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Chapter 1 Introduction An expressway is a controlled-access highway; it is a highway that controls entrances to it and exits from it by incorporating the design of the slip roads for entry and exit into the design of the highway itself. An expressway may be free to use and may not collect toll at all. Expressways are the highest class of roads in the Indian Road Network. These are six- or eight-lane highways with controlled-access. India has approximately 942 km expressways. National Highway system of India consists of approximately 10,000 km (6,200 mi) of four-laned highways, but do not fulfill the criterion expressways because they do not have control of access. Currently, a massive is underway to expand the highway network and the Government of India plans to add an additional 18,637 km (11,580 mi) of expressways to the network by the year 2022 [Daily News and Analysis]. These roads will be access-controlled roads and will feature between four and six lanes with 3,530 km (2,190 mi) km to come up by 2015. The Ministry of Road Transport and Highways is already in the process of preparing a draft for creation of a National Expressways Authority of India (NEAI) (Dash, 2009). Like people, most organizations are heavily dependent on roads to distribute their goods. The development of a wide range of transportation system in all over the world and safety of environment from its various activities is a global challenge. The Issue of transport activities and the environment is paradoxical in nature since transportation conveys substantial socioeconomic benefits, but at the same time transportation is also contribute pollutants to the environmental systems. From one 1 side, transportation activities support increasing mobility demands for people while on the other side transport activities grow levels of environmental externalities. This has reached a point where transportation system is a dominant source of emission of most pollutants poses detrimental impacts on the environment adversely affect health hazards to all living organisms including human beings (Kumar, 2013). The transport activities release several million tons of gases each year into the atmosphere, mostly they are toxic to living organisms (Zhu et al., 2002). These include chemicals, gases (CO, CO2, methane, NOx, SOx, CRCs, PFCs), carbon monoxide, carbon dioxide, methane, nitrogen oxides, nitrous oxide, heavy metals etc. The emission of some of these pollutants cause climate change. Some of these gases, particularly nitrous oxide, also participate in depleting the stratospheric ozone (O3) layer which naturally screens the earth’s surface from ultraviolet radiation. Vehicles on express highway are the source of pollution in the form of gases and particulate matters (SPM and RSPM) emissions affect air quality, causing damage to human health. Toxic air pollutants cause several diseases such as cancer, cardiovascular, respiratory and neurological diseases in human beings (Kumar, 2013. Carbon monoxide (CO) when inhale affects bloodstream, reduces the availability of oxygen and can be extremely harmful to public health (Seaton et al., 1995). An emission of nitrogen dioxide (NO2) from transportation sources reduces lung function and increases the risk of respiratory problems. The emissions of sulphur dioxide (SO2) and nitrogen oxides (NOx) in the atmosphere form various acidic compounds that when mixed in cloud water creates acid rain. Acid precipitation has detrimental effects on the soil conditions, reduces agricultural crop yields and causes forest loss (Pandey et al., 2008a). 2 Noise represents the general effect of irregular and chaotic sounds, that may affect the quality of life by its unpleasant and disturbing character. Long term exposure to noise levels above 75dB seriously hampers hearing and affects human physical and psychological wellbeing. Transport activities have an impact on hydrological conditions. Fuel, chemical and other hazardous particulates (Ukpong and Moses, 2001) discarded from various transport activities emitted heavy metals in the air, they settled down on the earth surface, and ultimately reach to the surface water bodies (Kisku et al., 2000; Epriewska and Bueior, 2001) and degrade their qualities (Pandey and Nautiyal, 2008). Can contaminate rivers, lakes, wetlands and other surface water bodies The environmental impact of transportation on soil consists of soil erosion and soil contamination (Kumar and Pandey, 2010). Coastal transport facilities have significant impacts on soil erosion. The removal of earth’s surface for highway construction has led to important loss of fertile and productive soils (Sukreeyapongse et al., 2002). Transportation also influences natural vegetation (Pilon-Smits, 2005). The need for construction materials and the development of land-based transportation. Many transport routes have required draining land, thus reducing wetland areas and driving-out water plant species. Major transport facilities can affect the quality of urban life (Quishlaqi et al., 2007) by creating physical barriers, increasing noise levels, generating odors, reducing urban aesthetic and affecting the built heritage (Hadjiliadis, 1997). 3 The metals are classified as “heavy metals” if, in their standard state, they have a specific gravity of more than 5 g/cm3. There are sixty known heavy metals. Heavy metals can accumulate over time in soils and plants and could have a negative influence on physiological activities of plants (e.g. photosynthesis, gaseous exchange, and nutrient absorption), causing reductions in plant growth, dry matter accumulation and yield (Devkota and Schmidt, 2000). Heavy metals in the atmosphere, soil and water, some are even in traces (Cd, Cr, Hg, As etc.) can cause serious problems to all living organisms, and their bioaccumulation in the food chain especially can be highly dangerous to human health (Afyoni et al., 1998; Pilon-Smits, 2005). The entry of pollutants in the environment and their adverse effects on living beings become uncontrolled due to extensive vegetation loss (Pandey, 2014). Chromium is toxic to plants and does not play any role in plant metabolism (Pandey et al., 2005). Accumulation of Cr by plants can reduce growth, induce chlorosis in young leaves, reduce pigment content, alter enzymatic function, damage root cells and cause ultrastructural modifications of the chloroplast and cell membrane (Paivoke, 2002; Morales et al., 2007; Zhang et al., 2007). During seed germination, hydrolysis of proteins and starch takes place, providing amino acids and sugars (Zied, 2001). Chromium when enter in plant parts can alter chloroplast and membrane ultrastructure in plants (Vajpayee et al., 2001; Pandey and Gautam, 2009a,b). Chromium can induce degradation of carotenoids in plants Choo et al., 2006). The increase in carotenoids content may act as an antioxidant to scavenge ROS generated as a result of Cr toxicity (Panda and Choudhury, 2004). In aquatic environments, 4 chromium will be present predominantly in a soluble form (Rout et al., 2000). Chromium have been shown to accumulate in many aquatic species, especially in bottom-feeding fish, such as the brown bullhead (Ictalujrus nebulosus); and in bivalves, such as the oyster (Crassostrea virginica), the blue mussel (Mytilus edulis) and the soft shell clam (Mya arenaria) (Scoccianti 2006; Seng and Bielefeldt, 2002). Chromium compounds are corrosive, and allergic skin reactions readily occur following exposure, independent of dose. Short-term exposure to high levels results in ulceration of exposed skin and irritation of the gastrointestinal tract (Horvath et al., 2008; Armienta et al., 2001). Chromium often accumulates in aquatic life, adding to the danger of eating fish that may have been exposed to high levels of chromium (Cifuentes et al., 1996; Epniewska and Bucior, 2001). Long-term occupational exposure to airborne levels of chromium higher than the natural environment leads to lung cancer (Akinola et al., 2008; Kumar and Pandey, 2010). Soil and water contamination with Ni has become a worldwide problem (Guo and Marschner, 1995). Ni is essential for plants (Brown et al., 1987; Salt et al., 1995), but the concentration in the majority of plant species is very low (0.05-10 mg kg-1 dw.). Further, with increasing Ni pollution, excess Ni rather than a deficiency, is more commonly found in plants (Ragsdale, 1998). Toxic effects of high concentrations of Ni includes inhibition of mitotic activities (Madhavrao and Shresty, 2000), reductions in plant growth (Molas, 2002) and adverse effects on fruit yield and quality (Gajewska et al., 2006) have been observed. Nickel is a dietary requirement for many organisms (Parida et al., 2003), but may be toxic at higher concentrations (Pandey et al., 2009b). Metallic nickel and some other nickel compounds are carcinogenic to mammals. The long-term exposure 5 of Ni can cause decreased body weight, heart and liver damage, and skin irritation (Kao et al., 2008). Nickel can accumulate in aquatic life, may adversely affect the aquatic ecosystem. Inhalation of nickel can result in chronic bronchitis, emphysema, and asthma and lung cancer. By ingestion, nickel has been associated with reduced body weight and reproductive and foetotoxic effects (Mulrooney and Hausinger, 2003). The average abundance of copper in the earth crust in 68 ppm, in soils, it is 933 ppm, in streams it is 4-12 g/L; and in ground water it is <0.1 mg/l (APHA, 2005). Copper is considered as essential trace element for plants and animals. At high concentration, Cu can become extremely toxic causing symptoms such as chlorosis and necrosis, stunting of plant growth, leaf discoloration and inhibition of root growth have been reported by several workers (Chen and Kao, 1999; Marschner, 1995). Copper is an essential substance to human life, but in high doses it can cause anemia, liver and kidney damage, and stomach and intestinal irritation. People with Wilson’s disease are at greater risk for health effects from overexposure to copper (Fritloff and Greger, 2006). Zinc occurs naturally in air, water and soil, but zinc concentrations are rising unnaturally, due to addition of Zn through various human activities (Naaz and Pandey, 2009), such as mining, waste combustion and effluent discharge (Pandey, 2006a, b; Pandey and Nautiyal, 2008). Some soils are heavily contaminated with zinc, and these are to be found in areas where zinc has to be mined or refined, or were sewage sludge from industrial areas has been used as fertilizer and vehicular exhaust Bunzl et al., 2001). 6 Zinc is an essential element for both plants and animals (Carroll and Lonearagan, 1968). It plays an important role in several plant metabolic processes; it activates several enzymes and is involved in protein synthesis and carbohydrate, nucleic acid and lipid metabolism (Cakmak and Marshner, 1993). However, like other heavy metals (Doncheva et al., 2001) when Zn is accumulated in excess in plant tissues, it causes alterations in vital growth processes such as photosynthesis and chlorophyll biosynthesis and membrane integrity (Doncheva et al., 2001). An excess of Zn have a negative effect on mineral nutrition (Baccouch et al., 1998a,b). Toxic levels of Zn for different varieties of crop have very wide limits in growth medium from 64 g L-1 Zn for sorghum to 2000 g L-1 Zn for cotton (Otte et al., 1995). Excess of Zn decrease growth and development, metabolic activity and induce oxidative damage in various plant species (Sekara et al., 2005). The chlorophyll content of green vegetables typically exceeds the levels of other bioactive pigments (Nagajyothi et al., 2009) such as carotenoids. Degradation of pigments has widely been used as an indicator of pollution. Chlorophyll, the green pigment is one of the main complex which influences photosynthesis. Decreases in chlorophyll content under stress (pollution or temperature stress) may be attributed to either its degradation or to reduced biosynthesis (Schutzendubel and Polle, 2002). Carotenoids belong to a large group of compounds called terpenoids. These compounds produce red orange, yellow and brown color in plants. They are further divided on the basis of presence and absence of oxygen into carotenes, which have formula C40H56, contain only C and H and xanthophylls contain oxygen along with C and H; common xanthophyll of leaves is lutein (C40H56O2). Over 600 carotenoids occurring in plants, fungi, bacteria and animal, including humans, are present 7 (Kenneth et al., 2000). Carotenoids content involve in photo protective functions in photosynthesis (Kenneth et al., 2000). Carotenoid pigments also have ecological significance. Marking flowers and fruits colored, they play an important role in ecosystems, attracting pollen-dispersing insects and fruit-eating animals. In humans, carotenoids normally occur in several types of tissues, e.g., muscles, liver, eye, blood and adipose tissue (Singh and Pandey, 2011). Proteins are made up of several nitrogen containing organic molecules called amino acids. The amino acid is the basic unit of protein. Protein dissociates to form amino acids and the energy produced is utilized for routine metabolic activities (Chandra et al., 2004). Proteins are the most important constituent of plant cells both from structural as well as functional point of view. Functionally, give rise to enzymes, which are responsible for regulating the cellular process (Rodriguez et al., 2007). Pollutants when enter into living organisms, degrade the structural and functional quality of the protein. During construction work of highway encroaches upon precious ecological resources. The most affected ecological resources are green vegetation and sunamps, this also disturbs the natural habitats of a lot of animals living in the catchment areas. The activities during the construction work of highways and post construction work some wild life away from their natural habitats, including migratory birds. Due to the road construction work, the destruction of vegetation occurs on the acquired land. On expresshighways, during operation, the traffic noise, traffic light at night and vehicle emissions may cause some adverse impacts on the wild life, growth and 8 flowering of plants around the road. The pollution by various chemical, heavy metals and dust particles pollutant, the possibility of loss of biodiversity around the road is highly possible. During the construction and operation of expresshighways a huge amount of carbondioxide, carbon monoxide, oxides of sulphur and nitrogen gases are released into the atmosphere. Therefore, gases, particularly sulphur dioxide, may pose a threat to ancient monuments which are made up of lime. Therefore, a high risk of ecological disturbances such as degradation of soil, water, plants, wild life etc., is possible during the construction and operational works of expresshighway. The best practice is to undertake the environmental impact assessment (EIA) before road is designed. The study areas are located in Unnao district (of Uttar Pradesh state, India) near expresshighway (NH 25) crossing the proposed Ganga expressway. The total length of proposed Ganga expressway is 1047 km link Noida to Ballia district of Uttar Pradesh. Therefore, ecological studies at proposed Ganga expressway area to environmental impact assessment (EIA) is necessary to protection of living organisms and planning to manage transport activities eco-friendly. 9 Aims and objectives The impact of transport activities to environmental degradation is a world wide problem. From one side, transportation conway substantial socio-economic benefits to the country and support increasing mobility demands for people, while on the other side, transport activities are a major source of pollution, and degrade natural resources (soil, water and plants). In India, about 3402 km expresshighway including about 1047 km of Ganga expresshighway have been proposed to be completed in near future. To reduce the negative consequences of transport activities on the environment, an eco-friendly policy should be made after a environmental assessment programme. Least research work is available on this aspect. Therefore, study was undertaken to explore the situation and possible way out of the problem in following objectives: Ecological studies of soil and water near the Ganga Expressway. To find out the effect of environmental changes due to construction of Ganga expressway on the wild species growing in the study area. To find out the impact of the water and soil on the agricultural fields. 10 Chapter 2 Review of Literature The transportation support the mobility demand of increasing population of human beings and pressure of vehicles. On other side, a huge net work of transportation system throughout the world is a dominant source of emission of most pollutants cause biosphere pollution. During the last few decades due to industrialization, civilization, vegetation loss due to road construction and other anthropogenic activities resulted ecological disturbances (Singh et al., 2008), health hazards of living organisms (Kumar and Pandey, 2010) and disruptions of natural ecosystems (He et al., 2005; Abida et al., 2009). Due to high degree of vehicular discharge the heavy metals are at higher concentrations, they enter the food chain (Barman et al., 2000)through uptake and accumulation in plants (Jinfang et al., 2008) posing a potential threat to human health. These metals can be transferred and concentrated into plant tissues from the soil (Karbassi et al., 2006) and brought significant reductions in plant growth (Naaz, 2012) et al., 2007). The various pollutants emitted from transportation enters in our delicate food web, the heavy metals are most injurious. Therefore, the elevated levels of various pollutants and altered concentration of heavy metals in soil and water (Singh and Pandey, 2011) is a matter of great concern to scientific workers (Moreno et al., 1994) to save the living beings from their effects. An assessment of the environmental risk due to soil pollution near highways with high vehicular load is of particular importance for agriculture and non-agricultural areas, because heavy metals, which are potentially harmful to human health (Kumar and Pandey, 2010), persist in soils for 11 a very long time (Pandey, 2006a,b) and they may enter the food chain in elevated amounts (Kabata et al., 1999; Grzebisz et al., 2001). 2.1 Indian Expressways An expressway is a controlled-access highway: it is a highway that controls entrances to it and exits from it by incorporating the design of the slip roads for entry and exit into the design of the highway itself are six or eight-lane highways with controlled-access. Expressways and the highest class of roads in the Indian road network. India has approximately 942 km expressways. National highway system of India consists of approximately 10,000 km of four-lane highways that collect toll from users but do not have control of access and cannot be called expressways. Currently, a massive project is underway to expand the highway network and the Government of India plans to add an additional 18,637 km (11,580 mi) of expressways to the network by the year 2011. These roads will be access-controlled roads and will feature between four and six lanes with 3,530 km (2,190 mi) km to come up by 2015. The Ministry of Road Transport and Highways is already in the process of preparing a draft for creation of a National Expressways Authority of India (NEAI) on the lines of NHAI. List of table 1 includes roads without access-control. Such a road cannot be called "expressway" though the name of the road may include the word "expressway" and may be a misnomer. Such a road should be excluded from this list. Eastern and Western Expresshighways in Mumbai are two examples of such roads. Ambala-Chandigarh NH is another such example as it does not have access control for entry and exit at predetermined points. As stated above, access-control is different from collection of toll. 12 Table 2.1: List of Expressways in India. Sl. Expressway Name Distance State *1. Ahmedabad Vadodara Expressway 95 km (59 mi) Gujarat *2. Mumbai-Pune Expressway 93 km (58 mi) Maharashtra *3. Jaipur-Kishanarh Expressway 90 km (56 ini) Rajasthan *4. Allahabad Bypass 86 km (53 mi) Uttar Pradesh *5. Durgapur Expressway 65 km (40 mi) West Bengal 6. Ambala-Chandigarh Expressway 35 km (22 mi) Haryana/Punjab 7. Chennai Bypass 32 km (20 mi) I Tamil Nadu 8. Delhi-Gurgaon Expressway 28 krn (17 mi) Delhi/Haryana 9. Noida-Greater Noida Expressway 24.53 km (15.24 mi) Delhi/Uttar Pradesh 10. Delhi Noida Direct Flyway 9.2 kin (5.7 mi) Delhi/Uttar Pradesh 11. Hyderabad Elevated Expressway 11.6 km (7.2 mi) Andhra Pradesh 12. Hosur Road Elevated Expressway (Bangalore) 9.985 km (6.204 mi) Karnataka 13. Kona Expressway 8 km (4.97 mi) West Bengal 14. Outer Ring Road (Hyderabad) 158 km (98 mi) Andhra Pradesh 15. Raipur-Bhilai-Durg Expressway 26 km (16 mi) Chhattisgarh *16. Yamuna Expressway 165 kni (103) mi) Uttar Pradesh 17. Bangalore Mysore. Infrastructure Corridor 111 km (69 mi) Karnataka 18. Lucknow Amar Shaheed Path, Elevated access controlled stretch 49 km (30 mi) Uttar Pradesh 19. Mumbai Nashik Expressway 150 km (93 mi) Maharashtra 20. City of Kanpur’s Elevated Bypass 25 km (16 mi) Uttar Pradesh 21. Bangalore-Outer Ring Road 62 km (39 mi) Karnataka 22. Bangalore-Nelamangala Elevated expressway on Tumkm Road 19.5 km (12.1 mi) Karnataka 23. Eastern Freeway 22 km (14 mi) Maharashtra Total 1,208.19 km (750.73 mi) *- Controlled-access expresshighway (4-6 lanes) 13 Table 2.2: Proposed National Express Highways under construction. Sl. Expressway Name Distance 1. Western Freeway Mumbai 25-33 km (15.7mi) Maharashtra 2. Eastern Freeway Mumbai 22 km (14 mi) Maharashtra 3. Sion Panvel Expressway 25 km (16 mi) Maharashtra 4. Nagpur-Aurangabad-Mumbai Expressway 700 km (430 mi) Maharashtra 5. Kundli Manesar Palwal Expressway (KMP) 135 km (8.3 mi) Haryana 6. Delhi Eastern Peripheral Expressway 135 km (84 mi) Uttar Pradesh / Haryana 7. Pathankot Ajmer Expressway 600 km (370 mi) Punjab/ Rajasthan 8. Ganga Expressway 1,047 km (651 mi) Uttar Pradesh 9. Bamroli Althan Expressway 12 km (7.5 mi) Gujarat 10. Upper Ganga Canal Expressway 150 km (93 mi) Uttar Pradesh 11. Chennai Port Maduravoyal Expressway 19 km (12 mi) Tamil Nadu 12. Hyderabad ORR 158 km (98 mi) Andhra Pradesh 13. Raipur-Bilaspur Expressway 126 km (78 mi) Chhattisgarh 14. Ganga Expressway 21.5 km (13.4 mi) Uttar Pradesh 15. Jaipur-Delhi Expressway 235 km (16 mi) Rajasthan/Haryana/Delhi 16. Pune-Solapur Expressway 110 km (68 mi) Maharashtra Total length of Expressways 3,401 km (2,113.86 mi) 14 State 2.2 Impacts of Transportation on the Environment The issue of transport activities and the environment is paradoxical in nature since transportation conveys substantial socioeconomic benefits, but at the same time transportation is also contribute pollutants to the environmental systems. From one side, transportation support increasing mobility demands for people while on the other side transport activities grow levels of environmental externalities. This has reached a point where transportation system is a dominant source of emission of most pollutants poses detrimental impacts on the environment including living organisms. These impacts are- (i) Direct (The immediate consequence of transport activities on the environment where the cause and effect relationship is generally clear and well understood), (ii) Indirect (The secondary) (or tertiary) effects of transport activities on environmental systems. They are often of higher consequence than direct impacts, but the involved relationships are often misunderstood and difficult to establish) and (iii) Cumulative impacts (The additive, multiplicative or synergetic consequences of transport activities. They take into account of the varied effects of direct and indirect impacts on an ecosystem, which are often unpredicted). First, transport activities contribute among other anthropogenic and natural causes, directly, indirectly and cumulatively to environmental problems. In some cases, they may be a dominant factor, while in others their role is marginal and difficult to establish. Transport activities contribute at different geographical scales to environmental problems, ranging from local (noise pollution and CO emissions) to 15 global (climate change), not forgetting continental / national / regional problems (smog and acid rain effects). 2.3 Pollutants into the environment from transport activities: Climate change The transport activities release several million tons of gases each year into the atmosphere (Muzyka et al., 1998) mostly which are toxic to living organisms (Zhu et al., 2002). These include lead, carbon monoxide, carbon dioxide, methane, nitrogen oxides, nitrous oxide chlorofluorocarbons (CFCs), perfluorocarbons (PFCs), silicon tetraflouride (SF6), benzene and volatile components, heavy metals (zinc, chrome, copper, lead, iron and cadmium) and particulate matters (ash, dust). The emission of some of these metals cause climate change and the role of anthropogenic factors. Some of these gases, particularly nitrous oxide, also participate in depleting the stratospheric ozone (O3) layer which naturally screens the earth’s surface from ultraviolet radiation. Air quality Vehicles on express highway are the source of pollution in the form of gases and particulate matters (SPM and RSPM) emissions affects air quality, causing damage to human health (Kumar, 2012). Toxic air pollutants cause several diseases such as with cancer, cardiovascular, respiratory and neurological diseases in human beings. Carbon monoxide (CO) when inhale affects bloodstream, reduces the availability of oxygen and can be extremely harmful to public health. An emission of nitrogen dioxide (NO2) from transportation sources reduces lung function and increases the risk of respiratory problems (Sagai et al., 1996). The emissions of 16 sulphur dioxide (SO2) and nitrogen oxides (NOx) in the atmosphere form various acidic compounds that when mixed in cloud water creates acid rain. Acid precipitation has detrimental effects on the soil conditions, reduces agricultural crop yields and causes forest loss (Pandey et al., 2008). The reduction of natural visibility by smog has a number of adverse impacts on the life of living organisms. Particulate emissions in the form of dust emanating from vehicle exhaust as well as from nonexhaust sources such as vehicle and road abrasion have an impact on air quality. The physical and chemical properties of particulates are associated with health risks (Kumar, 2013) such as respiratory problems, skin irritations, eyes inflammations, blood clotting and various types of allergies (Kumar, 2013). Noise represents the general effect of irregular and chaotic sounds, that may affect the quality of life by its unpleasant and disturbing character. Long term exposure to noise levels above 75dB seriously hampers hearing and affects human physical and psychological wellbeing. Transport noise emanating from the movement of transport vehicles and the operations of ports, airports and rail yards affects human health (Muzyka et al., 1998), through an increase in the risk of cardiovascular diseases. Increasing noise levels have a negative impact on the urban environment reflected in falling land values and loss of productive land uses. Water quality Transport activities have an impact on hydrological conditions. Fuel, chemical and other hazardous particulates (Ukpong and Moses, 2001) discarded from various transport activities emitted heavy metals in the air, they settled down on the earth surface, and ultimately reach to the surface water bodies (Kisku et al., 2000; Epriewska and Bucior, 2001) and degrade their qualities (Pandey and Nautiyal, 2008). 17 Can contaminate rivers, lakes, wetlands and other surface water bodies (Singh et al., 2008). These contaminated surface water bodies near expresshighways, often crops at nearby areas. Soil quality The environmental impact of transportation on soil consists of soil erosion and soil contamination (Pandey, 2014). Coastal transport facilities have significant impacts on soil erosion. The removal of earth’s surface for highway construction has led to important loss of fertile and productive soils (Sukreeyapongse et al., 2002). Soil contamination can occur through the use of toxic materials by the transport industry (Sims and Sklin, 1991). Fuel and oil spills from motor vehicles are washed on road sides and enter the soil. Heavy metals emited through human activities pollute soil depending upon properties of the soil (Meenakshi and Pandey, 2009). Gautam and Pandey (2008) reported effect of pollutants on loss of soil fertility. Hazardous materials and heavy metals have been found in areas contiguous to roads (Sahu et al. (2007). Biodiversity Transportation also influences natural vegetation (Pilon-Smits, 2005). The need for construction materials and the development of land-based transportation (Naaz, 2012). Many transport routes have required draining land, thus reducing wetland areas and driving-out water plant species. The need to maintain road and rail right-of-way or to stabilize slope along transport facilities has resulted in restricting growth of certain plants or has produced changes in plants with the introduction of new species different from those which originally grew in the areas (Hall, 2002). 18 Many animal species are becoming extinct as a result of changes in their natural habitats and reduction of ranges (Appelo and Postma, 2005). Land take Transportation facilities have an impact on the urban landscape. The development of transport infrastructure is significant features of the urban and peri-urban built environment. Social and economic cohesion can be severed when new transport facilities such as highway structures cut across an existing urban community (Han et al., 2002). Arteries or transport terminals can define urban borders and produce segregation. Major transport facilities can affect the quality of urban life (Quishlaqi et al., 2007) by creating physical barriers, increasing noise levels, generating odors, reducing urban aesthetic and affecting the built heritage (Hadjiliadis, 1997). 2.4 Important pollution parameters of water pH All phases of water and waste water treatment and waste quality management are pH dependent. The relative importance of these processes depends on their composition and pH (Pandey, 2014). Cation exchange reactions and complexation to organic matter are important in water, while specific adsorption and precipitation become more important at near-neutral to alkaline pH values in soil (Rout and Shaw, 2001). Under acidic and reducing conditions, most heavy metal salts are most likely to control its concentration in the soil solution (Aschman and Zasoski, 1987; Hausinger, 1997). 19 Temperature Temperature is an important indicator of water quality with regards to survival of organisms. The water temperature depends on the climatic conditions and activities in the water (Khan and Noor, 2002). Temperature affects the rate of all chemical reactions in the water and biochemical process of living beings (Leonard et al., 2004; Raffo et al., 2006). Electrical conductivity Conductivity varies both with number and types of ions in the solution, which in turn is related to the concentration of ionized substances in the water. Most dissolved inorganic substances in water are in the ionized form and hence contribute to conductance. Conductivity is a capacity of water to carry an electrical current and varies both with the number and type of ions and solution contains. Electrical conductivity is a function of total dissolved solids (TDS) known as ionic concentration, which determines the quality and suitability of water use for different purposes (Hem, 1989). Total solid Different forms of solids are defined on the basis of method applied for their determination. The term ‘solid’ refers to the matter either filterable or non-filtrable than remains as residue upon evaporation and subsequent drying at a defined temperature. Further categorization depends upon the temperature employed for drying the ignition. Solids may affect water or effluent quality adversely in number of ways (Pandey et al., 2011). Water with high dissolved solids may induce an 20 unfavourable physiological reaction in the transient consumer and generally are of inferior palatability (APHA, 2005). Residue left after the evaporation and subsequent drying in oven at specific temperature 103-105oC of a known volume of sample are total solids. Total solids include “total suspended solids” (TSS) and “total dissolved solids” (TDS) (APHA, 2005). Solids may affect water or effluent quality adversely in number of ways. High total dissolved solids indicate higher level of cations and anions (Han et al., 2002). High solids (suspended and dissolved) content in water cause salinity in water and soil which adversely affect various biological activities of aquatic life and growth of plants (Sharma et al., 2007). Hardness Water hardness is a traditional measure of the capacity of water to precipitate soap. Hardness of water is a variable (Pandey and Nautiyal, 2008) and complex mixture of cations and anions. It is caused by dissolved polyvalent metallic ions (Nagajyothi et al., 2009). Hardness is determined by the EDTA method in alkaline condition; EDTA and its sodium salts form a soluble chelated complex with certain metal ions. Calcium and magnesium ions develop wine red colour with eriochrome black T in aqueous solution. At this pH murexide (ammonium purpurate) indicator forms a pink colour with Ca++ ions remain in solution. When EDTA is added Ca++ gets complexed resulting in a change from pink to purple which indicates end point of the reaction (APHA, 2005). 21 Chloride The presence of chloride in natural waters can be attributed to dissolution of salt deposits, run-off water from agricultural field and discharges of effluents from chemical industries (Pandey, 2006). Each of these sources may result in local contamination of both surface and ground water (Barman et al., 2000). The salty taste produced by chloride depends on the chemical composition of the water (Chindah et al., 2004). A concentration of 250 ml/L may be detectable in some waters containing sodium ions. On the other hand, the typical salty taste may be absent in water containing 1000 mg/L chloride when calcium and magnesium ions are predominant. High chloride content may harm agricultural plants (Ahmed et al., 2006). Heavy metals Metals are defined as any element that has a silvery luster and is a good conductor of heat and electricity (Doncheva et al., 2001; Du Laing et al., 2009). The metals are classified as “heavy metals” if, in their standard state, they have a specific gravity of more than 5 g/cm3. There are sixty known heavy metals. Heavy metals can accumulate over time in soils and plants (Gautam and Pandey, 2008; Singh and Pandey, 2011) and could have a negative influence on physiological activities of plants (e.g. photosynthesis, gaseous exchange, and nutrient absorption), causing reductions in plant growth, dry matter accumulation and yield (Devkota and Schmidt, 2000). 2.5 Environmental effects near expresshighways High pollution of natural resources like soil and water (Singh and Pandey, 2011) and plant growth (Naaz and Pandey, 2009) affected due to the buildup of 22 soluble salts, sodium and heavy metals (Abida et al., 2009) near highways, have been reported. As trace elements, some heavy metals (copper, iron, zinc etc.) are essential to maintain the metabolism of the human body. However, at higher concentrations they can lead to poisoning (MacFarlane et al., 2003). Heavy metal poisoning could result, for instance, from drinking water contamination, high ambient air concentrations near emission sources, or intake via the food chain (Kumar and Pandey, 2010; Adeyeye, 2005). Heavy metals in the atmosphere, soil and water, even in traces can cause serious problems to all living organisms (Kannaujiya and Pandey, 2013), and their bioaccumulation in the food chain especially can be highly dangerous to human health (Afyoni et al., 1998; Pilon-Smits, 2005). The entry of pollutants in the environment and their adverse effects on living beings become uncontrolled due to extensive vegetation loss. 2.6 Effect of metals in plants Heavy metals accumulate in living organisms, circulate in the trophic chain and moreover their dangerous concentrations persist in ecosystems for a long time (Tiller, 1989; Pandey and Sharma, 2002; Pandey and Pathak, 2006; Tlustoš et al., 2006). For soil-plant system, heavy metal toxicity threshold is the highest permissible content in the soil (total or bioavailable concentration) that does not pose any phytotoxic effects or heavy metals in the edible parts of the crops does not exceed food hygiene standards (Adema and Henzen, 1989). Plant roots participate primarily in the heavy metal cation uptake (Lasat, 2002). The elevated concentration of heavy metals cause disturbances in cell membrane functioning in the photosynthetic and mitochondrial electron transport and in the inactivation of many enzymes active in the basic cell metabolism regulation, which as the result leads to diminishing energy 23 balance and disturbances in cell mineral nutrition (Kosobrukhov et al., 2004). All these changes in the plants leads to limited growth of roots (Gautam and Pandey, 2008), which are in direct contact with toxic substances in soil, limits adsorptive and conductive functions of these organs and results in limited growth of the top parts (Hara and Sonoda, 1979; Pandey et al., 2008, 2009). The accumulation of heavy metals in vascular plants provokes significant biochemical and physiological responses (Meenakshi and Pandey, 2009), modifying several metabolic processes (Vangronsveld and Clijsters, 1994; Macfarlane et al., 2003). Plants that grow in environments contaminated with traces of metals show strategies of escape or tolerance to metal toxicity that have been selected during evolution (Patra et al., 2004). Several plants species have developed tolerance to metals (Hall, 2002). Interaction between heavy metals and plants are based upon either heavy metals extraction of exclusion by plants (Keskinkan et al., 2004). 2.7 Occurrence of heavy metals in soil In some natural soils developed from metal rich parent materials, as well as in contaminated soils, upto 30 to 60% of heavy metals can occur in easily metals can occur in easily unstable forms (Kosobrukhov et al., 2004). In soil, metals are found in one or more of several “pools” of the soil, such as, metals are either dissolved in the soil solution or occupying exchange sites in inorganic soil constituents; associated with insoluble soil organic matter; present in the structure of secondary minerals; and/or present in the structure of primary minerals (Sims and Sklin, 1991). Natural and anthropogenically introduced concentrations of metals in near-surface soil can vary significantly due to different physical and chemical processes operating within soils across geographic regions (Sukreeyapongse et al., 2002). Migration of metals in 24 the soil is influenced by physical and chemical characteristics of each specific metal (Gautam and Pandey, 2013) and by several environmental factors. The most significant environmental factors appear to be (i) soil type, (ii) total organic content, (iii) redox potential, and (iv) pH (Sukreeyapongse et al., 2002). Although, heavy metals are generally considered to be relatively immobile in most soils, their mobility in certain contaminated soils may exceed ordinary rates and pose a significant threat to water quality (Ukpong and Moses, 2001). The elevated concentrations of heavy metals in runoff contaminate surface water bodies (Mehta and Gaur, 1999). Metal constituents of surface soil directly influence the movement of metals, especially in sandy soils towards the ground water (Moreno et al., 1994). Heavy metals influence ground and surface waters (Chandra et al., 2004; Boukhalfa, 2007), flora (Gautam and Pandey, 2008; Sen and Mukherjee, 2009), animals and humans (De Vries et al., 2007). The overall behaviour of heavy metals in soil is to be govern largely by their sorption and desorption reactions with different soil constituents, especially clay components (Appel and Ma, 2002). The chemical behaviour of heavy metals in soils is controlled by a number of processes, including metal cation release from contamination source materials (e.g., fertilizer, sludge), cation exchange and specific adsorption onto surfaces of minerals and soil organic matter (Malkowski et al., 2002). Oxidizing conditions generally increase the retention capacity of metals in soil while reducing conditions will generally reduce the retention capacity of metals (Ryan et al., 2008). Soil reduction has been shown to result in the coincident release of metals associated with minerals that are susceptible to reductive dissolution (Charlatchka and Cambier, 2000; Davranche and Bollinger, 2000). Toxic heavy metals and micronutrients utilized as metal ions exist in the soil as species with several types of mobility and take part in many interactions (Dowling and Doty, 25 2009). Depending on their solubility, heavy metals may eventually become associated to suspended particulate matter or accumulate in the bottom sediments (Alva et al., 2000; Pandey and Srivastava, 2002). The soil reaction (pH) is one of the most important factors in the control of the concentration of these metals in the soil solution (Sharma et al., 2005). Esakku et al. (2005) proposed that under acidic conditions the phenomenon of adsorption is more important in the control of metal bioavailability, while precipitation reactions and complexation have greater influence under neutral and alkaline conditions. As a general rule, the formation of complexes is favored at pH values next to neutrality, because, under acidic conditions the legends are protonated, whereas under alkaline conditions the metals can precipitate in the form of hydroxides (Peralta et al., 2001). However, metals have different soil behaviours. Therefore, soil conditions play very important role to metals availability, their absorption and effectiveness of the toxicity through altering biochemical constituents. 26 Table 2.3: Range of tissue accumulation of heavy metals in some plants. Heavy metals Nickel Range of tissue Responses accumulation 2.7-11.0 Puckering in old leaves 7.8.29.3 Chlorosis in young leaves 0.5-1.0 Normal growth Plants References Lettuce Beavington (1975) Sahu et al. (2007) 10-100 Toxicity symptoms Crops Chlorosis in young leaves Necrosis in older leaves Normal growth Vegetables Chromium 0.02-14.0 2.47-6.83 0.1-0.5 5-30 0.014-0.2 Zinc 32.5-78.80 20-100 Copper Vegetables Crops Plants Toxicity symptoms in Mustard test plants Smaller size Wheat grain Puckering in old leaves Normal growth in plants Spinach Crop plant 100-400 Toxicity symptoms in Tomato test plants 52-103.3 Intervenal chlorosis 2.8-21.8 Chlorosis in young Coriander leaves Browning of leaf tips Vegetables 9-93 5-20 20-30 Gourd Normal growth in Wheat plants Toxicity symptoms in Cabbage test plants Kabata-Pendias and Pendias (1991) Kabata-Pendias and Pendias (1991) Peterson (1983) Panda and Patra (2004) Kabata-Pendias and Pendias (1991) Kabata and Pendias (1991) Ouzounidou (1994) Barman et al. (2000) Kabata-Pendias and Pendias (1991) Kabata-Pendias and Pendias (1991) Chandra et al. (2004) Alva (2000) Barman et al. (2000) Cobb et al. (2000) Fakayode and Onianwa (2002) Bioaccumulation and bio-magnification are responsible for transforming concentrations considered normal into toxic concentrations for different species of 27 biota and man (Ghoreishi and Haghighi, 2003; Gomes and Asaeda, 2009). Once the heavy metal is mobilized in the environment, its total amount remains the same, whatever its form, whether ion, complex or precipitate (Khan and Patra, 2007). The assimilation of trace elements by plants varies greatly as a function of soil conditions (Albasel and Cottenie, 1985). Toxic metallic ions penetrate cells using the same absorption processes of essential micronutrient ions. The quantity absorbed by the plant depends on the concentration and speciation of the metal in the soil solution, together with its successive movement from the soil to the root surface and from the root to the aerial part (Nath et al., 2009). The translocation of these metallic ions to the aerial parts depends on the plant species, the metal involved and the environmental conditions (Liao et al., 2000). Their effects depend on the oxidation state of the metal, their concentration and the duration of exposition, and these are more pronounced at high concentrations (Alonso et al., 2006). Pollution with heavy metals depends on the properties of soil and on economic activities (Meenakshi and Pandey, 2009). The content of heavy metals in plants depends on their concentration and migration in soil. Their migration in calcareous soil decreases as follows: Zn > Cd > Pb > Cu (Alumaa et al., 2002). Heavy metals at elevated concentration are known to effect soil microbial population and their associated activities, which may directly influence the soil fertility (Smith, 1996; Hadjiliadis, 1997; Gautam and Pandey, 2008). Pollution due to heavy metals place human health at risk (Markert et al., 2008) and it is responsible for several environmental problems, including the decrease of microbial activity, soil fertility and crop yields (Cobb et al., 2000; Nriagu and Pacyna, 1988). 28 2.8 Chromium The particulate matter emitted due to transport activities may contribute metals including Cr into the environment (Kumar and Pandey, 2010). Chromium (Cr) is the seventh most abundant metal in the earth’s crust (Yu et al., 2004) and an important environmental contaminant released into the atmosphere due to its huge industrial use (Easton, 1992). Total amount of chromium release into the atmosphere from natural sources in 43,000 tonnes / year, compared with an estimated anthropogenic load of 30,400 tonnes/year (Kotas and Stasicka, 2000). In nature, Cr exists in two different stable oxidation states; trivalent (Cr3+) and hexavalent (Cr6+) chromium. Both Cr3+ and Cr6+ differ in terms of mobility, bioavailability and toxicity. Cr6+ is found to be more toxic than Cr3+ (Panda and Chaudhary, 2004). Both oxidized forms, however, have the capacity to form complexes with other species (Barlett and James, 1979). Chromium toxicity in plants Chromium is toxic to plants and does not play any role in plant metabolism (Pandey et al., 2005). Accumulation of elevated Cr concentration by plants can reduce growth, induce chlorosis in young leaves, reduce pigment content, alter enzymatic function, damage root cells and cause ultrastructural modifications of the chloroplast and cell membrane (Paivoke, 2002; Morales et al., 2007; Zhang et al., 2007). During seed germination, hydrolysis of proteins and starch takes place, providing amino acids and sugars (Zied, 2001). Cr can affect roots of plants causing wilting and plasmolysis in root cells (McGrath, 1982; Zayed et al., 1998a). Chromium can also inhibit the Hill reaction, affecting both the dark and light reaction (Zied, 2001). ROS that are common consequences of most biotic and abiotic 29 stresses are also formed as a result Cr toxicity (Panda et al., 2003). In many crop plants like rice, wheat, and pea significant increase in ROS production with concomitant increases in lipid peroxidation (Demirezen and Aksoy, 2004; Panda and Patra, 2004) have been observed. High production of H2O2 and O2– radicals in plant species exposed to Cr and the metal has been implicated in the generation of oxidative stress (Kannan et al., 2005; Pandey et al., 2009b). Chromium can degrade proteins (Dan et al., 2002). The amino acid cysteine in an important component of phytochelatins (Vajpayee et al., 2001). In aquatic environments, chromium present predominantly in a soluble form (USEPA, 1998; Rout et al., 2000). Chromium have been shown to accumulate in many aquatic species, especially in bottom-feeding fish, such as the brown bullhead (Ictalujrus nebulosus); and in bivalves, such as the oyster (Crassostrea virginica), the blue mussel (Mytilus edulis) and the soft shell clam (Mya arenaria) (Scoccianti et al., 2006; Seng and Bielefeldt, 2002). In soil, chromium is highly unstable and mobile, since it is poorly adsorbed onto soils under natural conditions (Pandey et al., 2008b; Sen and Mukherjee, 2009). Effects of chromium on human health Chromium compounds are corrosive, and allergic skin reactions readily occur following exposure, independent of dose. Short-term exposure to high levels results in ulceration of exposed skin and irritation of the gastrointestinal tract (Horvath et al., 2008; Armienta et al., 2001). Chromium often accumulates in aquatic life, adding to the danger of eating fish that may have been exposed to high levels of chromium (Cifuentes et al., 1996; Epniewska and Bucior, 2001). Chromium is widely distributed 30 in soil, water and biological material and occurs in the range of 5 to 1000 ppm in soils (Crawford, 1999; Devkota and Schmidt, 2000). Chromium is connected with the glucose tolerance factor (Garg and Chandra, 1990) and is important in animal and human nutrition for normal carbohydrate metabolism (Khan, 2007; Kimbrough, 1999). Long-term occupational exposure to airborne levels of chromium higher than the natural environment leads to lung cancer (Akinola et al., 2008; Kumar and Pandey, 2010). Table 2.4: Chromium concentration in growth medium and its uptake in some plants. Sl. Cr concentration No. 1. 0, 30 mg kg-1 Cr(III) and Cr(VI) 4. 50, 100 and 200 AM Cr(VI) 5. 6, 12 and 24 mg L-1 Cr Uptake and accumulation 2.8 Cr(III) and 3.14 Cr(VI) mg kg-1 Progressive increae with more Cr in roots than shoots Cr more in roots than shoots in A and more in shoots than roots in B References Spinach Singh (2001) Nelumbo nucifera Vajpayee et al. (2001) A. Dactylis glomerate B. Medicago sativa Cr shoot: 44 mg kg-1 dw Smart weed 1 mg L-1 for 10 days 7. 0, 50 and 100 mg L- Roots took up more than 1 Cr(III) shoots and not detected in fruits 8. 0-200 mg kg-1 Progressive increase with more Cr in roots than shoots 9. 0, 5, 50, 150 and 70-90% accumulation in 300 6, 12, 24 Cr roots -1 mL 10. Tannery effluent High Cr removal from 5%, 10% and 15% 10% and 15% 6. Crop/ plant 11. 0 and 8 mg L-1 Cr 6700 mg kg-1 in roots 12. 0, 100, 500 Cr(VI) and Cr(VI) 2.4 mg kg-1 shoot and 115.6 mg kg-1 in root sorghum A; 5.8 mg kg-1 shoot and 212 mg kg-1 in root in Sunflower 31 Tomato Shanker (2005) Mapanda et al. (2005) Morales et al. (2007) Sunflower, Mishra and maize and Vicia Tripathi (2009) faba Allium cepa Rai et al. (1995) Swiss chard Veronica beccabanga Sorghum and Sunflower Olayinka Nwachukwu (2008) Sinha et al. (2005) SinMishra et al. (2009) 2.9 Nickel Transportation activities may also contribute Ni contamination in the environment. Nickel (Ni) is a silvery-white, hard metal. Although it exists in several oxidation states, the divalent ion seems to be the most important for both organic and inorganic substances, but the trivalent form may be generated by redox reactions in the cell (Easton, 1992; APHA, 2005). Water-insoluble nickel compounds may dissolved in biological fluids. Particles of the same chemical entity (oxides and sulfides) have different biological activity depending on crystalline structure and surface properties (Eskew et al., 1984; Schutzendubel and Polle, 2002; Seregin and Kozhevnikova, 2005). Soil and water contamination with Ni has become a worldwide problem (Guo and Marschner, 1995). Ni is essential for plants (Brown et al., 1987; Salt et al., 1995), but the concentration in the majority of plant species is very low (0.05-10 mg kg-1 dw) (Demirezen et al., 2007). TheNi pollution, excess Ni rather than a deficiency, is more commonly found in plants (Ragsdale, 1998). Toxic effects of high concentrations of Ni includes inhibition of mitotic activities (Madhavrao and Shresty, 2000), reductions in plant growth and alter biochemical constituents such as chlorophylls and protein (Molas, 2002) and adverse effects on fruit yield and quality (Gajewska et al., 2006). Extremely high soil Ni concentrations have left some farmland unsuitable for growing crops, fruits and vegetables (Duarte et al., 2007). Nickel in plants The high Ni concentrations may turn toxic to plants (Singh and Pandey, 2011; Kao et al., 2008). There are approximately 70 species of plants that accumulate 32 extraordinarily high nickel concentrations. This may be upto 10,000 ppm (dry mass) (Singh and Pandey, 2011). For regular plant seed 0.5-2 ppm nickel in liquid substrates is considered toxic (Guo and Marschner, 1995). Liming of the soil may rapidly decrease nickel uptake. Sludge containing more than 200 ppm nickel (dry mass) is not be applicable to agricultural soils (Smejkalova et al., 2003). Effects of nickel on human health Nickel contamination due to transport activities may pose health hazards of living organisms inhabiting nearby areas of road. Traces of nickel are needed by the human body to produce red blood cells, however, in excessive amounts, can become mildly toxic. Short term overexposure to nickel is not known to cause any health problems, but long-term exposure can cause decreased body weight, heart and liver damage, and skin irritation (Kao et al., 2008). Nickel can accumulate in aquatic life, but its presence is not magnified along food chains. The route by which nickel enters the body is very important in assessing its impact on health. Inhalation of nickel can result in chronic bronchitis, emphysema, and asthma and lung cancer. By ingestion, nickel has been associated with reduced body weight and reproductive and foetotoxic effects (Mulrooney and Hausinger, 2003). 33 Table 2.5: Tissue accumulation of heavy metals by plants exposed to contaminated growth medium. Heavy Crop Tissue accumulation of heavy References metal metal (mg kg-1) in harvestable plant Min. – Max. Cu Wheat 3.8 – 6.2 Gerritse and Van Driel et al. (1984) Maize 1.9 – 7.0 Grass 6.4 – 21.5 Zn Wheat 33 – 94 Maize 28 – 174 Grass 0.1 – 4.3 Cu Maize, radish, 29.2 – 137.9 Groenenberg et al. Brassica napus (2003) Wheat, coriander 20.2 – 13.7 irrigated with industrial effluent in field (n=40) Zn Spinach, brinjal 23.6-34.2 Midrarul Haq et al. (2005) Cu Maize, radish, 14.7 – 18.6 Naaz (2012) Brassica napus Cr Linseed 1 – 4.2 Meenakshi and Pandey (2009) Ni Wheat, coriander 10 – 16 Gautam and Pandey irrigated with (2013) tubewell water (n=40) 2.10 Copper Copper is the first element in Group 1B in periodic table, it has an atomic number of 29, atomic weight of 63.54 and valences of 1 and 2. The average abundance of copper in the earth crust in 68 ppm, in soils, it is 9-33 ppm, in streams it is 4-12 g/L; and in ground water it is <0.1 mg/l (APHA, 2005). Copper is considered as essential trace element for plants and animals. The toxicity of Cu is mainly observe in acid sandy soils with low cation exchange capacities (Xiong et al., 2006). Due to anthropogenic and geogenic factors including transport activities, soils accumulate upto 3,200 mg kg-1 of copper in the top layer, a quantity several times higher in vine-growing areas throughout the world 34 (Wallace and Cha, 1999). Copper is an essential micronutrient for plant growth. Most soils contain adequate amounts of this nutrient for optimum crop yields (Yruela, 2005). Copper acts as a structural element in regulatory proteins and participates in photosynthetic electron transport, mitochondrial respiration, oxidative stress responses, cell wall metabolism and hormone signaling (Marschner, 1995). Cu ions act as cofactors in many enzymes such as Cu/Zn superoxide dismutase (SOD), cytochrome c oxidase, and amino oxidase. Thus, plants require Cu as an essential micronutrient for normal growth and development; when this ion is not available plants develop specific deficiency symptoms, most of which effect young leaves and reproductive organs (Sharma, 2006). Copper toxicity in plants Copper at excess levels can inhibit root elongation, block and photosynthetic electron transporter chain and degrade chlorophyll (Zyadah and Bakky, 2000). At high concentration, Cu can become extremely toxic causing symptoms such as chlorosis and necrosis, stunting of plant growth, leaf discoloration and inhibition of root growth have been reported (Chen and Kao, 1999; Marschner, 1995). At the cellular level, toxicity may result from: (i) binding to sulfhydryl groups in proteins, thereby inhibiting enzyme activity or protein function; (ii) induction of a deficiency of other essential ions; (iii) impaired cell transport processes; (iv) oxidative damage (Chen and Kao, 1999). At concentrations above required for optimal growth, Cu inhibit growth and interfere with important cellular processes such as photosynthesis and respiration (Marschner, 1995). At excess copper levels in plants, lower content of chlorophyll and alterations of chloroplast structure and thylakoids membrane composition have also been reported (Lidon and Henriques, 1991). For most of the 35 crop species, the critical toxicity level of copper in the leaves is 20 to 30 mg g-1 dr. wt. (Quartacci et al., 2000). Copper in soil The amount of Cu available to plants varies widely from soil to soil. Available Cu can vary from 1 to 200 ppm (parts per million) in both mineral and organic soils as a function of soil pH and soil texture. The finer-textured mineral soils generally contain the highest amounts of Cu (Marschner, 1995). The lowest concentration of Cu is associated with the organic or peat soils. Availability of Cu is related to soil pH (Bunzl et al., 2001). As soil pH increase, the availability of this nutrient decreases (Pandey, 2006a). Copper is not mobile in soils (Sharma, 2006). It is attracted to soil organic matter and clay minerals (Weyens et al., 2009). The amount of available Cu is measured by extracting the soil with a DTPA solution (Lindsay and Norwell, 1978). Effects of copper on human health Copper is an essential substance to human life, but in high doses it can cause anemia, liver and kidney damage, and stomach and intestinal irritation. People with Wilson’s disease are at greater risk for health effects from overexposure to copper (Fritioff and Greger, 2006). 2.11 Zinc Various anthropological activities contaminate the environment with high concentration of Zn. Zinc is a lustrous bluish-white metal. It is found in group IIb of the periodic table. It is brittle and crystalline at ordinary temperatures, but it becomes ductile and malleable when heated between 110oC and 150oC. It is a fairly reactive 36 metal than will combine with oxygen and other non-metals, and react with dilute acids to release hydrogen (APHA, 2005). Zinc is the 23rd most abundant element in the Earth’s crust. The dominant ore is zinc blende, also known as sphalerite. World production exceeds 7 million tonnes a year and commercially exploitable reserves exceed 100 million tones. More than 30% of the world’s need for xinc is met by recycling (Cezary and Singh, 2001). Effects on plants Zn is an essential macronutrient for plant growth but pose phytotoxic effects when in excess in growth medium. Phytotoxicity of Zn cause decrease in crop yield and quality and transfer into the food chain (Adriano, 2001). Zinc is an essential element for both plants and animals (Carroll and Lonearagan, 1968). It plays an important role in several plant metabolic processes; it activates several enzymes and is involved in protein synthesis and carbohydrate, nucleic acid and lipid metabolism (Cakmak and Marshner, 1993). However, like other heavy metals (Doncheva et al., 2001) when Zn is accumulated in excess in plant tissues, it causes alterations in vital growth processes such as photosynthesis and chlorophyll biosynthesis and membrane integrity (Doncheva et al., 2001). An excess of Zn have a negative effect on mineral nutrition (Baccouch et al., 1998a,b). Toxic levels of Zn for different varieties of crop have very wide limits in growth medium from 64 g L-1 Zn for sorghum to 2000 g L-1 Zn for cotton (Otte et al., 1995). To counteract this methabolic dysfunction caused by Zn toxicity stress, higher plants employ defense strategies. To protect themselves from heavy metal, plant cells must develop a mechanism by which the metal ion, entering the cytosol of the cell, is immediately complexed and inactivated 37 (Vysloužilová et al., 2003). This protection process is mediated by phytochelatins (Vogel-Mikus et al., 2005) and organic acids (Verma and Pandey, 2008). Zinc in the environment Zinc is a very common substance that occurs naturally. Many foodstuffs contain certain concentrations of Zn. Drinking water also contains zinc, which may be high when it is stored in metal tanks. Industrial sources or toxic waste sites may cause the Zn amounts in drinking water to reach levels that can cause health problems (Chapman, 1966; Deng et al., 2004). Zinc occurs naturally in air, water and soil, but zinc concentrations are rising unnaturally, due to addition of Zn through human activities (Naaz and Pandey, 2009), such as mining, waste combustion and effluent discharge (Pandey, 2006a,b; Pandey and Nautiyal, 2008). Some soils are heavily contaminated with zinc, and these are to be found in areas where zinc has to be mined or refined, or were sewage sludge from industrial areas has been used as fertilizer Bunzl et al., 2001). Effect of zinc on human health Zinc is a trace element that is essential for human health (Boon and Soltampour, 1992). When people absorb too little zinc they can experience a loss of appetite, decreased sense of taste and smell, slow wound healing and skin sores (Catlett et al., 2002). Zinc-shortages can even cause birth defects. Although humans can handle proportionally large concentrations of zinc, too much zinc can still cause eminent health problems, such as stomach cramps, skin irritations, vomiting, nausea and anaemia (Vousta et al., 1996). Very high levels of zinc can damage the pancreas and disturb the protein metabolism, and cause arteriosclerosis. Extensive exposure of 38 zinc chloride can cause respiratory disorders (Vogel et al., 2005). Water is polluted with zinc, due to the presence of large quantities of zinc in the effluent of industrial plants (Pandey, 2004). One of the consequences is that rivers are depositing zincpolluted sludge on their banks. Plants often have a zinc uptake that their systems cannot handle, due to the accumulation of zinc in soils and few plants could tolerate that level (Verma and Pandey, 2008). Thus, plant diversity deteriorates near zincdisposing factories (Pandey and Nautiyal, 2008). Zinc can interrupt the activity in soils; negatively influence the activity of microorganisms and earthworms (Marschner, 2003). 2.12 Biochemical constituents Chlorophyll Chlorophyll is the most ubiquitous of all natural pigments, reaching levels that can exceed 1000 to 2000 ppm wet weight in some species, and is responsible for the color of all green plants. considering the primary role of chlorophyll in photosynthesis and its close association with yellow/ orange carotenoid pigments well known for their bioactivity, these blue-green pigments have potential physiological impact (Poskuta et al., 1996). Structurally, chlorophyll is a substituted tetrapyrole with a centrally bound magnesium atom (Fig. 2.2). A chlorophyll molecule is a typical porphyrin derivative possessing a cyclic tetrapyrolic structure in which one pyrole ring is partially reduced. the tetrapyrolic nucleus contains a non ionic magnesium atom held by two covalent and two coordinated bonds. In addition to four pyrole rings, a fifth isocyclic ring is also present (Chandra et al., 2004). Both acid side chains are esterified, one as methyl ester and other as phytol ester. The presence of a long phytol tail along with the flat poryphyrin head gives the molecule an appearance 39 common to that of spatula. In chlorophyll b the methyl group at position three of second pyrole ring of chlorophyll a is replaced by formyl group (Gajewska et al., 2006). The chlorophyll content of commonly consumed green vegetables typically exceeds the levels of other bioactive pigments (Nagajyothi et al., 2009) such as carotenoids, by upto a 5-fold margin (Table 1). This relatively high concentration makes chlorophyll a significant contributor to the total dietary photochemical pool. The sensitivity of natural chlorophylls to extremes in pH and temperature allows for the formation of several distinct derivatives through processing of vegetable tissue and human digestion (Porra et al., 1989). Degradation of pigments has widely been used as an indicator of pollution. Chlorophyll, the green pigment is one of the main complex which influences photosynthesis. Decreases in chlorophyll content under stress (pollution or temperature stress) may be attributed to either its degradation or to reduced biosynthesis (Schutzendubel and Polle, 2002). Table 2.6: Representative chlorophyll and carotenoids content of common green vegetables. Fruit/Vegetable tissue Total chlorophyll content (mg g-1 fresh tissue) Total carotenoid content (mg g-1 fresh tissue) Green beans 52 8.6 Broccoli 79 42 Kale 1870 776 Peas 134 34 Spinach 1250 364 Nagajyothi et al. (2009), Poskuta et al. (1996), Singh et al. (2006). 40 Carotenoids Like chlorophyll, carotenoids content occur in their natural state as protein complexes. The ubiquitous presence of carotenoids in the photosynthetic tissue suggests a fundamental role in the photosynthetic process (Kenneth et al., 2000). Major role of carotenoids are protection against photodynamic destruction catalyzed by chlorophyll absorption and transfer of light energy to chlorophyll a, act as scavengers of free radicals and represent the antioxidative system of plants. Carotenoids belong to a large group of compounds called terpenoids. These compounds produce red orange, yellow and brown color in plants. They are further divided on the basis of presence and absence of oxygen into carotenes, which have formula C40H56, contain only C and H and xanthophylls contain oxygen along with C and H; common xanthophyll of leaves is lutein (C40H56O2). Over 600 carotenoids occurring in plants, fungi, bacteria and animal, including humans, are present (Kenneth et al., 2000). Carotenoids content involve in photo protective functions in photosynthesis (Kenneth et al., 2000). Under low light conditions, carotenoids may act as energetic antennae, harvesting light at the wavelengths not absorbed by chlorophylls and transferring electron excitation states towards photochemical reaction centers. In this way, they widen the range of light used in photosynthesis (Markert et al., 2008). Carotenoid pigments also have ecological significance. Marking flowers and fruits colored, they play an important role in ecosystems, attracting pollen-dispersing insects and fruit-eating animals (Pandey et al., 2005). In humans, carotenoids normally occur in several types of tissues, e.g., muscles, liver, eye, blood and adipose tissue (Singh and Pandey, 2011). Currently, about 25 carotenoids and their 41 metabolites have been found in serum. Carotenoid content of plants varies from species to species. Plants with high baseline value of carotenoids show that they are better equipped with antioxidative system (Raffo et al., 2006). Protein Protein is the basic constituent of the cell. These are complex substances of high molecular weight ranging up to several millions and contain nitrogen in addition to carbon, hydrogen and oxygen. Sometimes elements like phosphorus, sulphur, iron, zinc and iodine may also be present (Pandey et al., 2009a). However, elementary composition of most proteins is very similar (approximate percentages are C=50-55, H=6-8, O=20-23, N=15-18, and S=0-4). Proteins are made up of several nitrogen containing organic molecules called amino acids i.e., proteins are polymerized forms of organic molecules called amino acids. Thus amino acid is the basic unit of protein. Protein dissociates to form amino acids and the energy produced is utilized for routine metabolic activities (Chandra et al., 2004). Proteins are the most important constituent of plant cells both from structural as well as functional point of view. Functionally, give rise to enzymes, which are responsible for regulating the cellular process (Rodriguez et al., 2007). Many proteins carry out enzymatic activities and are vital for the rapid rate of biochemical reactions in the cell. Proteins are hydrogen ion buffers and structural component of cell. Heavy metals contamination reduces protein content in plants have been reported by several workers (Pandey et al., 2009; Sharma, 2006). 2.13 Some important parameters for Environmental Impact Assessment: The transport activities contribute various pollutants such as gases (Kumar and Pandey, 2010), heavy metals (Singh and Pandey, 2011; Pandey, 2008) and various 42 chemicals (Fakayode, 2005) in to the environment. The construction work of highways develop uneven areas cause water erosion, which make soil unfertile. Due to the loss of vegetation due construction of expresshighways affect local climate change (Abida et al., 2009). Heavy metals and pollutants emitted from transportation activities entered into food web and damage the ecosystem. The polluted environment due to transportation activities may cause loss of biodiversity, change the conditions of soil and water, affect the growth and metabolism of plants, ultimately affect the life of human beings, directly or indirectly. 2.14 Study area Study areas selected for the study were located near expresshighway (NH 25) at proposed Ganga expresshighway area in Unnao district of Utter Pradesh state (India). The expresshighway (NH 25) link Lucknow to Kanpur district (80 km distance). At this expresshighway study sites were selected to Environmental Impact Assessment to observe the ecological conditions after long operation of expresshighway (NH 25) for prediction of future ecological conditions after completion of proposed Ganga expressway. 43 Chapter 3 Materials and Methods 3.1 Cleaning of glasswares All glasswares used for analysis including pipettes, conical flasks, burettes, volumetric flaks were soaked overnight in diluted nitric acid (1:1) rinsed properly with deionised water and oven dried prior to analysis. Table 3.1: Locations of study area near Express highway (NH 25) in Unnao district. Sites Locations Distance from highway I Krishnapuram (500 m before Ganga river) 0 - 50 m II Dal Narayanpur 4.5 – 5 km III Sakalpur Nari 0 – 50 m IV Banthar 5 – 5.5 km 3.2 Soil samples collection Soil samples were collected from Unnao district (260 48’N latitude and 800 43’E longitude) from various study sites (I-IV) near expresshighway (NH 25) in proposed Ganga expressway area. The composite soils of Unnao district were collected following the method of Piper (1969) and used for physico-chemical studies. From study areas, surface soil (0-25 cm) was collected in an area of 20 x 20 m having level surface and soil of uniform texture. Before collection of the soil, it was ensured that the selected area had received no manure/fertilizer and does not have any industrial contamination. After scraping top 1 cm soil layer to remove surface vegetation, the areas were dug upto 25 cm depth. The collected composite soil was allowed to dry for a day. The soil was thoroughly mixed, filed in alkathene lined gunny bags and transported to the laboratory. 44 3.2.1 Soil samples preparation The bulk soil samples collected from the different study sites of Unnao district were air dried on alkathene sheet in shade for 2-3 days. After air drying, the soil clods were broken with clean wooden mallet and made free from the plant remains. The soil was thoroughly pulverized and stored in alkathene lined jute gunny bag. Composite soil were analysed for important physico-chemical properties (texture, bulk density, pH, electrical conductance, organic matter and heavy some potentially toxic metals). The methods of analysis are briefly summarized below. 3.3 Soil analysis Soil texture Weighing paper was placed on the pan of the balance and weighed. Data were used to subtract the mass of the paper for all soil measurements arranged the soil sieves so that the largest screen size is on the top (sieve 1), followed by decreasing screen size to the bottom. Balance was set to 100 g plus the mass of weighing paper. Weighed the mass of soil that has been broken up into loose particles. Soil sample was placed into sieve 1 and after shaking weighed, separately. The relative percent of sand, silt, and clay in the soil sample was determined as follows: % Sand = Mass of sand / Total soil mass x 100 % Silt = Mass of silt / Total soil mass x 100 % Clay = Mass of clay / Total soil mass x 100 45 Bulk density A preweighed 50 cc glass bottle was filled with dried soil and the bottle was tapped 15 to 20 times. The weight of the soil was determined by subtracting the weight of the empty bottle. The volume of the bottle in the sample was measured and bulk density was calculated as flows: W2-W1 Bulk density (g cc-1) = V Where: W2 = g weight of empty bottle (50cc) W1 = g weight of bottle + soil V = volume of water filling the bottle pH Pre-weighted oven dried soil sample (10 g) was taken in a clean and dry test tube. A pinch of barium sulphate and 25 ml distilled water (free of CO2) was added (soil : water 1:2.5). Then shaken vigorously for 5 minutes and then for 15 minutes. The test tubes are left as such for 1 hour to allow the soil to settle down. A small amount of clear supernatant was taken in watch glass. Its pH was tested with the pH meter. The electrode of pH-meter was standardized with the pH 4.0, 7.0 and 9.2 buffer solutions. Electrical conductivity The electrical conductivity of water extract of soil from 1:2.5 soil : water suspension was determined with the help of electrical conductivity meter in mS/cm. 46 Organic matter The soil organic matter of the samples was analyzed by the Walkley Black method (Piper, 1969). This method involved wet oxidation by a mixture of 1 N potassium dichromate (K2Cr2O7) solution and concentrated sulfuric acid. After 30 minutes, excess K2Cr2O7 was potentiometrically back-titrated with ferrous ammonium sulfate (FAS) using diphenylamine as indicator. The reduced dichromate produced during reaction with the soil is considered to be equivalent to the total organic carbon content in the soil. 0.003 Organic carbon (%) = {10-(10XS/B)}x 100 x 0.77 W Where: S = ml FAS used for titration of sample B = ml FAS used for titration of blank Organic matter (%) 1.72 4x Organic carbon (%) Calcium and Magnesium Calcium Calcium content was determined by vercenate method described by Piper (1969). Saturated soil extract (Soil and water ratio 1:2.5) was taken (5 ml) in a conical flask and made final volume 25 ml by adding distilled water. Added 5% NaOH (5 drops) into solution and shaken. Added 50 mg ammonium purpurate as an indicator. Colour of the solution turned orange red. The solution titrated against versenate (0.01 M). At end point, solution turned purple colour. 47 Magnesium Saturated soil extract (5 ml) was taken in a conical flask and diluted with distilled water to make final volume 25 ml. Added 10 drops of buffer (NH4Cl+NH4OH) and shaken. About 3-4 drops of Erichrome black-T indicator was added into the solution. Titrated the solution against versenate (0.01M). At the end point, solution turned greenish-blue colour. Calcium content calculated as follows: Versenate used (ml) X Strength of versenate X1000 Ca (meq./100g soil) = ----------------------------------------------------------------Volume of saturated soil extract (ml) Versenate used (ml) X Strength of versenate X 1000 Ca+Mg (meq./100g soil) = -------------------------------------------------------------Volume of saturated soil extract taken (ml) Mg (meq./100 g soil) = (Ca+Mg) – Ca Available phosphorus Available phosphorus content was determined by Olsen’s method (1954) as described by Piper (1969). For extraction of soil 0.5N sodium bicarbonate (pH 8.5) was used as extractant. 2.5g soil was taken in a conical flask with 50 ml 0.5N sodium bicarbonate solution (pH 8.5). After shaking, the solution was filtered with Whatman filter paper No. 1 and collected the filtrate in test tube. Taken 10 ml of filtrate (equivalent to 500 mg soil). For blank, in another test tube 10 ml 0.5 N sodium bicarbonate was taken. In test tubes, added 2-3 drops of para-nitrophenol and shaken the solution. The solution turned yellow. Then, added 5N sulphuric acid drop by drop till the disappearance of yellow colour and counted the number of drops of sulphuric 48 acid for the disappearance of yellow colour. Add the same no. of sulphuric acid in to the test tube having soil sample and shaken. Added 4 ml Murphy Reileye’ reagent and shaken. Final volume was made 25 ml with the help of distilled water. Measured the O.D. of the solution by colorimeter using red filter (660 nm). Compared the O.D. of samples with standard calibration graph of phosphorus. Calcium carbonate Calcium carbonate in soil samples determined by methods as described by Piper (1969). Oven dried 5g soil was taken in 100 ml 0.5 N HCl and after shaking extract was obtained. In a conical flask, taken 20 ml of aliquot, added 1 ml bromothymol blue and titrated the solution with 0.5 N NaOH. At end point, blue colour appeared. Calcium carbonate content calculated as follows: CaCO3 (%) = (Blank titration - Actual titration) X 2.5 Heavy metal analysis (soil) Lindsay Norwell (1978) method was used for determining trace elements in soil. DTPA (dithylenetriaminepentaacetic acid) extracting solution: The DTPA extraction solution was prepared to contain 0.005 M, DTPA, 0.01 M CaCl2, 0.1 M TEA (triethanolamine), and was adjusted to pH 7.30. For preparing 4 liters of this solution 59.68 g of reagent grade TEA, 7.868 of DTPA (diethylenetriamenepentaacetic acid), and 5.88 g of CaCl2.2H2O were dissolved in approximately 200 ml of deionised water. Allowed sufficient time for the DTPA to dissolve, and diluted to approximately 3.5 liters. Adjusted the pH to 7.30±0.05 with 1 N HCl, diluted to 4 liters and mixed well. This solution was stable for several months. 49 Procedure Air dried soil (20 g) was weighed and ground to pass a 2 mm sieve (nylon) into a 125 ml Erlenmeyer flask. Added 40 ml of DTPA extracting solution. Covered each flask with stretchable parafilm and secured uprights on a reciprocating shaker. Shaken at a speed of about 176 cycles/ minute for 2 hours. Filtered by gravity through Whatman filter paper No. 42. Analysed the filtrates for Zn, Cu, Ni, Cr and Cd by atomic absorption spectrophotometer (Perkin Elmer Aanalyst 700). Carbonate and Bicarbonate ions Taken 10 ml of saturated soil extract (Soil and Water 1:2:5 ratio) in conical flask. Added 5 drops of phenolphthalein indicator and shaken (appearance of pink colour indicated presence of carbonate). Solution was titrated with 0.1N sulphuric acid till the solution becomes colourless, note the burette reading (A) Add 5 drops of methyl red indicator in the above conical flask for determination of bicarbonate and shaken. The solution was titrated with 0.1N sulphuric acid till the colour changed from yellow to rose red. Recorded the burette reading (B). 2A X Normality of H2SO4 - CO3 (meq/l) = X 1000 Volume of water sample taken (ml) B-2A X Normality of H2SO4 - HCO3 (meq/l) = X 1000 Volume of water sample taken (ml) Where: A = The burette reading for carbonate using phenolphthaline indicator. B = The burette reading for bicarbonate using methyl red indicator. 50 3.4 Analysis of water samples The surface and ground water samples were collected from four study sites and analysed followed the standard methods (APHA, 2005). The characterization of water was accomplished by laboratory testing of pH, conductivity, hardness, chloride, solids carbonate, bicarbonate, calcium and magnesium and some potentially toxic heavy metals (Cu, Cr, Ni, Fe, Cd and Zn). pH pH was measured by immersing the electrode of pH meter (Systronics) in the sample after calibrating the instrument with standard with pH solutions of 4.0, 7.0 and 9.2, respectively. Conductivity Conductivity was measured by immersing the electrode of electrical conductivity meter (systronics) in the sample and the result expressed as µS/cm. Total solids Known volume of water sample in a beaker ignited to constant weight (W 1). Evaporated the sample to dryness at 103-1050C for 24 hours for total solid estimation. Beaker was cooled in desiccators and weighed (W2). The total solids are expressed as: W2-W1 Total solids (mg l-1) = x 1000 Sample (ml) 51 Where: W1= initial weight of beaker W2= final weight of beaker Known volume of water sample in a beaker ignited to constant weight (W 1). Ignited the beaker for 15-20 minutes in oven maintained at 180±50C with filtered known amount of sample for estimation of total dissolved solids. Beaker was cooler in desiccators, in a dry atmosphere and final weight (W3) was estimated. W3-W2 Total dissolved solids (mg l-1) = x 1000 Sample (ml) Where: W1 = initial weight of beaker W3 = final weight of beaker Chloride Procedure Well mixed sample (50 ml) adjusted to pH 7-8 was taken and added 1.0 ml K2CrO4. Titrated with standard AgNO3 solution till AgCrO4 starts precipitating as pale red precipitate. Standardized AgNO3 against standard NaCl. For better accuracy titrated distilled water (50 ml) in the same way to establish reagent blank. A blank of 0.2 to 0.3 ml in usual. -1 – (A – B) x N x 35.45 x 1000 Chloride (mg l ) as Cl = Sample (ml) Where: A= ml AgNO3 required for sample 52 B= ml AgNO3 required for blank N= Normality of AgNO3 used Total hardness Well mixed sample (50 ml) in porcelain dish or conical flask. To this added 12 ml buffer solution followed by 1 ml inhibitor. This was followed by addition of a pinch of Eriochrome black T and titrate with standard EDTA (0.01 M) till wine red colour changes to blue, noted down the volume of EDTA required (A). A reagent blank was also runned simultaneously. Noted the volume of EDTA (B). Calculated volume of EDTA required by sample, C = (A – B). C x D x 1000 Total hardness (mg l-1) as CaCO3 = Sample (ml) Where: C= Volume of EDTA required by sample B= mg CaCO3 equivalent to 1 ml EDTA titrant Carbonate and Bicarbonate ions Carbonate and bicarbonate ions in water samples were determined using phenolphthalein and methyl orange indicators, respectively. When pink colour of water sample with phenolphthalein disappeared, it was an indicative of half of the neutralization of carbonate resulting in the formation of bicarbonate. At this stage methyl red indicator was added and solution titrated with H2SO4 (0.1 N) till solution turned from yellow to rose red indicated value of bicarbonates. 53 Known volume of water sample taken in a conical flask and added 5 drops of phenolphthalein indicator and shaken solution turned pink colour titrated with H2SO4 (0.1 N) till the end point colourless. Noted the burette reading (4 ml). Thereafter, 5 drops of methyl red indicator added to this solution and titrated with 0.1 N H2SO4 till the end point from yellow to rose red colour. Carbonate and bicarbonate ions calculated as follows: 2A X Normality of H2SO4 CO3- (meq/l) = X 1000 Volume of water sample taken (ml) B-2A X Normality of H2SO4 HCO3- (meq/l) = X 1000 Volume of water sample taken (ml) Where: A = The burette reading for carbonate using phenolphthaline indicator. B = The burette reading for bicarbonate using methyl red indicator. Biochemical parameters Chlorophyll Leaves of harvested wild plants collected from different study sites were crushed finely in pestle and mortar in 10 ml cold 80% acetone centrifuged at 1000 rpm of 20 minutes in centrifuged tubes. Later supernatant was transferred to glass tubes and volume was made 10 ml by adding 80% cold acetone. Absorbance of the supernatant for chlorophyll a, chlorophyll b and carotenoids was taken at wavelengths 663.6, 646.6, 510.0 and 480.0 nm, respectively in spectrophotometer (Perkin Elmer 54 Lambda 40, USA). Chlorophyll a and b were calculated on leaf fresh weight basis, according to formula given by Porra et al. (1989) and the results were expressed on fresh weight basis in mg g-1. 12.25 (663.6) – 2.55 (A646.6) Chlorophyll a = xV D x W x 1000 20.31 (646.6) – 4.91 (A663.6) Chlorophyll b = xV D x W x 1000 17.76 (646.6) – 7.34 (A663.6) Total chlorophyll = xV D x W x 1000 Where: D = Distance of light path W = Weight (g) V = Volume (ml) Carotenoids Carotenoids content was calculated on leaf fresh weight basis according to formula: 7.6 (480) – 1.49 (A510) Carotenoids = xV D x W x 1000 55 Where: D = Distance of light path W = Weight (g) V = Volume (ml) Protein Protein content was estimated by method of Lowry et al. (1951). Four reagents were used in this process (Reagent A, B, C, D): Reagent A: Sodium potassium tartrate (2%) Reagent B: Copper sulphate (1%) Reagent C: Alkaline copper solution [Reagent A (50 ml) + Reagent B (1 ml)] Reagent D: Folin Ciocalteu’s phenol reagent. Plant material (250 mg) was crushed in 5 ml of trichloro acetic acid and centrifuged at 10000 rpm for 10 minutes. After decanting the supernatant, pellet was washed with 5 ml of 1 N NaOH twice, again centrifuged in 1 N NaOH and final supernatant was collected. Reagent C (1.0 ml) was added to final supernatant (0.5 ml) and kept for 10-15 minutes at 30oC. Reagent D was added finally and thoroughly mixed. After 30 min absorbance was recorded at 750 nm (Perking Elmer Lambda 40, USA). Bovine serum albumin was used as standard. Heavy metal analysis The concentration of heavy metals in different plant parts (roots and shoots) was determined in oven dried plant parts by Atomic Absorption Spectrophotometer 56 (AAnylist Perkin Elmer 700, USA). Wet digestion of plant samples was carried out in nitric acid: perchloric acid (HNO3 : HClO4) (3:1 v/v) by heating till insepeint dryness. These evaporated samples were diluted, filtered with Whatman No. 42 and finally volume was made upto 25 ml with distilled water and analyzed for heavy metals (Zn, Cu, Ni and Cr) respectively in Atomic Absorption Spectrophotometer (AAnylist Perkin Elmer 700, USA). Concentration of heavy metals in plant tissue is expressed as g g-1 dry weight. Reading (ppm) x Volume (ml) -1 Metal content (g g dw.) = Weight (g) 3.6 Study of vegetation The qualitative and quantitative study of vegetation was carried out near expresshighway (NH 25) in proposed Ganga expressway area in Unnao district. Qualitative study (Phytosociological method) was made by simple listing of the plant species in the year 2009, 2010 and 2011 in pre and post monsoon period. List of the plant species has been given in the chapter 4. The quantitative study was carried out following the quadrate method with respect to density, frequency and abundance of the wild plant species was growing near the study sites (I-IV). Quadrate method The numerical data of plant species in the study sites were calculated to find out density, frequency and abundance of the species following quadrate method as described by Sharma (2012). 57 The procedure was employed to count all the individual plant species on several quadrates of known size (1M X 1m). The sampling unit of quadrate was an area of 1m2. All the wild plant species in the study sites were recognized taxonomically. The sampling area of the quadrate was find out by ‘species-area-curve method’. quadrates were laid down by random sampling procedures. After the record of various plant species in a chart form, the values of density, frequency and abundance were determined as follows: Total number of individuals of the species Density (per m2) = Total number of quadrates studied Number of quadrates in which species has occurred Frequency (%) = x 100 Total number of quadrates studied Total number of individuals of the species in quadrates studied 2 Abundance (per m ) = Total number of quadrates in which species has occurred 3.7 Statistical analysis of data All experiments were conducted in three replicates (n=3) for each parameter. The data presented in the thesis represent mean of three replicates. The data were statistically analysed using student ‘t’ test of significance for all the parameters. 58 Chapter 4 Results Contents Page no. Field observations and vegetational studies. 60 - 76 4.1.1 Express highway (NH 25) at proposed Ganga expressway area in Unnao district. 60 - 61 4.1.2 Proposed Ganga express highway. 61 - 62 4.1.3 Study sites near expresshighway (NH 25) in Unnao district 63 - 63 4.1.4 Soil and water bodies. 63 - 64 4.1.5 Study of flora near express highway (NH 25) in Unnao district. 64 - 65 4.1.6 Quantitative analysis of plants by quadrate method 66 - 75 Biochemical constituents and tissue concentration of heavy metals in wild plant species near expresshighway (NH 25). 76 - 90 4.1: 4.2 4.2.1 Tissue concentration of heavy metals. 4.2.2 Biochemical constituents in plants. 4.3 Physico-chemical properties of soils at various sites of express highway (NH 25) at proposed Ganga expressway area in Unnao districts (U.P. state). 91 - 94 4.4 Analysis of surface and ground waters qualities at various locations of expresshighway (NH 25) at the area of proposed Ganga express-highway area (Unnao district). 95 - 106 59 4.1 Experiment Field observations and vegetational studies near expresshighway (NH 25) at proposed Ganga expressway area in Unnao district. The predicted environmental risk due to proposed Ganga expressway and their adverse effects on soil, water and plants and loss of biodiversity, this study was carried out. Field study was carried out for present environmental conditions near expresshighway (NH 25) at proposed Ganga expressway area in Unnao district, U.P. state, India. The study areas (Table 3.1) were selected at the proposed Ganga expressway area to observe the changes in the environment conditions at present after a long service given by expresshighway (NH 25) for transportation and prediction of environmental conditions, when proposed Ganga expresshighway will be completed and open for public use for transportation. The study on environmental impact assessment (EIA) may be helpful to make planning for proposed Ganga expressway eco-friendly 4.1.1 Express highway (NH 25) at proposed Ganga expressway area in Unnao district. In India, about 3402 km expresshighways in different states of India have been proposed to complete in near future. Ganga expresshighway has been proposed by U.P. Government, of about 1045 km from district Greater Noida to district Balia. Allahabad Lucknow 53 km Unnao Kanpur 35 km 185 km 208 km Jhansi 4.1 Map showing the distance from Lucknow – Kanpur Ganga expressway (NH 25). 60 Expresshighway (NH 25) a four laned highway linked Lucknow to Kanpur city (about 80 km length) passing through the Unnao district in Uttar Pradesh state (India). The study sites were selected near the expresshighway (NH 25) at proposed Ganga expressway area in Unnao district, just before the Ganga River. The ecological studies (field observations and analysis of soil, water and plants) were carried out in this area at various locations (Table 4.1.2). 4.1.2 Proposed Ganga expresshighway The various locations and related informations were observed regarding the study area. The Ganga expresshighway aimed to construct a 1047 km access controlled eight-lane express way running along the Ganga River in Unnao district. This proposed expressway will connect Greater Noida to Balia district. The proposed Ganga expressway has been divided in to four sectors (Table 4.1.1). Table 4.1.1: Distance and link of proposed Ganga expressway. Route Links Length (km) Greater Noida to Fatehgarh via Bulandshaher Farrukhabad 253 Fatehgarh to Dalmau (Raibareli) Unnao 305 Dalmau to Aurai (Bhadohi) Mirzapur 211 Aurai to Balia Varanasi / Ghazipur 278 The motivation to construct Ganga expressway was to mitigate flood problem in near by areas of river Ganga to large population and number of villages along river, to decongest the increasing traffic on the existing network of roads, reduction in accidents, employment opportunity to people, development of local industry and development of tourism. The Uttar Pradesh government awarded the Ganga 61 expressway contract to ‘Jaypee Infratech Pvt. Ltd. This project will require 26374 hectares of land from farmers. The environmental risk was that, this project may pose a large part of Uttar Pradesh to environmental hazards. The risk of environmental degradation and disturbances of ecological system including biodiversity of living organisms was observed to be possible due to construction and various activities including transportation on the Ganga expressway. The risk of environmental problems was observed, these were: Construction of the embankment would also required large amount of soil, which would be dug from the nearby agricultural field. This will create dug out holes, where rain water will collect permanently, promote water born diseases. Construction of expressway may cause unpredictable damage to trees, crops and wet lands. Due to industrial set up along the express way will dump all their industrial effluents and garbage directly into the surface water bodies and other component of ecosystem. This may lead the pollution problem in the environment. The various types of pollutants will be emitted from expressway transport activities such as various chemicals, gases and heavy metals. In future, these pollutants may damage the growth of plants by disturbances in their metabolic activities, and health hazards to various living organisms around express highway 62 4.1.3 Study sites near expresshighway (NH 25) in Unnao district Study sites were selected at proposed Ganga expressway area near (0-50 m) and about 5 km away from the NH 25 (Lucknow – Kanpur expresshighway). NH 25 expressway in Unnao district was selected for ecological studies because it is used for a long time for transportation and cross the area of proposed Ganga expressway. Sites I (Krishnapuram about 500 m from river Ganga) and III (Sakalpur Nari) were study areas located just near (0 – 50 m) to express way, and all the studies on soil, water and plants were carried out at these sites and compared with the sites II (Dal Narayanpur) and IV (Banthar) situated about 5 km away from the expresshighway. Table 4.1.2: Locations of study area near Express highway (NH 25) in Unnao district. Sites Locations Distance from highway I Krishnapuram (500 m before Ganga river) 0 - 50 m II Dal Narayanpur 4.5 – 5 km III Sakalpur Nari 0 – 50 m IV Banthar 5 – 5.5 km 4.1.4 Soil and water bodies The land area near expresshighway (NH 25) at location I (Krishnapuram) and III (Sakalpur Nari) was uneven, most of the area was observed eroded land. The plants grown at these sites (I and III) were poor in growth and land area occupied with scarced vegetation as compared to vegetation at sites II and IV (about 5 km away from the expresshighway). Water accumulation in different sizes of the pits was observed near the expresshighway (NH 25) were dug-up during the road construction (NH 25). The area just near to road observed (0 – 100 m) was uneven showed 10-15 m depth from highway, and at elevated areas a large number of channels ranged with 63 various widths were observed. These channels formed due to the flow of high velocity run-off rain water which would cut and carried soil particles away from its original place. Also, due to the uneven area and a large number of pits areas were observed as eroded soil which supported poor vegetation at growth. Some area near express highway (NH 25) was observed to use for agricultural purposes, but growing wild plants as well as crops growth indicated very poor fertility of the soil through their poor growth. Major portion of the land was observed barren, grazing animals were using the land for grazing and drinking the water from accumulated water in pits near the expresshighway (NH 25). The texture of the soil was sandy in nature with touch in between the finger. The accumulated water in pits near expresshighway (NH 25) was looking polluted, dirty (very poor transparency) and blackish in colour with decaying smell. 4.1.5: Study of flora near Express highway (NH 25) in Unnao district The flora observed near expresshighway (NH 25) was of various taxonomic groups and varying in size showed a wide range of plant diversity. The wild plant species observed in the year 2009, 2010 and 2011 in pre-monsoon period are listed in Table 4.1.3 and 4.1.4 and Post-monsoon period are listed in Table 4.1.5 and 4.1.6. In the middle line area of the road mainly planted with Nerium indica, Thevetia peruviana and Bougainvillea etc. The leaves of these plants were highly dusted and coated with black coloured smoke particles. Whereas, plants were washed with rain water and appeared new born leaves observed during post-monsoon period. Also, during pre-monsoon period plants were looking slow growing and leaves appeared necrotic. A large variety of other wild species were observed near the road; these were Parthenium, Croton, Ageratum, Sida, Amaranthus, Argimone mexicana,Casia, 64 Achyranthus etc. These wildly growing species showed very poor growth, leaves were small, chlorotic and necrotic and all plants showed stunted growth observed at sites I and III as compared to the same plants growing at sites II and IV (about 5 km away from the expresshighway). The vegetation observed at site I and III was scarce and poor as compared the sites away from the expresshighway (Site II and Site IV). Some winter crops grown at study sites were wheat, mustard, pea, potato, radish etc. observed during post-monsoon season (in year 2009, 2010 and 2011). The agricultural field was scattered near the expresshighway with poor growth observed at site I and III. Whereas, crop plants grown at site II and IV were healthy and with proper looking growth as compared to plants at site I and III were observed. The some common visible symptoms observed in plants at all the study sites were stunted plant growth, reduced size of leaf lamina, poor shoot length and branching, chlorosis and necrosis of young leaves and yellowing and dryness in older leaves. These symptoms were more severe at site I and III as compared to site II and IV. The uniform and continuous agricultural fields with healthy crops were observed at site II and IV. Some wild plants frequently grown and found healthy at site II and IV, but very lesser in number at site I and III, these plant species were Catheranthus roseus, Stelaria media, Lindenbergia, Eclipta, Ocimum sanctum, Oxalis, Solanum nigram etc. The appearance of these plants were healthy with normal leaves at site II and IV, while very poor in growth, scattered / rare and abnormal shape and size of leaves observed at site I and III. In post monsoon period, in the year 2009, 2010 and 20011 wild plants grow near express highway (NH 25) were observed and listed at sites I to IV (Table 4.1.5 and 4.1.6). In the visible field observation at location I and III; growth of wild species near the highway (NH 25) was very poor, their leaves were small, necrotic and 65 showed yellowing; stem was thin, weak, poorly branched, and shoot length was also reduced as compared to those plants who observed at locations II and IV. 4.1.6: Quantitative analysis of vegetation near Express highway (NH 25) by quadrate method. Vegetational study was carried out during Post-monsoon period (October – November) at site I, II, III and IV near and about 5 km away distance from the express- highway (NH 25) at proposed Ganga expresshighway area in district Unnao. Among the wild plant species studied the density, frequency and abundance was found to be poor at site I and III as compared to sites II and IV (Table 4.1.6, 4.1.7, 4.1.8 and 4.1.9). Vegetational study at site I and III near to expresshighway (NH 25) Quantitatively, Croton species showed maximum density (7/m2) and frequency (100%) at site I. Other wild species showed elevated density and frequency were Parthenium (density 6.5/m2 and frequency 75%), Phyllanthus (density 3.8/m2 and frequency 100%), Sida (density 5/m2 and frequency 75%) and Argimone mexicana (density 3.5/m2 and frequency 50%). The dominancy of species at site I was observed in order Croton > Parthenium > Sida > Phyllanthus > Argimone > Abutilon > Ageratum > Tridax > Malvastrum > Calotropis > Ameranthus > Acasia with respect to density and frequency. The Acacia, Amaranthus and Solanum nigram showed least dominancy as compared with density, frequency and abundance among other wild species studied at site I. Among the 15 wild species studied near Express highway (NH 25) in proposed Ganga Express way area, the most dominate species (Parthenium density 7.0/m2 and frequency 100%) and least dominant species (Gnephallium, density 66 0.75/m2 and frequency 25%) were observed. Other wild species dominated over the study area were Sida, Phyllanthus, Croton and Solanum species. Maximum density, Frequency and abundance was observed 7/m2, 100% and 8/m2, respectively, minimum values of these parameters were 0.25/m2, 25 and 2/m2 respectively were observed. The dominancy of the wild species with respect to their density was observed in order Parthenium > Ageratum > Croton > Sida > Phyllanthus > Amaranthus > Argimone > Solanum study showed some dominant species in the area were Parthenium, Ageratum and Croton. Quantitative study of vegetation at site II and IV about 5 km away from Express highway (NH 25), during October to November. Quantitatively, Majus species showed maximum density (20/m2) and frequency (75%) at site II. Whereas, croton showed maximum density (12/m2) and frequency (100%) at site IV. Comparatively, higer density and frequency showing species were Oxalis (11.3/m2), Eclipta (10/m2), Stelaria (12.5/m2) and Parthenium (8.8/m2) observed at site II; and Oxalis (20/m2), Parthenium (11.3/m2), Eclipta (12.5/m2) and croton (12/m2) were observed at site IV. The most abundant species grown at site II and IV were Eclipta, Oxalis and Majus (> 20/m2). At site II, low value of density and frequency showed by Cassia (density, 1.5/m2) and frequency 25%) and Ocimum species (density 0.3/m2 and frequency 25%). At site IV, the least value of density and frequency showed by Ocimum (density, 0.5/m2) and frequency 25%) and Solanum (density, 2.5/m2 and frequency 25%), were observed. Comparatively, most of the species were not observed during the study at site I and III, most of them were Solanum, Ocimum, Stelaria and Majus species. While, these species were abundant with more density and frequency at site II and IV. All the 67 wild species studied near the expresshighway (NH 25) at site I and III showed low density and frequency value as compared to site II and IV (about 5 km away from the expresshighway). Table 4.1.3: Field observations of common wild plants at site I and III just near to expresshighway (NH 25) at proposed Ganga expressway are in Unnao district (pre-monsoon period). Sl. No. Name of plants Family 1. Sida sp. 2. Year of study 2009 2010 2011 Malvaceae Phyllanthus sp. Ambliferae 3. Parthenium sp. Compositae 4. Calotropis sp. Euphorbiaceae NF 5. Ageratum sp. Compositae 6. Abutilon sp. Malvaceae NF 7. Argimone mexicana sp. Papavaraceae NF 8. Launeae sp. Compositae NF 9. Amarnathus sp. Amaranthaceae 10. Achyranthus sp. Achyranthaceae 11. Acacia nelotica sp. Fabaceae 12. Ricinus communis sp. Euphorbiaceae 13. Croton sp. Euphorbiaceae 14. Euphorbia hirta sp. Euphorbiaceae NF NF NF – not found during the observation; - observed. 68 Table 4.1.4: Field observations of common wild plants at site II and IV about 5 km away to expresshighway (NH 25) at proposed Ganga expressway are in Unnao district (pre-monsoon period). Sl. No. Name of plants Family 1. Sida sp. 2. Year of study 2009 2010 2011 Malvaceae Phyllanthus sp. Ambliferae 3. Parthenium sp. Compositae 4. Calotropis sp. Euphorbiaceae NF 5. Ageratum sp. Compositae 6. Abutilon sp. Malvaceae NF 7. Argimone mexicana sp. Papavaraceae NF 8. Launeae sp. Compositae NF 9. Amarnathus sp. Amaranthaceae 10. Achyranthus sp. Achyranthaceae 11. Acacia nelotica sp. Fabaceae 12. Ricinus communis sp. Euphorbiaceae 13. Croton sp. Euphorbiaceae 14. Euphorbia hirta sp. Euphorbiaceae NF NF 15. Achlypha sp. Euphorbiaceae 16. Eclipta sp. Euphorbiaceae 17. Ocimum sp. Labiatae 18. Lindenbergia sp. Scrophulariaceae 19. Majus sp. Scrophulariaceae 20. Salanum sp. Solanaceae NF – not found during the observation; - observed. 69 Table 4.1.5: Field observations of common wild plants at site I and III (Krishnapuram) (Sakalpur Nari) just near by areas of Lucknow – Kanpur express highway (NH 25) at proposed Ganga expressway area in Unnao district: (post-monsoon period). Sl. Name of plants No. Family 1. Parthenium sp. 2. Year of study 2009 2010 2011 Compositae √ √ √ Calotropis sp. Euphorbiaceae √ √ √ 3. Ageratum sp. Compositae √ √ √ 4. Achyranthus sp. Achyrantheceae √ √ √ 5. Croton sp. Euphorbiaceae √ √ √ 6. Solamum nigra sp. Solanaceae √ √ √ 7. Amaranthus sp. Amaranthaceae √ √ √ 8. Sida sp. Malvaceae √ √ √ 9. Abutilon sp. Malvaceae √ √ √ 10. Catherenthus roseus sp. Apocynaceae √ √ √ 11. Argimone mexicana sp. Papavaraceae √ √ √ 12. Malvestrum sp. Malvaceae √ √ √ 13. Casia occidentalis sp. Ceasalpiniaceae √ √ √ 14. Launeae sp. Compositae √ F √ 15. Phyllanthus nurii sp. Umbliferae √ √ √ NF- not found during the observation; - observed 70 Table 4.1.6: Field observations of common wild plants species at site II and IV (Dal Narain Pur) 5 km away from Lucknow – Kanpur expresshighway (NH 25): (post-monsoon period). Year of study Sl. No. Name of plants Family 1. Parthenium sp. 2. 2009 2010 2011 Compositae √ √ √ Phyllanthus nurii sp. Umbliferae √ √ √ 3. Achlypha sp. Euphorbiaceae √ √ √ 4. Euphorbia hirta sp. Euphorbiaceae √ √ √ 5. Eclipta sp. Euphorbiaceae √ √ √ 6. Amaranthus spinosus sp. Amaranthaceae √ √ √ 7. Ocimum sanctum sp. Labiatae N √ √ 8. Catheranthus roseus sp. Apacynaceae √ √ √ 9. Ageratum sp. Compositae √ √ √ 10. Vernonia sp. Compositae √ √ √ 11. Tridax sp. Compositae √ N √ 12. Malvestrum sp. Malvaceae √ √ √ 13. Sida sp. Malvaceae √ √ √ 14. Lindenbergia sp. Scrophulariaceae √ √ NF 15. Abutilon sp. Malvaceae √ √ √ 16. Achyranthus sp. Achyranthaceae √ √ √ 17. Cassia occidentalis sp. Ceaselbiniaceae √ √ √ 18. Canabis sativus sp. Canabinaceae √ √ √ 19. Majus rugosus sp. Scrophulareaceae √ √ √ 20. Croton sp. Euphorbiacea √ √ √ 21. Salanum nigram sp. Solanaceae √ √ √ 22. Gnephallium sp. Compositae √ √ √ 23. Argimon mexicana sp. Papavaraceae √ √ √ NF- not found during the observation; - observed 71 Table 4.1.7: Quantitative analysis of vegetation: density (per m2), frequency (%) and abundance (per m2) of wild plants near Lucknow-Kanpur expresshighway (NH 25) at proposed Ganga expressway area (Unnao) at site I during postmonsoon period Sl. Plants No. Total quadrates studied Total Quadrates in Density number of which /m2 individual species has species occurred Frequency % Abundance /m2 1. Sida sp. 4 20 3 5 75 6.7 2. Calotropis sp. 4 7 2 1.7 50 3.5 3. Parthenium sp. 4 26 3 6.5 75 8.6 4. Achyranthus sp. 4 6 1 1.5 25 6 5. Croton sp. 4 28 4 7 100 7 6. Ageratum sp. 4 12 2 3 50 6 7. Solanum sp. 4 1 1 0.25 25 4 8. Tridax sp. 4 10 2 2.5 50 5 9. Catherenthus sp. 4 6 2 1.5 50 3 10. Argimon sp. 4 14 2 3.5 50 2 11. Abutilon sp. 4 8 3 2 75 4 12. Ameranthus sp. 4 6 2 1.5 50 3 13. Malvestrum sp. 4 10 2 2.5 50 5 14. Phyllanthus sp. 4 15 4 3.8 100 3.8 15. Acassia sp. 4 2 1 0.5 25 2 72 Table 4.1.8: Quantitative analysis of vegetation by quadrate method near expresshighway (NH 25) at proposed Ganga Expressway area (Unnao district) at site II during post monsoon period, 2010). Sl. No. Plants Total Total quadrates number of studied individual species Quadrates Density FreAbun2 in which /m quency dance species has % /m2 occurred 1. Parthenium sp. 4 35 4 8.8 100 8.8 2. Phyllanthus sp. 4 24 4 6 100 6.0 3. Eclipta sp. 4 40 2 10 50 20.0 4. Tridax sp. 4 18 3 4.5 75 6.0 5. Achlypha sp. 4 12 3 3 75 4.0 6. Euphorbia hirta sp. 4 25 4 6.3 100 6.3 7. Ageratum sp. 4 30 2 7.5 50 15.0 8. Vernonia sp. 4 20 4 5 100 5.0 9. Oxalis sp. 4 45 2 11.3 50 22.5 10. Sida sp. 4 25 2 6.3 50 12.5 11. Malvestrum sp. 4 10 2 2.5 50 5.0 12. Achyranthus sp. 4 7 2 1.8 50 3.5 13. Amaranthus sp. 4 6 3 1.5 75 2.0 14. Majus sp. 4 80 2 20 75 40.0 15. Cassia sp. 4 6 1 1.5 25 6.0 16. Canabis sp. 4 4 3 1 75 1.3 17. Croton sp. 4 18 4 4.5 100 4.5 18. Gnephallium sp. 4 15 3 3.8 75 5.0 19. Argimone sp. 4 8 2 2 50 4.0 20. Abutilon sp. 4 6 3 1.5 75 2.0 21. Stelaria sp. 4 50 3 12.5 75 16.3 22. Ocimum sp. 4 1 1 0.3 25 1.0 23. Solanum sp. 4 7 2 1.8 50 3.5 73 Table 4.1.9: Quantitative analysis of vegetation by quadrate method near Express highway (NH 25) at proposed Ganga expresshighway (Unnao district) at site III (during post-monsoon period, 2010). Sl. Plant name No. Total Total quadrates number of studied individual species Quadrates in which species has occurred Density /m2 Frequen cy % Abundance /m2 1. Sida sp. 4 15 4 3.8 100 3.8 2. Calotropis sp. 4 12 2 3 50 6 3. Parthenium sp. 4 28 4 7 100 7 4. Achyranthus sp. 4 27 4 1.8 100 7 5. Croton sp. 4 18 4 4.5 100 6 6. Ageratum sp. 4 24 3 6 75 8 7. Solanum sp. 4 14 2 3 50 7 8. Malvestrum sp. 4 6 2 1.5 50 3 9. Catherenthus sp. 4 8 2 2 50 4 10. Argimon sp. 4 12 4 3 75 3 11. Abutilon sp. 4 1 1 0.25 25 4 12. Ameranthus sp. 4 12 2 3 50 6 13. Malvestrum sp. 4 4 2 1 50 2 14. Gnephallium sp. 4 3 1 0.75 25 3 15. 4 12 3 3 75 4 Phyllanthus sp. 74 Table 4.1.10: Quantitative analysis of vegetation by quadrate method at site IV near expresshighway (NH 25) in proposed Ganga Expressway area, Unnao district during post-monsoon period, 2010. Sl. No. Plants Total Total quadrates number of studied individual species Quadrates in which species has occurred Density FreAbun/m2 quency dance % /m2 1. Parthenium sp. 4 45 4 11.3 100 11.3 2. Phyllanthus sp. 4 30 4 7.5 100 7.5 3. Eclipta sp. 4 50 3 12.5 75 16.7 4. Tridax sp. 4 20 2 5 50 10 5. Achlypha sp. 4 15 3 3.5 75 5 6. Euphorbia sp. 4 20 2 5 50 10 7. Ageratum sp. 4 40 4 10 100 10 8. Vernonia sp. 4 30 2 7.5 50 15 9. Oxalis sp. 4 80 4 20 100 20 10. Sida sp. 4 18 2 4.5 50 9 11. Malvestrum sp. 4 15 2 3.8 50 7.5 12. Achyranthus sp. 4 10 1 2.5 25 10 13. Amaranthus sp. 4 10 2 2.5 50 5 14. Majus sp. 4 60 3 15 75 20 15. Cassia sp. 4 10 1 2.5 25 10 16. Canabis sp. 4 8 2 2 50 4 17. Croton sp. 4 48 4 12 100 12 18. Gnephallium sp. 4 20 2 5 50 10 19. Argimone sp. 4 12 1 3 25 12 20. Abutilon sp. 4 8 1 2 25 8 21. Stelaria sp. 4 40 3 10 75 13 22. Ocimum sp. 4 2 1 0.5 25 2 23. Solanum sp. 4 10 1 2.5 25 10 75 4.2: Experiment Biochemical constituents and tissue concentration of heavy metals in wild plant species near expresshighway (NH 25). Assessment of heavy metals (Zn, Cu, Fe, Cd, Cr and Ni) accumulation and some biochemical constituents such as pigments (chlorophyll a, b, total chlorophyll and carotenoids) and protein contents were determined out in some wild plants (Nerium, Bougainvillea and Croton). Plants were collected from nearby areas of expresshighway – NH 25 (0 to 10 m) exposed continuously to the vehicular environment; and same plant species were also collected from about 5 km away (site II and site IV). These wild plant species analyzed for metals accumulation in their tissues and for some biochemical constituents. Data are presented in tables (4.2.1A, 4.2.1B, 4.2.2A, 4.2.2B, 4.2.3 and 4.2.4). Plants were observed in Pre-monsoon as well as post-monsoon period in the year 2011). 4.2.1: Tissue concentration of heavy metals 4.2.1A: Heavy metals accumulation in plants just near to expresshighway (site I and III) Wild plants estimated for heavy metals accumulation in pre-monsoon period are presented in table 4.2.1A. These plants showed elevated levels of metal accumulation. The range of heavy metals accumulation was found to be 2.5 to 35.6 µg g-1 dry weight. These plants accumulated some toxic metals (such as Cd and Cr) which are not essential to the plant growth. The wild plants showed accumulated heavy metals at site I and III ranged from 0.5 to 125.6 µg g-1 dry weight. Maximum accumulation of Fe in wild plants was observed in both post and pre-monsoon period 76 (3.8 to 125.6 µg g-1 dry weight). The accumulation of most of the metals in wild plants was higher at post-monsoon period as compared to pre-monsoon period. Zinc Wild plants accumulated high content of Zn at sites I and III grown near (0-10 m) expresshighway observed in pre and post-monsoon period, ranged from 7.8 to 35.5 µg g-1 dry weight. Maximum accumulation of Zn was found in Bougainvillea 35.5 µg g-1 dr. wt. in pre-monsoon and 28.0 µg g-1 dr. wt. in post-monsoon period were observed at site I and III. The order of accumulation of Zn in wild plants was Bougainvillea > Calotropis > Croton > Parthenium > Ageratum > Euphorbia > Nerium in pre-monsoon and Bougainvillea > Parthenium > Calotrops > Nerium > Ageratum in post-monsoon were observed. The accumulation of Zn did not followed any regular pattern in between plants, and showed variable concentrations at different sites under different conditions of environment. Copper A high accumulation of copper (Cu) was determined in wild plants near express highway (NH 25) at site I and site III. The accumulation of Cu was ranged from 3.8 to 25 µg g-1 dr. wt. observed in pre-monsoon period, and from 3.3 to 15.4 µg g-1 dr. wt. in post-monsoon period. Maximum accumulation of Cu was found in Calotropis. In pre-monsoon period the order of Cu accumulation was Calotropis > Parthenium > Bougainvillea > Nerium > Ageratum > Croton > Euphorbia, whereas in post-monsoon period the order of Cu accumulation was Croton > Ageratum > Calotropis > Parthenium > Bougainvillea > Nerium > Euphorbia. The least 77 accumulation of Cu was determined in Euphorbia species grown near expresshighway (NH 25). Iron Iron accumulation in wild plants was found highest as compared to other metals studied at site I and III just near to ( 0 – 10 m) express highway (NH 25). Maximum tissue concentration of iron (Fe) was determined in Calotropis. The accumulation of iron was determined in post-monsoon period: Calotropis > Nerium > Croton > Euphorbia > Bougainvillea > Ageratum > Parthenium and at pre-monsoon period: Calotropis > Nerium > Croton > Euphorbia > Parthenium > Ageratum. Least accumulation of iron was determined in Ageratum species observed in pre and post-monsoon period both. The tissue concentration of iron was ranged from 56 to 125.6 µg g-1 dr. wt. in post-monsoon period and from 9.8 to 112.8 µg g-1 dr. wt. in pre-monsoon in wild plants grown near express highway (NH 25) iat proposed Ganga expressway area in Unnao district. Cadmium Cadmium (Cd) concentrations in wild plants determined in the range of 6.5 to 22.5 µg g-1 dr. wt. in pre-monsoon period. Croton, Parthenium and Bougainvillea showed higher accumulation of Cd. Comparatively, low accumulation of Cd was determined in Nerium, Euphorbia, Croton and Calotropis. In post-monsoon period, the concentration of cadmium in plants was low ranged from 0.2 to 16.5 µg g-1 dr. wt. 78 Chromium Chromium accumulation in wild plants grown near the express highway (NH 25) was ranged from 2.5 to 16.6 µg g-1 dr. wt. in pre-monsoon period; and from 0.5 to 12.6 µg g-1 dr. wt. in post-monsoon period. Bougainvillea, croton and Parthenium showed higher accumulation of Cr as compared to other plants studied. In postmonsoon period the accumulation of Cr was lower than in the pre-monsoon period. The order of accumulation of chromium in wild plants near expressway was Bougainvillea > Croton > Nerium > Parthenium > Ageratum and Euphorbia, observed in pre-monsoon period. Nickel A high accumulation of nickel (Ni) was quantified in wild plants at site I and III near expresshighway (NH 25). In pre-monsoon period, the nickel concentration of wild plants ranged from 6.3 to 27.6 µg g-1 dr. wt. In post-monsoon period, Ni content ranged from 1.8 to 15.6 µg g-1 dr. wt. The accumulation of Ni was low in postmonsoon period than the pre-monsoon period. The plants showed high accumulation of Ni were Ageratum, Bougainvillea and Nerium; whereas Euphorbia, Croton, Parthenium and Calotropis showed low Ni content in their tissues. 4.2.1B: Heavy metals accumulation in plants about 5 km away from the express highway (NH 25) at site II and site IV. Accumulation of heavy metals (Zn, Cu, Fe, Cd, Cr and Ni) are presented in table 4.2.1B. The wild plants studied and determined for heavy metal accumulation showed Zn, Cu and Fe, but most of them showed not detectable (ND) range of cadmium, chromium and nickel, observed in pre and post-monsoon period. 79 Zinc Wild plants showed accumulation of Zn ranged from 2.4 to 12.5 µg g-1 dr. wt. in pre-monsoon period; and from 11.8 to 20.5 µg Zn g-1 dr. wt. at post-monsoon period. The higher accumulation of Zn was observed in Nerium, Euphorbia, Calotropis and Ageratum and Croton ranged from 17.6 to 20.5 µg Zn g-1 dr. wt. at post-monsoon period (Table 4.2.2B). The most of the Zn-accumulator plants at sites II and IV showed lower value of accumulation as compared to plants at sites I and III. Copper Wild plants collected from sites II and IV showed accumulation of Cu ranged from 6.7 to 16 µg Cu g-1 dr. wt. at post-monsoon period; and 1.5 to 14.8 µg Cu g-1 dr. wt. in pre-monsoon period. Maximum of Cu is observed in Euphorbia, Parthenium, Ageratum and Calotropis. The accumulation of Cu content in wild plants was higher at sites II and IV as compared to sites I and III. Maximum 16 µg Cu g-1 dr. wt. was found in Calotropis plant, while least accumulation of Cu was found in Parthenium and Croton. Iron Some plants collected from sites II and IV showed high content of accumulation of iron (Fe). Comparatively, Fe accumulation in plants was higher at post-monsoon period as compared to pre-monsoon period. The Fe content in plant tissues ranged from 15.5 to 55.6 µg Fe g-1 dr. wt. determined at post-monsoon period; and 4.2 to 22.8 µg Fe g-1 dr. wt. in pre-monsoon period. Maximum accumulation of Fe determined in Euphorbia, Parthenium and Bougainvillea (22.8 to 55.6 µg Fe g-1 dr. wt.). The accumulation of Fe in wild plant species at sites I and III was found 80 higher (maximum 125.6 µg Fe g-1 dr. wt.) as compared to plants grown at about 5 km away from the expresshighway (NH 25) at sites II and IV. At sites II and IV, the order of accumulation of Fe was Euphorbia > Bougainvillea > Parthenium > Calotropis > Croton > Ageratum observed at post-monsoon period. Cadmium Some plants showed cadmium (Cd) accumulation at sites II and IV observed in pre and post-monsoon period ranged from 0.1 to 6 µg Cd g-1 dr. wt. while, a large no. of plants showed not detectable level of Cd. These plants were Euphorbia, Calotropis, Parthenium, Ageratum and Croton, observed at post-monsoon period. These wild plants showed accumulation of Cd, the tissue concentration was low at site II and IV as compared to sites I and III. Chromium Most of the plants showed chromium (Cr) at not detectable level. Only some plants accumulated Cr up to 2.4 µg Cr g-1 dr. wt., observed at post and pre-monsoon period. As compared to sites I and III, Cr accumulation in plants at sites II and IV was determined very much low and in some plants found not detectable. Nickel At sites II and IV, the accumulation of nickel (Ni) in wild plants was low as compared to sites I and III. In some plants Ni content was not detectable. The tissue concentration of Ni determined in some plants was ranged from 1.6 to 6.8 µg Ni g-1 dr. wt. at post-monsoon period, and 0.8 to 3.5 µg Ni g-1 dr. wt. at pre-monsoon period, were observed. 81 Table 4.2.1A: Heavy metals accumulation in wild plant species at various locations near (0-10 M) express highway (NH 25) in proposed Ganga expressway area (Unnao district) at Pre-monsoon period (2011). Heavy metals (µg g-1 dry weight) Sites I III Plants Zn Cu Fe Cd Cr Ni Euphorbia sp. 20.4±1.0* 4.7±0.5* 25.8±1.5* 7.1±0.1 1.2±0.1 6.8±0.1* Croton sp. 26.5±0.5* 3.8±0.2 75.0±0.5 8.2±0.1 12.4±0.1 11.6±0.2* Nerium sp. 15.8±0.5 8.7±0.1* 98.6±0.5** 10.8±0.1 9.5±0.1 Bougenvellia sp. 35.5±0.6 9.4±0.5 19.8± 15.5±0.1 16.6±0.1 17.2±0.1** Ageratum sp. 20.5±0.2 7.2±0.2 25.6±1.5 16.0±0.1* 14.8±0.2 6.3±0.5 Parthenium sp. 21.5±0.5* 14.8±0.1** 26.5±1.5 17.5±0.5 15.2±0.1 8.3±0.2 Calotropis sp. 30.0±0.5 21.5±1.0* 14.8±0.5** 7.0±0.1 7.3±0.1* 13.7±0.2 Euphorbia sp. 12.6±0.5 6.5±0.1 35.6±0.5 9.6±0.2 2.5±0.3* 7.6±0.1* Croton sp. 10.5±1.0* 5.2±1.0 46.7±1.5 22.5±0.5 15.6±0.1 9.8±0.1 Nerium sp. 8.0±1.0 10.5±0.5 60.8±1.2 6.9±0.5 15.7±0.1 18.8±0.1* 25.5±0.5* 12.4±0.5 25.0±1.0* 8.6±1.0 10.2±0.5* 20.5±0.5 9.8±0.1 6.5±0.1 5.5±0.1 27.6±0.2* Bougenvellia sp. Ageratum sp. 12.0±1.0** 8.5±0.5* 16.2±0.1 Parthenium sp. 7.50.5 18.5±0.5** 12.2±0.1 10.4±0.1 7.2±0.5* 6.8±0.2 Calotropis sp. 18.6±1.5 25.0±1.0 112.6±0.5** 7.5±0.1 12.5±0.1 8.6±0.1 82 Table 4.2.1B: Heavy metals accumulation in wild plant species at various locations about 5 km away from the express highway (NH 25) in proposed Ganga expressway area (Unnao district, U.P.) at Pre-monsoon period (2011). Heavy metals (µg g-1 dry weight) Sites II IV Plants Zn Cu Fe Cd Cr Ni Euphorbia sp. 12.5±2.0 14.8±1.0 16.5±1.5** 0.8±0.5 ND 2.1±0.1* Croton sp. 10.5±1.0 4.5±1.0 14.2±1.5 4.2±0.5* ND 0.8±0.1 Nerium sp. 6.0±0.1* 2.5±0.1* 10.5±1.0 2.9±0.2 0.6±0.1 2.5±0.1 Bougenvellia sp. 9.5±0.5 2.0±0.5 22.8±1.0 6.0±0.5 ND 3.5±0.1 Ageratum sp. 2.4±0.1 6.5±0.1 20.5±1.0 ND 1.2±0.1 2.0±0.1 Parthenium sp. 5.4±0.1 2.3±0.1 15.6±0.5 ND ND ND Calotropis sp. 6.5±0.5 1.5±0.1 10.2±0.1 ND 2.4±0.1 3.5±0.1 Euphorbia sp. 5.8±0.2 2.8±0.1** 8.6±0.2 ND ND 1.8±0.1 Croton sp. 8.6±0.5 7.6±0.2* 6.5±0.1 1.5±0.1 0.8±0.1 2.0±0.1 Nerium sp. 6.7±0.1 5.2±0.2 5.6±0.2 2.1±0.1 ND ND 7.2±0.2* 3.8±0.1* 5.2±0.1 ND ND ND Ageratum sp. 5.5±0.2 4.5±0.1 4.2±0.1 ND ND 0.8±0.1 Parthenium sp. 5.2±0.2 10.5±0.5* 10.2±0.5 ND ND 1.5±0.1 Calotropis sp. 2.5±0.1 4.6±0.2 8.6±0.1** 1.0±0.1 1.5±0.1 ND Bougenvellia sp. ND- not detectable; ± - S.E. (n=3); * - value significant at 0.05 level and **- value significant at 0.01 level. 83 Table 4.2.2A: Heavy metals accumulation in wild plant species at various locations just near to express highway (NH 25) at proposed Ganga expressway area (Unnao district, U.P.) at Post-monsoon period (2011). Heavy metals (Mg g-1 dry weight) Sites I III Plants Zn Cu Fe Cd Cr Ni Euphorbia sp. 7.8±0.3 4.5±0.2* 80.4±5.5 0.2±0.1* 8.0±0.1* 2.5±0.5 Croton sp. 15.8±0.8 11.4±0.6 110.5±15.6 11.5±0.1 4.0±0.1 5.6±0.5 Nerium sp. 17.3±1.0 7.8±0.5 118.1±18.5 16.5±0.1* 0.6±0.1 10.8±1.0 Bougainvillea sp. 28.0±0.5 12.5±1.0 56.8±3.5 10.8 5.8±1.0 15.6±1.0 Ageratum sp. 15.8±0.7* 9.1±0.4 71.2±5.5* 1.9±0.1 5.8±0.1 13.5±0.5* Parthenium sp. 22.6±0.5 11.4±0.6 125.6±25.5 8.0±0.05 1.8±0.1 1.8±0.1 Calotropis sp. 18.5±0.7* 3.3±0.5* 88.3±12.5** 0.5±0.01 1.5±0.1 6.5±0.1* Euphorbia sp. 8.6±0.2 11.4±0.4 95.7±10.0 11.5±0.5 6.5±0.1 5.6±0.5 Croton sp. 9.8±0.8 15.4±0.7 113.7±8.6* 14.0±0.1 0.5±0.1 4.5±0.2* Nerium sp. 11.8±0.5* 9.8±0.6 98.0±7.5** 8.6±0.1 1.5±0.1 8.6±0.2 Bougainvillea sp. 21.6±0.8 8.1±0.7 92.4±10.5 11.0±0.05 12.0±0.1 10.9±0.2 Ageratum sp. 16.5±0.8* 15.5±0.5 75.8±8.5 12.5±0.1 4.0±0.1 12.5±0.5 Parthenium sp. 18.5±0.7* 12.07±0.8 73.6±5.5* 6.1±0.1* 12.6±0.1 6.8±0.5 Calotropis sp. 22.2±0.6* 14.1±0.6 125.6±12.5** 2.0±0. 1 0.8±0.2 8.5±0.1* 84 Table 4.2.2B: Heavy metals accumulation in wild plant species at various locations about 5 km away from the express highway (NH 25) in proposed Ganga expressway area (Unnao district, U.P.) at Post-monsoon period (2011). Sites II IV Plants Heavy metals (µg g-1 dry weight) Zn Cu Fe Cd Cr Ni Euphorbia sp. 18.6±1.5* 12.5±2.0 55.6±5.0 0.08±0.1 ND ND Croton sp. 15.8±1.5 8.2±1.0 46.5±2.5** ND 1.0±0.1 2.5±0.5 Nerium sp. 20.5±2.0* 9.7±1.0 30.3±1.5* ND 0.8±0.1 1.8±0.2 Bougainvillea sp. 13.3±1.0 11.5±1.0* 55.5±5.0* 0.05±0.1 0.5±0.1 2.5±0.1 Ageratum sp. 12.1±1.0 15.8±1.0 30.5±2.5 ND ND ND Parthenium sp. 14.6±2.0* 6.7±1.0 41.8±3.8 ND 0.5±0.1 5.0±0.1 Calotropis sp. 15.8±1.5 8.8±0.5 51.1±2.8 ND ND 6.8±0.5 Euphorbia sp. 11.8±1.0* 14.4±1.6 40.4±5.0* ND ND ND Croton sp. 12.4±1.5 6.3±1.0 30.5±2.5 0.09±0.01 ND 1.6±0.1 Nerium sp. 11.8±1.0* 10.7±1.0 15.5±1.5* 0.06±0.01 ND 8.5±0.5 Bougainvillea sp. 14.6±0.8 13.5±1.5 38.6±2.5 ND ND 10.5±0.5 Ageratum sp. 19.4±2.0* 15.0±1.8 45.5±5.0** ND ND ND Parthenium sp. 14.5±1.5* 9.2±0.8 50.6±2.5** ND ND 6.0±0.1 Calotropis sp. 17.6±1.5 16.1±1.5 41.8±3.8** ND ND 2.4±0.1 ND- not detectable; ± - S.E. value; * - value significant at 0.05 level and **- value significant at 0.01 level. 85 4.2.2 Biochemical responses of wild plant species. Pigments Pigments content (chlorophyll a, b and total chlorophyll and carotenoids) in leaves of Nerium, Bougainvillea and Croton were estimated, grown at various sites of expressway (NH 25). The chlorophyll ‘a’ content in leaves was ranged from 0.18 to 0.60 mg g-1 fr. wt. at sites I and III; and from 0.31 to 0.98 at sites II and IV. Chlorophyll ‘a’ content in plants studied was high at site II and IV as compared to sites I and III. Maximum chlorophyll ‘a’ content was determined in Croton leaves as compared to Nerium and Bougainvillea leaves. Chlorophyll ,b, content in leaves was ranged from 0.56 to 1.45 mg g-1 fr. wt. at site I and III; and from 0.35 to 1.12 mg g-1 fr. wt. at site II and IV. The value of chlorophyll ‘b’ content was estimated more in plants grown just near to expresshighway (NH 25) as compared to plants away from the expresshighway determined in pre-monsoon period (Table 4.2.3). Chlorophyll ‘a’ and ‘b’ contents was determined higher in plants (test plants) at post-monsoon period as compared to premonsoon period. The chlorophyll a content in leaves at post-monsoon period ranged from 0.48 to 1.70 mg g-1 fr. wt. at site I and III, and 0.55 to 1.45 mg g-1 fr. wt. at site II and IV. Total chlorophyll content estimated in wild plant species ranged from 1.0 to 2.59 mg g-1 fr. wt. at site I and III; and 1.14 to 2.93 mg g-1 fr. wt. at site II and IV, were observed in pre-monsoon period. At post-monsoon period, total chlorophyll content ranged from 1.6 to 2.3 mg g-1 fr. wt. at site I; and III and from 1.5 to 2.8 mg g1 fr. wt. at site II and IV, were observed. The total chlorophyll content was found to elevated at post-monsoon period as compared to pre-monsoon period. The elevated total chlorophyll content (>2 mg g-1 fr. wt.) was observed in Croton sps. at site I and III, whereas in Nerium, Bougainvillea and Croton at site II, observed at post-monsoon 86 period. In over all studies, Croton species showed maximum chlorophyll content as compared to other wild species studied. Wild plants grown just near to express highway (NH 25) compared with wild plants grown at about 5 km away from the express highway (NH 25) did not showed a regular pattern of total chlorophyll content in their leaf tissues (Table 4.2.3). Carotenoids content showed higher value at site I and III in Croton (from 0.89 to 1.25 mg g-1 fr. wt.) than other species studied in pre-monsoon period. Whereas, differences in carotenoids content, plants were not significant at different locations of express highway (NH 25). The carotenoids content ranged from 0.84 to 1.25 at site I and III; and from 0.65 to 1.12 mg g-1 fr. wt. at site II and IV were observed at premonsoon period. At post-monsoon period carotenoids content slightly increased as compared to the estimation in pre-monsoon period. Maximum 1.25 mg g-1 fr. wt. in Croton in pre-monsoon period, and 1.65 mg g-1 fr. wt. in Croton in post-monsoon period were observed. In most of the cases, carotenoids content did not show any regular pattern at different sites (I to IV) near express highway (NH 25). Total protein contents estimated and data presented in the Table 4.2.2. In all over studies, protein content in wild species were more at site I and III (0-10 m from express way as compared to site II and IV (about 5 km away from the expressway). At sites I and III, total protein content in leaves of Croton, Bougainvillea and Nerium ranged from 416 to 6.8 µ g g-1 fr. wt.; and at sites II and IV from 398 to 560 µ g g-1 fr. wt. were observed at pre-monsoon period. Protein content showed higher values in plants determined in post-monsoon period as compared the values at pre-monsoon period. The elevated levels in protein content (above 650 µ g g-1 fr. wt.) was observed 780.5 µg g-1 fr. wt. in Nerium at site IV, 665 µg g-1 fr. wt. in Nerium at site III and 651 µg g-1 fr. wt. in croton at site IV, were observed at post-monsoon period. Also, protein 87 content was found to be higher at sites I and III as compared to sites II and IV. Least value of protein content 398 µ g g-1 fr. wt. in Bougainvillea in pre-monsoon and410 µg g-1 fr. wt. in Bougainvillea in post-monsoon period was determined. Therefore, in most of the cases Bougainvillea plants showed low value of carotenoids and protein content in their tissues as compared to Croton and Nerium. 88 Table 4.2.3: Biochemical constituents (carotenoids (mg g-1 fresh weight) and protein contents (µg g-1 fresh weight)) in wild plant species grown at various locations near express highway (NH 25) in proposed Ganga expressway area (Unnao district) observed at pre-monsoon and post-monsoon period (2011). Pre-monsoon Sites I II III IV Post-monsoon Plants Carotenoids Protein Carotenoids Protein Nerium sp. 0.84±0.1* 415.9±0.2 0.92±0.3 488.6±2.1* Bougainvillea sp. 0.89±0.5 536.3±0.1 0.85±0.2 610.2±5.6* Croton sp. 1.25±0.1 525.9±5.5 1.12±0.1* 600.0±20.8 Nerium sp. 0.85±0.1* 428.5±11.4 0.94±0.5 490.8±3.9** Bougainvillea sp. 0.65±0.1 398.0±10.5 0.80±0.5 410.5±15.5 Crotom sp. 0.89±0.2 475.8±10.0** 1.5±0.2 525.5±20.5 Nerium sp. 1.12±0.2* 576.8±15.5 0.98±0.5* 665.6±20.8** Bougainvillea sp. 0.98±0.1 608.2±10.0 1.10±0.2 598.8±20.0 Croton sp. 1.15±0.2 510.8±10.5 1.50±0.5 475.6±18.5 Nerium sp. 0.95±0.2 560.0±15.0 1.54±0.5* 780.5±35.5 Bougainvillea sp. 0.76±0.1 480.5±8.5* 1.12±0.2 610.8±28.5 Croton sp. 0.98±0.1 498.0±10.0 1.65±0.1* 650.5±30.6 ± - S.E. value; * - value significant at 0.05 level and **- value significant at 0.01 level. Site I and III – near (0-10 m) express highway and Site II and IV – away (about 5 km) from the express highway. 89 Table 4.2.4: Biochemical constituents (chlorophylls content mg g-1 fresh weight) in wild plants species grown at various locations near expresshighway (NH 25) at proposed Ganga expressway area (Unnao district) at pre-monsoon and postmonsoon season (2011). Pre-monsoon Sites I II III IV Post-monsoon Plants Chl. a Chl. b Total Chl. Chl. a Chl. b Total Chl. 0.39±0.1* 0.72±0.1* 1.0±0.1 0.56±0.2 0.97±0.5* 1.86±0.5** Bougainvillea sp. 0.31±0.1 0.56±0.1 2.2±0.1 0.42±0.1 0.48±0.2 1.60±0.2 Croton sp. 0.51±0.1 0.92±0.2 2.59±0.5 0.72±0.1 1.20±0.2 2.32±0.5 Nerium sp. 0.46±0.1 0.85±0.1 2.53±0.5 0.62±0.2** 0.78±0.2 2.65±0.5 Bougainvillea sp. 0.31±0.1 0.57±0.1 2.93±0.5 0.48±0.1 0.75±0.2* 2.76±0.5 Crotom sp. 0.75±0.1 0.35±0.1 2.15±0.1 0.43±0.5 0.55±0.2* 2.25±0.5 Nerium sp. 0.62±0.1 1.45±0.2 1.34±0.2 0.85±0.1 1.30±0.5 1.65±0.5 Bougainvillea sp. 0.18±0.1 1.0±0.1 1.60±0.2 0.66±0.1 1.25±0.5* 1.86±0.5* Nerium sp. Croton sp. 0.60±0.1* 1.45±0.2** 1.69±0.2 0.92±0.2 1.70±0.5* 2.25±0.5 Nerium sp. 0.57±0.1 1.10±0.1 1.29±0.1 0.76±0.2 1.45±0.2 1.89±0.2** Bougainvillea sp. 0.68±0.2 0.98±0.2 1.42±0.1 0.86±0.1 0.90±0.5 1.68±0.5 Croton sp. 1.12±0.1* 1.14±0.1 0.77±0.2 1.30±0.2* 1.50±0.1** 0.98±0.1 ± - S.E. value; * - value significant at 0.05 level and **- value significant at 0.01 level. Site I and III – near (0-10 m) express highway and Site II and IV – away (about 5 km) from the express highway. 90 4.3: Experiment Physico-chemical properties of soils at various sites near expresshighway (NH 25) at proposed Ganga expressway area (Unnao district). A composite soil sample was collected from each study sites in the year 201112 (at three periodical times). The site I and III was located just near to express highway (NH 25) at proposed Ganga expressway area in Unnao district of U.P. state (India), and site II and IV was located at about 5 km away from the expresshighway (NH 25). These composite soils were sampled, prepared and analyzed for their physical (texture and density) and chemical (pH, E.C., Ca, Mg, Carbonate, bicarbonate, chloride, phosphorus and DTPA extractable available Zn, Cu, and Fe contents) properties. Data are presented in Table 4.3.1 and 4.3.2. 4.3.1: Physico-chemical properties of soils at sites I and III. The site I and III located just near (0-50 m) to express highway (NH 25) in Unnao district, data are presented in table 4.3.1. The soil pH was moderately to highly alkaline in range (7.5-8.6). At on average data soil in close vicinity to expressway (NH 25) was moderately alkaline in reaction (pH < 0.0). Electrical conductance was high ranged from 1.5 to 1.8 mS/cm. Soil was sandy loam with low organic matter content (average 0.2 to 0.25%) and calcareous in nature. Calcium content in soil at site I and III was determined <6.0 meq./100 g soil; and magnesium content showed the value 12.5 to 15.6 meq./100 g soil. The presence of carbonate and bicarbonate ions was observed in the soil near the expressway, average carbonate ions 0.4; and bicarbonate ions 0.3 meq./l) were determined. At these sites (site I and III) iron content in soil showed high values. The Fe content ranged 150-280 µg g-1 soil at site I, and 60 to 86 µg g-1 soil at site II were observed. The DTPA extractable zinc and copper determined 0.58-0.81 ppm Zn and 0.15 to 0.26 ppm Cu, respectively in the 91 soil collected from site I and III. Near express highway, soil contained high content of chromium (Cr) and nickel (ranged from 0.12 to 0.15 ppm Ni, and 0.04 to 0.13 ppm Cr). 4.3.2 Physico-chemical properties of soil at site II and IV The soils from site II and IV (located at about 5 km away from the express highway (NH 25). The soils of these sites were sandy to silty loam in texture and alkaline in pH. The soil pH was moderately alkaline (pH < 8.0) at site II and IV. Also, soil showed high value of E.C. (1.28 to 1.36 mS/cm), calcium carbonate (1.32 to 1.46%) with low bulk density (Table 4.3.2). The value of calcium (2.8 to 3.2 meq./100 g soil) and magnesium (8.5 to 9.8 meq./100 g soil) observed in soil at site II and IV. The high value of carbonate and bicarbonate ions were observed 0.5 to 0.8 and 0.36 to 0.6 meq./l, respectively. The phosphorus content in soil also determined in soils (<50 µ g g-1 soil). Iron content in soil ranged minimum 60 and maximum 280 µ g g-1 soils at site II and IV. The Zn content (0.8 to 1.35 ppm) was high as compare to Cu (0.52 to 0.40 ppm) content in soil. A very low concentrations of Ni was observed (up to 0.12 ppm), whereas at some places the Ni concentration was not detectable in range. The Cr content in all the soil samples was found not detectable in the samples of sites II and IV, while determined at site I and III. Comparatively, E.C. value in soil at site I and III was high (1.8 mS/cm) as compared to site II and IV (1.3 mS/cm). At all the study sites, soil texture was determined sandy loam to silty loam. Organic matter content was two to three times more at site II and IV (< 0.3%) as compared to site I and III (> 0.81%). There was no major difference was observed in calcium carbonate content in soil at all study sites. Calcium and magnesium content was higher in soils near expresshighway (NH 25) as compared to soils away from the expresshighway at study sites. Carbonate and 92 bicarbonate ions are present high in soils at sites I and III as compared to sites II and IV. Iron and phosphorus content is essential for plant growth, these nutrients determined is soil was higher at site I and III as compared to site II and IV. Zinc and Cu content also showed similar results as by Fe and P. Whereas, Ni and Cr content was less at sites II and IV as compared to sites I and III located near expresshighway (NH 25). Table 4.3.1: Physico-chemical properties of soils collected from various sites just near to express highway - NH 25 (0-50 m) at proposed Ganga expressway area (Unnao district). Site I Site III Parameters Min. Max. Average Min. Max. Average pH (1: 2.5, soil: water ratio) 7.9 8.6 7.8 7.5 8.0 7.6 E.C. (mS/cm) 1.2 2.1 1.8 1.0 1.8 1.5 Texture Sandy loam Sandy loam Bulk density (g/m3) 1.2 1.6 1.4 1.5 1.8 1.6 Organic matter (%) 0.16 0.36 0.25 0.18 0.29 0.20 Calcium carbonate (%) 1.21 1.45 1.28 0.96 1.5 1.35 Calcium (meq./100 g soil) 3.8 5.5 4.5 2.9 42 3.8 Magnesium (meq./100 g soil) 12.5 16.5 14.5 13.8 18.0 15.6 Phosphorus (µg/g) 14.8 28.0 19.5 20.0 30.0 27.5 Carbonate (meq./l) 0.4 0.6 0.45 0.2 0.6 0.4 Bicarbonate (meq./l) 0.1 0.3 0.20 0.15 0.4 0.30 Iron (ppm) 150.0 280.0 110.0 60.0 86.0 70.0 Available zinc (ppm) 0.42 0.86 0.58 0.38 89 0.81 Available copper (ppm) 0.08 0.25 0.15 0.1 0.36 0.26 Nickel (ppm) 0.08 0.15 0.12 0.06 0.18 0.12 Chromium (ppm) 0.006 0.08 0.04 0.08 0.16 0.12 93 Table 4.3.2: Physico-chemical properties of soils collected from various sites about 5 km away from express highway (NH-25) at proposed Ganga expressway area, Unnao district. Site II Site IV Parameters Min. Max. Average Min. Max. Average pH (1: 2.5, soil: water ratio) 7.2 7.9 7.5 7.4 8.5 7.8 E.C. (mS/cm) 0.73 1.6 1.28 0.93 1.78 1.36 Texture Sandy loam Sandy loam Bulk density (g/m3) 0.98 1.2 1.0 0.88 1.3 0.90 Organic matter (%) 0.46 1.34 1.15 0.69 1.60 0.83 Calcium carbonate (%) 0.72 3.6 1.46 1.29 1.50 1.32 Calcium (meq./100 g soil) 2.5 4.8 3.2 2.0 4.2 2.8 Magnesium (meq./100 g soil) 7.8 10.5 8.5 5.6 13.8 9.8 Carbonate (meq./l) 0.3 1.5 0.5 0.5 1.2 0.8 Bicarbonate (meq./l) 0.2 0.6 0.36 0.08 0.9 0.6 Phosphorus (µg/g) 6.0 20.0 18.0 12.0 25.0 18.5 Iron (ppm) 40 60 55 60 80 75 Available zinc (ppm) 0.58 1.4 0.8 0.75 1.86 1.35 Available copper (ppm) 0.28 0.8 0.52 0.31 0.58 0.40 Nickel (ppm) ND 0.08 0.08 0.06 0.12 0.08 Chromium (ppm) ND ND ND ND ND ND ND - not detectable. 94 4.4: Experiment Physico-chemical properties of surface and ground waters near expresshighway (NH 25) at various study sites at proposed Ganga expressway area, (Unnao district). Physico-chemical properties of surface water and ground water was analysed and data presented in Tables (4.4.1A, 4.4.1B, 4.4.2A, 4.4.2B, 4.4.3A, 4.4.3B, 4.4.4A and 4.2.4B). The samples of surface water and ground water were collected around 10 am for 15 consecutive days during the summer season (April to June, 2011). These collected samples were analyzed for pH, E.C., solids, hardness, chloride, carbonate, bicarbonate, calcium and some potentially toxic heavy metals content (Cr, Fe, Ni and Zn), following the standard methods described in Chapter-3. The surface water and ground waters were collected near the expresshighway (0 – 50 m, site I and III) and about 5 km away from the Express highway (site II and III). 4.4.1 Physico-chemical properties of surface water At site I and III, surface waters were alkaline in reaction (pH>8.5), showed high value of electrical conductance (>10.5 mS/cm) and contained high solids content. At site I, the value of total solids was about three times more (7760 mg/l) than the site III (2500 mg/l). Hardness of water estimated as CaCO3 showed maximum value 430 and 450 mg/l at site I and III, respectively. Surface water was more hard with respect to presence of CaCO3 at site III as compared to site I. The chloride (363 mg/l) and carbonate (0.5 meq./l) content was found to higher at site I than site III (chloride 220 mg/l and carbonate nil) in surface waters. Both the sites I and III was located just near to expresshighway (NH 25) showed higher values of bicarbonate (9.0 meq./l at site I and 6.5 meq./l at site III). The value of calcium content in water was found in the range of 50-66.8 mg/l at study sites I and III 95 adjacent (0-50 m) to expresshighway (NH 25). Some potentially toxic heavy metals (Cr, Fe, Ni and Zn) also showed high values ranged from 0.35 ppm to 40.5 ppm in surface water samples near the expresshighway (0-50 m distance). The concentration of these metals was observed in the order Fe>Zn>Ni>Cr. Table 4.4.1A : Physico-chemical properties of surface water near expresshighway (NH 25) at site I at proposed Ganga expressway area in Unnao district. Value Minimum Maximum Average ISI Standards (1999)* 8.4 8.9 8.5 5.5-8.0 EC (mS/cm) 10.30 12.56 11.28 - Total solids (mg/l) 6560 7760 7110 2000 Hardness (mg/l) 290 430 360 600 Chloride (mg/l) 580 760 680 600 Carbonate (meq./l) 0.4 0.6 0.5 - Bicarbonate (meq./l) 6.5 10.5 9.0 - Iron (ppm) 28.6 40.5 33.2 0.5 Calcium (mg/l) 51.30 66.8 55.5 <0.05 Chromium (mg/l) 0.35 0.48 0.45 0.05 Nickel (mg/l) 0.55 0.78 0.65 <0.01 Zinc (mg/l) 5.0 6.5 5.5 5-15 Parameters pH * Indian Standard Institution: Discharge limit of polluted water in surface water bodies. 96 Table 4.4.1B: Physico-chemical properties of ground water near expresshighway (NH 25) at site I at proposed Ganga expressway area in Unnao district. Value Minimum Maximum Average ISI Standards (1999)* pH 7.6 8.2 7.8 5.0-8.0 EC (mS/cm) 0.4 1.8 0.8 - Total solids (mg/l) 1540 1850 1625 2000 Hardness (mg/l) 186 295 160 600 Chloride (mg/l) 52.54 65.5 60.5 600 Carbonate (meq./l) Nil nil nil - Bicarbonate (meq./l) 4.0 6.0 4.5 - Iron (ppm) 20.0 28.0 25.0 5.0 Calcium (mg/l) 36.0 46.5 42.8 - Chromium (mg/l) 0.008 0.002 0.01 <0.05 Nickel (mg/l) 0.081 0.096 0.088 <0.01 2.5 6.5 4.8 5-15 Parameters Zinc (mg/l) * Indian Standard Institution: Discharge limit of polluted water in surface water bodies. 97 The surface waters collected from sites II and IV (about 5 km away from the expresshighway (NH 25) were in alkaline range (pH 7.5 to 8.6). The electrical conductance was measured in the range of 7.5 to 5.0 mS/cm, also showed high values. But, pH and E.C. values of surface waters at site II and IV was much lower as compared to site I and III. At these study sites (II and IV), the maximum average value of total solids, hardness and chloride were observed 185, 110 and 95.5 mg/l, respectively. The presence of bicarbonate in water samples was observed in surface waters on both the sites II and IV, the carbonate content was found nil at these sites. The value of bicarbonate content was higher at site II (maximum 12.5 meq./l) as compared to site IV (maximum 0.8 meq./l) in surface waters. Surface waters away from the expresshighway showed high value of calcium as compared to site I and III (which was near the express highway). Surface waters at site II and IV also contained heavy metals content (Cr, Zn, Fe and Ni), although concentration was much lower than in surface waters of site I and III. Maximum value of heavy metals in surface waters at site II and IV was observed in the range 0.01 to 18.6 mg/l. The concentration of heavy metals was found in the order Fe>Zn>Ni>Cr. The surface waters analyzed for some pollution parameters at site I and III (near to the Express highway) compared with surface waters naturally collected in pits at site II and IV (about 5 km away from the Express highway (NH 25). Surface waters near Express highway (NH 25) was more alkaline (pH>8.5) with high electrical conductance (>12 mS/cm), contained high value of solids (>7760 mg/l) and heavy metals (ranged 0.46 to 28 ppm) as compared to surface waters collected about 5 km away from the expresshighway. Chloride content at its elevated levels cause salinity to the water, it was also high (>200 mg/l) at site I and III as compared to II and IV. 98 The heavy metals such as iron, nickel, zinc and chromium contents was also showed higher concentration in surface waters at site I and III than the sites II and IV. 4.4.2: Physico-chemical properties of ground water The ground water quality was estimated at sites I to IV, and data presented in the Table 4.4.1B, 4.4.2B, 4.2.3B and 4.2.4B). The ground watrers at site I and III (Just near (0-50 m) to expresshighway (NH 25) was alkaline in range (pH>7.0). Also showed high E.C. value (ranged 0.8-3.6 mS/cm), total solids (ranged 1250-1625 mg/l), hardness (ranged 65-90 mg/l) and chloride values (ranged 60.5 to 80 mg/l). The value of hardness, chloride and total solids was higher at site I as compared to site III. Whereas, the value of chloride in ground water was higher at site III than the site I. Carbonate content was determined nil at sites I and III, whereas showed presence of bicarbonate (ranged 1.8 to 4.5 meq./l). The value of bicarbonate was more in ground water at site I as compared to site III. The ground water also contained elevated content of heavy metals ranged 0.025 to 28 mg/l, observed at both sites I and III just near expressway. Maximum value of Fe was observed 25 ppm at site I and 12.5 ppm at site III. The average value of heavy metals was found in order Fe>Zn>Ni>Cr in ground water. 99 Table 4.4.2A: Physico-chemical properties of surface water about 5 km away from the expresshighway (NH 25) at site II in proposed Ganga expressway area in Unnao district. Value Minimum Maximum Average ISI Standards (1999)* pH 8.5 9.5 8.6 5.5-8.0 EC( mS/cm) 6.0 8.5 7.5 - Total solids (mg/l) 1500 2450 1850 2000 Hardness (mg/l) 102 125 110 600 Chloride (mg/l) 92.3 113.0 95.5 1000 Carbonate (meq./l) Nil nil nil - Bicarbonate (meq./l) 7.0 12.5 10.5 - Iron (ppm) 12.0 22.5 18.6 - Calcium (mg/l) 35.0 70.0 56.8 - Chromium (mg/l) 0.06 0.15 0.08 0.05 Nickel (mg/l) 0.05 0.25 0.10 <0.01 Zinc (mg/l) 0.48 0.56 0.50 5-15 Parameters * Indian Standard Institution: Discharge limit of polluted water in surface water bodies. 100 Table 4.4.2B: Physico-chemical properties of ground water about 5 km away from the express-highway (NH 25) at site II at proposed Ganga expressway area in Unnao district. Value Minimum Maximum Average ISI Standards (1999)* pH 7.6 7.8 7.8 5.0-8.0 EC (mS/cm) 0.6 1.3 0.8 - Total solids (mg/l) 440 618 565 2000 Hardness (mg/l) 130 145 140 600 Chloride (mg/l) 5.6 9.5 6.8 600 Carbonate (meq./l) 0.6 0.8 0.6 - Bicarbonate (meq./l) 2.4 4.2 3.8 - Iron (ppm) 1.8 4.6 2.1 0.5 18.43 20.4 19.2 - Nil 0.064 0.003 <0.05 0.028 0.028 0.065 <0.01 1.8 6.2 3.25 5-15 Parameters Calcium (mg/l) Chromium (mg/l) Nickel (mg/l) Zinc (mg/l) * Indian Standard Institution: Discharge limit of polluted water in surface water bodies. 101 The physico-chemical properties of ground water collected from about 5 km away from expresshighway (NH 25) at proposed Ganga expressway area in Unnao district presented in Table 4.4.2B and 4.4.4B. The ground water moderately alkaline (pH 7.6-7.8) and showed high value of E.C., bicarbonate and solids at site I. Hardness value was found within limit of prescribed standard value ISI (1974). The heavy metals was found below the prescribed limit of Central Pollution Control Board adopted ISI (1974) value. Whereas, nickel and chromium contents showed higher values at certain places than the prescribed standard norms. The heavy metals was found in the order Fe>Zn>Ni>Cr. Ground water quality at site IV was also found not suitable with respect to some parameters such as E.C., total solids, and chromium content. At this site water was alkaline (slightly) contained high content of solids (800-1200 mg/l) and chloride (8 to 12 meq./l). Carbonate content was nil in ground water at site IV. Also water contained some heavy metals such as iron (2.8 to 3.5 ppm), chromium (0.004 to 0.009 ppm), nickel (0.011 to 0.05 ppm) and zinc (5 to 8.2 ppm) contents in ground water. The average range of heavy metals was observed in order Fe>Zn>Ni>Cr. At site II carbonate content was present from 0.6 to 0.8 meq./l, whereas found nil at site IV. The heavy metals content was more at site II as compared to site IV, while these ground waters were not affected with activities of expresshighway( NH 25). The parameters of ground water quality were compared from sites I and III (just near expresshighway) to sites II and IV (about 5 km away from the Expressway). Ground water at all studied sites were alkaline in nature but alkalinity was more at site III (Table 4.4.3B) than sites II and IV. Other values such as total solids, E.C., hardness, chloride and calcium contents was higher at sites I and III as compared to 102 sites II and IV. Heavy metals content (Fe, Zn, Ni and Cr) in ground water was found also very high at sites I and III than the sites II and IV. Table 4.4.3A: Physico-chemical properties of surface water near expresshighway (NH 25) at site III at proposed Ganga expressway area in Unnao district. Value Parameters ISI Standards (1999)* Minimum Maximum Average pH 7.2 7.6 7.4 5.5-8.0 EC (mS/cm) 8.0 13.5 13.5 - Total solids (mg/l) 2100 2800 2500 2000 Hardness (mg/l) 320 800 710 600 Chloride (mg/l) 185 265 220 600 Carbonate (meq./l) Nil Nil nil - Bicarbonate (meq./l) 5.6 8.0 6.5 - Iron (ppm) 20 28 21 0.5 Calcium (mg/l) 50.10 60.5 55.0 - Chromium (mg/l) 0.46 0.50 0.48 <0.05 Nickel (mg/l) 0.48 0.69 0.50 <0.01 Zinc (mg/l) 2.8 4.6 3.71 5-15 * Indian Standard Institution: Discharge limit of polluted water in surface water bodies. 103 Table 4.4.3B: Physico-chemical properties of ground water near expresshighway (NH 25) at location III at proposed Ganga expressway area in Unnao district. Value Minimum Maximum Average ISI Standards (1999)* pH 8.0 8.5 8.2 5.0-8.0 EC (mS/cm) 3.5 4.0 3.6 - Total solids (mg/l) 950 1800 1250 2000 Hardness (mg/l) 42 80 65 600 Chloride (mg/l) 60 110 80 600 Carbonate (meq./l) Nil Nil nil - Bicarbonate (meq./l) 1.6 2.5 1.8 - Iron (ppm) 10 0.15 12.5 0.5 Calcium (mg/l) 26.45 30.0 28.0 - Chromium (mg/l) 0.045 0.052 0.040 <0.05 Nickel (mg/l) 0.038 0.059 0.048 <0.01 5.4 6.5 5.5 5-15 Parameters Zinc (mg/l) * Indian Standard Institution: Discharge limit of polluted water in surface water bodies. 104 Table 4.4.4A: Physico-chemical properties of surface water about 5 km away from expresshighway (NH 25) at site IV in proposed Ganga expressway area in Unnao district. Value Minimum Maximum Average ISI Standards (1999)* pH 7.8 8.2 7.5 5.0-8.0 EC (mS/cm) 4.5 6.5 5.0 - 1500 2200 180 2000 Hardness (mg/l) 80 120 18.0 600 Chloride (mg/l) 17 20 18.0 600 Carbonate (meq./l) Nil nil nil - Bicarbonate (meq./l) 0.4 0.8 .06 - Iron (ppm) 15 10 8.0 0.50 Calcium (mg/l) 20.8 40.6 32.8 - Chromium (mg/l) 0.068 0.25 0.056 <0.05 Nickel (mg/l) 0.328 0.859 0.560 <0.01 1.2 5.8 3.2 5-15 Parameters Total solids (mg/l) Zinc (mg/l) * Indian Standard Institution: Discharge limit of polluted water in surface water bodies. 105 Table 4.4.4B: Physico-chemical properties of ground water about 5 km away from expresshighway (NH 25) at site IV in proposed Ganga expressway area in Unnao district. Value Minimum Maximum Average ISI Standards (1999)* pH 7.2 7.9 7.5 5.0-8.0 EC (mS/cm) 1.4 2.8 1.8 - Total solids (mg/l) 800 1200 850 2000 Hardness (mg/l) 30 60 40 600 Chloride (mg/l) 8 12 9.5 600 Carbonate (meq./l) Nil nil nil - Bicarbonate (meq./l) 0.24 2.4 1.8 - Iron (mg/l) 2.8 3.5 2.8 0.5 Calcium (mg/l) 2.6 4.0 3.2 - Chromium (mg/l) 0.004 0.009 0.005 <0.05 Nickel (mg/l) 0.011 0.05 0.03 <0.01 5.0 8.2 6.0 5-15 Parameters Zinc (mg/l) * Indian Standard Institution: Discharge limit of polluted water in surface water bodies 106 Chapter 5 Discussion The study was undertaken to find out the current status of environment (soil, water and plants) in the area under the vicinity of expresshighway (NH 25) used for transportation since 20 years. This highway (NH 25) is crossing the proposed Ganga expressway area (Unnao district) just before (about 500 m) the Ganga river (in Kanpur district). Therefore, study showed a high risk of ecological disturbances due to transportation. In the study area, the pollution in Ganga river due to industrial activities have been reported (Singh and Pandey, 2011; Sinha and Pandey, 2003). Also, in the vicinity of industrial setup in Unnao district, the accumulation of heavy metals in soil and plants has been reported (Sahu et al., 2007). The accumulation of these pollutants in environment pose detrimental effects on living organisms (Baker, 1990; Barman et al., 2001). They enter into food web and biomagnified cause ecological disturbances (Basta et al., 2005). The field observation results 4.1 and 4.1.4, showed valuable importance of transportation as in consonance with Kumar (2012). On the other hand, adverse effects of operations of express highway on soil, water and plants in nearby areas described earlier (Kumar, 2012) to adverse effects of transportation on the environment. Observations showed that the proposed Ganga expressway will run through the area of a large agricultural field they may be affected with transport activities in future (Kumar and Pandey, 2010). The proposed Ganga expressway area may also be develop uneven and degraded due to the construction work near the expresshighway (NH 25). Due to the construction work, a large nearby areas of Ganga expressway may be converted into a unfertile land (Adeyeye, 2005). The 107 chance of soil erosion increased in uneven area, which make it eroded and unfertile (Brady and Weil, 1996), as in this study a large uneven area observed with a large number of different sizes of pits at expresshighway (NH 25). These eroded land supported poor vegetation could be due to the loss of macro (Brady, 1996) and micronutrients (Sharma, 2006). The development of poor vegetation in the study areas due to the deficiency of minerals and soil conditions was also found in accord with Pandey (2014), Gimmler et al. (2002) and Marschner (2003). The animals and human beings living near the expresshighway (NH 25) was observed, they may be under danger of health due to pollutants emitted from transport activities (Kumar, 2012). The heavy metals from the automobile exhaust accumulate in living organisms and pose health risk have been observed (Bunzl et al., 2001; Cobb et al., 2000). The poor growth of wild plant species as well as crop plants could be due to the loss of soil fertility (Adeyeye, 2005; Alumaa et al., 2002) and pollutants emitted due to transport activities (Barman et al., 2000). Observations (Results 4.1.4) showed poor vegetation and crop growth with uneven land at site I and III. The most of the pits near express highway (NH 25) were filled with water, and grazing animals were using this water for drinking. These waters in pits accumulated by run-off water and drainage of human activities near expresshighway (NH 25), use of these water may pose health hazards to grazing animals (Albasel and Cotteme, 1985). Contamination of water bodies might lead to a change in their trophic structure and render them unsuitable for aquatic life (Vajpayee et al., 2001). The leaves of plants in the study area were highly dusted and coated with black coloured smoke particles. These appearance was mainly due to the settled dust particles on the leaves as well as due to exposure of emitted pollutants from the light and heavy vehicles on the expresshighway (NH 25) as observed earlier by Rogge et 108 al. (1993). The wild plants near the expresshighway (NH 25) were poor in growth, leaves were small in size, chlorotic and necrotic, comparatively the wild plants away from the expresshighway (NH 25) showed better growth. These visible effects could be developed due to continuous exposure of automobile exhaust to the plants near the road (Kumar, 2012) and toxicity of air born pollutants to the plants (Mckenzie et al., 2005). The dust particles and smokes coated leaves alter exposure of light intensity may interfere the photosynthetic activity by interrupting the light incidence on leaf surface, a cause of low growth (Sharma, 2006; Pandey and Sharma, 2002). The scattered agricultural crops with poor growth and adverse visible effects were observed in study area were compared to properly growing crops away from the expresshighway (NH 25). These poor growth parameters of crops observed, could be attributed by the poor fertility of soil near expresshighway (Sharma, 2012) and poor nutritional status of the soil (Brady and Weil, 1996). The crops showed poor growth may also be due to the polluted water accumulated in pits near expresshighway (NH 25) and their use for irrigation of these crops, in accord with some workers (Barma et al., 2001). These results also in consonance with observations of several other workers (Pandey, 2004; Pandey and Nautiyal, 2008; Toze, 2006). In experiment 4.2, the accumulation of some important heavy metals in some commonly growing wild plant species was determined, their concentrations in tissues alter metabolic activities have been reported (Rodriguez et al.,2007). Also, some important biochemical constituents (chlorophylls and protein contents) were determined in wild plants (Nerium, Bougainvillea and Croton). The status of these biochemical constituents are indicative of plant health has been reported ( Poskuta et al., 1996). The wild plants just near (0-50m) to expresshighway (NH 25) showed higher concentration of heavy metals (Zn, Cu, Fe, Cd, Cr and Ni) accumulation than 109 the plants away from the transportation activities it could be due to the continuous exposure of pollutants loaded with these heavy metals discharged by the vehicles (Kumar, 2012), plants in exposure with automobile exhaust near road side accumulate elevated levels of heavy metals such as Fe, Cd, Pb, Cr and Ni have also been reported (Kumar and Pandey, 2010; Pandey, 2006a). Some accumulated metals in wild plants, are essential elements (Zn, Cu, Fe and Ni) promoted growth and metabolic activities in plants (Sharma, 2006). But some metals are not essential to plant growth (Cd and Cr), the metals pose adverse effects on growth and metabolism of plants even at low concentrations described earlier (Guo and Marschner, 1995). A high accumulation of heavy metals in plants have been reported either through exposure of automobile exhaust (Kumar, 2010) or pollution in soil near highways (Albasel and Cotteme, 1985) and irrigation with polluted water (Scancar et al., 2000; Schmidt, 2003). The accumulation of heavy metals in plants depend on various factors (Adriano, 2001). Due to the accumulation of heavy metals in plants from a long time may pose risk of loss of biodiversity (Sharma, 2012). Maximum accumulation of Fe in wild plants studied was observed, it could be due to higher Fe content in air and soil near expresshighway (NH 25). Similar results also reported the pollution by some workers due to transport activities (Vousta et al., 1996; Kumar and Pandey, 2010). Accumulation of Cr, Cd and Ni at high concentration in plants may leads various physiological disorders (Deng et al., 2004; Khan, 2007). These metals if accumulated in food crops may pose health hazards to plants (He et al., 2005) and animals (O’Dell et al., 1996). Wild plants used to determination of biochemical constituents showed low chlorophylls content (0.18 to 0.6 mg g-1 fresh weight) at site I and III just near to expresshighway (NH 25), while at sites II and III (away from expresshighway NH25, about 5 km) showed increased 110 chlorophylls (a and b) content. The decrease in chlorophylls content in wild species near expresshighway (NH 25) could be due to effect of pollutants by automobile exhaust (Kumar, 2012), or could be due to the phytotoxicity of elevated heavy metals accumulation in plants (Chatterjee and Chatterji, 2000; Poskuta et al., 1996). The decrease in chlorophylls content (a, b, and total chlorophyll) in wild plants may lead photosynthetic disorder and growth (Pandey and Gautam, 2009b) and cause failure of defence system as reported earlier (Pandey and Pathak, 2006). The high accumulation of toxic metals inhibited seed germination may cause loss of biodiversity (Pandey, 2008). The chlorophyll ‘b’ content was more in plants as compared to chlorophyll ‘a’ The chlorophylls content was determined more in plants in post monsoon period, it could be due to availability and accumulation of some essential elements which promoted more sysnthesis of pigments in post monsoon period (Brown et al., 1987; Pandey et al., 2008). The carotenoids content in plants may also an indicative of antioxidative responses (Pandey and Gautam, 2009b). The total chlorophyll content was not found much decreased at study sites, it could be due to the concentration of essential elememts in plants responsible for synthesis of chlorophyll a (Pandey et al., 2008b), carotenoids content showed higher values at site I and III near to expresshighway (NH 25), than in plants at site II and IV. The result of study could be due to environmental conditions and heavy metals content to strengthen defence system (Baccouch et al., 1998a; Cakamak, 1993). Total protein contents estimated and data presented in the Table 4.2.2. Protein content in wild plant species shoed higher values near (0.50 m) expresshighway than in plants away from the expresshighway (NH 25). The increase in protein content could be attributed due to the availability of essential elements to the plants (Samantarary et al., 1998) or could be due to the protein synthesized in response to heavy metals stress conditions 111 (Sharma, 2006; Schutzendubel and Polle, 2002). Plants adapted themselves to stress conditions by the production of specific proteins has been reported (Peterson, 1983). Heavy metals when accumulated in plants they form metallo-protein (Naaz and Pandey, 2010). In experiment 4.3, physico-chemical properties of soils at various study sites at expresshighway (NH 25) at proposed Ganga expressway area (Unnao) were determined. The soil pH was moderately alkaline with high range of electrical conductance at site I and III, these conditions of soil make it unsuitable for crop growth (Crawford, 1999), only some salt tolerant plants can grow in such soils (Dey et al., 2009). Soil was sandy with low organic matter content it could be due to the soil erosion on uneven area near the expresshighway (NH 25) at site I and III, as described by Brady and Weil (1996). The presence of carbonate ions may pose sodicity in the soil and presence of bicarbonate cause salinity and alkalinity to the soil, contributed the high alkaline pH and high E.C. value in study soil. The results in accord with Sharma (2012) and Brady and Weil (1996). The iron content was found higher in soils near expresshighway (NH 25) as compared to the soil away from the expresshighway (NH 25), it could be due to the transportation activities released high content of Fe in to the environment (Kumar and Pandey, 2010; Gerritse and Dneel, 1984). A high concentration of heavy metals (nickel and chromium) in soil at site I and III could attributed phytotoxic effects in plants (Foy et al., 1978). Low levels of Ni along with Zn and Cu may induce plant growth has been reported (Gerendas et al., 1999; Nath et al., 2009). Therefore, the physico-chemical properties of soil which showed high content of available heavy metals near the expresshighway make the soil unfit for plant growth (Foy et al., 1978). These degradation in soil may be due to the 112 construction work and transportation activities of expresshighway (NH 25) (Marschner, 2003; Sharma, 2012; Kumar and Pandey, 2010). In experiment 4.4, physico-chemical properties of surface and ground water at various study sites near expresshighway (NH 25) have been analysed. At site I and III the surface and ground water showed alkaline pH value was more than the site II and IV. A high level of solids (dissolved and suspended solids) determined in surface water at all study sides. The value of most pollution parameters was more in surface and ground waters collected from just near to the expresshighway (NH 25) as compared to the waters away (about 5 km) from the expresshighway (NH 25). High value of total solids may cause salinity in water and also in soil after their long irrigational use (Pandey, 2006a). The high level of solids in waters could be due to the high content of salts such as sodium chloride, and other inorganic salts present in the soil near surface water bodes (Karbassi et al., 2006) and pollutants emitted from transportation vehicles (Sharma, 2012). The heavy metals content was very high at site I and III, while low at sites II and IV. These elevated contents of heavy metals in water could be due to the automobile exhaust (Kumar and Pandey, 2010) or/and due to the activity of human population living near the road side (Kumar, 2012). If the surface waters contain elevated levels of heavy metals, pose a chance to pollute ground water, particularly, in the area where soil is coarse textured (Brar et al., 2000). The low levels of heavy metals content (Zn and Cr) was determined in ground water at site II and IV, but Fe and Ni content was found to be high above ISI (1999) standard prescribed the discharge limit of waste waters in inland surface water. These could be due to their concentration in soil and their percolation in ground water (Blowes, 2002; Kannan, 2005). The heavy metals content such as Fe, Cr, and Ni content was high in surface 113 waters and also the concentration of Fe and Ni was higher in ground water at site I and III, the values were higher than the ISI (1999) standards. The plants irrigated with such contaminated waters (surface water as well as ground water) accumulate high content of heavy metals (Ensley, 2000; Kao et al., 2008) have been reported. These metals enter in food web and ultimately it affects health of animals and human beings (Kumar, 2012). Elevated concentration of Ni and Cr in aquatic plants (Pandey et al., 2008) and crop plants (Naaz and Pandey, 2010; Deng et al., 2004), adversely effect metabolites and metabolic activities have also been reported. A particular critical concentration of Zn, Fe and Ni, specific to plant species, are essential to plant growth (Gautam and Pandey, 2008) cause positive effects to increase biomolecules content (Kumar and Pandey, 2010) and enzymes activity (Gajewska et al., 2006) in plants. The physico-chemical properties estimated, showed the presence of bicarbonate in almost all samples of surface and ground water. The presence of carbonate observed only at site I in surface water. The presence of these ions along with chloride ions pose risk to make water sodic and saline (Toze, 2006). The ground water showed alkaline pH range, high solids and presence of carbonate and bicarbonate ions, it could be attributed due to their leaching from surface waters to ground water (Boukhalfa, 2007). The results was also in consonance with Kannan et al. (2005). The long term use of such saline waters pose salinity into soil (Aijamal et al., 2000) which adversely affects nutrients uptake and plant growth (Upadhyay et al., 2012). The ground water as well as surface waters near (0-50 m) expresshighway (NH 25) showed higher values of pollution parameters than the water samples collected away from the transport activities, results also in accord with Blowes (2002) and Kumar (2012). At sites I and III near expresshighway (NH 25), the maximum 114 content of Fe and Ni was observed in soil and water both and observed beyond the limit of ISI standards. The presence of these metals including Cr and Ni may contaminate environment, biomagnified through accumulation in vegetables and may pose human health risk (Barman et al., 2000; Bunzl et al., 2001; Fritloff and Greger, 2006; Kumar, 2012). Therefore, transport activities and construction work of expresshighways may pose risk to environmental degradation including degradation of soil, water and plants. These results also indicated that, the proposed Ganga expressway in this area and nearby areas of its total length may pose environmental degradation and vegetational loss as earlier also described and supported various observations (Hussainj et al., 2001; Nriagu and Pacynaj, 1988; Horvath, 2008) in other countries. 115 Chapter 6 Summary The issue to the environmental impacts of transportation’ is a great concern to the sociologist, economist and environmentalist throughout the world. Transport activities conveys substantial socio-economic benefits to the country, but at the same time transport activities also contribute a huge loss of natural resources (soil, water and living organisms) and degradation of environmental system. From one side, transportation activities support increasing mobility demands for people, while on the other side, transport activities associated with growing levels of environmental externalities. It has been very clear that, the transportation is a major source of pollution cause impacts on the environment. In India, about 3402 km expresshighway including Ganga expressway in different states have been proposed to complete in near future. Transport activities contribute among other anthropogenic and natural causes, directly, indirectly and cumulative to environmental problems. Also, contribute at different geographical scales to environmental problems, ranging from local (such as noise pollution and CO emissions and global (climate change and green house effect). The regional to continental problems are smog and acid rain effects. Therefore, the environmental impacts of the net work, traffic and modes, technology, economic process (industrial and other activities) sustaining the transport system, must be considered to make the policies. Without environmental impact assessment, the various problems have led to much controversy in environment policy and in the role of transportation. Therefore, study was undertaken to assessment of environmental problems on soil, water and plants due to transport activities at expresshighway (NH 25) in district 116 Unnao of U.P. state under the proposed Ganga expressway area. The study was aimed to predict the possible environmental problems in the area from the proposed Ganga expressway through the study near expresshighway (NH 25) taken as a standard in the same area. In experiment 4.1, vegetational studies and field observations were carried out. The vegetation studies near the transport activities at site I and III (expresshighway, NH 25), quantitative as well as qualitative were studied. The study of these sites (I and III) compared with study of sites II and IV away from the expresshighway (NH 25). The study of land and water bodies also observed at these study sites. The Ganga expressway has been proposed by U.P. Government of about 1047 km from district Greater Noida to district Balia. The study areas were located at expresshighway (NH 25) in Unnao district of Uttar Pradesh state (India) of about 35 km distance from Unnao to Kanpur district (just before Ganga river), this expresshighway is already in working since 20 years. The proposed Ganga expressway passing through this area needs evaluation to predict extent of environmental problems after completion and operations of proposed Ganga expressway. The motivation to construct Ganga expressway to mitigate flood problems in nearby areas of Ganga to large population and number of villages along river, to decongest the increasing traffic, to reduction in accidents, development of local industry and development of tourism, In the same area, at the express highway (NH 25) the land was uneven and eroded. The land in study areas just near expresshighway (NH 25), observed unfertile and unproductive on the basis of vegetational studies. The growth and distribution of wild plants was found poor near the expresshighway (NH 25) as compared to wild plants away from the expresshighway (NH 25). The prominent species growing in the vicinity of expressway (NH 25) and away from the transportation activities were Croton, 117 Nerium, Parthenium, Bougainvillea, Sida, Euphorbia sps. etc. The density, frequency and abundance level of wild plants was less near expresshighway (NH 25) as compared to wild plants away from the road. The maximum density of wild plants Parthenium (6.5/m2) at site I, and Majus (15/m2) and Parthenium (11.3/m2) at site IV were observed. The other dominant species with respect to density. at site I and III were Croton (4.5/m2), Phyllanthus (3/m2), Ageratum (6/m2) and Sida (3.8/m2) were observed (Table 4.1.6 to 4.1.9). The land away from the expressway was (NH 25) leveled and showed proper growth of crop plants. Whereas, crop growth was poor in area near to expresshighway (NH 25). The texture of soil was sandy loam, and land area near road side showed a large number of pits filled with or without water. The water accumulated in these pits were drinking by grazing animals was observed. In experiment 4.2 accumulations of some potentially toxic heavy metals (Zn, Cu, Fe, Cd, Cr and Ni) was determined in wild species collected from different study sites. The accumulation of Zn and Cu was found under the critical limits of toxicity in plants just near to expresshighway (NH 25). These limits were 7.8 to 28 µg g -1 dr. wt. for Zn and 3.3 to 15.4 µg g-1 dr. wt. for Cu. These heavy metals are essential to promote plant growth and metabolism. Whereas, a high content of heavy metals such as Cd and Cr was determined in wild plants ranged 0.2 to 16.5 µg g-1 dr. wt. for Cd and 0.6 to 12.6 µg g-1 dr. wt. for Cr. These metals may pose toxic effects in plants and animals even at its low concentrations. Maxcimum concentration of Fe (56 to 125.6 µg g-1 dr. wt.) determined in wild plant species at post monsoon period exposed to pollutants near NH 25. The concentration of Ni was excess (upto 13.5 µg g-1 dr. wt.) in some plants. The Bougainvillea, Parthenium, Croton and Nerium accumulated elevated concentration of Ni, Cd and Fe as compared to other plant species studied. In most of the cases, tissue accumulation of heavy metals was found more in post- 118 monsoon period as compared to pre-monsoon period. At study sites away from the epresshighway, the accumulation of heavy metals was very low, even some heavy metals was not detectable, in wild plant species. Some biochemical constituents (chlorophyll a, b and total chlorophyll, carotenoids and protein contents) determined in wild plant species grown just near (0-50 m) and away (about 5 km) from the expressway (NH 25). At all the sites, chlorophyll ‘a’ content was found low as compared to chlorophyll ‘b’. Total chlorophyll content was found in normal range (from 1.0 to 2.59 mg g-1 fr. wt. in leaves). The variations in pigments content was not found at a regular pattern, because soil conditions or other factors, rather than transportation factors, may be involved. The, pigments content was determined more in leaves at post-monsoon period as compared to pre-monsoon period. The carotenoids and protein contents showed slightly higher values at site I and III as compared to site II and IV in pre-monsoon period. A high values of protein content was estimated at site III and IV as compared to I and II, observed at post monsoon period. In experiment 4.3, physico-chemical properties of soils analysed at various study sites of expresshighway (NH 25). The soil was alkaline in reaction (pH > 7.6) with high electrical conductance at each study sites. Most of the soil properties determined showed poor fertility of the land at all study sites. Comparatively, sites ( I and III) just near expresshighway (NH 25) was found more degraded with respect to soil texture, topography, pH, electrical conductance, presence of carbonate, bicarbonate and chloride ions and high content of potentially toxic heavy metals such as Fe, Cd, Cr and Ni than sites (II and IV) away from the road side. The calcium carbonate content was found high at all study sites (I to IV) at 1.28 to 1.46%. The calcium and magnesium content determined high at site I and III (maximum 4.5 119 meq./100g soil Ca and 15.6 meq./100 g soil Mg) than the sites II and IV (maximum 3.2 meq./100g soil Ca and 9.8 meq./100g soil of Mg). A high content of Fe (upto 210 ppm) in soil just near to expresshighway was determined, it was comparatively low (upto 75 ppm) at sites II and IV. At sites I and III, the maximum heavy metals content (available Zn, 0.8; Cu, 0.26; Ni, 0.12 and Cr, 0.13 ppm) determined in soil. At sites II and IV, the maximum heavy metals content (available Zn, 1.35; Cu, 0.52; Ni, 0.08 ppm and Cr, not detectable in the soil) was observed. In experiment 4.4, various physico-chemical properties of surface and ground water were determined at sites I to IV at expresshighway (NH 25) at proposed Ganga expressway in Unnao district collected during April to June, 2011. The quality of water determined in surface as well as ground water to study the impact of transportation . The water quality was determined and found more degraded at site I and III (just near to expresshighway, NH 25 in Unnao district). The surface and ground waters at all the study sites were alkaline in pH (pH 8.4 to 8.9 in surface water; and 7.6 to 8.2 in ground water ). The pH values (8.6 at site II and 7.5 at site IV in surface water) and (7.8 at site I and 7.5 at site III in surface water) were determined. The values of total solids were found high at all study sites, but solids content was more in waters at sites I and III. Most of the values of total solids were under the permissible limits of ISI (1999) standards, but at some places at sites I and III the limit was above the values of ISI standards. At sites I and III, the water (surface and ground water) was more hard than the waters from sites II and IV. At site III, the value of hardness (710 mg/l) was higher than the ISI standard, while at other study sites hardness was low with prescribed ISI standard value (600 mg/l). The minimum value of hardness observed at site IV (40 mg/l) in ground water. At all study sites, the carbonate ions was not detectable except 120 at site II in ground water. The carbonate ions were present in waters at all study sites. Maximum 10.5 meq./l bicarbonate ions were determined in surface water at site II. The presence of calcium determined in surface and ground water, showed its higher values at all sites. The maximum calcium content (56.8 mg/l) was determined in surface water at site II. Maximum value of hardness (42.8 mg/l) in ground water was determined at site I. The presence of calcium carbonate, bicarbonate, chloride ions make water unsuitable for drinking, bathing as well as irrigational purposes. After prolonged application, such water may create salanity to the soil which pose adverse effects on plant growth. In surface and ground water, a high concentration of heavy metals (Fe, Cr, Ni and Zn) was determined. The iron content showed maximum value (21-25 ppm) in surface water at site I and III, the value was observed above the prescribed ISI (1999) standard. The maximum content of Ni (0.6 ppm in surface water and 0.09 ppm in ground water at site I), Cr (0.5 ppm in surface water and 0.09 ppm in ground water at site I) was determined. In some water samples the value of Ni and Cr was found higher as compare to ISI standards. The presence of these heavy metals in waters and their use for various purposes may pose risk to their entry in to food web. Therefoe, water quality was not found in good quality in these study areas. Conclusion Therefore study concluded that, the ecological effects of transport activities on expresshighway (NH 25) may be indicative on ecological adverse effects, in future, when proposed Ganga expressway will be in operation. The findings of ecological studies on expresshighway (NH 25) were: The land was eroded, uneven and poor fertile, which supported poor growth and distribution of plant vegetation. 121 A high content of heavy metals (Zn, Cu, Fe, Ni and Cr) was determined in soil just near to expresshighway (NH 25). 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