally negative impacts on biodiversity. On the other hand, increased soil organic matter will be likely to enhance populations of soil micro-organisms and invertebrates; and applications of organic manures can provide an important food resource for coprophagous organisms (e.g. species of Basidiomycete fungi and scarab beetles) – which, in turn, will be a source of food for species higher in the food chain. Manures may also be the source of weed seeds. The supply of N in manures, slurries or sewage sludge will increase crop growth rates, producing more and ‘softer’ foliage with greater susceptibility to grazing insects. It is important, of course, to consider all possible direct and indirect ecological effects of waste applications to bioenergy crops within the context of standard management practices for these crops. Neither the effects of weed suppression nor nutrient enrichment referred to above are likely to have particular ecological significance, if the alternative is increased inputs of herbicides and inorganic N. Indeed, it could be that organic waste utilisation could provide a net positive effect in these circumstances. 59 BIOENERGY CROPS AND BIOREMEDIATION 2.4.2 Agricultural and municipal wastes Vegetation As has been clearly shown previously in this review, there is plentiful evidence that applying organic ‘waste’ materials such as sewage sludge, municipal wastewater or farm slurries can provide very useful sources of nutrients to biomass crops – with consequent positive growth responses. Perttu (1999), for example, has produced figures to illustrate how municipal wastewater in Sweden provides macronutrients in a ratio that almost exactly matches the calculated requirements of SRC willow crops, as well as supplying potentially useful quantities of water. Although nutrients and water supplied in sewage, wastewater, farm manures, animal slurries and dirty water will boost plant growth generally, the results will not necessarily be beneficial to wildlife. There are four main factors to consider. 1. Direct ‘mulching’ effects of solid manures on ground vegetation: Irregular distribution of manures will lead to suppression of ground vegetation beneath thicker patches. This is likely to be a particular problem where application rates are high. However, it could be considered less of a problem if the ground flora is already very sparse, because of efficient weed control, or where flora consists primarily of agricultural weed species of low conservation value. 2. Effects of nutrient supply on ground flora species composition: The supply of high levels of N and P in organic wastes will favour undesirable, highly competitive weed species (e.g. nitrophilous species such as common nettle, U. dioica) at the expense of more desirable, less competitive woodland species (e.g. herb robert, Geranium robertianum; wood sorrel, Oxalis acetosella; primrose, Primula vulgaris; common dog violet, Viola riviniana; bugle, Ajuga reptans; and hedge woundwort, Stachys sylvatica). This factor can be considered less significant if the alternative to organic waste application is the supply of equivalent quantities of nutrients in inorganic fertilisers – as would normally be the case. 3. The indirect effects of increased nutrient supply to the crop: The application of additional water and nutrients to any crop, if either was previously available at below optimum levels, will produce a positive growth response – commonly exhibited in higher shoot numbers, greater shoot lengths, and increased leaf size and area. Taller, leafier energy crops will have high light interception levels earlier in the season, and more effectively shade out plant species in the ground and field layers. 4. The introduction of new species to the planted area: There is a strong possibility that the wastes being disposed of in biomass crops may include plant seeds and fragments of stolons or rhizomes, and consequently be responsible for the introduction of additional species to the flora. Sewage sludge applications are commonly responsible for the introduction of tomato plants, farmyard manure may contain seeds of grassland weeds (e.g. common 60 BIOENERGY CROPS AND BIOREMEDIATION chickweed and docks), and composted green household wastes may have viable seeds of garden weed species. Lynn et al. (1992) studied the short-term effects of applying different rates of sewage sludge on the ground flora of a mixed-species woodland in Powys, Wales. High rate applications (400 m3 ha-1, equivalent to a 4 cm deep mulch) in March greatly reduced ground flora species diversity and vegetative cover, and delayed plant emergence. The emergence of some species was delayed by as much as 10 weeks. Lower rate sludge applications (100 or 200 m3 ha-1) had less of an effect on diversity, cover and emergence date. For this reason, Lynn et al. (1992) suggest that lower rate applications might be more acceptable. Late season applications are likely to have less of an impact on species diversity, as the sludge mulch will be able to break down during the autumn and winter period. Another, potentially negative effect of animal manure applications on biodiversity (although positive in terms of crop production), is the possible inhibition of seed germination by phenolic acids in the manures (Marambe & Ando, 1992). Mastrota et al. (1989) reported that areas of a mixed oak forest in central Pennsylvania that had been irrigated with chlorinated sewage effluent had significantly lower densities of understorey trees and shrubs, but significantly higher densities of herbs in the ground flora. A Canadian study (Vasseur et al., 2000) of the effects of sewage sludge applications to agricultural lands, on ground flora composition, found no significant differences in species number between sludge-treated and untreated sites. Vasseur et al. (2000) concluded that, although sewage sludge produced little direct effect on ecological parameters, longer term effects on soil chemical factors are likely to be reflected in plant community composition in the longer-term. Soil biodiversity The long-term effects of applying municipal wastewater on the spatial distribution and biomass of soil micro-organisms were studied by Filip et al. (2000). Although the soil samples were not taken from under a biomass crop, the results are still of some relevance to this review. One plot at the grassland study site, on a sandy soil in Germany, had been regularly irrigated with wastewater for almost 100 years. Another plot had been irrigated with wastewater, until about 20 years earlier. There was also a control plot that had never received wastewater. During the sampling period, the irrigated plot received approximately 2,000 mm wastewater (primary effluent) per annum. The typical chemical composition of the wastewater was estimated to be 115 mg l-1 total C, 80 mg l-1 organic N, 71 mg l-1 NH4-N, 0.8 mg l-1 NO3-N, 14.2 mg l-1 P, 0.06 mg l-1 Cu, 1.1 mg l-1 Fe, 0.06 mg l-1 Mn, and 0.34 mg l-1 Zn. The pH of the wastewater was around 8.0. Results showed that numbers of bacteria, actinomycetes and fungi were generally higher in the long-term irrigated soil. Organic particles in soil from this plot had the highest microbial counts overall. However, less nutrient-dependent 61 BIOENERGY CROPS AND BIOREMEDIATION oligotrophic bacteria were mostly associated with the silt + clay soil fraction, irrespective of wastewater treatment. Slightly enhanced microbiological activity was still detectable 20 years after the cessation of wastewater irrigation. At a higher taxonomic level, earthworms grown in pots containing acid mine spoils showed a positive growth response (increased surface area and mass) where the spoil had been treated with sewage sludge (Pallant & Hilster, 1996). Ground-dwelling invertebrates Slater et al. (1997) found no effects of fertiliser or sludge treatment, or willow cultivar, on ground-dwelling invertebrate populations, in terms of numbers of taxa or individuals, within an SRC willow plantation in mid-Wales. However, studies in other habitats have indicated that the application of organic manures or sewage sludge can, in some instances, lead to increased invertebrate populations. A field study by Larsen et al. (1996), in Ohio, USA, examined the effects of 11 years of inorganic fertiliser or sewage sludge applications on the ground beetle (Carabidae) fauna of an ‘old-field’ grassland community. Results showed that plots treated with sewage sludge or inorganic fertiliser had significantly more carabid species than untreated control plots. Another study (Kielhorn et al., 1999), on a sandy mine spoil in Germany, showed that higher numbers of ground beetle species and individuals were captured in plots ameliorated with sewage sludge (compared with catches from plots treated with compost, mineral fertiliser or untreated plots). All plots had been sown with a grass cover (Secale multicaule) and planted with pine seedlings, and increases in ground beetle populations were closely correlated with the amount of vegetative cover. Areas of a mixed oak forest in Pennsylvania that had been irrigated with chlorinated sewage effluent had a greater biomass of invertebrates (gastropods and annelids) than non-irrigated areas (Mastrota et al., 1989). In some situations, livestock slurries may contain insecticides – a fact that is likely to significantly affect their value as a substrate for invertebrates. A move away from organophosphate-based sheep dips, towards synthetic pyrethroid-based chemicals has been followed by new procedures for the disposal of these dips. Pyrethroid sheep dips must now be diluted in animal slurry or water before application to land. The negative environmental effects of slurries treated with sheep dip may go beyond the direct impacts on pyrethroid-sensitive invertebrate species. There may also be indirect effects on species higher in the food chain through a) loss of invertebrate prey species and/or b) increased numbers of faecal coliforms and pathogenic bacteria in slurries mixed with pyrethroid-based dips (Semple et al., 2000). Larsen et al. (1996) also showed that two of the most common carabid species, Harpalus pensylvanicus and Poecilus lucublandus, accumulated Cd, Pb and Zn on sewage-treated plots. However, concentrations recorded in these beetles were lower than those present in the soil, indicating that they do not ‘bioconcentrate’ heavy metals. These are important findings, in relation to the potential 62 BIOENERGY CROPS AND BIOREMEDIATION bioaccumulation of heavy metals in the food chain and resultant risks that sewage sludge applications pose to wildlife (see later discussion). Canopy invertebrates In contrast to their results for ground-dwelling invertebrates, Slater et al. (1997) found that arboreal species were more abundant and taxa richness increased on willow plots that were treated with P and K fertilisers and magnesian limestone, when compared with a sewage sludge treatment. They suggest that this may be a consequence of increased leaf areas and decreased foliar phenolic concentrations. It has previously been shown that the phenolic compound salicylic acid is more concentrated in leaves of willows grown in soils of low nutrient status, and that high concentrations are toxic and a deterrent to leaf-feeding invertebrates (Pasteels & Rowell-Rahier, 1992). Of course, it does not necessarily follow that inorganic fertilisers will produce larger leaf areas than organic wastes. The opposite situation may frequently apply e.g. if slurries apply optimum (or higher) rates of N and other nutrients and useful additional water. Birds No relevant information was found on the effects of waste applications to bioenergy crops on bird populations. However, manures and other bulky organic wastes will provide an important source of food for many invertebrates and thus, indirectly, for some birds. Most bird species, even those that are herbivorous or granivorous throughout most of their adult lives, have a predominantly invertebrate diet prior to fledging. The availability of an abundant source of dung-feeding insects, and their insect predators, during the spring and early summer nesting period may improve the breeding success of birds in the vicinity of SRC or Miscanthus plantations. Late winter or early spring manure applications to recently harvested crops may provide a source of invertebrate food at the right time, although the high density of most energy crops and their rapid rates of regrowth will probably make access very difficult for birds feeding second broods in June or July. Manures from cattle or sheep dosed with ‘worming’ insecticides such as ivermectin may be toxic to invertebrates and, consequently, of little indirect benefit to birds and insectivorous mammals. Wastes containing high concentrations of heavy metals or organic toxins pose a potential risk to birds, particularly raptors, through bioaccumulation of toxins in animal tissues. Mammals Lynn (cited by Slater et al., 1997), in an unpublished PhD study, found that sewage sludge applications had no significant effects on populations of wood mice or bank voles in broadleaved woodland. However, any resultant increase in invertebrate populations as a result of manure, sludge or slurry applications is likely to be beneficial to insectivorous mammals. 63 BIOENERGY CROPS AND BIOREMEDIATION A detailed study of small mammal populations within a mixed-oak forest in central Pennsylvania (Mastrota et al., 1989) showed that wastewater (chlorinated sewage effluent) had generally beneficial effects on small mammal populations. Small mammal species richness was higher in areas of the forest that had been irrigated with sewage effluent. Although Mastrota et al. recorded higher invertebrate densities in sewage-treated areas, they suggest that the increased small mammal diversity were more likely to be a consequence of the increased herb density in these areas. This supposition appears to be supported by the findings that numbers of white-footed mice (Peromyscus leucopus) and southern red-backed voles (Clethrionomys gapperi) were significantly higher in sewage-treated areas, whereas there were no significant differences between the numbers of two insectivorous species (masked shrew, Sorex cinereus, and northern short-tailed shrew, Blarina brevicauda) numbers in irrigated and non-irrigated zones. Populations of white-footed mice also had significantly greater proportions of adults on sites irrigated with sewage effluent. The pollution of watercourses resulting from improper application of organic wastes to energy crops is likely to have negative environmental effects overall, although Vaughan et al. (1996) have shown how river eutrophication from sewage pollution can have mixed effects on bat species foraging along that river. Their results suggested that sewage discharges into the river had negative effects on pipistrelles (Pipistrellus pipistrellus, 45 kHz phonic type), but positive effects on Daubenton’s bats (Myotis daubentonii). Another issue of considerable concern, however, is the possible accumulation of heavy metals, and other toxins, in the internal organs of mammals when sewage sludge or other wastes are repeatedly applied. This has been the subject of research in a number of studies – although not in energy crops. Sewage sludge can contain relatively high levels of heavy metals, such as Cd, Cr, Cu, Pb, Ni and Zn – as well as organic pollutants and pesticides. Pig and poultry manures have relatively high concentrations of Cu and Zn (Jongbloed & Lenis, 1998; Nicholson et al., 1999). The toxicological effects of sewage sludge on young rats have been studied, under laboratory conditions, by Bag et al. (1999). These include significant reductions in levels of several enzymes, including liver alanine aminotransferase, liver succinate dehydrogenase (SDH), serum lactate dehydrogenase (LDH) and muscle SDH; and increases in liver and muscle LDH, serum and liver aspartate aminotransferase, serum and muscle alkaline phosphatase, and brain and muscle acetylcholinesterase activities. The potentially toxic effects of heavy metal accumulation in the organs of mammals inhabiting plantations regularly treated with sewage sludge or pig slurry are likely to be greatest in predatory species – a point made by Bag et al. (1999) and reinforced by various field studies. In one US study, Hegstrom & West (1989) found higher levels of Cd, Cu, Pb and Zn in the livers and kidneys of insectivorous Trowbridge’s shrews (Sorex trowbridgii) from forest plots treated with sewage sludge. Despite high levels of heavy metals in shrews, no evidence was found of 64 BIOENERGY CROPS AND BIOREMEDIATION heavy metal-induced lesions in these tissues. Hegstrom and West also found that sewage sludge applications increased levels of Cd and Pb in organs of insectivorous shrew-moles (Neurotrichus gibbsii) and Cd in organs of granivorous deer mice. In another US study (Woodyard et al., 1987), a single application of municipal sewage to forest stands in Michigan (10-year-old aspen, 50-year-old pine, 50-yearold mixed broadleaves and 70-year-old oak) produced no gross changes in concentrations of Cd, Cr, Cu, Ni and Zn in the tissues of herbivorous small mammals (or plants), when these were analysed 1-2 years later. Nickelson & West (1996) examined the longer-term effects of sewage sludge applications in forests on the Cd levels in kidneys of insectivorous shrews (Sorex sp.) and omnivorous mice (Peromyscus sp.). Animals were captured at 15 sites in western Washington, USA. Eight sites had been treated with anaerobically digested biosolids (from mixed domestic and industrial sources), applied at varying rates; 4, 11 or 15 years previously. The other seven, control, sites had not received sewage applications. Results showed that Cd concentrations were higher in the kidneys of shrews than mice, at all treatment sites. Except for sites receiving the lowest rate applications, shrews from all sewage-treated forests had significantly higher kidney Cd levels than shrews from controls. Cd concentrations in kidneys of shrews trapped at the 11-year post-application sites were approximately equivalent to those recorded from the same sites two years after sludge application. These elevated levels of Cd in shrews did not, however, appear to be biologically significant. The results for Peromyscus showed some significantly increased Cd concentrations in mice from some sewage-treated sites, but this was not a consistent trend, and Nickelson and West concluded that it was doubtful if sewage biosolids had a significant long-term effect on these omnivores. Campa et al. (1987) found that white-tailed deer (Odocoileus virginianus) and elk (Cervus elaphus canadensis) in a clear-felled forest area browsed more heavily on sludge-treated (9,980 kg ha-1) vegetation than in control areas. The vegetation in sludge-treated areas had a higher crude protein content. Deer grazing predominantly in sludge-treated areas had higher heavy metal concentrations in their tissues, but level were not considered to be high enough to threaten the health of the deer or humans who might eat them. There are numerous other publications relating to the accumulation of heavy metals in mammals (e.g. Babish et al., 1982; Maly, 1984; Telford et al., 1984; Anderson, 1985; Bray et al., 1985; Dressler et al., 1986; Alberici et al., 1989; Brueske & Barrett, 1991). The presence of pathogens (e.g Salmonella) in animal manures, improperly treated sewage sludge (Wray & Callow, 1985) or slurries mixed with sheep dip (Semple et al., 2000) may also pose a potential disease risk to grazing mammals and their predators. 65 BIOENERGY CROPS AND BIOREMEDIATION 2.4.3 Industrial wastes There is a very wide range of waste materials from industrial processes that could potentially be disposed of on bioenergy crops – or on sites intended subsequently for bioenergy crop production. Those of greatest potential interest probably include wastes from the paper industry, such as de-inked sludges. Paper industry wastewater sludges are relatively low in heavy metals. Some fresh sludges may be toxic (e.g. to bacteria and plants), but composting can readily reduce toxicity making them suitable for application to agricultural land (Rantala et al., 1999). Vegetation No relevant information was found on the effects of industrial waste applications to bioenergy crops on ground vegetation. However, the direct effects of mulching on the ground flora and indirect effects of nutrient enrichment on the crop and ground flora described for agricultural manures will also be applicable for large volume applications of industrial wastes. Soil biodiversity Erstfeld & Snow-Ashbrook (1999) studied the effects on soil-dwelling invertebrates of low levels of polycyclic aromatic hydrocarbons (PAHs), in soils from an abandoned industrial site. Their results suggested that higher PAH concentrations (in the range 5.28 to 80.46 mg kg-1) were associated with increased taxonomic diversity of nematodes, increased abundance of omnivorous and predatory springtails (Collembola) and increased growth rates in earthworms (Eisenia andrei). The same study showed a negative effect of increased PAH concentration on the total abundance of mites (order Acarina). It is not known what effects, if any, the much lower rates of PAHs found in animal manures and slurries (see Section 2.3.2) might have on soil-inhabiting invertebrates. Krogh & Pedersen (1997) reported the results of a Danish study, which examined the effects of applying dried and sterilised waste-water sludge, from a pesticide factory, on the microarthropod fauna in the soil of a mature Norway spruce (Picea abies) forest. The granulated sludge had a dry matter content of 93%, 8-10% organic matter and contained 9.2% P and 25.5% CaCO3. The sludge granules also contained small residues of phosphate triesters, pesticide residues, pyrimidines and organic phosphoric acids. Results showed that the sludge significantly reduced populations of springtails and mites. However, applications of inorganic fertiliser had similar effects, although springtails were less affected by this treatment. Both direct toxicity and changes in the microbial community as a result of increased nutrient availability were suggested as probable causes of these population declines. Overall, springtail populations were 30-35% lower in fertiliser and sludge treatments than in control plots, one year after treatment application. One springtail species, however, Isotoma notabilis, actually increased in numbers in 66 BIOENERGY CROPS AND BIOREMEDIATION response to sludge applications; to 2.6 times the control level (60% of total springtail numbers) in the high dosage treatment (1.5 t ha-1). Krogh & Pedersen had expected that the uneven distribution of sludge would result in ‘protective sub-habitats’ - where soil fauna populations were unaffected – but found no evidence of this. Ground-dwelling invertebrates No relevant information was found on the effects of industrial waste applications to bioenergy crops on populations of ground-dwelling invertebrates. However, the indirect effects on invertebrates resulting from changes in the ground flora crop, due to mulching or nutrient enrichment – as described for agricultural manures will also be applicable for large volume applications of industrial wastes. Some industrial wastes may contain chemicals that are directly toxic to invertebrates. Canopy invertebrates No relevant information was found on the effects of industrial waste applications to bioenergy crops on populations of canopy invertebrates. However, wastes that are nutrient-rich may increase the susceptibility of the energy crop to herbivorous insects, although (as described for agricultural wastes and sewage sludge) this will be largely irrelevant if the alternative is the application of similar quantities of nutrients in inorganic fertilisers. Birds No relevant information was found on the effects of industrial waste applications to bioenergy crops on bird populations. However, certain types of organic waste will be attractive to birds, which either utilise the waste directly as a food source or feed upon the invertebrate fauna foraging and breeding within the waste. For example, Gabrey (1997) surveyed bird populations at different waste management facilities in the USA and collected data that clearly demonstrates the attractiveness of putrescible waste to gulls and other birds. In total, about 350 times more birds were recorded at a putrescible landfill site than at five other sites, which included a ‘yard-waste compost facility and a ‘trash-transfer station. Mammals No relevant information was found on the effects of industrial waste applications to bioenergy crops on mammal populations. However, as has been illustrated for sewage sludge and animal manures, whenever wastes containing high levels of heavy metals are applied there are risks of toxic materials accumulating in the organs of animals that are resident within or utilise treated areas. Similar risks will exist where energy crops are used to stabilise or bioremediate metal contaminated sites, although the risks are likely to have been present prior to the establishment of the energy crop. The dangers of toxicological damage are greatest to predatory species, with the risks of ‘bioconcentration’, in the food chain. Relevant studies include the work of Koeck et al. (1989), on heavy metal accumulation in small mammals at a waste disposal site, and Andrews et al. (1984) on cadmium 67 BIOENERGY CROPS AND BIOREMEDIATION accumulation in the tissues of field voles and common shrews on grassland over metalliferous mine waste. Meyn et al. (1997) modeled the exposure risk of three representative bird and mammal species, to assess the levels of hazard posed by 2,3,7,8tetrachlorodibenzo-p-dioxin (TCDD) in land applications of pulp and paper mill sludge. Their results suggest that TCDD may, in some circumstances, pose significant risks to terrestrial wildlife. Shrews were considered to be at most risk because of their high rates of consumption of invertebrates that are expected to accumulate TCDD from soils and vegetation at application sites. Other research has looked at the potential toxicological effects in small mammal species from exposure to hazardous materials, including petrochemical wastes (McBee, 1991; Feuston & Mackerer, 1996), organochlorine insecticides (Rowley et al., 1983) and even radioactive wastes (Arthur et al., 1986). 2.4.4 References Alberici, T M; Sopper, W E; Storm, G L & Yahner, R H (1989). Trace metals in soil, vegetation, and voles from mine land treated with sewage sludge. Journal of Environmental Quality 18 (1). 115-120. Anderson, D A (1985). Influence of sewage sludge fertilization on food habits of deer in Western Washington. Journal of Wildlife Management 49 (1). 91-95. Andrews, S M; Johnson, M S & Cooke, J A (1984). Cadmium in small mammals from grassland established on metalliferous mine waste. Environmental Pollution. Series A: Ecological and Biological 33 (2). 153162. Arthur, W J; Markham, O D; Groves, C R; Keller, B L & Halford, D K (1986). Radiation dose to small mammals inhabiting a solid radioactive waste disposal area. Journal of Applied Ecology 23 (1). 13-26. Babish, J G; Johnson, B; Brooks, B O; Lisk, D J (1982). Acute toxicity of organic extracts of municipal sewage sludge in mice. Bulletin of Environmental Contamination and Toxicology 29 (4). 379-384. Bag, S; Vora, T; Ghatak, R; Nilufer, I; D’Mello, D; Pereira, L; Pereira, J; Cutinho, C & Rao, V (1999). A study of toxic effects of heavy metal contaminants from sludge-supplemented diets on male Wistar rats. Ecotoxicology and Environmental Safety 42 (2). 163-170. Bray, B J; Dowdy, R H; Goodrich, R D & Pamp, D E (1985). Trace metal accumulations in tissues of goats fed silage produced on sewage sludgeamended soil. Journal of Environmental Quality 14 (1). 114-118. Brueske, C C & Barrett, G W (1991). Dietary heavy metal uptake by the least shrew, Cryptotis parva. Bulletin of Environmental Contamination and Toxicology 47 (6). 845-849. 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Distribution of microorganisms, biomass ATP, and enzyme activities in organic and mineral particles of a longterm wastewater irrigated soil. J. Plant Nutr. Soil Sci. 163. 143-150. Feuston, M H & Mackerer, C R (1996). Developmental toxicity study in rats exposed dermally to clarified slurry oil for a limited period of gestation. Journal of Toxicology and Environmental Health 49 (2). 207-220. Fuller, R J & Green, G H (1998). Effects of woodland structure on breeding bird populations in stands of coppiced lime (Tilia cordata) in western England over a 10-year period. Forestry 71(3). 199-218. Gabrey, S W (1997). Bird and small mammal abundance at four types of wastemanagement facilities in northeast Ohio. Landscape and Urban Planning 37 (3-4). 225-235. Hegstrom, J & West, S D (1989). Heavy metal accumulation in small mammals following sewage sludge application to forests. Journal of Environmental Quality 18 (3). 345-349. Jongbloed, A W & Lenis, N P (1998). Environmental concerns about animal manure. In: Proceedings of ‘Nutrient Management Procedures to Enhance Environmental Conditions’ symposium. July 1997; Nashville, USA. Journal of Animal Science 76 (10). 2641-2648. Kielhorn, K –H; Keplin, B & Hüttl, R F (1999). Ground beetle communities on reclaimed mine spoil: Effects of organic matter application and revegetation. In: Proceedings of the international symposium Organic matter application and turnover in disturbed terrestrial ecosystems (Ed. D Vetterlein). Cottbus, Germany. November 1997. Plant and Soil 213 (1-2). 117-125. Koeck, M; Schaffler, R; Sixl, W; Pichler-Semmelrock, F P; Kosmus, W & Marth, E (1989). Accumulation of heavy metals in animals. III: Heavy metal accumulation in small mammals at a waste disposal site. Journal of Hygiene, Epidemiology, Microbiology and Immunology 33 (4). 536-541. Krogh, P H & Pedersen, M B (1997). Ecological effects assessment of industrial sludge for microarthropods and decomposition in a spruce plantation. Ecotoxicology and Environmental Safety 36. 162-168. Larsen, K J; Purrington, F F; Brewer, S R & Taylor, D H (1996). Influence of sewage sludge and fertilizer on the ground beetle (Coleoptera: Carabidae) fauna of an old-field community. Environmental Entomology 25 (2). 452459. 69 BIOENERGY CROPS AND BIOREMEDIATION Lynn, S F; Slater, F M & Randerson, P F (1992). The ecological impact of sewage sludge applications on woodland vegetation. Aspects of Applied Biology 29, Vegetation management in forestry, amenity and conservation areas. 383-388. Maly, M S (1984). Survivorship of meadow voles, Microtus pennsylvanicus, from sewage sludge-treated fields. Bulletin of Environmental Contamination and Toxicology 32 (6). 724-731. Marambe, B & Ando, T (1992). Phenolic acids as potential seed germinationinhibitors in animal waste composts. 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Renal cadmium concentrations in mice and shrews collected from forest lands treated with biosolids. Journal of Environmental Quality 25 (1). 86-91. Pallant, E & Hilster, L M (1996). Earthworm response to 10 weeks of incubation in a pot with acid mine spoil, sewage sludge and lime. Biology and Fertility of Soils 22 (4). 355-358. Pasteels, J M & Rowell-Rahier, M (1992). The chemical ecology of herbivory on willows. In: Proceedings of Botanical Society of Edinburgh symposium on Willow; Edinburgh, UK; 27-29 September 1991. Proc. Royal Society of Edinburgh (Sect B) 98 (0). 63-73.. Perttu, K (1999). Environmental and hygienic aspects of willow coppice in Sweden. Biomass and Bioenergy 16. 291-297. Rantala, P-R; Vaajasaari, K; Juvonen, R; Schultz, E; Joutti, A & MäkeläKurtto, R (1999). Composting of forest industry wastewater sludges for agricultural use. In: Forest Industry Wastewaters VI (Ed. A Luonsi). Selected proceedings of the 6th IAWQ Symposium on Forest Industry Wastewaters, Tampere, Finland; 6-10 June 1999. Wat. Sci. Tech. 40 (11-12). 187-194. Reddersen, J (2001). SRC-willow (Salix viminalis) as a resource for flowervisiting insects. Biomass and Bioenergy 20. 171-179. 70 BIOENERGY CROPS AND BIOREMEDIATION Rowley, M H; Christian, J J; Basu, D K; Pawlikowski, M A & Paigen, B (1983). Use of small mammals (voles) to assess a hazardous waste site at Love Canal, Niagara Falls, New York. Archives of Environmental Contamination and Toxicology 12 (4). 383-397. Sage, R B & Robertson, P A (1996). Factors affecting songbird communities using new short rotation coppice habitats in spring. Bird Study 43. 201-213. Sage, R B & Tucker, K (1997). Invertebrates in the canopy of willow and poplar short rotation coppices. Aspects of Applied Biology 49, Biomass and energy crops. 105-111. Sage, R B & Tucker, K (1998a). The distribution of Phratora vulgatissima (Coleoptera: Chrysomelidae) on cultivated willows in Britain and Ireland. Eur. J. For. Path. 28. 289-296. Sage, R & Tucker, K (1998b). Integrated crop management of SRC plantations to maximise crop value, wildlife benefits and other added value opportunities. ETSU/B/W2/00400/REP. Semple, K T; Hughes, P; Langdon, C J & Jones, K (2000). Impact of synthetic pyrethroid sheep dip on the indigenous microflora of animal slurries. FEMS Microbiology Letters 190 (2). 255-260. Šlapokas, T & Granhall, U (1991). Decomposition of willow-leaf litter in a short-rotation forest in relation to fungal colonization and palatability for earthworms. Biology and Fertility of Soils 10. 241-248. Slater, F M; Hodson, R W; Randerson, P F & Lynn, S F (1997). Some environmental impacts of short rotation willow coppice. In: Making a Business from Biomass in Energy, Environment, Chemistry, Fibres and Materials (Eds R P Overend & E Chornet). Proceedings 3rd Biomass Conference of the Americas, Montreal, Quebec, Canada, August 24-29, 1997. Pergamon, Elsevier Science. 29-37. Telford, J N; Babish, J G; Johnson, B E; Thonney, M L; Currie, W B; Bache, C A; Gutenmann, W H & Lisk, D J (1984). Toxicological studies with pregnant goats fed grass-legume silage grown on municipal sludge-amended topsoil. Archives of Environmental Contamination and Toxicology 13 (5). 635-640. Vasseur, L; Cloutier, C & Ansseau, C (2000). Effects of repeated sewage sludge application on plant community diversity and structure under agricultural field conditions on Podzolic soils in eastern Quebec. Agriculture, Ecosystems & Environment 81 (3). 209-216. Vaughan, N; Jones, G & Harris, S (1996). Effects of sewage effluent on the activity of bats (Chiroptera: Vespertilionidae) foraging along rivers. Biological Conservation 78 (3). 337-343. Vaughan, N; Jones, G & Harris, S (1997). Habitat use by bats (Chiroptera) assessed by means of a broad-band acoustic method. Journal of Applied Ecology 34. 716-730. Woodyard, D K; Campa, H & Haufler, J B (1987). The influence of forest application of sewage sludge on the concentration of metals in vegetation and small mammals. In: The forest alternative for treatment and utilization of 71 BIOENERGY CROPS AND BIOREMEDIATION municipal and industrial wastes (Eds. D W Cole, C L Henry & W L Nutter). University of Washington Press, Seattle. 199-205. Wray, C & Callow, R J (1985). A note on potential hazards to animals grazing on pasture improperly treated with sewage sludge. Journal of Applied Bacteriology 58 (3). 257-258. 72 BIOENERGY CROPS AND BIOREMEDIATION CHAPTER 3 BIOREMEDIATION OF CONTAMINATED SITES PETER NIXON 3.1 INTRODUCTION The public welfare concern over the effects of environmental pollution has increased substantially since the Industrial Revolution, mainly as a consequence of an enhanced understanding of the risk to human health. Much concern has concentrated on the visible effects of pollution, but the hidden effects are also of great importance. Large concentrations of inorganic pollutants (i.e. trace elements) in soils can be of either natural or anthropogenic origin. Natural sources are the result of weathering processes, volcanic activity and natural fires in which plants or wood burn. The main sources of pollution are from the burning of fossil fuels, mining and smelting of metalliferous ores, inorganic and organic municipal waste and sewage, and residues from materials used in agriculture. The necessity to decontaminate polluted sites is recognised, both socially and politically, because of the increasing importance placed on environmental protection and human health. As the number of sites and levels of contamination rise, so does the need to develop effective and affordable methods for decontamination (Lombi et al., 1998). 3.2 PHYTOREMEDIATION The use of plants for soil reclamation has begun to be looked at as an alternative to physical or chemical processes. The term ‘phytoremediation’ is used to describe techniques in which plants are used for the in situ treatment of soils polluted by chemicals or radioactivity. There are four fundamental processes by which plants can be used to remediate soils contaminated with trace elements (Salt et al., 1996): 1. 2. 3. 4. Phyto-immobilisation – where plants are used to prevent the movement and transportation of dissolved contaminants within the soil. Phyto-stabilisation - where pollutant-tolerant plants are used to mechanically stabilise polluted soils, and to prevent bulk erosion and airborne transport to other environments. In addition, both phyto-immobilisation and phytostabilisation may reduce contaminated run-off, due to higher evaporation rates relative to bare soil. Phyto-extraction – where plants are used to extract metallic and organic compounds from the soil into plant tissue. Phyto-volatilisation – where specialised enzymes transform and volatilise contaminants in a plant-microbe-soil system. Phytoremediation is particularly suited to large sites that have relatively low levels of contamination, but still sufficiently contaminated to restrict their use and limit development potential. This low-cost option can also be used to stabilise the site 73 BIOENERGY CROPS AND BIOREMEDIATION until funds and/or techniques have become available to remediate the contaminated land. The production of biomass fuel crops on degraded or contaminated land has several advantages as a means of site remediation. Biomass crop production will not only bring such sites into economic use, but may also improve their aesthetic value, and potentially lead to long-term remediation, through heavy metal removal in the harvested crop (D. RiddellBlack - pers. com.). Some current biomass crops, such as Salix, Populus and Miscanthus are pioneer species and are adapted to harsh growing conditions - which typify derelict sites. For example, Miscanthus invades derelict, abandoned and volcanic areas in its native Japan (personal observation). Establishment and management costs are low compared with chemical washing. The contaminates are contained on site, rather than being removed and transported to another site. The energy conversion offers a method for concentrating the metals contained within the biomass. Lastly the production of a low risk, non-food crop on otherwise unproductive land can play a role in the rejuvenation of the local economy (D. RiddellBlack - pers. com.). In Britain, Punshon et al. (1995) found that species of willows accumulated cadmium in the leaves and stems. In Sweden, again with willows, it was found that the average stem contained 2.1 mg dm kg-1, and it was estimated that 21 g ha-1 annually could be removed from the soil, with an annual crop yield of 10 t DM ha-1. This corresponded to the removal of 3-4% of the plant available cadmium (Ostman, 1994). Under controlled conditions in the laboratory it was observed that Betula pendula was able to accumulate high concentrations of cadmium (Goransson, 1994). It was estimated that up to 1.5 kg Cd ha-1 yr-1 could be removed from the soil with no detrimental effect on the growth of the plant. The vast majority (95%) of this Cd was found to be in the roots. This study estimated that there was also a significant annual uptake of other metals, such as Al (25 kg ha-1 yr-1), Cu (1.5 kg ha-1 yr-1), Fe (5 kg ha-1 yr-1), and Mn (50 kg ha-1 yr-1). Again in Sweden, Ericson (1994) found that Salix spp. are capable of accumulating 1.3 g Cd ha-1 annually. Punshon et al. (1995) also investigated the resistance to copper toxicity in various British willows and found that there were significant differences in root length, the number of lateral roots and the pattern of metal uptake between species, hybrids and clones. This study also found that Salix caprea, S. cinerea and their hybrids, and S. viminalis appear to grow best in elevated copper solution. Other studies have also demonstrated the ability of Salix species to tolerate polluted and hostile environments. For example, S. caprea will grow on lead mine spoils (Eltrop et al., 1991) and the hybrids of S. caprea, S. cinerea and S. viminalis will grow on metal contaminated river silt (Mang & Reher, 1992). 74 BIOENERGY CROPS AND BIOREMEDIATION The uptake of heavy metals by twenty different varieties of willow showed that there were significant differences in the concentrations of Pb, Cr, Cu and Ni in the above ground tissues (Riddell-Black, 1997). However, the varieties tested appeared to fall into two distinct groups: Those that accumulated Cu or Ni, in the above ground biomass, and as a consequence suffered substantially reduced yields. Those that did not accumulate Cu or Ni, and so suffered no yield reduction. The willow varieties tested seemed able to tolerate high concentrations of Zn or Cd in bark tissue, without yields being influenced. From this study, there appears to be two scenarios for the use of willow SRC on contaminated land: 1. Planting of non-accumulating varieties (e.g. Salix ‘Rosewarne White’ and S. spaethii), to stabilise the metal losses via wind and water erosion. This option allows only limited heavy metal removal, but does permit the use of the wood for fuel. 2. Planting a variety such as S. burjatica ‘Germany’, which appears to remove Cu, Zn, Cd and Ni from the soil in significant quantities, having both high metal concentrations in the above ground tissue and high yields. However, it may not be possible to use this wood for fuel with the current technology. Carol et al. (1996) used grey alder (Alnus incana), silver birch (Betula pendula) and Scots pine (Pinus sylvestris) in the phytoremediation of an area of steelworks’ slag, in Lanarkshire - contaminated with copper, nickel and zinc. They found that these three tree species could all be successfully grown on the moderately alkaline slag. Trees grew best on slag that had been treated with sewage cake prior to planting. Grey alder had the best growth rates. However, the trees only managed to accumulate less than 1% of the top soil metal contamination in the above ground biomass. This could have been due to the high pH and the chemical form of the metals in the waste. The scheme was, nonetheless, judged a success as it greened the site, created a wildlife habitat, improved soil fertility and stabilised the metal contaminants. Scott et al.. (1995) looked at the uptake of copper, chromium, lead and zinc in birch, willow and sycamore (Acer pseudoplatanus) growing on two sites; one an ex-chromium works and the other an ex-ammunition factory. These trees were not able to significantly accumulate the metals under investigation, although zinc was the most mobile and was detected at reasonably high concentrations in the above ground biomass, the concentrations being highest in the bark and leaves. Changes in zinc concentration in different parts of the tree took place over the growing season, with the suggested optimum time for harvest being the winter period. The other metals were only found at high concentrations in the tree roots; a positive correlation between zinc uptake and zinc concentration in the soil was found. Steer & Baker (1997) investigated the use of three trees - Populus deltoides x trichocarpa (P. x interamericana) ‘Beaupré’, common alder (Alnus glutinosa) and grey willow (Salix cinerea) – grown as SRC biomass crops, to re-generate two old reclaimed coal tips in Wales. All three species showed very high rates of survival. On the better of the two sites, poplar produced mean yields of up to 6 t DM ha-1 yr1 (excluding dead trees), at the first harvest, in plots pre-treated with an application of 300 mm raw sewage sludge (which was applied below the top 1 m of colliery 75 BIOENERGY CROPS AND BIOREMEDIATION spoil). The authors suggest that their results indicate the potential for SRC poplar grown on old coal spoil to produce yields comparable to those from some lowland agricultural sites, but only under the right conditions i.e. with ready availability of primary nutrients and a lack of compaction. The other two species (and poplar in other treatments) were less impressive in terms of yield, but alder was capable of becoming established and growing with little if any pre-treatment. This work (Steer & Baker, 1997) highlighted the criteria for successful establishment of trees on this type of land: choice of the correct species or clones for local site conditions thorough site preparation and, especially, avoidance of compaction availability of nutrients good site management, especially the use of effective weed control and fencing. Steer & Baker (1997) concluded that the use of SRC on re-claimed coal sites had real potential, both for the production of biomass, and also for the benefits of the environment. A pot study by Wilkins (1997) on the uptake of copper, arsenic and zinc by Miscanthus sp. grown on metalliferous soils, showed little difference in uptake on polluted or unpolluted soils. This study did demonstrate that Miscanthus was able to survive and to grow on highly polluted soils, and that the use of inorganic fertiliser and/or lime greatly improved the yield of Miscanthus with very little influence on the uptake of metals. The work concluded that there was little cause for concern in the growing of Miscanthus as a biofuel on soil polluted by mining. Another pot study followed the uptake and mobilisation of cadmium, zinc and copper in Miscanthus x giganteus. It showed that levels of Cu and Zn were mainly confined to the roots and rhizomes, but concentrations in plants did not show any increase in uptake with increased soil supply. This is in complete contrast to cadmium concentrations, which did increase with increases in soil supply. Levels of cadmium were found to be highest in the roots and rhizomes (S. Wilson - pers. comm.). The use of Miscanthus in Portugal as an accumulator of heavy metals was studied, using pots. It was found that the level of heavy metal concentration in the soil negatively affected growth and productivity. The results indicated that Miscanthus was able to accumulate and remove heavy metals (Cd, Cr, Cu, Ni, Pb & Zn) from the soil in the below-ground fraction of the plant, but that the aerial fraction did not significantly accumulate the heavy metals (Fernando et al., 1996). The work was repeated in the field, comparing fields with different levels of contaminants. Results showed that productivity, plant height and stem numbers all increased with increased levels of sewage sludge (up to 100 t ha-1), but levels of heavy metals (Cd, Cr, Cu, Ni, Pb & Zn) in the above-ground biomass did not significantly increase with increased metal concentrations in the soil. The ash, nitrogen and phosphorus content of the above ground biomass did increase with increased levels of contaminants. However, the below-ground material did accumulate higher concentrations of heavy metals with increased levels of contaminants (A Fernando - pers. com.). 76 BIOENERGY CROPS AND BIOREMEDIATION The suitability of perennial grasses from the botanical garden of the Plant Breeding and Acclimatization Institute (PBAI), Bydgoszcz, Poland was assessed, in order to see if they could be used to phyotremediate a coal dump in Poland (Majtkowski & Majtkowska, 1997). The use of grasses are favoured in Poland as there is concern that deep rooting trees may cause oxidation of pyrite, resulting in spontaneous combustion. Two of the most promising species were found to be Miscanthus sacchariflorus and Spartina michauxiana. It was also found that the incorporation of biogel (water absorbent product) into the soil increased the absorption ability of plants in poor soils and improved the plants' ability to utilise moisture. The study concluded that the use of grass offered one of the most effective methods of remediating the coal-mine spoils of the Katowice region. AEA Technology, in collaboration with Border Biofuels and Derbyshire County Council, have investigated the use of SRC as a low cost route to the remediation of coal spoil sites - and identified a number of benefits. These include the rapid capture and greening of a site, site stabilisation due to the extensive network of fibrous roots, high productivity, leaf fall leading to increased organic matter and the creation of soil, and the control of contaminated run-off. The use of trees may benefit the local area in other ways such as improving the visual amenity of the area and the creation of new woodlands with increased public access and recreational activities. Their methods are based on the incorporation of organic material (e.g. sewage and other organic sludges) into the surface of the contaminated soil, prior to the planting of SRC (F Dumbleton - pers. com.). The recently completed, EC-funded, BIORENEW project examined the potential for the use of biomass fuel crops in the bioremediation and economic renewal of industrially degraded land. The main objective of the BIORENEW project was to develop a system for the rehabilitation of heavy metal contaminated land using biomass fuel crops, which brings a net environmental benefit and contributes to the economic regeneration of areas suffering industrial decline. A number of field screening trials of Salix, Phalaris and Eucalyptus in the UK, Sweden and Spain were undertaken; along with the screening of 150 Salix clones, 20 Phalaris clones and 10 Miscanthus for the uptake of heavy metals. The use of vegetation on completed landfill sites has been studied for a number of years. Ettala (1988), in southern Finland, concluded that SRC could be successfully established on sanitary landfills. Of the five Salix spp. planted, the most productive was S. aquatica. The SRC on the landfill was managed in the same way as a conventional SRC crop on an arable soil. The study also found that the best results were achieved when the substrate had a high humus content and a thickness of at least 0.2-0.3 m. The planting of SRC on landfill notably improved the landscape of the site but also increased evapotranspiration. The use of vegetation on landfill sites restoration is also important, in that it provides shelter and reinforcement for the capping system and contributes to its basic functioning. However, it may cause a potential risk, due to the damage that some tree species roots may cause by penetrating some types of sealers (McDonald et al., 1997). Work in Hong Kong on the restoration of completed landfill sites (Chan, 1997), using Acacia confusa and Casuarina equisetifolia, found that these 77 BIOENERGY CROPS AND BIOREMEDIATION two N2-fixing trees had a rooting system which was confined to the upper 15 cm of the topsoil and did not penetrate any deeper; thus the risk of penetrating the 1 m cap was very low. It was also possible to establish these species without the need for imported planting medium, which could increase the cost of remediation. 3.3 POTENTIAL ENVIRONMENTAL PROBLEMS The two main areas of concern with regard to planting a biomass crop for the phytoremediation of contaminated land are the possible contamination of the wider environment via the food chain and of the release of metals following combustion. 3.3.1 Bioaccumulation of pollutants Studies on the transfer of heavy metals by animals feeding on contaminated sites and the subsequent toxic hazards to birds and other top predators is incomplete, but the main route is likely to be via soil feeding animals (Punshon & Dickinson, 1997). The ability of earthworms to accumulate cadmium is a good example, and thus earthworm-eating birds may be susceptible. However, most birds feed over a large area and on a range of invertebrates, and unless they feed exclusively on a contaminated site their chances of suffering from toxic effects are unlikely (Furness, 1996). Small mammals may also accumulate high levels of cadmium and lead, so top predators feeding exclusively on small mammals from contaminated sites may be exposed to elevated levels. These issues are discussed more fully in Section 3.5 of this review. 3.3.2 Release of metals during combustion Besides the possible threat to the environment from the combustion of biomass, there are also additional operating costs due to the higher specification of the flue gas cleaner to ensure satisfactory metal removal. It has been estimated that the cost of flue gas cleaning is between 5 and 15% of the capital cost of a combined heat and power (CHP) system, but less than 1% of the annual running costs when using conventional fuel sources. Therefore, any increase in the level of metals in biomass fuel material may have an implication in the capital and annual costs for systems using such fuels. There is a further problem, as the resulting ash may have to be disposed of in a landfill site at a cost of between £5 and £15 per tonne, rather than being used as a good source of P and K for biomass and other crops. If the ash was designated as hazardous or as special waste this would further increase the disposal cost to between £50 and £150 per tonne (Riddell-Black et al., 1996). Narodoslawsky et al. (1996) have suggested some solutions to this problem. Firstly the mixing of fly-ash and bottom in equal quantities to produce a blended ash which may be recycled on to the land. Secondly (the long-term solution) the use of different temperatures in a biomass combustion plant to concentrate volatile metals, such as cadmium, in a very small portion of the ash. This small portion of ash could then be disposed of safely in a landfill site, and the remaining clean ash used in agriculture. 78 BIOENERGY CROPS AND BIOREMEDIATION 3.4 REFERENCES Carol, S A; Hipkin, A J & Davidson, B (1996). Phytoremediation - a feasible option at Lanarkshire steelworks? In: Heavy Metals and Trees (Ed. I Glimmerveen). Institute of Chartered Foresters, Edinburgh. 51-62. Chan, Y. S. G (1997). Root growth patterns of two nitrogen-fixing trees under landfill conditions. Land Contamination & Reclamation 5. 55-62. Eltrop, L; Brown, G; Hinchee, R E & Olfenbuttel, R F (1991). Lead tolerance of Betula and Salix in mining areas of Mechernich, Germany. Plant Soil 131. 275-285. Ericson, S-O (1994). Salix can remove cadmium from arable land - technical and infra-structural implications. Proceedings of a study tour, conference and workshop in Sweden, June 1994. Eds. P Aronsson & K Perttu. Swedish University of Agricultural Sciences Report 50. 173-174. Ettala, M O (1988). Short-rotation tree plantations at sanitary landfills. Waste Management & Research 6. 291-302. Fernando, A; Duarte, P & Oliveira, J F S (1996). Bioremoval of heavy metals from soil by Miscanthus sinensis giganteus. Biomass for Energy and the Environment. Proceedings of the 9th European Bioenergy conference. 531536. Furness, R W (1996). Transfers of heavy metals from contaminated land to top predators; implications for birds for growing trees for soil amelioration. In: Heavy Metals and Trees (Ed. I Glimmerveen). Institute of Chartered Foresters, Edinburgh. 107-122. Goransson, A & Philippot, S (1994). The use of fast growing trees as ‘metalcollectors’. Proceedings of a study tour, conference and workshop in Sweden, June 1994. Eds. P Aronsson & K Perttu. Swedish University of Agricultural Sciences Report 50. 129-132. Lombi, E; Wenzel, W W & Adriano, D C (1998). Soil contamination, risk reduction and remediation. Land Contamination & Reclamation 6. 183-197. Majtkowski, W & Majtkowska, G (1997). Suitability of ornamental perennial grasses to reclamation of coal-mining dump. Ecological aspects of breeding fodder crops and amenity grasses. Plant Breeding and Acclimitization Institute, Bydgoszcz, Poland. 249-252. Mang, F W C; Reher, R (1992). Heavy metal resistance clones of willows from polluted areas useful for land restoration programmes. Proceedings of the Royal Society, Edinburgh 98B. 244. McDonald, C; Meggyes, T & Simmons, E (1997). Landfill capping: engineering and restoration, 1. Structure and function of landfill capping system. Land Contamination & Reclamation 5. 89-97. Narodoslawsky, M & Obernberger, I. (1996). From waste to raw material - the route from biomass to wood ash for cadmium and other heavy metals. Journal of Hazardous Materials 50. 157-168. 79 BIOENERGY CROPS AND BIOREMEDIATION Ostman, G (1994). Cadmium in Salix - a study of the capacity of Salix to remove cadmium from arable soils. Proceedings of a study tour, conference and workshop in Sweden, June 1994. Eds. P Aronsson & K. Perttu. Swedish University of Agricultural Sciences Report 50. 153-155. Punshon T; Lepp, N W & Dickinson, N M (1995). Resistance to copper toxicity in some British willows. Journal. of Geochemical Exploration 52, 259-266. Punshon, T & Dickinson, N M (1997). Mobilisation of heavy metals using shortrotation coppice. Aspects of Applied Biology 49. Biomass and Energy Crops. 285-292. Punshon, T; Dickinson, N M & Lepp, N W (1996). The potential of Salix clones for bioremediating metal polluted soil. In: Heavy Metals and Trees (Ed. I Glimmerveen). Institute of Chartered Foresters, Edinburgh. 93-104. Riddell-Black, D M; Pulford, I D & Stewart, C (1997). Clonal variation in heavy metal uptake by willow. Aspects of Applied Biology 49. Biomass and Energy Crops. 327-334. Riddell-Black, D M; Rowlands, C & Snelson, A (1996). The take up of heavy metals by wood fuel crops - implications for emission and economics. Biomass for Energy and the Environment. Proceedings of the 9th European Bioenergy conference 3. 1754-1759. Salt, D E; Blaylock, M; Kumar, N P B A; Dushenkov, V; Ensley, B D; Chet, L & Raskin, L (1996). Phytoremediation: a novel strategy for the removal of toxic metals from the environment using plants. Biotechnology 13. 468-474. Scott, McG D; Duncan, H J; Pulford, I D & Wheeler, C T (1996). Uptake of heavy metals from contaminated soil by trees. In: Heavy Metals and Trees (Ed. I Glimmerveen). Institute of Chartered Foresters, Edinburgh. 171-176. Steer, P & Baker, R M (1997). Colliery spoil, sewage and biomass - potential for renewable energy from wastes. Aspects of Applied Biology 49. Biomass and Energy Crops. 300-305. Wilkins, C (1997). The uptake of copper, arsenic and zinc by Miscanthus environmental implications for use as an energy crop. Aspects of Applied Biology 49. Biomass and Energy Crops. 335-340. 80 BIOENERGY CROPS AND BIOREMEDIATION CHAPTER 4 WASTE UTILISATION AND BIOREMEDIATION: THE ROLE OF GENETICALLY MODIFIED ORGANISMS NATASHA SMITH 4.1 INTRODUCTION The genetic modification of biomass crops to facilitate increased pollutant uptake, transport, accumulation and tolerance offers the potential to dramatically increase the effectiveness of phytoremediation of organic and metal pollutants from contaminated sites. Optimisation of performance using traditional plant breeding techniques is limited to the genetic diversity found within the species itself. Recent advances in molecular biology, in particular genetic engineering, make it possible, in theory, to introduce virtually any gene into any organism. In order to achieve such a goal, it is necessary to understand the mechanisms employed by organisms to prevent poisoning, as a result of exposure to high levels of metals and organic compounds. This chapter will examine the current understanding of these mechanisms, their application to phytoremediation and the progress of transformation technology with respect to biomass crops. The differing nature of inorganic and organic pollutants requires different mechanisms to facilitate remediation. Heavy metals and radionuclides are classed as inorganic pollutants and examples include cadmium, cesium, chromium, lead, mercury, strontium and uranium. These pollutants are immutable in terms of biological or physical processes, excluding nuclear fission and fusion. Effective phytoremediation of such substances are therefore, limited to (i) the extraction, translocation and sequestration of the toxic ions to plant tissues; facilitating the prevention of leaching from the site, or removal from the site by harvesting the plant material, or (ii) conversion of the element to a less toxic chemical species, for example: transformation of chromium VI to chromium III (James, 1996). In contrast, many organic pollutants can be reduced to relatively non-toxic compounds and even CO2 and water. Targets for phytoremediation include polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), trichloroethylene (TCE) and trinitrotoluene (TNT). Plants have many endogenous genetic, biochemical and physiological systems that make them ideal for the remediation of soils and water. Pollutants are often taken up by the same mechanisms that adsorb, transport and translocate nutrients. Once adsorbed and transported, pollutants can either hyperaccumulate or undergo some form of transformation. 81 BIOENERGY CROPS AND BIOREMEDIATION 4.2 PHYTOREMEDIATION OF INORGANIC POLLUTANTS 4.2.1 Root uptake Plant root systems offer a large surface area with which to adsorb nutrients from the surrounding soil matrix (Boyajian & Carreira, 1997). Inorganic pollutants also become bound to the root surface during adsorption. In liquid media, the roots of Brassica juncia have been shown to contain 500 times more Cd(II), Ni(II), Pb(II) and Sr(II) than the surrounding liquid (Salt et al., 1995; Salt & Kramer, 1999). Similarly, sunflower roots can accumulate uranium such that it is 30,000 times more concentrated than the surrounding liquid media (Dushenkof et al., 1997). These levels of accumulation from water are dramatic; in soils, however, much of the metal is bound to the soil material itself and must be mobilised into the soil solution before it can be adsorbed. To achieve this the plant can secrete metalchelating molecules known as phytosiderophores which once in the rhizosphere chelate and solubilise soil-bound metals. Two compounds, mugenic acid and avenic acid behave as phytosiderophores in graminaceous species (Kinnersely, 1993). They are released from the roots in response to Fe and Zn deficiency and mobilise Cu, Zn and Mn (Romheld, 1991). Plants can also solubilise metals by lowering the pH of the soil environment, via the release of protons from the roots. Citrate, an organic acid is secreted from plants and is able to chelate numerous metals. It also plays a role in resistance to aluminium toxicity through its ability to complex with Al(III) and prevent uptake by the plant (De la Fuente et al. 1997). It is also possible to increase plant metal uptake by the addition of synthetic chelators to the soil. Ethylene diamine tetra-acetic acid (EDTA) is a popular choice for addition to lead contaminated soils, which can facilitate a 100 fold increase in the uptake and transport of lead-EDTA-chelate complexes into stem and leaves (Huang et al., 1997; Vassil et al., 1998). By increasing the secretion of appropriate organic acids, it may be possible to increase uptake of metals. Metal ions generally enter plant cells via specific or generic transmembrane carriers or channels. Non-essential heavy metals can compete for the same transmembrane carriers used by essential metals. This has been demonstrated with Cu and Zn, essential metals, and Ni and Cd, non-essential metals (Clarkson & Luttge, 1989). Specialised carriers may also transport chelated metal complexes across the plasma membrane. An example of this is Fe-phytosiderophore transport in graminaceous species (Crowley et al., 1991). More recent research has identified four proteins from Arabidopsis thaliana (a model plant) - ZIP1, ZIP2, ZIP3 (Eng et al., 1998) and ITR1 (Eide et al., 1996) – that all appear to be involved in facilitating pathways for the active uptake of toxic metal ions, as a result of nutrient deficiency and stress. ZIP1 and ZIP3 mRNA has been shown to accumulate in Arabidopsis roots as a result of zinc deficiency (Grotz et al., 1998) and all contain an extra-membranal metal-binding motif HXHXH. The ITR1 protein has been shown to transport both Cd(II) and Zn(II) (Yi & Guerinot, 1996; Cohen et al., 1998). Once genes like these have been identified, 82 BIOENERGY CROPS AND BIOREMEDIATION they can be introduced into plants, using constitutive promoters enabling high levels of expression, to examine whether or not metal uptake can be increased. 4.2.2 Transport within plants Once metal ions have entered the root, they can either be stored or exported to the shoot. Transport to the shoot probably occurs via the xylem, but may also occur via the phloem (Stephan & Scholz, 1993). Movement of ions into the xylem may be a rate limiting step because the metal ions must move from the cell cytoplasm to the xylem by transversing the cell wall, symplastically. Xylem cell walls have a high cation exchange capacity, therefore retarding the movement of cations. Noncationic chelated metal complexes such as Cd-citrate therefore, probably facilitate the movement of metals via this route (Senden et al., 1992). 4.2.3 Hyperaccumulation Some plants are able to concentrate metals in their tissues to levels far exceeding those present in the surrounding soil or by other non-accumulating plants. A generally accepted definition of a hyperaccumulator is one that is able to accumulate more than 0.1% of Ni, Co, Cu, Cr and Pb or 1% of Zn in its leaves (dry weight) (Baker, 1999). At these levels it has been proposed that the recovery of metals would be economical (Baker, 1999), however, for the purposes of land remediation accumulation of metal pollutants does not need to be so high. Hyperaccumulators can be divided into groups according to the metals they accumulate: i) Cu/Co accumulators ii) Zn/Cd/Pb accumulators and iii) Ni accumulators (Raskin et al., 1994). Mostly they are restricted to only a few geographical locations and tend to be small plants, not suited to biomass production. The majority of hyperaccumulators are found in the cabbage family, Brassicaceae (Baker et al., 1994). One of the best examples of a hyperaccumulator is Alyssum lesbiacum. It is able to accumulate Ni(II) in the shoots and leaves to >3% of the dry weight (Krämer et al., 1996). When this plant is exposed to nickel, a large and proportional increase in the level of free histidine occurs which can be shown to be coordinated with nickel in vivo. The molecular mechanisms involved in hyperaccumulation are poorly understood, however when the genes involved are identified and isolated, their over-expression in biomass crops could lead to great enhancement in metal tolerance and accumulation. Plants are able to protect themselves from the damaging effects of metal ions by compartmentalization, chelation and precipitation of metal complexes. The two most extensively studied systems employed by plants to achieve this, involve cysteine-rich peptides; metallothioneins and phytochelatins. 83 BIOENERGY CROPS AND BIOREMEDIATION 4.2.4 Metallothioneins Metallothioneins (MTs) are low molecular weight cysteine-rich metal-binding peptides. They have been found in animals, fungi, cyanobacteria and higher plants. They are thought to provide housekeeping functions for the regulation of intracellular concentrations of metals, ensuring, for example, that enzymes are supplied with appropriate metal ions. In animals they have been shown to bind copper, cadmium, zinc and silver and to detoxify normally lethal concentrations of cadmium and copper in yeast (Tohayama et al., 1995). Numerous MT genes/proteins have been identified in plants; wheat (Kawashima et al., 1992), maize (De Framond, 1991), barley (Okumura et al., 1991), pea (Evans et al., 1990) and soybean (Kawashima et al., 1991). A whole family of MT genes has been identified in Arabidopsis thaliana (Zhou & Goldsbrough, 1995). The plant MTs are usually 60-80 amino acids long with around 9-16 cysteine residues. Several experiments have been carried to over-express various MT genes in plants in the hope of conferring increased heavy metal tolerance and accumulation. One of the first experiments of this nature to be reported is that of Misra & Gedamu, (1989). They demonstrated that transgenic Brassica napus L. and tobacco, containing a human metallothionein gene, were able to grow unaffected on 100M CdCl2, in contrast to control seedlings which displayed severe inhibition of shoot and root growth and chlorosis of leaves. In 1993 Pan et al. reported the expression of a mouse metallothionein gene in transgenic tobacco plants. The MT gene was fused to the cauliflower mosaic 35S (CaMV 35S) promoter which facilitates constitutive expression of the gene in all cell types. Of the plants recovered from the transformation, 20% displayed high levels of expression from the inserted gene and were able to grow unaffected on media containing 200 M cadmium, whereas the health of control plants was severely affected on just 10 M cadmium. Much work has been done to characterise MT genes. They appear varied in the metals they bind and in the manner with which their expression is regulated (Robinson et al. 1993). Further research to examine their potential for phytoremediation is required. 4.2.5 Phytochelatins Phytochelatins (PCs) are cysteine-rich proteins that have the ability to complex metals (Ag(I), Hg(II), Pb(II), Zn(II), Cd(II) and Cu(II)) and sequester them in the vacuole. They have been found in most higher plants, algae and in some yeasts. PCs are synthesised via an enzymatic pathway that converts cysteine to glutamylcysteine, -glutamylcysteine to glutathione and glutathione to phytochelatin. The three enzymes catalysing these steps are -glutamylcysteine synthetase (-ECS), glutathione synthetase (GSH synthetase) and phytochelatin synthase respectively. Although involved in PC synthesis glutathione (GSH) also plays an important role in countering oxidative stress and may be conjugated to many different xenobiotic compounds to facilitate their sequestration in the vacuole. PCs are synthesised in response to heavy metals and can cause both a reduction in the amount of GSH in the cell (Grill et al., 1987; Meuwley & Rauser, 1992) and an increase in GSH synthetase activity (Schneider & Bergmann, 1995). Arabidopsis plants mutant in the synthesis of PCs or glutathione have been shown to be hypersensitive to cadmium (Howden et al., 1995a, 1995b) demonstrating the role of PCs in protecting plants from cadmium and possibly other toxic metals. 84 BIOENERGY CROPS AND BIOREMEDIATION By over-expressing the enzymes involved in the pathway, cellular levels of phytochelatin might be increased with a corresponding increase in metal sequestration. Two of the genes, -glutamylcysteine synthetase and glutathione synthetase, in the pathway have been identified and cloned from several organisms including Arabidopsis thaliana (May & Leaver, 1994; Rawlins et al., 1995) and E. coli (Gushima et al., 1984; Watanabe et al., 1986). A further gene, HMT1, isolated from the fission yeast Schizosaccharomyces pombe, is thought to be involved in the compartmentalisation of heavy metals in the vacuole (Ortiz et al., 1992). The deduced protein sequence of the gene displays homology with ABC (ATP-binding cassette) type transport proteins suggesting that it may be an integral membrane protein. It has also been found associated with vacuolar membranes (Ortiz et al., 1995). Over-expression of the HMT1 gene in yeast confers enhanced cadmium tolerance and increased levels of intracellular cadmium. However, attempts to over-express the yeast gene in plants have been unsuccessful so far (Ow, 1993). Whether homologues from plants can be identified remains to be seen. Most recently, plant, animal and fungal genes encoding phytochelatin synthase have been identified (Clemens et al., 1999; Vatamaniuk et al., 1999). Exposure to cadmium has been shown to increase the synthesis of phytochelatin synthase several fold and over-expression of plant phytochelatin synthase in yeast increases tolerance and accumulation of cadmium (Vatamaniuk et al., 1999). Two rate-limiting steps have been identified in PC synthesis; the supply of cysteine and the step catalysed by -ECS. This was deduced from experimental evidence resulting from the over-expression of -ECS in transgenic poplar (Noctor et al., 1996). Transgenic plants over-expressing -ECS were found to contain 10 and 3 times more -glutamylcysteine and glutathione respectively and the application of exogenous cysteine increased the glutathione content further in both transformed and untransformed control plants. Based on the evidence from Noctor et al., 1996 it would be useful to examine whether or not increased levels of PCs are obtained as a result of over-expressing -ECS and whether the poplar plants are able to tolerate and accumulate higher levels of metals than non-transformed control plants. In addition to the above, glutathione synthetase is also suggested as being rate limiting. When the E. coli glutathione synthetase gene was over-expressed in Brassica juncia, transformed plants were found to have higher concentrations of GSH and PC and increased tolerance to cadmium than the control plants (Zhu et al., 1999). It appears therefore, that the manipulation of glutathione and phytochelatin concentrations offers good potential for increasing metal tolerance and accumulation in plants. 4.2.6 Transformation of toxic elements The transformation of toxic elements into relatively non-toxic forms using plants has been demonstrated using transgenic Arabidopsis thaliana to convert organic mercury to much less toxic elemental mercury (Bizily et al., 2000). The Arabidopsis thaliana plants were transformed with two bacterial genes: (i) merA which encodes mercuric reductase and (ii) merB encoding organomercurial lyase. 85 BIOENERGY CROPS AND BIOREMEDIATION These genes have been cloned from bacteria isolated from organic-mercury contaminated soils. The enzymes they encode are capable of converting methylmercury and other organomercurials to elemental mercury, which is volatile and diffuses out of the bacterial cells. Arabidopsis plants expressing both of the genes were found to grow on 50 times higher concentrations of methylmercury than control plants and were shown to transpire elemental mercury. This work demonstrates the potential for phytoremediation using genes from other organisms. It now requires scaling up from the model plant, Arabidopsis thaliana, to a biomass crop followed by field trials. Selenium is a particular problem in the wetlands of western USA. Transgenic expression of the Arabidopsis plastid ATP sulfurylase (APS1) in Brassica juncia has been shown to increase the uptake of selenate and its reduction to selenite (Pilon-Smits et al., 1999). Plants were also more able to tolerate selenate. It has also been shown that the volatilization of selenium by plants is enhanced by bacterial activity in the rhizosphere (De Souza et al., 1999). It is thought that many phytoremediation processes are enhanced in this way, which suggests that further research examining plant/bacterial co-existence would be of value. The major form of iron found in soils is Fe(III), which is relatively unavailable to plants and fairly toxic. Arabidopsis thaliana is an example of a plant that is able to reduce Fe(III) to Fe(II) fairly well. It is able to do this via a ferric chelate reductase (FRO2) at the root surface (Yi & Guerinot, 1996; Robinson et al., 1999). Fe(II) is then transported into the root cells by a ZIP transporter (discussed earlier). Plants that are mutant for FRO2 are iron deficient, but over-accumulate Cu(II), Zn(II) and Mn(II) as they try to extract more iron from the soil. However, Arabidopsis mutants for a manganese accumulator, show co-ordinate deregulation for Fe(II), Cu(II), Zn(II), Mn(II) and Mg(II) and also for FRO2 activity (Delhaize, 1996). 4.3 PHYTOREMEDIATION OF ORGANIC POLLUTANTS Strategies employing the use of microorganisms for the detoxification of organic waste products have been used for many years. More recently however, microorganisms, that have evolved catabolic genes enabling them to detoxify toxic chemicals, have been utilised for the bioremediation of polluted land. Using natural bacterial gene transfer systems it has been possible to transfer certain degradative processes to certain bacteria, increasing the potential for remediation under differing circumstances. Recent advances in molecular biology have taken this concept further by making it possible, in theory, to introduce virtually any gene into any organism. However, experiments to evaluate the effectiveness of bio-augmentation, the addition of microorganisms for the purposes of remediation, have been disappointing. The reasons for this include: (i) competition from endogenous microorganisms causing rapid decreases in populations of introduced microorganisms and (ii) there is often a need for induction of the degradative genes by the application of undesirable inducers to the land. The biodegradative capability of plants is much less impressive than those of adapted bacteria and fungi. The alternative, therefore, is to use the array of 86 BIOENERGY CROPS AND BIOREMEDIATION catabolic bacterial genes for expression in biomass plants, combining the best elements from two systems for effective remediation. In this way genes can be designed without the need for induction using undesirable compounds. The mechanisms controlling the uptake and sequestration of toxic organic compounds in plants are relatively unknown. One system we do know something about is the pumping out of the cell or sequestration into the vacuole of glutathione conjugates of organics and other conjugated-toxic organic complexes. This is facilitated by a glutathione-S-conjugate pump, the gene for which has been isolated from Arabidopsis thaliana (Tommasini et al., 1998). Halogenated compounds like TCE are difficult to metabolize and are generally toxic and carcinogenic. They are widespread contaminants and pose significant risks to animals and humans. Plants are known to produce aliphatic dehalogenases that degrade TCE (Anderson et al., 1993; Anderson & Walton, 1996). They have been shown to extract TCE from polluted sites and transpire it. Root exudate has been shown to enhance degradation of TCE in the rhizosphere. Axenically grown hybrid poplar has been demonstrated to remove TCE from the surrounding medium and convert it to trichloroethanol, chlorinated acetates and finally CO2 (Gordon et al., 1998). In one experiment, using poplar cells in tissue culture, more than 10% of the TCE present was converted to CO2 within 10 days. More recently, TCE remediation has been investigated using transgenic plants expressing a mammalian cytochrome P450 2E1 (Doty et al., 2000). This enzyme oxidises many compounds including; TCE, dibromoethane (EDB), carbon tetrachloride, benzene, styrene, chloroform, 1,2-dichloropropane and vinyl chloride. Transgenic tobacco plants expressing the introduced gene displayed a 640-fold increase in the metabolism of TCE as compared to control plants. Transgenic plants also displayed increased uptake and debromination of ethylene dibromide. Cytochrome P450 monooxygenases have been found in plants, bacteria and animals and catalyse many types of chemical transformations, including aliphatic hydroxylations, expoxidations, dealkylations, dehalogenations and many deactivations (Guengerich & MacDonald, 1990). Several hundred P450 enzymes have been characterised at the primary sequence level and other natural P450s have yet to be isolated and characterised. In addition to the natural P450s available, much work is being carried to specifically alter substrate specificity and catalytic efficiency of P450s using protein engineering (Kellner et al., 1997). The ability to degrade xenobiotics and their wide range of substrate specificities makes cytochrome P450s very good candidates for exploitation in phytoremediation. A combination of bacterial gene in a plant system has been tested to evaluate whether or not explosive residues could be remediated. A gene encoding pentaerythritol tetranitrate reductase, isolated from an explosive-degrading bacterium, was introduced into tobacco. Seeds from the transgenic plants expressing pentaerythritol tetranitrate reductase were able to germinate and grow on 1 mM glycerol trinitrate (GTN) or 0.05 mM trinitrotoluene; concentrations that inhibited the growth of control seeds (French et al., 1999). Transgenic seedlings grown in liquid medium containing 1 mM GTN displayed more rapid and complete denitration of GTN than control seedlings. Again, these results offer promise for the enhancement of remediation by plants. 87 BIOENERGY CROPS AND BIOREMEDIATION 4.4 PLANT TRANSFORMATION The availability of appropriate genes and strategies for the over-expression of these in biomass crops, relies on the ability to transfer such genes to the target plant in such a way that a copy of the foreign DNA is stably integrated into every cell of the plant. To achieve this, it is necessary to have a tissue culture system, specific to the plant, that enables the regeneration of whole plants from single totipotent cells. Once a successful tissue culture system has been developed, a plant transformation system must be established to enable the transfer of foreign DNA to those single cells capable of regeneration into whole plants. It is widely recognised that monocotyledonous plants are generally more difficult to transform than dicotolyedonous plants - and grasses (Poaceae) in particular have proven problematic. The totipotent cells can be generated either as embryogenic suspension cultures or as embryogenic callus tissue on solid media. Plant cell suspension cultures able to regenerate plants through somatic embryogenesis were first reported in carrot in 1958 (Steward et al., 1958). It was not until the early 1980s that embryogenic suspension cultures were achieved for species from the Poaceae family, of which Miscanthus is a member (Lu & Vasil, 1981; Vasil & Vasil, 1981; Ho & Vasil, 1983). Efficient embryogenic suspension culture systems have now been established for the most commercially important members of the Poaceae family, including Miscanthus (Holme, 1996). Callus induction and plant regeneration on solid media has also been achieved using various explant types from Miscanthus x ogiformis Honda ‘Giganteus’ (Holme & Petersen, 1996; Petersen et al., 1999). We are not aware of any published literature describing the stable transformation of Miscanthus tissues, however, our own unpublished work has shown that Miscanthus callus tissue can be transformed via microprojectile bombardment resulting in transient expression of a foreign reporter gene, GUS, driven by a ubiquitin promoter. It is not unreasonable to expect that the development of a stable transformation system for Miscanthus is underway. Combined with the extensive work, carried out at the Danish Institute of Plant and Soil Science, to establish a successful tissue culture system, it is likely that the generation of transgenic Miscanthus will not be long in the making. In contrast to Miscanthus, poplar species have received much greater attention with respect to the introduction and over-expression of foreign genes. The first reports of poplar transformation were published in the late 1980s (Parsons et al., 1986; Fillatti et al., 1987). Since then, a number of different poplars have been successfully transformed using the two most popular transformation methods available; microprojectile bombardment and Agrobacterium tumefaciens-mediated transformation. These include, amongst others, white poplar (Populus alba) (Confalonieri et al., 2000); the hybrid poplars, Populus tremula x alba (Gallardo et al., 1999; Franke et al., 2000) and Populus x canadensis (Confalonieri et al., 1997; Liang et al., 1999); and eleven different hybrid cottonwood genotypes, several of which were previously difficult to transform and are economically important (Han et al., 2000). Surprisingly, willow appears to have received little attention with respect to the development of tissue culture or plant transformation systems. Amo-Marco & 88 BIOENERGY CROPS AND BIOREMEDIATION Lledo (1996) report the development of in vitro culture techniques for the propagation of an endangered willow, Salix tarraconensis. In addition, Xing & Maynard (1995) report the production of transgenic shining willow (Salix lucida). In this experiment, shoot explants were transformed using Agrobacteriummediated transformation, the introduced gene encoding -glucuronidase, a protein that can be used as a marker for gene expression. Willow is being examined for its potential to phytoremediate land and water. The EC-funded BIORENEW project is developing a rapid screening system to facilitate the assessment of metal tolerance from large numbers of willow varieties (Watson et al., 1999). Preliminary results suggest that plant responses tested in a hydroponic system mimic responses seen under field conditions. Willow has also been shown to facilitate the transformation of perchlorate into chloride (Nzengung et al., 1999). Two phytoprocesses were identified as being responsible for decontamination: (i) uptake and phytodegradation in tree branches and leaves and (ii) rhizodegradation. Although willow is being examined for phytoremediation purposes, it appears that the step to enhancing phytoremediation processes using genetically modified willow has not been taken. Progress in the development of transformation systems for biomass willow is necessary to ensure that transgenic varieties can be developed in the near future. Transgene integrations are essentially random, and are therefore, subject to position effect variation which affects the level of transgene expression. A general approach to overcoming these limitations usually involves generating a large number of transgenic plants, from which the ideal transformant can be selected. This type of approach can be time consuming and costly and particularly difficult when dealing with plants whose transformation efficiency is low. One method that can help to alleviate positional effects is the use of matrix attachment regions (MARs), which are lengths of DNA added to either side of the transgene and shield the transgene from surrounding influence. This method has been used successfully in poplar transformation, enhancing transformation frequency and transgene expression (Han et al., 1997). 4.5 RISKS In addition to the many potential benefits of genetic modification, it is recognised that the introduction of foreign genes into plant genomes and the expression of foreign proteins in plants have the potential to result in undesirable effects on the environment and human health. The release of genetically modified organisms (GMOs) into the environment, is, therefore, controlled in the UK by domestic and European legislation as defined by Directive 90/220/EEC and implemented by the Genetically Modified Organisms (Deliberate Release) Regulations 1992 (amended 1995 and 1997). This serves to prevent or minimise damage to the environment via a statutory system of risk assessment and prior consent before any GMO may be released or marketed. All applications are received by the Department for Environment, Food and Rural Affairs (DEFRA), and the Advisory Committee on Releases to the Environment (ACRE) advise on whether or not release should be granted. This decision is arrived at, after careful consideration of the required 89 BIOENERGY CROPS AND BIOREMEDIATION information supplied with each application. Central to this is an environmental risk assessment that considers the potential risk to the environment and human health posed by the release of the organism in question. The accumulation of high levels of metals in plant tissues raises issues over the toxicity of these plants to other organisms. It may be useful in eliminating pest damage, for example, but could also adversely affect beneficial insects and other organisms. The potential for the generation of unexpected toxic compounds during the conversion of organic or other pollutants to less toxic forms is also an important consideration that would need to be monitored closely. Changing just one step in a metabolic process can have a dramatic effect on the production of many different compounds. For these reasons it is important that the dispersal of transgenes conferring pollutant remediation abilities should be minimised or prevented. This can be achieved in a number of ways and is the subject of much investigation (Advisory Committee on Releases to the Environment, 2000). The most obvious solution might be to use plants that are genetically sterile. Sterility can of course occur naturally, but can also be conventionally bred into suitable plant lines and the trait can even be genetically engineered into plants. Poplar has been genetically engineered for both sterility and accelerated flowering (Meilan and Strauss, 1997). Another possible solution might be to insert the required DNA sequences into the chloroplast genome rather than the nuclear genome. Most plants inherit the chloroplast DNA maternally, avoiding dispersal in pollen. 4.6 DISCUSSION The development of genetically modified plants for phytoremediation is still in its infancy, but investigations to date suggest great potential for increasing the ability of plants to remove pollutants from contaminated land and water. Many genes involved in facilitating tolerance, accumulation and detoxification of metal and organic pollutants have been identified in plants and other organisms, but there are still a great many to be discovered. For many of the pathways facilitating the detoxification of pollutants in plants, it will be possible to enhance detoxification by modifying rate limiting steps by the over-expression of the corresponding genes. In addition to modifying plant pathways, it has also been shown to be possible to express genes from other plant species and even genes from animals, bacteria, yeast and fungi. Much of this work has been performed in model plant species such as Arabidopsis thaliana and tobacco. Poplar transformation has been the subject of much study. There are many publications reporting the transformation of poplar species, including examples specifically investigating phytoremediation potential. Other biomass crops have not been extensively studied and therefore require further development. Genetic engineering facilitates a choice of promoter to drive the expression of the required gene. By choosing promoters that are induced by metals, organics or other compounds, expression of the gene may be controlled on a demand basis. In addition, the use of tissue-specific promoters may enable the accumulation of metals in specific parts of the plant; for example, in the harvestable tissues, such as the shoots and leaves, and not in the below ground tissues. It is possible to introduce numerous genes to the plant genome during plant transformation and 90 BIOENERGY CROPS AND BIOREMEDIATION also to re-transform an already transgenic plant. This means that in instances where several genes are required to facilitate the remediation of several pollutants, a suitable plant can be generated. The most successful remediation strategies might be those using a combination of genetically engineered or conventional plants and associated rhizosphere bacteria. Careful consideration must be given to the development of plants for phytoremediation, because of the potential for adverse environmental effects. The prevention of transgene flow to conventional plants and possible effects on other organisms are important issues. 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Genetic evidence that induction of root Fe(III) chelate reductase activity is necessary for iron uptake under iron deficiency. Plant Journal 10. 835-844. Zhou, J & Goldsbrough, P B (1995). Structure, organisation and expression of the metallothionein gene family in Arabidopsis. Molecular and General Genetics 248. 318-328. Zhu, Y L; Pilon-Smits, E A H; Jouanin, L & Terry, N (1999). Overexpression of glutathione synthetase in Indian mustard enhances cadmium accumulation and tolerance. Plant Physiology 119. 73-79. 97 BIOENERGY CROPS AND BIOREMEDIATION CHAPTER 5 LEGISLATION AND CODES OF PRACTICE AFFECTING THE RECYCLING AND LAND APPLICATION OF ORGANIC WASTES GORDON HICKMAN 5.1 INTRODUCTION This chapter provides an overview of the main legislation and codes of practice affecting the disposal and recycling to land of farmyard manures, sewage sludge and other organic wastes that might be applied to biomass crops. It is based on desk-based research and expert knowledge, but does not purport to be a definitive listing of legislation which would impact on all wastes that might be used for bioremediation. The main aim is to highlight the key issues and potential barriers. 5.2 UK WASTE REGULATIONS AND EC DIRECTIVES The possible adverse environmental effects of agricultural activities have been a concern in many European countries since the mid-1970's. Concerns were initially related to water pollution and odours, but more recently have included emissions of ammonia and greenhouse gases and protection of the soil. In 1999, agriculture caused 4,254 (14% of the total) substantiated water pollution incidents in England and Wales (EA, 2001). Of these, some 238 (24% total) were regarded as major or significant incidents (category 1 & 2). Although this figure includes incidents due to pesticides, fuel oil and other contaminants, 2,161 (7% total) substantiated incidents were caused by livestock excreta, silage effluent or other organic wastes such as sewage sludge or raw sewage. Legislation has often been aimed at reducing such point source water pollution incidents. Table 5.1 summarises some of the key legislation relating to the actual disposal or recycling to land of organic wastes. Further commentary is provided in the following sections. 98 BIOENERGY CROPS AND BIOREMEDIATION Table 5.1. Summary of legislation and voluntary codes of practice affecting disposal of farm manures and organic wastes. Title Type of waste affected What does it regulate? Control of Pollution Regulations 1991/1997 Manures, slurry, silage effluent and fuel New storage structures. Requirements for impermeability, structural stability and siting in relation to watercourses. Nitrate Vulnerable Zone Regs. (EC Nitrate Directive) Animal manures, sludges, wastes high in available N Manure N application rate and timing. MAFF/WOAD Water Code 1998 All types All aspects of avoiding pollution from manures, including odours, ammonia and gaseous emissions. Waste Management Licensing Regs, 1994 (EC Framework Directive on Waste), HMSO, 1994. Industrial, household and commercial wastes “controlled wastes” Licensing requirements and Duty of Care. (Beneficial wastes exempted subject to limits on application rate). Integrated Pollution Prevention and Control (IPPC) Regs. (2000). (EC Directive 96/1) All types All types of emissions. To apply from 2004 to disposal sites for non-hazardous wastes, food and drink installations from 2004 (typically) and from 2007 to units for the intensive rearing of poultry or pigs (with >40,000 poultry, 2000 pig places, or 750 sow places). Sludge (Use in Agriculture) Regs. (1989) and Code of Practice for the Agricultural Use of Sewage Sludge (1996) (EC Sewage Sludge Directive 86/278/EEC) (Currently under revision) Sewage sludge and sludge products Application rates and timing, management of sludge; limits on soil metal levels and metal additions. “Safe Sludge Matrix” Voluntary agreement currently being incorporated into Sludge (Use in Agriculture) Regs. Sewage sludge of all types Guidance on minimum standards for application practice and minimum acceptable level of treatment for any sludge product (biosolids) to be applied to any crop or rotation. 99 BIOENERGY CROPS AND BIOREMEDIATION 5.2.1 Codes of practice The Department for Environment, Food and Rural Affairs (DEFRA), formerly MAFF, and National Assembly for Wales Agriculture Department (NAWAD), formerly WOAD, publish three separate Codes of Good Agricultural Practice for the Protection of Water, Air and Soil - which give guidance on ‘best practice’ for avoiding pollution. These Codes were first published in 1991-3 and were revised and re-issued in 1998 (MAFF, 1998a, 1998b & 1998c). These Codes are statutory codes under Section 97 of the Water Resources Act 1991 (HMSO, 1991). This means that although failure to comply with the Codes is not an offence, it would be taken into account in any legal action taken as a result of a pollution incident. Furthermore, many Farm Assurance Schemes for both crops and livestock include compliance with the codes as part of their protocols. All the Water Operators have also signed up to complying with these codes. The Codes also detail the limits to application rates for organic materials, namely 250 kg ha-1 total N per year, or a single application of 500 kg ha-1 total N of solid materials with limited N availability in non sensitive catchments every two years. The Codes also give guidance on the identification of areas where manures should not be spread, or where rates or timing should be restricted to minimise the risk of surface runoff. A 10 m wide no-spreading zone is recommended adjacent to watercourses and a minimum of 50 m radius adjacent to springs, wells or boreholes supplying water. Other areas can be classified for water pollution risk according to slope, soil characteristics and proximity to watercourses. Farmers are encouraged to produce a farm waste management plan, which takes account of these factors. Such plans normally incorporate a map of the farm with field areas annotated in different colours, to indicate restrictions on timing of spreading or amounts to be spread. The Water Code also differentiates between low and high available N organic manures, viz.: Low available N High available N - sludge cake, thermally dried and lime-treated sludge (straw-based farmyard manures) liquid digested sludge (cattle/pig slurry and poultry manures) The Code also recommends that where practically possible, manures containing a high proportion of available N should not be applied to arable land in the autumnearly winter period, to minimise the risk of nitrate leaching. Impact and implications for bioremediation The Codes of Practice provide a framework of guidance within which most operators work when applying organic wastes to agricultural land. They are not likely to limit or hinder the use of wastes on bioenergy crops for bioremediation. 100 BIOENERGY CROPS AND BIOREMEDIATION 5.2.2 Odours and gaseous emissions General guidance and legislation Justifiable complaints from the public about odours from agriculture totaled 66,658 in 1994. Half of these were due to land spreading of manures and the majority of complaints concerned odours arising from pig farms. Guidance on minimising odours from buildings, manure stores and land spreading is given in the Code of Good Agricultural Practice (COGAP) for the Protection of Air. Under the Environmental Protection Act 1990, an abatement notice can be served on the owner of a unit, which causes a statutory nuisance, which includes odours (HMSO, 1990a). Integrated Pollution Prevention and Control (IPPC) legislation. The system of IPPC comes under the Pollution Prevention and Control (England & Wales) Regulations 2000, as amended (SI, 2002). It is a means of applying an integrated environmental approach to the regulation of certain industrial activities. This means that emissions to air, water and land, plus a range of other environmental effects, must be considered together. IPPC aims to prevent emissions and waste production and, where that is not practicable, to reduce them to acceptable levels. Waste reduction should be treated as a priority and energy must be used efficiently. Installations falling under the requirements of IPPC will be given permits to operate, providing certain conditions are met. These permits will require that best available techniques (BAT) are used in meeting the requirements of the conditions. In order to apply for a permit under IPPC, the applicant must provide detailed information regarding the operation of the installation in question. For example, an intensive livestock farmer will be expected to supply, amongst other information, the following: Details of the management structure and staffing arrangements, including details of training and emergency planning. A raw materials inventory. Records of current water and energy use. Confirmation that a water and energy audit will be undertaken within 18 months of permit issue date. Manure management plans. Under the PPC Regulations, installations fall under one of three classifications dependant upon the activities that take place within them: The Environment Agency regulates Part A(1) installations. Part A(2) installations are regulated by the relevant local authority. However, the local authority will always be the statutory consultee where the Environment Agency is the regulator and vice versa. The two will work together in the permitting process. Part B installations come under the regime of Local Air Pollution Prevention and Control (LAPPC). LAPPC is similar to IPPC from a 101 BIOENERGY CROPS AND BIOREMEDIATION procedural point of view but focuses on controlling air emissions only. Local authorities are the regulators. Impact and implications for bioremediation The impacts are difficult to assess, since the implementation timetable is still some way off. The regulations should result in less waste being produced, as well as more responsible and informed disposal/recycling of any waste that is produced. The inclusion of disposal sites treating more than 50 tonnes/day of non-hazardous wastes, such as food and livestock waste, by biological or physio-chemical means may provide some barriers to development of composting and anaerobic digestion sites. However, they are not likely to limit or hinder the use of wastes on bioenergy crops for bioremediation, unless it was decided that the BAT for application of liquids was injection. If this were the case, it would limit applications to pre-establishment in the case of SRC and possibly Miscanthus. 5.2.3 Water pollution General Under the Water Resources Act 1991 (HMSO, 1991) it is an offence to cause pollution of groundwater, lakes, ponds, rivers, streams and canals. Farmers and others may be prosecuted for causing such pollution and fined up to £20,000 in a local court. In addition the Act contains provisions for secondary legislation which aims to prevent pollution occurring in the first instance. Storage of Slurry and Manure The Control of Pollution (Silage, Slurry and Agricultural Fuel Oil) Regulations 1991 (SI, 1991), introduced minimum requirements for the size and siting of new or substantially modified slurry storage structures and laid down minimum constructional standards based on BS 5502 (Buildings and Structures for Agriculture). No specific controls exist for solid manures temporarily stored in field areas without a structured base, although there are recommendations on siting. The EA is able to regulate and control the use of long-term field stockpiles. Groundwater Protection Zones Up to 35% of public drinking water is derived from groundwater sources such as aquifers, and the protection of such sources is an important function of the EA. Source protection zones have been established throughout England and Wales and in most cases waste materials (does not currently apply to farm wastes) are not permitted in Zone 1 (most sensitive) catchment areas or around private boreholes. 102 BIOENERGY CROPS AND BIOREMEDIATION Impact and Implications for Bioremediation The regulations provide a framework of guidance, within which most operators work when applying organic wastes to agricultural land. They are not likely to limit or hinder the use of wastes or bioenergy crops for bioremediation. 5.2.4 Land application, materials including bioremediation, The recycling of nutrients in livestock manures is excluded regulations on the use of ‘waste materials’ spread to land. specified geographical areas, such as in Nitrate Vulnerable Special Scientific Interest (SSSIs), there are no regulations timing or rate of application of manures. of organic from EC and UK Except in certain Zones or Sites of which govern the DEFRA have recently updated and revised its guidance on fertiliser use in Fertiliser Recommendations for Agricultural and Horticultural Crops (RB209) 7th Edition (2000). Although primarily aimed at providing guidance on giving the best financial return to farmers, the guidelines should also help to minimise nutrient losses to the water and air. Recommendations, which encourage the optimum use of nutrients in manures, also help to minimise diffuse water pollution risks from land spreading. Literature, like booklet RB209, is available to give typical analyses of manures and slurries and details of how to calculate optimum application rates for particular crops, soils and application methods. Computer programs to aid decision-making, such as MANNER (ADAS, 2000), are also available. Whilst farmers are encouraged to take full account of the nutrients available in manures when planning their fertiliser applications, many do not. Impact and implications for bioremediation The codes of practice and guidance provide a framework within which most operators work when applying organic wastes to agricultural land. They are not likely to limit or hinder the use of wastes on bioenergy crops for bioremediation. There is, however, relatively little available information on nutrient requirements of bioenergy crops under a range of soil types and climatic conditions; although the apparently low requirement of some may limit the use of ‘nutrient rich’ organic wastes, if application rates are limited to probable crop uptake. 5.2.5 Draft Soil Strategy – England DEFRA has published a consultation draft of a Soil Strategy for England (DETR, 2001). The document was produced as a direct response to recommendations made by the Royal Commission on Environmental Pollution. The overall strategy is to ensure that soil is used and protected in such a way that it is sustainable in its own right and that it contributes to sustainable development generally. One of the key objectives is to address the issue of soil erosion, through R&D on the extent and causes of soil erosion on agricultural land. It also refers to the 103 BIOENERGY CROPS AND BIOREMEDIATION promotion of good practice for the land treatment of industrial wastes and the extension of waste management controls to agricultural wastes. Impact and implications for bioremediation The strategy is still in a draft form but gives an indication as to the likely direction of future policy. The proposals will be incorporated into the relevant legislation and codes of practice discussed elsewhere in this section and are not likely to limit or hinder the use of wastes on bioenergy crops for bioremediation. The possible inclusion of farm wastes under waste management controls may have an impact, depending on how the regulations are introduced. The focus on erosion could present some limitations to bioenergy crops, since there is some limited evidence to suggest that the reduced ground cover and relatively slow establishment of short rotation coppice and other bioenergy crops can lead to increased risk of erosion on some sites. 5.2.6 Nitrate The UK government has established 32 Nitrate Sensitive Areas (NSAs), where farmers are paid compensation to voluntarily join a scheme which aims to reduce nitrate loss from the soil to groundwater. In these areas there is a maximum limit of 250 kg total N ha-1 in any 12 month period from applied organic manures, limitations on timing of slurry and poultry manure applications and various other control options affecting the way in which farmers may use fertilisers and their land. The rules have been successful in reducing nitrate loss from the soil zone. Under the EC Nitrate Directive (EC, 1991b) 68 Nitrate Vulnerable Zones (NVZs) in England and Wales, totalling 600,000 ha, were designated in April 1996; where compulsory limits on the application, and deposition by grazing animals, of total nitrogen are enforced. An upper limit of 250 kg ha-1 yr-1 of organic N is allowed on grassland and 210 kg ha-1 on arable land [reducing to 170 kg ha-1 yr-1 after four years (from 19 December 2002)]. In addition, the application of slurries and poultry manures are banned on sandy and shallow soils during the late summer and early autumn, when there is little crop growth removing available nitrogen from the soil. Within Nitrate Vulnerable Zones, the rules are compulsory and no compensation is paid to farmers, as they are regarded as ‘good agricultural practice’. Impact and implications for bioremediation The regulations provide a framework of guidance within which most operators work when applying organic wastes to agricultural land. They are not likely to limit or hinder the use of bulky solid wastes on bioenergy crops for bioremediation, but the ‘closed periods’ limit the use of slurries and liquid wastes in the autumn. Biennial cropping of short rotation coppice (instead of the usual three-yearly harvesting) may provide an opportunity to take advantage of the option for ‘double’ applications, of 500 kg ha-1 total N, every two years. Applications could be made in late winter or spring, after harvest. Such applications would not, however, be permitted in NVZs, where there will be an annual limit of 250 kg ha-1 total N from organic sources. 104 BIOENERGY CROPS AND BIOREMEDIATION The number of NVZs is expected to significantly increase in the near future and depending on the extent of the new areas covered, may impose some additional constraints on the use of organic wastes in these areas. 5.2.7 Phosphate The Environment Agency has identified nutrient enrichment as one of the ten priority issues requiring attention, in order to achieve a more sustainable balance between the needs of society and the needs of freshwater ecosystems. The issue of phosphate enrichment of water is becoming increasingly important, given the link between phosphate and eutrophication of lakes, reservoirs and slow moving rivers. The Environment Agency has identified agriculture as being a particular problem, since most of the pollution occurs via diffuse pollution. The Environment Agency estimates that livestock (34% of phosphates), human and household waste (24%), fertiliser (16%) and detergents (10%) represent the major sources of phosphate inputs to surface waters. The Soil and Water Codes (MAFF, 1998a & b) provide guidance on limiting phosphate losses and recommend that, at soil index 3 and above, care should be taken to avoid total phosphorous inputs exceeding the amount removed by crops in the rotation. The guidance is however not statutory and the Environment Agency have stated that unless inputs can be controlled by voluntary means, regulatory restrictions may be required. Some restrictions are possible under the provisions of the Urban Wastewater Directive (EC, 1990). The UK has designated 80 Sensitive Areas (Eutrophic) covering both running and standing water. The number and size of such areas may be increased in the future. There are some 140 sites currently being monitored and a review of areas is to be undertaken during 2001. The Environment Agency has stated that it does not intend to apply for powers to establish water protection zones under Section 93 of the Water Resources Act (HMSO, 1991), although it will do so in catchments where other control initiatives have failed. Impact and implications for bioremediation The issue of phosphate will undoubtedly become even more important and may eventually give rise to the creation of ‘phosphate sensitive zones’. These could have an effect on high phosphate materials such as sewage sludge. However, the current regulations and codes are not likely to hinder the use of wastes on bioenergy crops for bioremediation - although the apparently low phosphate requirements of bioenergy crops could limit the total amount of organic matter that can be applied to soils, since the limiting constituent may, in some cases, be the total phosphate content. 105 BIOENERGY CROPS AND BIOREMEDIATION 5.2.8 Sewage sludge Sewage sludge applications to land are regulated by European Community Directive 86/278/EEC (EC, 1986) which has been implemented in England, Scotland and Wales by The Sludge (Use in Agriculture) Regulations 1989 (SI, 1989) and as amended by The Sludge (Use in Agriculture) Amendment Regulations 1990 (SI, 1990). These are complemented by the DEFRA Code of Practice for Agricultural Use of Sewage Sludge (DoE, 1996) and Codes of Good Agricultural Practice for the Protection of Water (MAFF, 1998a), Air (1998b) and Soil (1998c). In operational practice, sludge use on agricultural land needs to comply with the DEFRA Water Code recommendation that applications should not supply more than 250 kg total N ha-1 per annum, which in most situations will provide the working limit on major nutrient and heavy metal loading rates. The revised DEFRA Water Code (MAFF, 1998a) does, however, allow low available N manures, such as composts and sludge cakes, to be applied at 500 kg total N ha-1 every other year. The above regulations are currently being revised and a consultation draft is expected later in 2001. The regulations will adopt the provisions of the Safe Sludge Matrix and for the first time introduce standards for pathogen content in sludges. The regulations will also enforce a ban on the use of untreated sludges on food crops, and will include septic tank liquor within the regulations as untreated sludge – thereby preventing its application to grassland and other food crops. (The Waste Management Licensing Regulations will also be amended accordingly). Impact and implications for bioremediation The regulations provide a framework of guidance within which all operators work when applying sewage to agricultural land. They are not likely to limit or hinder the use of wastes on bioenergy crops for bioremediation. The revised regulations will however ban the use of untreated sewage sludge to all crops from 31 December 2005. 5.2.9 Safe Sludge Matrix Negotiations involving the UK Water Industry, the British Retail Consortium (BRC) representing the major retailers and ADAS, aimed to secure a sustainable route for recycling sludge to agricultural land that was acceptable to the food industry, water industry, regulators and farmers and growers were concluded in July 1998. The ‘Safe Sludge Matrix’ agreement (ADAS, 2001a), commonly known as the ADAS Matrix, came into being on 31 December 1998, and has been accepted as the minimum standard for sustainable sludge recycling to agricultural land. The Matrix consists of a table of crop types, together with clear guidance on the minimum acceptable level of treatment for any sludge based product (commonly referred to as biosolids) which may be applied to that crop or rotation (Table 5.2). All UK outdoor crops are covered from grass for grazing and silage making, maize for silage, combinable crops and animal feed crops, through to horticultural crops, vegetables, salads and fruit. 106 BIOENERGY CROPS AND BIOREMEDIATION Since December 1999, untreated sludge has no longer been allowed on land used to grow food crops although, under the agreement, industrial crops grown under contract for non-food use (e.g. industrial crops grown on set-aside) continued to receive untreated sludges until December 2001. The surface spreading of treated sludge on grazed grassland was banned from 31 December 1998. Treated sludge can only be applied to grazed grassland where it is deep injected. More stringent treatment processes are required where sludge is applied to land growing vegetable crops and in particular those crops that may be eaten raw (e.g. salad crops). Treated sludge can be applied to agricultural land, which is used to grow vegetables - provided that at least 12 months have elapsed between application and harvest of the following field vegetable crop. Where the crop is a salad, which might be eaten raw, the harvest interval must be at least 30 months. Table 5.2. The ‘Safe Sludge Matrix’ “Safe Sludge Matrix” Crop Group Fruit Salads Untreated sludges x x Treated sludges Enhanced treated sludges x x (30 month harvest interval applies) Vegetables x x (12 month harvest interval applies) Horticulture Combinable & Animal Feed Crops Grass - GRAZED & Forage- HARVESTED x x x x x (Deep injected or ploughed down only) x 10 month harvest interval applies) 3 week no grazing and harvest interval applies 3 week no grazing and harvest interval applies All applications must comply with the Sludge (Use in Agriculture) Regulations 1989 and DoE Code of Practice 1996 x Applications not allowed (except where stated conditions apply) An ‘Enhanced Treated Sludge’ (previously known as ‘Advanced’) category has been included in the Matrix, to describe treatment processes that are capable of virtually eliminating any pathogens that may be present in the original sludge (e.g. thermal drying). This is the first time in the UK that there has been recognition of differences in the effectiveness of treatment methods. Parallels can be drawn with the US Environment Protection Agency (EPA) standards, which differentiate between Class A and Class B sludge products. The current Code of Practice and Regulations will be amended by DEFRA to take account of the ‘Safe Sludge Matrix’. 107 BIOENERGY CROPS AND BIOREMEDIATION Following discussions between the major stakeholders in 2000, agreement was reached to allow untreated sewage sludge to continue to be applied to land used to grow specified industrial crops that do not normally have a food use, until 31 December 2005. After that date untreated sewage sludge can no longer be applied to agricultural land (ADAS 2001b). The specified crops include willow and poplar, for coppicing; Miscanthus, for biomass; hemp, for fibre; and high erucic acid rape (HEAR), for the production of high erucic acid rape oil (HERO). Impact and implications for bioremediation The Safe Sludge Matrix provides a framework of guidance within which all operators work when applying sewage to agricultural land. The Matrix is not likely to limit or hinder the use of wastes on bioenergy crops for bioremediation. The agreement will, however, ban the use of untreated sewage sludge to all crops from 31 December 2005. The ‘Industrial Crops Agreement’ permits the use of untreated sewage sludge, including sludges that fail the end product standards for pathogens, on crops such as Miscanthus and short rotation coppice until 31 December 2005. This may actually be helpful in stimulating interest in these crops, as sewerage operators look for secure outlets in the wake of the foot and mouth disease crisis. 5.2.10 Sewage sludge use in forestry and on restored land The use of sewage sludge on non-agricultural land, including forestry, and for land restoration is outside the scope of the Sludge (Use in Agriculture) Regulations (as amended) (SI, 1989 & 1990)and the Code of Practice for Agricultural Use of Sewage Sludge (DoE, 1996). These activities are however controlled by the Waste Management Licensing Regulations 1994 (SI, 1994b). Applications are exempt from the waste licensing, provided that the sludge provides ecological benefit and does not exceed the metal limits as set out in Schedule 2 of the Sludge (Use in Agriculture) Regulations (SI, 1990). It is, however, necessary to register the activity with the Environment Agency. Further guidance can be found in a Forestry Commission Bulletin – A Manual of Good Practice for the Use of Sewage Sludge in Forestry (Forestry Commission, 1992). Impact and implications for bioremediation The regulations and information bulletin are discussed in other sections and provide a framework of guidance within which all operators work when applying sewage to non-agricultural land. They are not likely to limit or hinder the use of wastes on bioenergy crops for bioremediation. The Industrial Crops Agreement will however affect the use of sludge on short rotation coppice, but is not thought to limit uptake in any way. 108 BIOENERGY CROPS AND BIOREMEDIATION 5.2.11 Waste Management Licensing Regulations The Waste Management Licensing Regulations (SI, 1994) deal with all aspects of the storage, treatment, and recovery or disposal of controlled wastes, including the licensing of facilities and operators. Wastes arising from agriculture, as defined by the 1947 Agricultural Act (HMSO, 1947), are excluded from this Regulation. However, under these regulations, certain controlled wastes for use in agriculture are exempt from the requirements of the licensing regulations. These wastes are listed in Schedule 3 of the Regulations (Table 2 of paragraph 7), and include waste soil or compost; waste wood, bark or other plant material; waste food, drink or materials used in or resulting from the preparation of food or drink; blood and gut contents from abattoirs; waste lime; lime sludge from cement manufacture or gas processing; waste gypsum; paper waste sludge, waste paper and de-inked paper pulp; dredgings from inland waterways; textile waste; sludge from biological treatment plants; and waste hair and effluent treatment sludge from a tannery. Such exempt activities must however result in benefit to agriculture or ecological improvement and are subject to an annual maximum application of 250 tonnes ha-1, or in the case of dredgings from inland waters, 5,000 tonnes ha-1, and are subject to pre-notification to the regulator. The regulations also require that waste is recovered or disposed of without endangering human health and without using processes or methods which could harm the environment and in particular without: risk to water, air, soil, plants or animals; or causing nuisance through noise or odours; or adversely affecting the countryside or places of special interest. The Regulations are due to be revised in the near future. Impact and implications for bioremediation The regulations provide a framework of guidance within which all operators work when applying controlled wastes to agricultural land. They are not likely to limit or hinder the use of appropriate wastes on bioenergy crops for bioremediation although, due to the definitions used in the regulations, there can be difficulties in categorising certain wastes. The revised regulations will, however, ban the use of septic tank wastes and are likely to require more pre-notification and auditing of applications. The meanings of the terms “benefit to agriculture” and “ecological improvement” are not clearly defined in the regulations and are open to interpretation by waste producers and their contractors, and by the Environment Agency. As a result, a waste can sometimes be accepted in one region and prohibited in another. It is hoped that clearer definitions, and an indication of what evidence is required to demonstrate ‘benefit’, will be included in future guidance. 109 BIOENERGY CROPS AND BIOREMEDIATION 5.3 UK LEGISLATION, NOT PRIMARILY AIMED AT WASTE DISPOSAL, WHICH MAY AFFECT ORGANIC WASTES 5.3.1 Animal By-Products Order The Animal By-Products Order 1999 (ABPO) (SI, 1999), amending earlier orders in 1992 and 1996, controls all material derived from animals which is not intended for human consumption. It applies to any unprocessed animal by-products (animal carcasses and parts of animal carcasses which are not intended for human consumption) and to catering waste which contains meat or other products of animal origin. It does not apply to BSE suspects, ‘specified risk material’ (SRM), animals suspected of carrying a notifiable disease or stock slaughtered under the ‘over thirty month’ scheme. The ABPO requires that such animal by-products be consigned to rendering, incineration, or other permitted routes. It permits landfilling and/or treatment of catering waste, but requires that ruminant animals, pigs and poultry do not gain access to the waste. Catering waste, by definition, appears to include domestic waste and, therefore, would also include non-source segregated municipal solid waste (MSW). Guidance notes indicate that ‘poultry’ includes wild birds and that composting and biodigestion are not permitted disposal routes for animal by-products under the ABPO. Impact and implications for bioremediation The regulations were designed to control the spread of animal diseases and are necessary, particularly given recent outbreaks of swine fever and foot and mouth disease. However, current interpretation of the regulations effectively prevents the use of open air composting of MSW - since it is virtually impossible to prevent access by wild birds. Furthermore, recent guidance from DEFRA would suggest that, even after treatment, access by wild birds would need to be prevented, thereby effectively preventing the recycling of any MSW to agricultural or restored land. If this interpretation were correct then the ABPO would limit the use of such wastes on bioenergy crops. The UK ABPO regulations are due to be superceded by new EU regulations currently being agreed by the Commission and it is understood that land application of certain, lower risk wastes will be permitted as long as they undergo ‘pre-treatment’ to specified conditions of temperature and pressure. The Environment Agency are due to publish the results of a risk assessment of such wastes and it is understood that the findings will be used as the basis of any new guidance on the treatment of wastes that may contain animal by-products. 110 BIOENERGY CROPS AND BIOREMEDIATION 5.3.2 Plant Health (Great Britain) Order 1993 Under Article 20 of the Plant Health (Great Britain) Order 1993 (as amended) (SI, 1996) growers are required to notify Agricultural Departments of the known or suspected presence of any quarantine pest or harmful organism not normally found in Great Britain. Guidance on minimising the risks of transmitting organisms or disease through the recycling or land application of waste material, including soil or growing media, wash water, trimmings and peelings and outgrades or other plant material can be found in PB3580 (MAFF/WOAD/SOAEFD, 1998). Rhizomania and potato brown rot, as well as more common pests and diseases such as Phytophthora, Fusarium, Verticillium and stem and bulb nematodes, can all be potentially spread through waste products and care should be taken to ensure appropriate treatment of wastes is undertaken. Impact and implications for bioremediation The Code of Practice, and associated regulations, provide a framework of guidance within which operators work when applying potentially infected wastes to agricultural land. They are not likely to limit or hinder the use of appropriate wastes on bioenergy crops for bioremediation. 5.3.3 Planning controls The Town and Country Planning Act 1990 (as amended by the Planning and Compensation Act 1991) controls the use of land for mineral excavation. Assuming consent is granted, detailed planning conditions will be established which control the working, restoration and aftercare of such sites. The Environment Act 1995 provides for a review and updating of permissions granted between 1950 and 1980, with periodic reviews afterwards. Impact and implications for bioremediation The regulations provide a framework of guidance within which all operators work. The planning conditions are often very detailed and may define the type of products that should be used in the restoration process. This may limit the use of alternative wastes or products that can be used. Detailed conditions will only apply to more recent consents; older consents may have little or no aftercare requirements. Many derelict former industrial sites fall into this latter category. 5.3.4 Contaminated land Contaminated land may be a statutory nuisance under the Environmental Protection Act 1990 (HMSO, 1990) and the local authority may have the power, in some circumstances, to issue a notice requiring the person responsible, or the owner or occupier, to prevent the nuisance. 111 BIOENERGY CROPS AND BIOREMEDIATION Under the Environment Act 1995 (HMSO, 1995) local authorities are required to identify contaminated land which poses a significant risk of harm to human health or the environment generally. The owner or person responsible may then be required to remedy the problem. Local authorities may also, under certain circumstances, fund the clean-up of contaminated land. Impact and implications for bioremediation The regulations provide a framework of guidance and may well be a driver for identifying sites, which require bioremediation. 5.4 EC LEGISLATION: PROPOSALS THAT MAY AFFECT ORGANIC WASTES 5.4.1 Introduction A range of measures are currently proposed at an EU level that may also impact on the use of organic wastes for bioremediation, as the proposals - if adopted – will, in time, have to be adopted into UK legislation. 5.4.2 EC Sludge Directive – Working Document 3rd Draft Sewage sludge applications to land are currently regulated by the EC Sludge Directive (EC, 1986). During 1999 and 2000, a series of national expert meetings were held, with a view to revising the Sludge Directive. The Commission has issued its 3rd Working draft although it is understood that this may not represent the final proposals. Revised proposals, which will be subject to consultation and formal agreement with Commissioners and Member States, are expected in spring 2003. The draft proposals of the EC Sludge Directive include: significant reductions in the maximum levels for heavy metals in soils (this would cause major difficulties in the UK) inclusion of septic tank, food wastes, paper wastes and other manufacturing wastes within the remit of the directive inclusion of treatment standards, based on the degree of pathogen reduction achieved (similar to the proposed UK regulations) introduction of maximum levels of heavy metals in the sludge itself, and introduction of limits on organic compounds (based on the limits in Annex III, all UK sewage sludges would fail the limits for PAH). The EU are keen to progress with the concept of sustainability, based on the idea of zero addition of contaminants; whereas the UK is proposing limits based on scientific studies and a precautionary principle approach. There is also concern about the absence of agreed analytical methods and protocols. 112 BIOENERGY CROPS AND BIOREMEDIATION Impact and implications for bioremediation The Directive is still far from agreed and it is likely that further compromises will be made before it is finalised. In any event it is not likely to be ratified until 2003 and is, therefore, unlikely to be enacted in UK legislation until about 2005. It would, however, severely restrict the recycling of sewage sludges to agricultural land and may lead to problems with other ‘wastes’ that will fall within the scope of the directive - since even less is known about the level of organic contaminants, for example, that may be present in other sludges. It also confirms the principle that if you mix any sludge with other non-regulated wastes, then the entire combined waste stream becomes affected by the Directive. This potentially limits some treatment sites where sewage sludge is either used as a feedstock or is treated in order to receive a gate fee, to subsidise the treatment of other wastes. 5.4.3 EC Biological Treatment of Biowaste Directive – Working Document 2nd Draft Often referred to as the ‘Compost Directive’ this new directive is still being discussed and debated by the various national experts. The current working document is intended as a basis for preliminary discussions. Its objectives are to promote the treatment of bio-waste, and to reduce any negative impact of such wastes. It also seeks to protect the soil and ensure human and animal health is not affected by the use of treated or untreated bio-waste. The draft promotes the waste hierarchy: Prevent or reduce production Re-use Recycle Compost or anaerobically digest and use in agriculture Mechanical/biological treatment of bio-waste Use bio-wastes as a source for generating energy The document promotes home composting; on-site composting and anaerobic digestion; ‘community composting’; separate collection schemes (targets are proposed) and promotes the mechanical/biological treatment of any residual municipal waste. The document also proposes the introduction of end-product standards and suggests two classes of compost/digestate, as well as figures for stabilised biowaste. There appears to be a reasonable ‘read-across’ to the sludge directive for organic contaminant limit values – but, based on a quick review of the figures for heavy metals, it would appear that the proposed levels may be difficult to achieve from mixed waste streams and appear to have been based on green waste composting. There are also some concerns about the choice of indicator organisms i.e. Salmonella senftenberg and Clostridium perfringens. Although both are under review, the UK has historically used E. coli and Salmonella spp. as indicator organisms. 113 BIOENERGY CROPS AND BIOREMEDIATION The draft also reinforces the need for any reuse on land to result in agricultural benefit or ecological improvement. It also proposes the following limits on application rates: Class 1 – no limit, but used according to best agronomic practice. Class 2 – 30 t dry matter ha-1 (three year average) Stabilised bio-waste – no repeat application in 10 years and a limit of 200 t dry matter ha-1 in a single application. All applications must be made in accordance with the record keeping and soil sampling requirements of the Sludge Directive (EC, 1986). Impact and implications for bioremediation The Directive is still far from agreed and it is likely that further amendments will be made, in light of negotiation over the Sludge Directive. It would, however, if implemented in the draft form, severely restrict the treatment of many mixed waste streams and industrial wastes. It also confirms the principle that if you mix any digestate with other non-regulated materials, such as peat or inorganic fertilisers, then all of the combined material becomes affected by the Directive. This potentially limits some treatment sites where wastes are either used as a feedstock or are treated in order to receive a gate fee, to subsidise the treatment of other wastes, and also limits the opportunity to add value. As they currently stand, however, the proposed application rate limits would severely restrict the use of biowastes in land reclamation and bioremediation since current practice is to apply much higher rates than are proposed in the second working draft. 5.4.4 EC Landfill Directive (EC 1993/31) The recently adopted Landfill Directive 1993/31/EC (EC, 1993) requires that significant reductions must be made in the amount of putrescibles being disposed of to landfill sites by 2006, further targets also apply for 2009 and 2016. Government estimates that 3.2 m tonnes per year must be diverted from landfill to meet initial targets. While the paper ‘Limiting Landfill’ (DEFRA, 1999) recognises that a significant part will be played by waste minimisation initiatives, it remains focused on treatment of waste as an alternative to disposal to landfill. Surveys of household waste (MSW) identify 20% as organic food, and 33% paper/card, giving a potential 53% of MSW material that can be composted. It is estimated that 5 million tonnes per annum of organic material is disposed of within the MSW stream, which could be realistically diverted if a suitable product and market could be found. The Directive is driving waste management companies and waste producers to look at land spreading and bioremediation as a possible route to divert material away from landfill. The Directive was implemented in England and Wales under the Pollution Prevention and Control (England and Wales) Regulations 2000, as amended (SI, 2002). 114 BIOENERGY CROPS AND BIOREMEDIATION Impact and implications for bioremediation This directive is acting as a driver for change and is encouraging the development of alternative routes for recycling, either in the form of waste treatment or the establishment of new markets for recycling of wastes. Recent concern from retailers and food processors has led many waste management operators to look at the use of organic wastes for bioremediation and biomass production, since they are seen as more ‘secure, than traditional outlets. 5.5 REFERENCES ADAS (2000). MANNER User Guide. ADAS, ADAS Gleadthorpe Research Centre, Mansfield, Nottingham. ADAS (2001a). The Safe Sludge Matrix – Guidelines for the Application of Sewage Sludge to Agricultural Land. ADAS, Wolverhampton. ADAS (2001b). Guidelines for the Application of Sewage Sludge to Industrial Crops. ADAS, Wolverhampton. DEFRA (1999). Limiting Landfill: A consultation paper on limiting landfill to meet the EC Landfill Directive’s targets for the landfill of biodegradable municipal waste. Originally published by DETR. http://www.defra.gov.uk/ environment/waste/strategy/landfill/index.htm. DETR/MAFF/WO (1997). Draft regulations establishing the action programme measures to apply in Nitrate Vulnerable Zones in England and Wales Consultation Document. EC Nitrate Directive (91/676/EEC): Department of the Environment, Transport and the Regions, Ministry of Agriculture, Fisheries and Food, Welsh Office. December 1997. DoE (1990). Environmental Protection Act, 1990. Environment. HMSO, London. Department of the DoE (1993). Sludge Use in Agriculture 1990/91. UK report to the EC Commission under Directive 86/278/EEC. Department of the Environment, London. DoE (1996). Code of Practice for the Agricultural Use of Sewage Sludge. Department of Environment. HMSO, London. EA (2000). Aquatic Eutrophication in England and Wales – a management strategy. Environment Agency. EA (2001). Water pollution incidents in England and Wales 1999. Environment Agency. EC (1991a). Council Directive on Waste (91/156/EEC) of 18 March 1991 amending Directive 75/442/EEC on waste. Official journal of the European Community L 078, 26/03/1991. pp 0032–0037. EC (1991b). Nitrate Directive (91/676/EEC). Official Journal of the European Community. 115 BIOENERGY CROPS AND BIOREMEDIATION EC (1991c). Urban Waste Water Treatment Directive (91/271/EEC). Official Journal of the European Community, L13530.5.91, p 40. As amended by Council Directive 98/15/EC, February 1998. Summary of Measures Implemented by Member States COM (1998). EC (1993). Landfill Directive (1993/31/EEC). Official Journal of the European Community. Forestry Commission (1992). A Manual of Good Practice for the Use of Sewage Sludge in Forestry. Bulletin 107. Forestry Commission Publications, Wetherby, West Yorkshire. HMSO (1947). Agriculture Act 1947. Chapter 48. HMSO, London. HMSO (1990a). London. Environmental Protection Act 1990. Chapter 43. HMSO, HMSO (1990b). Town and Country Planning Act 1990. Chapter 8. HMSO, London. HMSO (1991). Water Resources Act 1991. Chapter 57. HMSO, London. HMSO (1995). Environment Act 1995. Chapter 25. HMSO, London. MAFF (1998a). The Water Code. Code of Good Agricultural Practice for the Protection of Water. Ministry of Agriculture, Fisheries and Food/Welsh Office Agriculture Department. MAFF Publications, London. (PB0585). MAFF (1998b). The Air Code. Code of Good Agricultural Practice for the Protection of Air. Ministry of Agriculture, Fisheries and Food/Welsh Office Agriculture Department. MAFF Publications, London (PB0618). 74 pp. MAFF (1998c). The Soil Code. Code of Good Agricultural Practice for the Protection of Soil. Ministry of Agriculture, Fisheries and Food/Welsh Office Agriculture Department. MAFF Publications, London. (PB0617). MAFF (2000). Fertiliser Recommendations for Agricultural and Horticultural Crops. 7th edition. MAFF Reference Booklet 209. HMSO, London. MAFF/WOAD (1998). Guidelines for Farmers in NVZs. MAFF Publications, London (PB3277). MAFF/WOAD/SOAEFD (1998). Code of Practice for the Management of Agricultural and Horticultural Waste. Ministry of Agriculture, Fisheries and Food/Welsh Office Agriculture Department/Scottish Office Agriculture Environment and Forestry Department. MAFF Publications, London. (PB3580). SI (1989). United Kingdom Statutory Instrument No. 1263. The Sludge (Use in Agriculture Regulations, 1989. HMSO, London. SI (1990). United Kingdom Statutory Instrument No. 880. The Sludge (Use in Agriculture) (Amendment) Regulations, 1990. HMSO, London. SI (1991). United Kingdom Statutory Instrument No. 324. The Control of Pollution (Silage, Slurry and Agricultural Fuel Oil) Regulations 1991. HMSO, London. SI (1992). United Kingdom Statutory Instrument No. 3293. The Animal Health Act 1981 (Amendments) Regulations 1992. HMSO, London. 116 BIOENERGY CROPS AND BIOREMEDIATION SI (1994a). Statutory Instrument No. 2841. Urban Waste Water Treatment (England & Wales) Regulations 1994. HMSO, London. SI (1994b). Statutory Instrument No. 1056. The Waste Management Licensing Regulations 1994. HMSO, London. SI (1996). United Kingdom Statutory Instrument No. 3242. Plant Health (Great Britain) (Amended) (No 3) Order 1996. HMSO, London. SI (1998). Statutory Instrument No. 1202. The Action Programme for Nitrate Vulnerable Zones (England and Wales), 1998. HMSO, London. SI (1999). Statutory Instrument No. 646. The Animal By-Products Order 1999. HMSO, London. SI (2002). United Kingdom Statutory Instrument No. 275. The Prevention and Control (England and Wales) (Amendment) Regulations 2002. HMSO, London. 117 BIOENERGY CROPS AND BIOREMEDIATION CHAPTER 6 CONCLUSIONS AND RESEARCH RECOMMENDATIONS 6.1 CONCLUSIONS The planned expansion of the area in the UK planted with energy crops, to partially facilitate the requirement that 10% of all electricity generation in 2010 must come from renewable sources, provides an opportunity for the safe disposal of organic wastes and for the bioremediation of polluted, ex-industrial land. The safe disposal of organic wastes, such as sewage sludge, animal manures and slurries, paper sludge and abattoir wastes can be problematic. Many of these materials contain heavy metals, organic pollutants or pathogens that are potentially damaging to human health; and their utilisation on food crops causes public concern, despite stringent safeguards embodied in various articles of legislation and codes of practice. The opportunity to apply such wastes to vigorous non-food crops, with extensive and rapidly developing root systems, would appear to have obvious attractions – offering an environmentally-friendly alternative to food crop applications, landfill or incineration. The facts that energy crops are regularly harvested, with the above-ground parts being removed from the site and burnt, and that the postharvest period (in late winter and early spring) offers ready access to the crop for waste spreading, would appear to further increase the ‘bioremediation’ potential. One further, but very important, benefit that might accrue to landowners exploiting the waste utilisation or bioremediation potential of energy crops is improved financial viability. Current levels of government grants and anticipated income from crop sales are proving insufficiently attractive to entice more than a small minority of landowners into energy crop production. Any opportunity to ‘add value’ to energy crops, such as providing a regular disposal point or organic wastes, will increase their attractiveness to many potential growers. This review has confirmed many of the potential opportunities for waste utilisation/bioremediation that energy crops offer. However, it has also highlighted certain environmental risks that need to be considered. For example… Repeated applications of organic wastes with a high P content (e.g. pig or poultry manures), at rates which apply the maximum permitted quantity of N (i.e. 250 kg N ha-1 yr-1), are likely to supply P in excess of the maximum permitted under the relevant code of practice. This will increase the risks of eutrophication of water-bodies. Similarly, repeated applications of organic manures (livestock manure, biosolids, industrial waste) will be likely to add heavy metals in excess of crop uptake, leading to an accumulation in the soil. Heavy metals can also 118 BIOENERGY CROPS AND BIOREMEDIATION be carried through to the ash left after burning, so disposal of ash must be made with due regard to its chemical composition. Slurry applications might lead to excessive ammonia volatilisation, and local odour problems – as the most effective way of reducing volatilisation is to inject the slurry into the soil, but this is not an option in energy crops. Very little is known about the ecological impacts of waste applications to energy crops. From the limited amount of information that is available, it can be surmised that the generally low conservation value of actively managed energy crops is unlikely to be significantly reduced or increased. However, the potential bioaccumulation of heavy metals in predator species would be an issue of some concern – if large areas of energy crops around, for example, a single power station were to be regularly treated with wastes containing high metal concentrations. The genetic modification of biomass crops to facilitate increased pollutant uptake, transport, accumulation and tolerance offers the potential to dramatically increase the effectiveness of phytoremediation of organic compounds and metals from contaminated sites. Although the genetics of poplars have been extensively studied, very little research has so far been undertaken on the genetic modification of the primary candidates for energy cropping in the UK - willows and Miscanthus. Consequently, the realisation of this potential is still some distance away. 6.2 RESEARCH RECOMMENDATIONS The following subjects should be considered as priority areas for research: 1. Nutrient and CO2 losses to the environment on removal of biomass crops and return of land to conventional agriculture. 2. The nutrient requirements of energy crops under a range of conditions. 3. Investigation of the potential use of leaf analysis in the assessment of nitrogen requirements for energy crops. 4. Further research on species or varietal sensitivity of SRC and Miscanthus crops to landfill leachate and the maximum conductivity (ec) of leachates for ‘safe’ irrigation. 5. N2O efflux from sites without organic amendments, but with and without inorganic nutrient fertilisation, and in the presence of normal litter quantities, to establish a baseline flux balance. 6. N2O and CH4 flux after organic amendment application in normal SRC conditions i.e. after the passage of harvest machinery and in the presence of litter and stools. 7. Contribution of both applied organic wastes and annual litter fall to medium and long-term soil organic matter carbon sequestration. 8. Longevity and distribution of carbon sequestered in woody root systems of SRC, and the turnover of root biomass in Miscanthus. 119 BIOENERGY CROPS AND BIOREMEDIATION 9. Longer-term N2O and CH4 flux from Miscanthus sites receiving annual applications of organic/inorganic fertilisers. 10. Potential of the mineralisation efflux of CO2 from frequently applied organic wastes to result in carbon fertilisation of C3 plants such as willow and poplar, by a CO2-enriched micro-climate. 11. Further screening of willow, poplar and energy grass varieties to evaluate their levels of tolerance to inorganic and organic pollutants, and assessment of the extent to which pollutants are either inactivated (e.g. certain organic compounds) or accumulated in harvestable plant parts. 12. Assessment of the types and quantities of organic contaminants in organic manures and rates of uptake and breakdown by energy crops. 13. The genetic modification of energy crops to increase their bioremediation potential. 14. The agronomic and ecological impacts of applying different waste materials (at different rates) to energy crops. 15. Environmental fate of pollutants in contaminated soils after establishment of energy crops. 16. Environmental fate of pollutants in waste materials after their application to energy crops. 17. The nature and environmental fate of pollutants in flue emissions and ash after combustion/pyrolysis for power generation. 120