A study on phytogenotoxicity induced by biogenic amines: cadaverine and putrescine.

Among the compounds present in necro-leachate, a liquid released during the process of decomposition of the human body, are the biogenic amines cadaverine and putrescine. Although some studies on necro-leachate have indicated a potential ecotoxicological and public health risk associated with it, the research on this type of contamination is still rather limited. This study presents information about the phytotoxic and cytogenotoxic potential of cadaverine and putrescine, evaluated separately and within a mixture. Phytotoxicity was evaluated through a germination test, the initial growth of seedlings with Lactuca sativa, and cytogenotoxicity through chromosomal aberration and micronucleus tests with Allium cepa. The L. sativa results showed a phytotoxic effect for the evaluated amines, by reducing root (> 90%) and hypocotyl (> 80%) elongation. The co-exposure of cadaverine and putrescine potentiated cytogenotoxic activity by aneugenic action in the meristematic cells of A. cepa. From this result, it is possible to infer the eco-toxicogenic potential of cadaverine and putrescine. This study not only highlights the importance of the phytotoxic and cytogenotoxic effects of these amines but also emphasizes the urgent need for further investigation into contamination originating from cemetery environments. By evaluating the risks associated with necro-leachate, this research is aimed at informing global efforts to protect ecological and public health.

Evaluation of the ecotoxicological effects of biogenic amines derived from cadaverous putrefaction on springtails Folsomia candida.

Necro-leachate, a liquid released during cadaveric decomposition, is considered the main culprit for impacts on cemetery environments. The biogenic amines cadaverine and putrescine make up part of the composition of necro-leachate and have a certain toxicity to different organisms. Springtails are among the most used bioindicators to assess the impacts of soil contaminants. As there are no data on the acute and chronic toxicity of springtails exposed to cadaverine and putrescine, the objective of this study was to evaluate the toxic potential of both amines, under the behavioral effect of avoidance and reproduction in the species Folsomia candida. Springtails were exposed to soils contaminated with different concentrations of cadaverine and putrescine, and different mixtures of these amines. To evaluate the avoidance and reproduction test, the individuals were exposed for periods of 48 h and 28 days, respectively. The results obtained in the avoidance test showed that springtails exhibited a preference for the treated soil in both isolated and mixed treatments. The chronic evaluation assays showed that the reproduction was affected, particularly in the treatments with combined amines, resulting in a reduction in the total number of juveniles. From the results, it is possible to infer that the methods applied in this research have provided information that will contribute to a better understanding of the toxicity of putrefactive biogenic amines, since there exist few ecotoxicological studies carried out with these amines, and especially with those from cemetery environments.

Amino Acid and Biogenic Amine Adductions Derived from Reactive Metabolites.

Reactive metabolites (RMs) are products generated from the metabolism of endogenous and exogenous substances. RMs are characterized as electrophilic species chemically reactive to nucleophiles. Those nucleophilic species may be nitrogen-containing bio-molecules, including macro-biomolecules, such as protein and DNA, and small biomolecules, i.e., amino acids (AAs) and biogenic amines (BAs). AAs and BAs are essential endogenous nitrogen-containing compounds required for normal development, metabolism, and physiological functions in organisms, through participating in the intracellular replication, transcription, translation, division and proliferation, DNA and protein synthesis, regulation of apoptosis, and intercellular communication activities. These biological amines containing an active lone pair of electrons on the electronegative nitrogen atom would be the proper N-nucleophiles to be attacked by the abovementioned RMs. This review covers an overview of adductions of AAs and BAs with varieties of RMs. These RMs are formed from metabolic activation of furans, naphthalene, benzene, and products of lipid peroxidation. This article is designed to provide readers with a better understanding of biochemical mechanisms of toxic action.

Biogenic amines: formation, action and toxicity - a review.

Biogenic amines (BA) are organic compounds commonly found in food, plants and animals, as well as microorganisms that are attributed with the production of BAs. They are formed as an effect of a chemical process: the decarboxylation of amino acids. Factors determining the formation of BAs include the availability of free amino acids and the presence of microorganisms that show activity with respect to carrying out the decarboxylation process. On the one hand, BAs are compounds that are crucial for maintaining cell viability, as well as the proper course of the organism's metabolic processes, such as protein synthesis, hormone synthesis and DNA replication. On the other hand, despite their positive effects on the functioning of the organism, an excessive content of BAs proves to be toxic (diarrhea, food poisoning, vomiting, sweating or tachycardia). Moreover, they can accelerate carcinogenesis. Amines are a natural component of plant and animal raw materials. As a result of the proven negative effects of amines on living organisms, the reduction of these compounds should be the subject of scientific research. The present review aims to synthesize and summarize the information currently available on BAs, as well as discuss the interpretation of the results. © 2020 Society of Chemical Industry.

New insights into the impacts of huangjiu compontents on intoxication.

Although huangjiu is a popular alcoholic beverage in China, the occurrence of quick-intoxication suppresses huangjiu consumption and impedes development of the huangjiu industry. In this study, the Cryprinus carpio intoxication model was used to compare the differences in intoxication effect of alcoholic beverages and to assess the impacts of huangjiu components on intoxication for the first time. Exposure to huangjiu led to the most rapid physical imbalance of C. carpio, followed by red wine and Western liquor. Higher alcohols, biogenic amines and aldehydes could cause physical imbalance of fish by themselves, and synergistic effects were observed when combined with ethanol. 2-Phenylethanoland and isopentanol had the greatest positive effect on huangjiu intoxication, followed by histamine and phenethylamine. No synergistic effect was observed between individual aldehydes and ethanol. Identification of these impactful huangjiu components provides a new perspective on the establishment of more rigorous control on the quality and flavor of huangjiu.

Biogenic Amines and Free Amino Acids in Traditional Fermented Vegetables-Dietary Risk Evaluation.

Biogenic amines (BAs) are low molecular weight organic bases. BAs occurring naturally in living organisms are responsible for a number of vital functions, including (in humans) secretion of gastric acids, controlling body temperature, differentiation and growth of cells, immune reactions, and brain activity. However, if oversupplied with food, BAs may cause food poisoning and produce undesirable effects. Nine BAs and eight free amino acids (FAAs) were determined in 85 samples of 19 different varieties of fermented vegetables available on the Polish retail market. Both BA and FAA levels differed significantly among various varieties of the studied fermented vegetables. Averages for the sum of all tested BAs ranged from 30.29 ± 16.43 mg·kg-1 in fermented olives to 612.1 ± 359.33 mg·kg-1 in fermented Brussels sprout. BA profiles were dominated by putrescine (42%), tyramine (20%), cadaverine (18%), and histamine (8%); jointly, the four amines amounted to 88% of all nine studied BAs. The combined level of the latter four BAs was calculated for each vegetable variety as the so-called BA index (BAI). On that basis, the risk of BA-related adverse health effects has been assessed as high/medium/low in 6/3/10 of all 19 studied varieties of fermented vegetables. Brussels sprout and broccoli turned out to be the most risky vegetables from that point of view (BAI above 400 mg·kg-1). FAA levels ranged from 54.8 ± 12.76 (fermented olives) to 3917.42 ± 1528.73 mg·kg-1 (fermented garlic). The high content of FAAs may increase the risk of forming toxic amounts of BAs, depending on characteristics of the current and added microflora as well as on environmental and technological conditions the product is subjected to.

The biogenic amines putrescine and cadaverine show in vitro cytotoxicity at concentrations that can be found in foods.

Putrescine and cadaverine are among the most common biogenic amines (BA) in foods, but it is advisable that their accumulation be avoided. Present knowledge about their toxicity is, however, limited; further research is needed if qualitative and quantitative risk assessments for foods are to be conducted. The present work describes a real-time analysis of the cytotoxicity of putrescine and cadaverine on intestinal cell cultures. Both BA were cytotoxic at concentrations found in BA-rich foods, although the cytotoxicity threshold for cadaverine was twice that of putrescine. Their mode of cytotoxic action was similar, with both BA causing cell necrosis; they did not induce apoptosis. The present results may help in establishing legal limits for both putrescine and cadaverine in food.

The dietary biogenic amines tyramine and histamine show synergistic toxicity towards intestinal cells in culture.

Tyramine and histamine are the biogenic amines (BA) most commonly found at high concentrations in food; they may even appear together at toxic concentrations. The present work examines, via real-time cell analysis, whether histamine and tyramine show synergistic toxicity towards intestinal cell cultures. Employing a constant equipotency ratio, their interaction was examined via the combination index (CI) method of Chou & Talalay. Co-treatment with tyramine and histamine was associated with a stronger cytotoxic effect than was treatment with either BA or on its own. Indeed, a synergistic interaction (CI<1) was observed in the range of concentrations found in foods. The results also show that histamine, at concentrations below the legal limit, increases the cytotoxicity of tyramine at concentrations frequently reached in some foods. The synergistic cytotoxicity of tyramine and histamine should be taken into account when establishing legal limits designed to ensure consumer safety.

Comparative analysis of the in vitro cytotoxicity of the dietary biogenic amines tyramine and histamine.

Tyramine and histamine, the most toxic biogenic amines (BA), are often found in high concentrations in certain foods. Prompted by the limited knowledge of BA toxicity, and increasing awareness of the risks associated with high intakes of dietary BA, the in vitro cytotoxicity of tyramine and histamine was investigated. Tyramine and histamine were toxic for HT29 intestinal cell cultures at concentrations commonly found in BA-rich food, as determined by real-time cell analysis. Surprisingly, tyramine had a stronger and more rapid cytotoxic effect than histamine. Their mode of action was also different, while tyramine caused cell necrosis, histamine induced apoptosis. To avoid health risks, the BA content of foods should be reduced and legal limits established for tyramine.

Safety assessment of the biogenic amines in fermented soya beans and fermented bean curd.

To evaluate the safety of biogenic amines, high performance liquid chromatography (HPLC) was used to evaluate the levels of biogenic amines in fermented soya beans and fermented bean curd. In fermented soya beans, the total biogenic amines content was in a relatively safe range in many samples, although the concentration of histamine, tyramine, and ß-phenethylamine was high enough in some samples to cause a possible safety threat, and 8 of the 30 samples were deemed unsafe. In fermented bean curd, the total biogenic amines content was more than 900 mg/kg in 19 white sufu amples, a level that has been determined to pose a safety hazard; putrescine was the only one detected in all samples and also had the highest concentration, which made samples a safety hazard; the content of tryptamine, ß-phenethylamine, tyramine, and histamine had reached the level of threat to human health in some white and green sufu samples, and that may imply another potential safety risk; and 25 of the 33 samples were unsafe. In conclusion, the content of biogenic amines in all fermented soya bean products should be studied and appropriate limits determined to ensure the safety of eating these foods.

Simultaneous determination of selected biogenic amines in alcoholic beverage samples by isotachophoretic and chromatographic methods.

A simple and useful method for the determination of biogenic amines in beverage samples based on isotachophoretic separation is described. The proposed procedure permitted simultaneous analysis of histamine, tyramine, cadaverine, putrescine, tryptamine, 2-phenylethylamine, spermine and spermidine. The data presented demonstrate the utility, simplicity, flexibility, sensitivity and environmentally friendly character of the proposed method. The precision of the method expressed as coefficient of variations varied from 0.1% to 5.9% for beverage samples, whereas recoveries varied from 91% to 101%. The results for the determination of biogenic amines were compared with an HPLC procedure based on a pre-column derivatisation reaction of biogenic amines with dansyl chloride. Furthermore, the derivatisation procedure was optimised by verification of concentration and pH of the buffer, the addition of organic solvents, reaction time and temperature.

Biodiversity and safety aspects of yeast strains characterized from vineyards and spontaneous fermentations in the Apulia Region, Italy.

This work is the first large-scale study on vineyard-associated yeast strains from Apulia (Southern Italy). Yeasts were identified by Internal Transcribed Spacer (ITS) ribotyping and bioinformatic analysis. The polymorphism of interdelta elements was used to differentiate Saccharomyces cerevisiae strains. Twenty different species belonging to 9 genera were identified. Predominant on the grape surface were Metschnikowia pulcherrima, Hanseniaspora uvarum and Aureobasidium pullulans, whereas M. pulcherrima and H. uvarum were dominant in the early fermentation stage. A total of 692 S. cerevisiae isolates were identified and a number of S. cerevisiae strains, ranging from 26 to 55, was detected in each of the eight fermentations. The strains were tested for biogenic amines (BAs) production, either in synthetic media or grape must. Two Pichia manshurica, an Issatchenkia terricola and a M. pulcherrima strains were able to produce histamine and cadaverine, during must fermentation. The production of BAs in wine must was different than that observed in the synthetic medium. This feature indicate the importance of an "in grape must" assessment of BAs producing yeast. Overall, our results suggest the importance of microbiological control during wine-making to reduce the potential health risk for consumer represented by these spoilage yeasts.

Chromatographic methods for biogenic amines determination in foods of animal origin / Métodos cromatográficos para determinar aminas biogênicasem alimentos de origem animal

Biogenic amines (BAs) are formed as a result of specific free amino acid decarboxylation. Analysis of these metabolitesmay be of great importance to determine food quality and for monitoring the levels of biogenic amines such as histamineand tyramine related to intoxication episodes in humans. Chromatography is a chemistry separation technique usedto characterize biogenic amines in foods. Variations of this technique (liquid, thin layer and gas chromatography)have been widely applied; however, the food matrix complex requires that changes in the methodology of extraction,derivatization and detection must be performed according to each group of foods. High-performance liquidchromatography is the most widely used chromatographic method applied for biogenic amines in foods. However, dueto the current importance of biogenic amines in quality control and consumer safety, researchers try to develop newmethods for a fast, reliable analysis of foods in the market. This review presents some chromatographic techniquesapplied to monitoring BAs in different foods of animal origin...

Aminas biogênicas são formadas como resultado da descarboxilação de aminoácidos livres específicos. A análise dessesmetabólitos é de grande importância na determinação da qualidade e monitoramento de biogênicas como histamina etiramina relacionadas com episódios de intoxicação em humanos. A cromatografia é uma técnica de separação químicausada para caracterizar aminas biogênicas. Variações da técnica (cromatografia líquida, em camada delgada e gasosa)têm sido amplamente usadas, porém a complexidade da matriz alimentar faz com que sejam realizadas mudanças nosprocessos de extração, derivatização e detecção em concordância com cada grupo de alimento. A cromatografia líquidade alta eficiência (CLAE) é o método mais utilizado na determinação de aminas biogênicas em alimentos. Contudo,devido à importância das aminas biogênicas no controle da qualidade e a segurança do consumidor, os pesquisadorestentam desenvolver novos métodos com o intuito de uma análise mais rápida e precisa para o controle de alimentos nomercado. O objetivo da revisão é apresentar algumas técnicas cromatográficas aplicadas no monitoramento de aminasbiogênicas em produtos de origem animal...

Aminas biogênicas nos alimentos: revisão de literatura / Biogenic amines in foods: a literature review

Muitos estudos têm sido realizados acerca da presença e quantificação de aminas biogênicas nos diversos alimentos devido ao seu potencial tóxico. Histamina, putrescina, cadaverina, tiramina, triptamina, espermidina e espermina se destacam como as principais, sendo a análise da presença na matriz alimentar uma importante ferramenta tanto para indicar o grau de frescor ou de deterioração dos alimentos como para prevenir efeitos deletérios à saúde humana. Esta revisão aborda a importância da presença das aminas biogênicas nos alimentos, os efeitos tóxicos e os métodos analíticos.

Biogenic amines in dairy products.

Biogenic amines (BA) are organic, basic, nitrogenous compounds with biological activity, mainly formed by the decarboxylation of amino acids. BA are present in a wide range of foods, including dairy products, and can accumulate in high concentrations. In some cheeses more than 1000 mg of BA have been detected per kilogram of cheese. The consumption of food containing large amounts of these amines can have toxicological consequences. Although there is no specific legislation regarding the BA content in dairy products, it is generally assumed that they should not be allowed to accumulate. Greater knowledge of the factors involved in the synthesis and accumulation of BA should lead to a reduction in their incidence in foods. This article focuses on the factors that affect BA production, in particular environmental conditions, the microorganisms that produce them, the genetic organization and regulation of the biosynthetic pathways involved, and the available methods for detecting the presence of BA or BA-producing microorganisms in dairy products.

A new method for the determination of biogenic amines in cheese by LC with evaporative light scattering detector.

This paper presents a new LC method with evaporative light scattering detection (ELSD), for the separation and determination of the biogenic amines (histamine, spermidine, spermine, tyramine, putrescine and ß-phenylethylamine) which are commonly present in cheese, as their presence and relative amounts give useful information about freshness, level of maturing, quality of storage and cheese authentication. The LC-ELSD method is validated by comparison of the results with those obtained through LC-UV determination, based on a pre-column dansyl chloride derivatisation step. The obtained data demonstrate that both methods can be interchangeably used for biogenic amines determination in cheese. The new LC-ELSD method shows good precision and permits to achieve, for standard solutions, limit of detection (LOD) values ranging from 1.4 to 3.6 mg L(-1) and limit of quantitation (LOQ) values ranging from 3.6 to 9.3 mg L(-1). The whole methodology, comprehensive of the homogenization-extraction process and LC-ELSD analysis, has been applied in the analysis of a typical Calabria (Southern Italy) POD cheese, known as Caciocavallo Silano. The most aboundant amine found was histamine, followed, in decreasing order, by tyramine, spermine, putrescine, ß-phenylethylamine and spermidine, for a total amount of 127 mg kg(-1). This value does not represent a possible risk for consumer health, according to the toxicity levels reported in literature and regarded as acceptable.

Influence of fertilisation with foliar urea on the content of amines in wine.

Amines are substances that could cause toxic effects in the consumer. The concentration of amines in wine depends on different factors such as grape variety, vinification conditions and nitrogen fertilisation of the vines. The aim of this work was to study the influence of the application of foliar urea on the concentration of amines in wine. To carry out the study, grapevines of Tempranillo variety were used. These grapevines were treated with foliar urea at two different concentrations: 2 and 4 kg N ha(-1). Treatment with foliar urea significantly increased (p < 0.05) the concentration of histamine in the wines compared with the control sample (65% in the treatment with 2 kg N ha(-1) and 93% in the treatment with 4 kg N ha(-1)), reaching higher concentrations than the threshold level where it could provoke toxic effects in the consumer (8-20 mg l(-1)). On the other hand, treatment with foliar urea did not increase the concentrations of other amines which could be toxic such as tyramine or phenylethylamine, nor amines such as putrescine which could enhance the toxic effect of histamine. In the case of the volatile amines containing secondary amine groups, the concentration of pirrolidine increased by 37% after treatment with 2 kg N ha(-1) and 61% after treatment with 4 kg N ha(-1).

Biogenic amines in fermented foods.

Food-fermenting lactic acid bacteria (LAB) are generally considered to be non-toxic and non-pathogenic. Some species of LAB, however, can produce biogenic amines (BAs). BAs are organic, basic, nitrogenous compounds, mainly formed through decarboxylation of amino acids. BAs are present in a wide range of foods, including dairy products, and can occasionally accumulate in high concentrations. The consumption of food containing large amounts of these amines can have toxicological consequences. Although there is no specific legislation regarding BA content in many fermented products, it is generally assumed that they should not be allowed to accumulate. The ability of microorganisms to decarboxylate amino acids is highly variable, often being strain specific, and therefore the detection of bacteria possessing amino acid decarboxylase activity is important to estimate the likelihood that foods contain BA and to prevent their accumulation in food products. Moreover, improved knowledge of the factors involved in the synthesis and accumulation of BA should lead to a reduction in their incidence in foods.

Biogenic amines in fish: roles in intoxication, spoilage, and nitrosamine formation--a review.

Biogenic amines are non-volatile amines formed by decarboxylation of amino acids. Although many biogenic amines have been found in fish, only histamine, cadaverine, and putrescine have been found to be significant in fish safety and quality determination. Despite a widely reported association between histamine and scombroid food poisoning, histamine alone appears to be insufficient to cause food toxicity. Putrescine and cadaverine have been suggested to potentiate histamine toxicity. With respect to spoilage on the other hand, only cadaverine has been found to be a useful index of the initial stage of fish decomposition. The relationship between biogenic amines, sensory evaluation, and trimethylamine during spoilage are influenced by bacterial composition and free amino acid content. A mesophilic bacterial count of log 6-7 cfu/g has been found to be associated with 5 mg histamine/100 g fish, the Food and Drug Administration (FDA) maximum allowable histamine level. In vitro studies have shown the involvement of cadaverine and putrescine in the formation of nitrosamines, nitrosopiperidine (NPIP), and nitrosopyrrolidine (NPYR), respectively. In addition, impure salt, high temperature, and low pH enhance nitrosamine formation, whereas pure sodium chloride inhibits their formation. Understanding the relationships between biogenic amines and their involvement in the formation of nitrosamines could explain the mechanism of scombroid poisoning and assure the safety of many fish products.

HPLC determination of free nitrogenous compounds of Centaurea solstitialis (Asteraceae), the cause of equine nigropallidal encephalomalacia.

Centaurea solstitialis (yellow star thistle) has been proven to cause equine nigropallidal encephalomalacia in horses. Over the last fifty years, nigropallidal encephalomalacia has been of interest to human medicine due to the possible connection with Parkinson's disease. Previous studies indicated the presence of neurotoxic nitrogenous compounds in polar extracts of the plant. In order to give a more detailed description of the nitrogen-containing fraction of C. solstitialis, various samples were collected at different development stages. Different aliquots of the same aqueous extract were directly derivatized with o-phthaldialdehyde and dansyl chloride and analyzed separately by reversed-phase HPLC. A complete profile of the free nitrogenous fraction of C. solstitialis was given and results obtained with the two derivatization procedures were compared. No particularly high level of free aspartic and glutamic acids, two potent neuroexcitotoxic amino acids, were found in polar extracts of the plant. Tyramine resulted to be the most important biologically active amine present in C. solstitialis (with a mean concentration of 2.0 mg/100 g of dry weight).