Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Critical Reviews in Food Science and Nutrition, 44:489–499 (2004) C Taylor and Francis Inc. Copyright ISSN: 1040-8398 DOI: 10.1080/10408690490489341 Biogenic Amines in Meat and Meat Products DR. CLAUDIA RUIZ-CAPILLAS and FRANCISCO JIMÉNEZ-COLMENERO Instituto del frı́o (CSIC), José Antonio Novais 10, Ciudad Universitaria, Madrid, Spain It has been recognized for some time that biogenic amines occur in a wide range of foods, among them meat and meat products. Meat is an important component of the diet in developed countries. The presence of these amines in food is of interest for two reasons: firstly, for toxicological reasons, in the sense that high levels of dietary biogenic amines can be toxic for certain consumers, and secondly, for their role as possible quality indicators. Based on these two premises, the present article offers a new analysis on aspects of toxicology and on the use of biogenic amines as a quality control method, as well as on their presence in meat and meat products. The article focuses particularly on factors affecting the production of biogenic amines, with reference to various parameters relating to microorganisms, meat raw materials, and processing conditions. A better understanding of the factors determining their formation (i.e., microorganisms, raw materials used, and technological processes applied) and their effects could lead to ways of controlling their production, limiting their presence in the end product, and hence, rendering them less toxic. Keywords Biogenic amines, formation factors, meat products, quality index, toxicological aspects INTRODUCTION Amines are naturally present in living organisms and, hence, in foods. Dietary amines are classified in three categories, according to their chemical structure: aromatic amines (histamine, tyramine, serotonin, β-phenylalanine, and tryptamine), aliphatic diamines (putrescine and cadaverine), and aliphatic polyamines (agmatine, spermidine, and spermine) (Smith, 1980). They also have been classified as “natural polyamines” and “biogenic amines,” depending on their synthesis (Bardózc, 1995). Natural polyamines are formed during de novo polyamine biosynthesis. They are produced naturally by animal, plant, and microorganism metabolism. This group consists chiefly of spermidine and spermine along with the putrescine diamine, and also cadaverine and agmatine in the case of plants and microorganisms. These compounds play an important role in nucleic acid regulation and protein synthesis, and possibly in the stabilization of membranes (Smith, 1980; Bardócz, 1995). Biogenic amines are produced by decarboxylation of free amino acids (FAAs) mediated by amino acid decarboxylase enzymes. Amino acid decaboxylation occurs through removal of the α-carboxyl group to give the corresponding amines (Smith, 1980; Halázs et al., 1994; Bardócz, 1995). This article deals exclusively with biogenic amines. These compounds have been known for some time and are found Address correspondence to Claudia Ruiz-Capillas, Department of Meat and Fish Science and Technology, Instituto del Frı́o (CSIC), C/ Jose Antonio Novais 10, Ciudad Universitaria, 28040 Madrid, Spain. E-mail: [email protected] in varying concentrations in a wide range of foods, including fish, cheese, meat, wine, beer, vegetables, and chocolate (Smith, 1980; ten Brink et al., 1990; Halász et al., 1994). Biogenic amines are important for two reasons. Firstly, the intake of foods containing high concentrations of biogenic amines can present a health hazard through the direct toxic effect of these compounds and their interaction with some medicaments (Bardócz, 1995; Shalaby, 1996). Secondly, they may have a role as indicators of quality and/or acceptability in some foods (Miet and Karmas, 1977; Hernandez-Jover et al., 1997; Ruiz-Capillas and Moral, 2001). There have been a number of studies along these lines, particularly in cheese and fish, the two foods associated with most cases of biogenic amine poisoning (histamine poisoning and cheese reaction) (Smith, 1980; Taylor, 1985; Stratton et al., 1991). Nevertheless, biogenic amines are also present, in widelyvarying concentrations, in meat and meat products (Table 1). Levels tend to be highest in fermented meat derivatives (also the type most researched), but considerable concentrations also have been detected in fresh meat and cooked meat products. Meat is an important component of the diet in developed countries. In the USA and most European countries, it is a major component of total expenditure on food. It accounts for about 35% of expenditure in Denmark, France, and Belgium; 30% in Italy, Spain, and Ireland; and 25% in the UK, the Netherlands, and Greece (Tuley, 1996). The fact that a major component of diet contains varying concentrations of compounds like biogenic amines, which, in certain circumstances, can put consumers at risk of food poisoning, surely merits special consideration. This 489 490 4.7 nd Nd-1.1 Nd-8 31.8 85.4 9.9 0.5 — 3 16 — — — nd — — 5.9–16.1 Nd-6 Nd-108.3 2.35–87.3 1.95–128.40 7.10–78.10 3.10–14.25 7.5–40.50 <1–200 4–16 15.2 Nd-4.8 Nd 1.40 0.50–0.70 Pork raw Pork raw Beef raw Minced beef and pork Raw ground beef at 4◦ C for 12 days Cooked ground beef at 4◦ C for 12 days Pork storage at 5◦ C for 15 days Pork storage at −20◦ C for 15 days Leg Lamb storage at 5◦ C for 5 days Vacuum packed beef at 1◦ C for 7 week Beef Fresh pork (CO2 ) at −1.5◦ C for 13 weeks Pork storage in CO2 /air at 2 ± 1◦ C for 21 days Vacuum packed sterile beef at 1◦ C for 8 wks Vacuum packed beef at 1◦ C for 7 wks Fresh vacuum packed beef at 1◦ C for 120 days Bacon Cooked Spanish meat products “Morcilla” Hamburger Bologna sausage Ripened meat products “Chorizo” Ripened meat products “Salchichón” Ripened meat products “Jamón serrano” Ripened meat products “Lomo embuchado” Ripened meat products “Sobrasada” Egyptian dry sausages Dry sausages Spanish ripened sausage “Mini-salami” Spanish ripened sausage “Fuet” Mortadella Cooked ham Cooked meat products “Chopped” Cooked meat products “Butifarra catalana” Nd: not detected. Histamine Levels of biogenic amines in meat and meat products Products Table 1 — Nd-3.5 Nd Nd-39 12.4 25.1 — — — 6 60 0.7 — — 286 — Nd-8.4 1.5–35.5 Nd-29 76.5–477.8 67.5–465.2 0.45–69.50 60.50–99.25 14.15–77.55 9.5–52.8 3–320 3–12 156.9 Nd-66.0 Nd-11.9 17.60 14.95–151.8 Tyramine 13.3 Nd-0.7 Nd Nd-96 Nd Nd 43.0 41.2 1.3 54 68 39.6 0.3 90–158 — Nd-5 — 1.3–10.2 Nd-57 3.9–34.9 2.1–68.5 — — — 5.6–38.5 <1–790 Nd 367 0.6–7.0 Nd-0.9 — — Cadaverine 7.8 Nd-0.6 Nd-1.75 Nd-69 74.1 85.4 18.9 11.2 3.3 18 20 6.6 1 22–110 — Nd-1 — 0.3–12.3 Nd-29 31.6–361.9 85.9–184.5 — — — 12–102 <1–580 42–139 64.7 Nd-3.9 Nd-3.9 — — Putrescine — — — — — — — — — — — Nd — — 49 — — 2.3–13.5 — 5.7–65.1 12.9–47.4 — — — 2.5–33.2 <10–91 — 10.0 Nd-1.0 nd — — Tryptamine Biogenic amines (mg/kg) — — — — — — — — — — 40 — — — Nd — — Nd-2.5 — Nd-7.7 2.7–34.7 — — — 1.5–80.7 <1–48 — 10.1 Nd-1.4 nd — — Phenylalanine 7.0 2.2–4.1 1.9–4.2 Nd-5 113.3 189.0 3.1 4.3 — 3 9 3.2 — — — 1–6 — 1.6–5.1 1.5–4 1.4–7.9 2.5–3.9 — — — 5.3–11.7 <1–14 5–1010.3 1.9–8.9 1.7–3.0 — — Spermidine 67.1 25.5–38.6 28.7–44.6 14–39 331.3 382.1 31.2 42.8 — 25 600 26.5 — — — 20–49 — 2.1–6.1 15–36 15.4–37.8 8.9–19.4 — — — 1.5–5.2 19–48 25 30.6 7.8–32.2 18.1–25.4 — — Spermine Halász et al., 1994 Hernandez-Jover et al., 1996a Hernandez-Jover et al., 1996a Wortberg and Woller, 1982 Sayen-El-Daher et al., 1984 Sayen-El-Daher et al., 1984 Halász et al., 1994 Halász et al., 1994 Edwards et al., 1983 Edwards et al., 1987 Nadon et al., 2001 Ordoñez et al., 1991 Edwards et al., 1985 Edwards et al., 1985 Smith et al., 1993 Nakamura et al., 1979 Santos et al., 1985 Durlu-Ozkaya et al., 2001 Wortberg and Woller, 1982 Hernandez-Jover et al., 1997 Hernandez-Jover et al., 1997 Vidal-Carou et al., 1990 Vidal-Carou et al., 1990 Vidal-Carou et al., 1990 Shalaby, 1993 Eerola et al., 1998 Treviño et al., 1997 Bover-Cid et al., 1999 Hernandez-Jover et al., 1996a Hernandez-Jover et al., 1996a Vidal-Carou et al., 1990 Vidal-Carou et al., 1990 References BIOGENIC AMINES IN MEAT is particularly true for the meat industry, where health-related incidents have been all too frequent in recent years. Today, society is increasingly aware of the importance of diet for health, and hence, any issue relating to food safety has a considerable impact on consumer behavior and official policy. At the same time, consumers increasingly prefer high-quality products that are minimally processed, safe, etc., and the meat industry is, therefore, looking for emerging technologies that can achieve this in processing and storage. However, the conditions in which traditional or emerging technologies are applied affect the characteristics of the products, and such modifications may produce changes in the formation of different compounds, some of which may be toxic and/or mutagenic, with implications for consumer health. There is, therefore, a clear interest in the study of toxicity and/or mutagenicity of biogenic amines and the factors determining their formation in the context of food processing conditions and preservation. Although there are such studies on biogenic amines in meat products, these are generally based on fermented products, and much less work is available in relation to other types of products, such as boiled food or fresh meat. In most of the available reviews (Smith, 1980; Halász et al., 1994; Silla, 1996; Shalaby, 1996), many of the aspects most closely related to myosystems are addressed only tangentially in broad analyses, focusing on other foods more generally associated with biogenic amine poisoning. As far as we know, no detailed, specific analysis on the formation of biogenic amines in different types of processed meat has been done yet, despite their outstanding presence in this kind of food. What is original about this study is that it is the first such analysis and should help to gain a better understanding of the factors entailed in the production of biogenic amines and to indicate possible ways of limiting their presence in the product and reduce dietary biogenic amines. Given their health implications, the achievement of these aims could be of considerable benefit to consumers. The aim of this study, then, is to analyze, on the one hand, toxicological aspects and the use of biogenic amines as a quality control method in meat and meat products, and on the other hand, their formation, with particular emphasis on factors related to microorganisms, raw meat materials, and processing conditions. TOXICOLOGICAL ASPECTS OF BIOGENIC AMINES: IMPORTANCE IN MEAT AND MEAT PRODUCTS The intake of foods with high concentrations of biogenic amines can cause migraine, headaches, gastric and intestinal problems, and pseudo-allergic responses, chiefly brought about by the toxic action of histamine and tyramine, known respectively as “histamine poisoning” and “cheese reaction” (Smith, 1980; Taylor, 1985; Stratton et al., 1991). The most common symptoms of histamine poisoning are due to the effect on the cardiovascular system, producing low blood pressure, reddening of the skin, and headaches, oedemas and rashes typical of allergic reactions (Stratton et al., 1991; Van Gelderem et al., 1992; 491 Bardócz, 1995). Typical symptoms of tyramine poisoning are migraines, headaches, and increased blood pressure (Joosten, 1987). Tyramine also causes the release of noradrenaline from the sympathetic nervous system (Bardócz, 1995). Other amines, such as spermidine or spermine, have also been associated with the development of food allergy (Lux et al., 1980). In normal circumstances, the human body is able to rapidly detoxify histamine and tyramine absorbed from foods by acetylation and oxidation mediated by the enzymes monoamine oxidase (MAO; EC 1.4.3.4), diamine oxidase (DAO; EC 1.4.3.6), and polyamine oxidase (PAO; EC 1.5.3.11) (Rice et al., 1976; Bardócz, 1995). However, if these detoxifying mechanisms are upset, either because of high amine intake or because the individual is allergic or is deficient in aminooxidases due to consumption of or treatment with oxidase enzymes (e.g., monoamine oxidase inhibitor—MAOI), biogenic amines may build up in the body and could cause serious toxicological problems (McCabe, 1986; Halász et al., 1994). The toxic potential of these dietary amines is even more alarming if we consider that approximately 20% of the European population regularly consume MAOI drugs as antidepressants, which inhibit aminooxidase activity (Sattler et al., 1988; Maijala and Eerola, 1993). Putrescine and cadaverine, although not considered toxic individually, can enhance the effect of histamine and tyramine by interacting with the aminooxidases and interfering with the detoxifying mechanism (Rice et al., 1976; Taylor, 1985; Sattler et al., 1988). Similarly, other compounds, like alcohol and acetaldehyde, also can augment the toxic potential of biogenic amines, since they promote the transportation of these through the intestinal wall. It is generally very difficult to establish limits of toxicity of biogenic amines in a given product, because their effect does not depend on their presence alone (type of amine and levels present), but is also influenced by other compounds (modulating their effect) and by the specific efficiency of the detoxifying mechanisms in different individuals. Hence, the toxicity of biogenic amines will depend on factors associated with the food itself (quantitative and qualitative) and also on factors associated with the consumer (individual susceptibility and state of health). There are large gaps in our understanding of the toxicological importance of these compounds and the mechanics of their toxicity. Various international studies of epidemiological outbreaks (Lipp and Rose, 1997) have failed to establish a clear dose-effect relationship (Clifford et al., 1989). The role of various substances that enhance the toxicity of biogenic amines and the existence of synergic effects have been demonstrated (Leigh et al., 2000), and therefore the determination of the amine concentrations in each case is not enough to assess their toxic potential. In recent years, there have been some studies on the in vitro toxicity and mutagenicity of biogenic amines (Badolo et al., 1998), in which the effect has been analyzed on the basis of the cytotoxicity of individual compounds. The most prevalent biogenic amines in meat and meat products are tyramine, cadaverine, putrescine, and also histamine. 492 C. RUIZ-CAPILLAS AND F. JIMÉNEZ-COLMENERO The concentrations of these amines tend to vary depending on the different products (Table 1). The concentrations of some of these amines, such as histamine, are usually quantitatively lower than those found in other foods (e.g., fish) and their toxicological effects, therefore, appear to be severely limited. However, there are some meat products where this is not the case. It has been observed that histamine concentrations in cured products can reach 100 mg/kg or considerably higher, for example, in dry sausages (Table 1). Given that 100 mg/kg is the accepted threshold for the appearance of food poisoning symptoms in a normal person with no health problem (ten Brink et al., 1990; Stratton et al., 1991), such products should be excluded from the diet of persons also ingesting MAOIs. In an analysis of biogenic amine concentrations in commercial processed foods, Vidal-Carou et al. (1990) determined that 63% of salchichon samples and 64% of chorizo samples (both ripened meat products) contained enough tyramine to poison MAOI consumers; in this case, it was considered that 6 mg/kg of tyramine would be toxic if ingested with MAOIs (McCabe, 1986). In “normal” persons, 125 mg/kg would be required for tyramine to be toxic. Similarly, Santos et al. (1985) suggested that an intake of 100 g of any of a number of meat products (chorizo, salami, lomo embuchado, Spanish meat products, blood sausage, or hamburger) containing up to 8.4 mg/kg could be toxic in the event of interaction with MAOIs. In the absence of MAOIs, on the other hand, none of these processed meats would be toxic if ingested in normal amounts, but always depending on the susceptibility of the individual. Apart from the toxic effects described above, biogenic amines can play a role in other kinds of processes harmful to human health. Various biogenic amines (spermidine, spermine, tyramine, putrescine, and cadaverine), when subjected to heat, can give rise to the formation of secondary amines, and in the presence of nitrites, these can generate nitrosamines, chemical agents considered to possess major carcinogenic properties (Patterson and Mottram, 1974; Warthesen et al., 1975). This is particularly important in some meat products with high biogenic amine levels and added nitrates and nitrites (Table 1). Evidently there are difficulties and limitations in determining true levels of toxicity, but considering the figures produced Table 2 by various authors and the restrictions that they imply, there is undoubtedly a large number of meat products containing biogenic amines that present some potential risk. Despite all the foregoing, there is a general absence of specific legislation setting limits on biogenic amines in meat products; at best, there is reference to histamine in some foods, but there is no reference at all to maximum concentrations of other biogenic amines. For instance, there is European legislation (Directive 91/439/EEC) that limits permitted histamine levels in fishery products from the scombrid or cupleoid family to 100–200 mg/kg for fresh fish and up to 400 mg/kg for cured products. Similarly, the Food and Drug Administration (FDA, 1990) sets the limit of histamine tolerance in fresh fish at 100 mg/kg. Some laboratories have made more general recommendations; for example, the Netherlands Institute of Dairy Research (ten Brink et al., 1990) sets a limit of 100 mg/kg on histamine in foods. As noted earlier, some authors (Smith et al., 1993; Hernandez-Jover et al., 1996a; Eerola et al., 1996; Eerola et al., 1998; Bover-Cid et al., 2001a) have used this limit to establish the potential health risk in certain sectors of society where some products are consumed beyond a given storage period. BIOGENIC AMINES AS A QUALITY INDEX The concentrations of some biogenic amines (tyramine, putrescine, and cadaverine) normally increase during the processing and storage of meat and meat products, whereas others (spermidine and spermine) decrease or remain constant (ten Brink et al., 1990; Halázs et al., 1994; Bardózc, 1995). There have, therefore, been attempts to establish a relationship between meat quality and changes in the content (individual or combined) of such compounds in different meat derivatives (Table 2). The usefulness of biogenic amines as a quality index will depend on the nature of the product; results tend to be more satisfactory in fresh meat and heat-treated products than in fermented products. In the latter, they seem to be of very limited use. Biogenic amines have been used as quality indexes and indicators of unwanted microbial activity in meat and cooked meat products (Table 2). A combination of putrescine and cadaverine Index of biogenic amines in meat and meat products Products Biogenic amines References Fresh beef meat Bologna sausage Minced beef and pork Pork meat at 6–8◦ C Raw and cooked ground beef Hamburger Vacuum packed beef at 1◦ C Vacuum packed beef at 1◦ C Wrapped and unwrapped fresh meat (pork, beef and rabbit) Cooked Spanish products “Jamón york,” “chopped” and “butifarra” Dry sausages Putrescine and cadaverine BAI = putrescine + cadaverine + histamine + tyramine Slemr, 1981 Wortberg and Woller (1982) BAI = putrescine + cadaverine + histamine + tyramine Putrescine tyramine and 1,3 diaminopropane Tyramine, histamine, putrescine and cadaverine Putrescine and cadaverine Tyramine Putrescine and cadaverine Hernandez-Jover et al., 1996b Sayem-El-Daher et al., 1984 Durlu-Ozkaya et al., 2001 Edwards et al., 1985 Edwards et al., 1987 Guerrero-Lejarreta and Chavez-gallardo, 1991 Putrescine and cadaverine Vidal-Carou et al., 1990 Tyramine, histamine, putrescine and cadaverine Eerola et al., 1996–98 BIOGENIC AMINES IN MEAT has been suggested as an index of acceptability in fresh meat, because their concentrations increase prior to spoilage and correlate well with the microbial load (Slemr, 1981; Edwards et al., 1985). Wortberg and Woller (1982) found that a high cadaverine concentration was clearly indicative of spoilage. These authors proposed a biogenic amine index (BAI) consisting of the sum of putrescine, cadaverine, histamine, and tyramine and established 500 mg/kg as the limit for Bologna sausage, minced beef and pork. Using the same index, Hernandez-Jover et al. (1996b) suggested the following limits: BAI <5 mg/kg for good quality fresh meat; between 5 and 20 mg/kg for acceptable meat, but with initial spoilage signs; between 20 and 50 mg/kg for low meat quality; finally, >50 mg/kg for spoiled meat. It generally has proven more difficult to apply similar quality criteria to fermented products. This is at least partly because biogenic amine concentrations vary much more widely in fermented products than in fresh meat and cooked meat products (Table 1), because of the number of different factors involved in their formation. These factors include the type and degree of contamination of raw materials, which are promoted by structural breakdown, manufacturing practices, certain processing stages, and the use of starters. All these factors vary according to the nature of the product and, in some cases, can mask changes in the type and concentration of biogenic amines through the different phases of treatment and storage, delaying visible signs of spoilage and/or off-odor development (Edwards et al., 1983). The following example illustrates the problem. Not all spoilage or starter microorganisms can decarboxylate FAAs. Even within the same species, not all strains develop the same decarboxylating capacity, so that a low biogenic amine concentration does not always signal good microbiological quality (Santos et al., 1985, Dainty et al., 1987, Halász et al., 1994). It is, therefore, no simple matter to establish a biogenic amine index that reliably predicts quality for products of this kind (Eerola et al., 1998). Figure 1 493 FACTORS INFLUENCING THE FORMATION OF BIOGENIC AMINES As noted earlier, biogenic amines are produced by enzymatic decarboxylation of certain free amino acids (Figure 1). Bacterial decarboxylases are not very specific, but their activity varies according to the bacterial species and strain (Bardócz, 1995). Thus, biogenic amine formation requires the presence of free amino acids, the decarboxylase enzyme and suitable environmental conditions. Hence, all the factors bearing on production of the substrate (FAAs), the enzyme, and also their level of activity, effect the type and amount of biogenic amines present in each case. Factors associated with the raw material (meat composition, pH, handling conditions, etc.) as the substrate source and reaction medium directly affect the availability of FAAs, whereas the presence of the enzyme is closely tied to microbiological aspects (bacterial species and strain, bacterial growth, etc.). It is also apparent from the literature that diamine oxidase formation by histaminolytic bacteria can influence final histamine levels (Ienistea, 1971). These factors are obviously interdependent and are further influenced by the technological processes associated with types of meat derivative (steak, roast, ham, ground, restructured, comminuted, fresh, cooked, smoked, fermented, etc.) and storage conditions (time/temperature, packaging, temperature abuses, etc.). The combined action of these factors is what chiefly determines final concentrations of biogenic amines, because it either directly or indirectly determines the presence and the activity of substrate and enzyme (Figure 2). More knowledge of these factors would, therefore, help to better understand the mechanics of biogenic amine formation and find means of keeping them at nontoxic levels. Following is a specific analysis of the most important factors. We chose this approach for the sake of clarity of exposition, at the risk of occasional simplification. Many of these factors are Formation of biogenic amines. 494 C. RUIZ-CAPILLAS AND F. JIMÉNEZ-COLMENERO Figure 2 Factors influencing the formation of biogenic amines. interrelated and would otherwise have to be addressed in several sections requiring more complex analyses. Microorganisms Bacterial amino acid decarboxylase enzymes play a fundamental role in the formation of biogenic amines, and therefore, the microorganisms that produce them are an extremely important element (Figure 2). Slemr (1981), and Slemr and Beyermann (1985) reported that there was no formation of biogenic amines in sterile meat, and that when concentrations increased in meat, they did so along with microorganisms. Biogenic amine production is influenced by the microbial load in the product and the type of microbiot constituting that load (bacterial species and strain) (Bardócz, 1995). This, in turn, depends on various factors: characteristics of the meat raw material, use of thermal treatments, use of additives, etc. (Figure 2). Numerous researchers have sought to establish a relationship between the formation of biogenic amines in meat and meat products and the activity of various types of microorganisms (Table 3). Eitenmiller et al. (1978) attributed the formation of biogenic amines in dry sausages to a bacterial response that maintains cellular viability, when this is threatened by acidity in the medium. In general, decarboxylase activity in meat products is attributed chiefly to Enterobacteriaceae, Pseudomonadaceae, Micrococcaceae, and lactic bacteria (Table 3). In fresh meat, Enterobacteriaceae have been identified as the main producers of cadaverine (Edwards et al., 1983; Guerrero-Lejarreta and Chavez-Gallardo, 1991; Ordoñez et al., 1991, Bover-Cid et al., 2001a), while putrescine has been associated with high total aerobic viable counts (TAVCs) (Edwards et al., 1983, Bauer et al., 1996). Lactic bacteria appear to be the main producers of biogenic amines in fermented products; high Lactobacillus counts have been associated with the formation of high histamine concentrations. However, amine formation seems to be a characteristic only of some Lactobacillus strains, as other strains do not produce them (Maijala and Eerola, 1993; Straub et al., 1994; Roig-Sagués and Eerola, 1997). Differences in concentrations and types of lactic bacteria could help to explain the high variation of biogenic amine concentrations found in products of this kind (Table 1). Masson et al. (1996) found that tyramineproducing strains in dry sausage belonged to Carnobacterium, L. curvatus and L. plantarum, whereas Micrococcaceae and L. sake did not produce tyramine. It has been suggested that the use of amino-negative starters comprised of L. sakei or Pediococus pentosaceus could prevent the formation of biogenic amines in dry sausage (Maija et al., 1995; Bover-Cid et al., 2001b), although any reduction would always depend on other factors influencing formation, especially the raw material (Figure 2). Meat Raw Materials Meat raw material is the natural source of the substrate (FAAs) from which biogenic amines are produced. It also is the largest component of the matrix in which the decarboxylation reactions take place and is, therefore, basic in determining essential factors in enzymatic activity: pH, ionic strength, substrate 495 BIOGENIC AMINES IN MEAT Table 3 Production of biogenic amines by microorganisms in meat and meat products Products Microorganisms Biogenic amines References Fresh meat Fresh beef Raw Pork Pseudomonads Putrescine Carnobacterium Lactobacillus curvatus Lactobacillus plantarum Enterobacter cloacae Klebsiella pneumoniae Enterobacteriaceae Total aerobic viable counts (TAVC) Enterobacteriaceae Pseudomonads Enterobacteriaceae Tyramine Bauer et al., 1996 Edwards et al., 1987 Masson et al., 1996 Raw pork at 5◦ C Beef, pork lamb at 5◦ C Wrapped and unwrapped fresh meat (pork, beef and rabbit) Ground meat and processed meat Vacuum packed beef at 1◦ C Vacuum packed beef at 1◦ C Pork stored in CO2/air at 2◦ C Dry products (sausages) Dry sausage Ripened sausages Fermented sausages Lactobacillus divergens Lactobacillus carnis Hafnia alvey Serratia liquefaciens Brochothrix thermosphacta Lactobacilli Enterobacteriaceae Lactobacilli Carnobacterium Lactobacillus carvatus Lactobacillus plantarum Enterococci Lactic acid bacteria (LAB) LAB Enterobacteriaceae concentration, inhibitors, and their mobility, etc. Therefore, any conditions that alter its nature and characteristics will influence the formation of biogenic amines in one way or another (Figure 2). Some biogenic amines (e.g., putrescine and cadaverine) also are formed and broken down during normal cell metabolism (Bardócz, 1995) and are, therefore, naturally present in muscle and meat. However, when an amine exceeds these natural concentrations, it clearly must have some other origin. Thus, the amount of biogenic amines in the meat raw material will contribute to the pool of biogenic amines in the end products. It is vital to achieve guaranteed use of suitable raw materials in order to limit the presence of amines in the end product and, hence, assure better quality. Free amino acids play a fundamental role in the formation of amines in meat, in that they are their precursors and, moreover, constitute a substrate for microbial growth (Smith 1980; Arnold and Brown, 1978). Concentrations increase in association with proteolytic events taking place in the course of treatment and storage, due essentially to many of the microorganisms present. However, in meat such as fish, it has not been possible to establish a clear relationship between FAA concentrations and formation of the corresponding biogenic amines (Eerola et al., 1996; Ruiz-Capillas and Moral, 2001). Various studies have shown that fat content influences the formation of biogenic amines (Hernandez-Jover et al., 1997; Kebary et al., 1999). This phenomenon has been most intensively studied in cheese, where Putrescine Cadaverine Cadaverine Putrescine Cadaverine Putrescine Putrescine, tyramine, Histamine Tyramine Halász et al., 1994 Cadaverine Putrescine Cadaverine Putrescine Dainty et al., 1986 Histamine Tyramine Maijala and Eerola, 1993 Masson et al., 1996 Tyramine Roig-Sagués et al., 1997 Tyramine Cadaverine Bover-Cid et al., 2001a Edwards et al., 1983 Guerrerolejarreta and Chavez-Gallardo, 1991 Durlu-Ozkaya et al., 2001 Edwards et al., 1987 Ordoñez et al., 1991 it has been observed that the concentration of biogenic amines decreases along with fat content. Banwart (1981) attributes this fact to changes in the water activity (Aw) rather than the actual amine content; Aw causes some inhibition of growth of proteolytic bacteria and, hence, a fall in the concentration of free amino acids in the medium. It has been noted that the meat source can influence the formation of biogenic amines. Some authors (Wortberg and Woller, 1982; Vidal-Carou et al., 1990) have reported that histamine and tyramine formation tends to be found less in packed meats made only with pork (e.g., cooked ham and cured ham) than in other meat derivatives containing mixtures of beef and pork (salami, salchichon, chorizo or Bologna sausages), where there was a greater tendency to form tyramine. The conditions in the reaction medium are a key factor in enzymatic activity, quite apart from their effect on the actual production of the enzyme through their impact on microbial growth. Histidin decarboxylase activity increased in acid media, with an optimum pH range of 4 to 5.5 (Arnold and Brown, 1978; Halázs et al., 1994). The final pH of the meat, which can vary depending on a variety of factors, could, therefore, have a considerable influence. High pHs, for instance in DFD (dark, firm and dry) meat, favor microbial proliferation, which promotes FAAs, but limits decarboxylase activity. The opposite is true of PSE (pale, soft, exudative) meat. We are not aware of any studies on the influence of meat type (DFD, PSE) on biogenic amine formation, but such an influence cannot be ruled 496 C. RUIZ-CAPILLAS AND F. JIMÉNEZ-COLMENERO out, given that the meat type affects factors directly involved in the activity of amino descarboxylases enzymes and the final presence of biogenic amines in these meat. Meat Processing Meat processing and storage conditions influence the formation of biogenic amines, because they affect some of the elements implicated in biogenic amine production (Figure 2). We know that any processing step, directly or indirectly, affects the concentration of substrate and enzyme and determines the presence of other compounds or conditions that modulate decarboxylase activity; therefore, there are many factors to be considered, especially in connection with the technology applied (fresh, cooking, curing, fermenting, etc.). Thus, to limit the formation of biogenic amines, it is not enough to have suitable raw materials; it also is necessary to optimize the processing conditions. The production of biogenic amines is affected by the conditions in which meat raw materials are handled. For example, after the decontamination of beef carcasses (washing with chlorine and lactic acid), followed by vacuum packing of cuts, of all the amines monitored (histamine, phenylethylamine, tryptamine, and tyramine), only tyramine was consistently detected over 120 d storage at 1◦ C (Smith et al., 1993). Any process entailing structural breakdown (grinding, chopping, sectioning, slicing, etc.) favors microbial contamination. Santos et al. (1985) attributed tyramine formation in hamburgers to increased contamination, as a result of grinding. Chilling storage conditions considerably affect amine production. Unsuitable storage temperatures (i.e., temperatures >5◦ C), prolonged storage, or temperature abuses during storage have a two-fold effect: on proteolysis due to increased microbial growth promoting penetration in the muscle; on amino decarboxylase activity (Maijala and Nurmi, 1995). There have been numerous studies on the formation of biogenic amines in the course of fresh meat storage, frequently with conflicting results. Nakamura et al. (1979) detected no changes in biogenic amine concentrations (<100 g/kg) in pork stored at 4◦ C for 16 d, even after rejection for the onset of decomposition; however, other authors (Wortberg and Woller, 1982; Edwards et al., 1983) reported increased concentrations, especially of cadaverine, which is possibly associated with the onset of spoilage. Halász et al. (1994) reported increased putrescine and cadaverine concentrations in pork meat over 15 d storage at 5◦ C. Sayem El Daher et al. (1984) found that putrescine, spermidine, spermine, cadaverine, and tyramine concentrations correlated with both time (12 d) and temperature (4, 7, and 10◦ C) of storage, while histamine concentration was apparently unaffected by storage conditions. In addition to storage temperature and time, biogenic amine production is dependent on packaging conditions (vacuum or modified atmosphere), which decisively influence microbial flora (Wortberg and Woller, 1982; Dainty et al., 1986; Edwards et al., 1987). Vacuum or modified atmosphere packaging, commonly used to prolong the shelf life of these kinds of products, entails altering O2 levels and, hence, the growth and selection of some microorganism strains that can affect the production of biogenic amines. For example, Enterobacter cloacae produces half the amount of putrescine in an aerobic atmosphere and Klebsiella pneumoniae synthesizes significantly less cadaverine and produces more putrescine in anaerobic conditions (Halász et al., 1994). Nadon et al. (2001) reported that by using CO2 atmospheres it was possible to prolong the shelf life of pork stored at −1.5◦ C by up to 13 wk, as this limited the formation of biogenic amines, chiefly during the first 2 wk. Freezing and frozen storage can cause chemical and structural changes in meat, depending on species and storage conditions (temperature, duration, temperature fluctuations, etc.). Most decarboxylases are unstable to freezing; for example, histidine decarboxylase becomes inactive after 8–15 d at −20◦ C (Halász et al., 1994; Silla, 1996; Chen et al., 1994). Although dependent on conditions, the effect of freezing and frozen storage on microbial growth and enzymatic activity does not generally favor the formation of biogenic amines. The existing concentrations must, therefore, come mainly from the fresh product and will be strongly influenced by prefreezing treatment and storage conditions. In this connection, Hernandez-Jover et al. (1996b) found that the proportion of biogenic amines remained stable in pork stored for 12 d at −18◦ C. On the other hand, other authors (Halász et al., 1994) have reported an increase of cadaverine and putrescine and a decrease of histamine, spermidine, and spermine after 8 d at −20◦ C. There are various meat products that are usually prepared in the meat industry with frozen meat rather than fresh meat. However, there are few studies that attempt to assess the impact of frozen meat on the formation of biogenic amines in processed products. Maijala et al. (1995) reported that the thawing time of raw materials affected microbial flora and the amounts of free amino acids available as precursors and, hence, the formation of biogenic amines. On the other hand, Bover-Cid et al. (2001b) found that the formation of biogenic amines during ripening of dry sausage made with thawed pork was dependent on the use of a starter culture rather than the thawing time of the raw materials. Processing of Meat Products The different types of processing and storage conditions that are applied to meat products can influence the formation of biogenic amines in them. It has generally been found that cooking processes reduce biogenic amine concentrations to between 4 and 10 times less than in cured and ripened products, mainly due to the elimination of biogenic amines in cooking loss (Lakritz et al., 1975). Given that cooking does not favor amine formation, its presence in cooked meat products (Table 1) would appear to be closely linked to the quality of the meat raw material used; high concentrations of biogenic amines indicate “low quality” meat containing high concentrations of these compounds at origin (Paulsen et al., 1997; Bover-Cid et al., 2001b). Another factor to be considered is the thermal treatment conditions (i.e., BIOGENIC AMINES IN MEAT final temperature or heating rate); these influence decarboxylase, which in some cases, becomes inactive (Maija et al., 1995; Kebary et al., 1999) at over 65◦ C, a temperature reached in most cooked products. The cadaverine and putrescine detected in these meat products (Table 1) merit special attention, in that during heating they can undergo transformation to pyrrolidine and piperidine, respectively; if they react with the nitrites in the meat, they can form nitrosamines, which are highly carcinogenic (Patterson and Mottram, 1974; Wathensen et al., 1975). Generally, although initially all boiled food contains low levels of biogenic amines, the storage system applied to these kind of products should be taken into account, because it could increase the final level of biogenic amines. Such an increase implies a heightened toxicological risk in the product. Fermentation processes also generally promote the formation of biogenic amines, and in fact, this is the group of meat products that presents the greatest amount and diversity of these compounds (Table 1). Fermented products contain large quantities of microorganisms, accompanied by proteolysis giving rise to high concentrations of the amino acids constituting the nutrient required by the bacteria and the substrate on which decarboxylase enzymes work. However, these circumstances are affected by several factors, such as the temperature of the medium, the microorganisms present, and the presence of additives. The temperature at which fermentation takes place (usually between 7–28◦ C), influences the formation of biogenic amines; indeed, it has been suggested that temperature could be a very useful parameter for preventing tyramine formation in dry sausage, chiefly by assuring conditions favorable to starter growth (Maijala et al., 1995; Eerola et al., 1998). One explanation for the influence of processing temperature is that the higher fermentation temperature gives the starter culture the opportunity to outgrow nonstarter lactic acid bacteria. Maijala et al. (1995) detected low levels of amines in the final processing phase at a higher temperature (24◦ C). However, Kranner et al. (1991) also found that histamine formation was reduced when the ripening temperature was reduced (7–18◦ C). These results suggest that low temperatures can make for improved quality and longer shelf-life. In some cases, the presence of biogenic amines in fermented products has been attributed to poor quality of raw materials and defective processing. The quality of raw meat material is an essential factor in the formation of these products (Maijala et al., 1995). Nevertheless, while the source of biogenic amines in ripened meat products may derive partly from the raw materials, the largest proportion appears to arise in the various processing stages (Maijala et al., 1995; Vidal-Carou et al., 1990; Bover-Cid et al., 2001a). However, not all processing stages influence amine formation equally. Some authors (Maijala and Eerola, 1993; Santos-Buelga et al., 1986) have reported higher biogenic amine concentrations during ripening than during drying and salting. Zee et al. (1983) and Hernandez-Jover et al. (1997) reported similar results, also noting a reduction in the concentration of biogenic amines. This might be explained by a decrease of Aw, which, combined with fat, could inhibit biogenic 497 amine formation to some extent (Eerola et al., 1996; Treviño et al., 1997). On the other hand, Spinelli, Labritz, and Wasserman (1974) reported no significant change in the presence of these amines during the drying and smoking of pork bellies. The additives used in preparation of fermented products also are important factors in the formation of biogenic amines during this process, and their effect appears to depend on several factors (concentration, processing conditions, and others). However, the results in this respect have been contradictory. Vandekerckhove (1977) found that none of the various types of sugar used for ripening dry fermented sausages influenced the formation of biogenic amines, while other authors (Bover-Cid et al., 2001a) have reported that the sugar limited biogenic amine formation in fermented sausages. It has been demonstrated that the acidification produced by these sugars in the fermentation process influences the formation of biogenic amines (Arnold and Brown, 1978; Smith, 1980; Masson et al., 1996). The optimum glucose concentration for the formation of decarboxylase enzymes is .5–2%, whereas concentrations above 3% inhibit it. This is related to the effect of reduced pH on microorganism growth and the activity of enzymatic systems (Kranner et al, 1991; Masson and Montel, 1995; Hernandez-Jover et al., 1997). Bover-Cid et al. (2001a) reported that sodium sulphite in sausages inhibited cadaverine production and promoted production of tyramine and putrescine. Potassium sorbate, a microbial inhibitor, may be used to limit the formation of biogenic amines in foods (Shalaby, 1996). Various authors (Santos et al., 1985; Straub et al., 1994) also have reported that neither potassium nitrate nor potassium nitrite affected the production of biogenic amines in ripened meat products, such as salchichón, but that the presence of these additives in soy hamburger texturizers produced appreciable concentrations of tyramine. Besides, salt (NaCl) significantly affects biogenic amine production, mainly through its role in reducing Aw. It has been reported that 30 g NaCl/l in fermented sausages enhanced tyramine production by promoting the growth of Lactobacillus curvatus (Straub et al., 1994). Other factors, like sausage diameter in fermented products, also have been reported to affect the formation of biogenic amines during ripening (Bover-Cid et al., 1999). These authors found that the concentrations of biogenic amines in largerdiameter sausages were greater than in sausages of smaller diameter; moreover, the concentration was higher in the central part than in the ends. This could be related to Aw, which influences the growth of microorganisms in products of this type (Eerola et al., 1996). CONCLUSIONS Biogenic amines are very frequently involved in human pathologies the world wide: neurological disorders, gastrointestinal diseases, abnormal immune responses, cancer, etc. They are potentially toxic. They could be used as quality indexes in food, and they are known to occur in commonly-consumed 498 C. RUIZ-CAPILLAS AND F. JIMÉNEZ-COLMENERO foods, like meat and meat products. There is, therefore, a need for thorough research into their formation and their possible effects on consumers, in order to avoid new food safety problems. More research is needed to evaluate the impact of the factors discussed, regarding biogenic amine formation and to shed some light on how toxic compunds of this kind could affect consumers. Such research would be useful, with regard to technological and biochemical aspects and aspects relating to toxicology and/or mutagenicity. We need to expand our knowledge of technological factors relating to meat raw materials and meat processing that influence the amount and proportion of biogenic amine formation. Their importance varies according to the type of product and the technological treatment used. Many of the factors that promote or inhibit biogenic amine formation can be considerably altered by the application of emerging technologies (e.g., vacuum cooking, high pressure, and modified atmospheres) or meat product reformulation (incorporation of new ingredients/additives, changes in cooking temperature, reduction of fat content, salt, and so on). Such knowledge would be helpful in introducing compositional and/or technological changes that reduce the formation of these amines. Further research also is needed for the activity of the decarboxylase enzymes responsible for their formation and the factors that modulate this activity. At the same time, research into toxicity and/or mutagenicity is required to gain new and useful information about the toxicity of biogenic amines and possible synergisms. And again, we need to expand our understanding of the relationship between dose and effect, in order to explain some of the outbreaks that have occurred after the intake of foods containing different concentrations of biogenic amines. In recent years, there have been some studies of in vitro toxicity and mutagenicity of biogenic amines, but these have basically addressed the effects of compounds taken individually. All the proposed lines of research would make it possible to limit the presence of biogenic amines in raw materials and/or end products and would help health authorities to set safe, legal limits for consumers on biogenic amines in general and not only on histamine. Regarding the presence of biogenic amines in meat, it also would be useful to study their role in the formation of certain flavor precursors in meat and meat products and their connection with antioxidant action (Lovaas, 1991). There are a number of possibilities to be explored here. We, therefore, think it is essential to combine efforts in the technological, biochemical, and toxicological spheres, with a clear end in view—namely, to improve and guarantee consumer safety, an issue that is of increasing concern today. ACKNOWLEDGEMENTS Claudia Ruiz-Capillas acknowledges funding support from the Ministry of Science and Technology (Project Ramon y Cajal and AGL2003-00454). REFERENCES Arnol, S.H. and Brow, W.D. 1978. Histamine toxicity from fish products. Adv. Food Res., 24:113–154. Badolo, L., Gelbcke, M., Dubois, J., and Hanocq, M. 1998. Synthesis and cytotoxicity studies of new analogs of polyamines. Pharmazi., 53:15–19. Baley, G.S. and Williams, D.E. 1993. Potential mechanisms for food-related carcinogens and anticarcinogens. Food Technol., 47:105–118. Banwart, G.J. 1981. Basic Food Microbiology. AVI publishing Company Inc., Westport, CN. Bardócz, S. 1995. Polyamines in food and their consequences for food quality and human health. Trends Food Sci. Tech., 6:341–346. Bauer, F., Potzelberge, D., Hellwing, E., and Paulsen, P. 1996. Formation of biogenic amines in fresh meat packed in oxygen permeable foil. In: Proceedings of the 42nd International Congress of Meat Science and Technology., pp. 552–553. Lillehammer, Norway. Bover-Cid, S., Schoppen, S., Izquierdo-Pulido, M., and Vidal-Carou., M.C. 1999. Relationship between biogenic amine contents and the size of dry fermented sausages. Meat Sci., 51:305–311. Bover-Cid, S., Izquierdo-Pulido, M., and Vidal-Carou M.C. 2001a. Changes in biogenic amines and polyamine contents in slightly fermented sausages manufactured with and without sugar. Meat Sci., 57:215–221. Bover-Cid, S., Izquierdo-Pulido, M., and Vidal-Carou., M.C. 2001b. Effectiveness of a Lactobacillus sakei starter culture in reduction of biogenic amine accumulation as a function of the raw material quality. J. Food Protect., 64:367– 373. Chen, C.M., Lin, L.C., and Yen, G.G. 1994. Relationship between changes in biogenic amine contents and freshness of pork during storage at different temperatures. J. Chin. Agri. Chem. Soc., 32:47–60. Dainty, R.H. Edwards, R.A., Hibard, C.M., and Ramantanis, S.V. 1987. Amines in fresh beef of normal pH and the role of bacteria in changes in concentration observed during storage in vacuum packs at chill temperatures. J. Appl. Bacteriol., 63:427–434. Directive 91/439/EEC. Directive of 22 July 1991 establishing standards to be applied to the production and commercialization of fishery products. Official Journal of the European Communities L268:15–34. Durlu-Ozkaya, F., Ayhan, K., and Vural, N. 2001. Biogenic amines produced by Enterobacteriaceae isolated from meat products. Meat Sci., 58:163–166. Edwards, R.A., Dainty, R.H., and Hibard, C.M. 1985. Putrescine and cadaverine formation in vacuum packed beef. J. Appl. Bacteriol., 58:13–19. Edwards, R.A., Dainty, R.H., Hibard, C.M., and Ramantanis, S.V. 1987. Amines in fresh beef of normal pH and the role of bacteria in changes in concentration observed during storage in vacuum packs at chill temperatures. J. Appl. Bacteriol., 63:427–434. Edwards, R.A., Daintry, R.H., and Hibard, C.M. 1983. The relationship of bacterial numbers and types to diamine concentration in fresh and aerobically stored beef, pork, and lamb. J. Food Technol., 18:777–788. Eerola, H.S., Roig-Sagués, A.X., and Hirvi., T.K. 1998. Biogenic amines in Finnish dry sausages. J. Food Safety., 18:127–138. Eerola, S., Maijala, R., Roig Sagués, A.X., Salminen, M., and Hirvi, T. 1996. Biogenic amines in dry sausages as affected by starter culture and contaminant amine-positive Lactobacillus. J. Food Sci., 61:1243–1246. Eitenmiller, R.R., Koehler, P.E., and Reagan, J.O. 1978. Tyramine in fermented sausages: Factors affecting formation of tyramine and tyrosine decarboxylase. J. Food Sci., 43:689–693 Food and Drug Administration (FDA). 1990. Decomposition of histamines; raw, frozen tuna and malu malu, canned tuna, and related species. Revised compliance policy guide, Availability-Federal Register, 60, (149), 39574– 39756. Guerrero-Legarreta, I. and Chavez-Gallardo, A.M. 1991. Detection of biogenic amines as meat spoilage indicators. J. Muscle Foods., 2:263–278. Halász, A., Baráth, A., Simon-Sarkadi, L., and Holzapfel, W. 1994. Biogenic amines and their production by microorganisms in food. Trends Food Sci. Tech., 5:42–49. Hernández-Jover, T., Izquierdo-Pulido, M., Veciana-Nogues, M.T., and VidalCarou, M.C. 1996a. Ion-pair high performance liquid chromatographic BIOGENIC AMINES IN MEAT determination of biogenic amines in meat and meat products. J. Agr. Food Chem., 44:2710–2715. Hernández-Jover, T., Izquierdo-Pulido, M., Veciana-Nogues, M.T., and VidalCarou, M.C. 1996b. Biogenic amine sources in cooked cured shoulder pork. J. Agr. Food Chem., 44:3097–3101. Hernández-Jover, T., Izquierdo-Pulido, M., Veciana-Nogues, M.T., MarinéFont, A., and Vidal-Carou, M.C. 1997. Effect of starter cultures on biogenic amine formation during fermented sausage production. J. Food Protect., 60:825–830. Ienistea, C. 1971. Bacterial production and destruction of histamine in foods, and food poisoning caused by histamine. Nahrung., 15:109–113. Kebary, K.M.K., El-Sonbaty, A.H., and Badawi, R.M. 1999. Effects of heating milk and accelerating ripening of low fat ras cheese on biogenic amines and free amino acids development. Food Chem., 64:67–75. Kranner, P., Bauer, F., and Hellwig, E. 1991. Investigation on the formation of histamine in raw sausages. In: Proceeding of the 37 International Congress of Meat Science and Technology. pp. 889–895. Kulmbach, Germany. Lakritz, L., Spinelli, A.M., and Wasserman, A.E. 1975. Determination of amines in fresh and processed pork. J. Agr. Food Chem., 23:344–346. Leigh, L, Lehane L., and Olley J. 2000. Histamine fish poisoning revisted. Int. J. Food Microbiol., 58:1–37. Lipp, E.K. and Rose, J.B. 1997. The role of seafood in foodborne diseases in the United States of America. Rev Sci Tech OIE (Off Int Epizoot), 16:620–640. Lovaas, E. 1991. Antioxidative effects of polyamines. J. Amer. Oil Chem Soc., 68:353–358. Lux, G.D., Marton, L.J., and Baylin, S.B. (1980). Ornithine decarboxylase is important in intestinal mucosal maturation and recovery from injury in rats. Science., 210:195–198. Maijala, R. and Nurmi, E. 1995. Influence of processing temperature on the formation of biogenic amines in dry sausages. Meat Sci., 39:9–22. Maijala, R. and Eerola, S. 1993. Contaminant lactic acid bacteria of dry sausages produce histamine and tyramine. Meat Sci., 35:387–395. Maijala, R., Eerola, S., Lievonen, S., Hill, P., and Hirvi, T. 1995. Formation of biogenic amines during ripening of dry sausages as affected by starter culture and thawing time of raw materials. J. Food Sci., 60:1187–1190. Masson, F. and Montel, M.C. 1995. Les amines biogènes dans les produits carnés. Viandes et Produits Carnés., 16:3–7. Masson, F., Talon, R., and Montel, M.C. 1996. Histamine and tyramine production by bacteria from meat products. Int. J. Food Microbiol., 32:199–207. McCabe, B.J. 1986. Dietary tyramine and other precursor amines in MAOI regimens: A review. J. Am. Diet Assoc., 86:1059–1064. Mietz, J.L. and Karmas, E. 1977. Polyamine and histamine content of rockfish, salmon, lobster and shrimp as an indicator of decomposition. J. Assoc. Offic Anal. Chem., 61:139–145. Nadon, C.A. Ismond, M.A.H., and Holley, R. 2001. Biogenic amines in vacuumpackaged and carbon dioxide-controlled atmosphere-packaged fresh pork stored at −1.5◦ C. J. Food Protec., 64:220–227. Nakamura, M., Wada, Y., Sawaya, H., and Kawabata, T. 1979. Polyamine content in fresh and processed pork. J. Food Sci., 44:515–523. Ordoñez, J.A., De Pablo, B., Perez de Castro, B., Asensio, M.A., and Sanz, B. 1991. Select chemical and microbiological changes in refrigerated pork stored in carbon dioxide and oxygen enriched atmospheres. J. Agr. Food Chem., 39:668–672. Patterson, R.L. and Mottram, D.S. 1974. The occurrence of volatile amines in uncured and cured pork their possible role in nitrosamine formation in bacon. J Sci Food Agric., 25:1419–1425. Paulsen, P. Bauer, F., and Vali, S. 1997. Biogenic amines in fermented sausage. 1. Methods for the determination of biogenic amines. Fleischwirtschaft., 77:450–452. Rawles, D.D., Flick, G.J., and Martin, R.E. 1996. Biogenic amines in fish and shellfish. In: Advances in Academic Press., pp. 329–365 Rice, S.L., Eitenmiller, R.R., and Koehler, P.E. 1976. Biologically active amines in food: A review. J. Milk Food Technol., 39:353–358. 499 Roig-Sagués, A.X. and Eerola, S. 1997. Biogenic amines in meat inoculated with Lactobacillus sake starter strains and an amine-positive lactic acid bacterium. Z. Lebensm. Unters. F. A., 205:227–231. Ruiz-Capillas, C. and Moral, A. 2001. Production of biogenic amines and their potential use as quality control indices for hake (Merluccius merluccius, L.) stored in ice. J. Food Sci., 66:1030–1032. Santos, C., Jalón, M., and Marine, A. 1985. Contenido de tiramina en alimentos de origen animal. I. Carne, derivados cárnicos y productos relacionados. Rev. Agroquim Tecnol. Aliment., 25:362–368. Santos-Buelga, C., Peña-Egido, M.J., and Rivas-Gonzalo, J.C. 1986. Changes in tyramine during chorizo-sausage ripening. J. Food Sci., 51:518–519. Sattler, J., Hafner, D., Klotter, H.J., Lorenz, W., and Wagner, P.K. 1988. Foodinduced histaminosis as an epidemiological problem: Plasma histamine elevation and haemodynamic alterations after oral histamine administration and blockade of diamine oxidase (DAO). Agents and Actions, 23:361–365. Sayem-El-Daher, N., Simard, R.E., and Fillion, J. 1984. Changes in the amine content of ground beef during storage and processing. Lebensm-Wiss. Technol., 17:319–323. Shalaby, A.R. 1993. Survey on biogenic amines in Egyptian foods: Sausage. J. Sci. Food Agri., 62:291–293. Shalaby, A.R. 1996. Significance of biogenic amines to food safety and human health. Food Res. Int., 29:675–690. Silla, M.H. 1996. Biogenic amines: Their importance in foods. Int. J. Food Microbiol., 29:213–231. Slemr, J. and Beyermann, K. 1985. Concentration profiles of diamines in fresh and aerobically stored pork and beef. J. Agr. Food Chem., 33:336–339. Slemr, T. 1981. Biogenic amine als potentioller chemescher qualitatsindikator fur fleish. Fleischwirtsschaft, 61:921–924. Smith, J.S., Kenney, P.B., Kastner, C.L., and Moore, M.M. 1993. Biogenic amines formation in fresh vacuum-packaged beef during storage at 1◦ C for 120 days. J. Food Protect., 56:497–500, 532. Smith, T.A. 1980. Amines in food. Food Chem., 6:169–200. Spinelli, A.M., Lakritz, L., and Wasserman, A.E. 1974. Effects of processing on the amine content of pork bellies. J. Agr. Food Chem., 22:1026–1029. Stratton, J.E., Hutkins, R.W., and Taylor, S.L. 1991. Biogenic amines in cheese and other fermented food: A review. J. Food Protect., 54:460–470. Straub, B.W., Tichaczek, P.S., Kicherer, M., and Hammes, W.P. 1994. Formation of tyramine by Lactobacillus curvatus LTH 972. Z. Lebensm. Unters. F. A., 1:9–12. Taylor, S.L. 1985. Histamine poisoning associated with fish, cheese, and other foods. pp. 17. Monografia (VPH/FOS/85.1), W.O.H., Geneva, Switzerland. ten Brink, B., Damink, C., Joosten, H.M.L.J., and Huis in’t Veld, J.H.J. 1990. Occurrence and formation of biologically active amines in foods. Int. J. Food Microbiol., 11:73–84 Treviño, E., Beil, D., and Steinhart, H. 1997. Formation of biogenic amines during the maturity process of raw meat products, for example of cervelat sausage. Food Chem., 60:521–526. Tuley, L. 1996. The search is on for low fat meat. International Food Manufacture., 1:29–32. Vandekerckhove, P. 1977. Amines in dry fermented sausage. J. Food Sci., 42: 283–285. Vidal-Carou, M.C., Izquierdo, M.L., Matı́n, M.C., and Mariné, A. 1990. Histamina y tiramina en derivados cárnico. Rev. Agroquim Tecnol. Aliment, 30:102–108. Warthensen, J.J., Scanlan, R.A., Bills, D.D., and Libbely, L.M. 1975. Formation of heterocyclic N-nitrosamines from the reaction of nitrite and selected primary diamines and amino acids. J. Agr. Food Chem., 23:898–902. Wortberg, B. and Woller, R. 1982. Quality and freshness of meat and meat products as related to their content of biogenic amines. Fleischwirtsch, 62:1457– 1463. Zee, J.A., Simard, R.E., and Heureux, L.L. 1983. Evaluation of analytical methods for determination of biogenic amines in fresh and processed meat. J. Food Protect., 46:1044–1049.