Potential of wine-associated lactic acid bacteria to degrade biogenic amines

Some lactic acid bacteria (LAB) isolated from fermented foods have been proven to degrade biogenic amines through the production of amine oxidase enzymes. Since little is known about this in relation to wine micro-organisms, this work examined the ability of LAB strains (n = 85) isolated from wines and other related enological sources, as well as commercial malolactic starter cultures (n = 3) and type strains (n = 2), to degrade histamine, tyramine and putrescine. The biogenic amine-degrading ability of the strains was evaluated by RP-HPLC in culture media and wine malolactic fermentation laboratory experiments. Although at different extent, 25% of the LAB isolates were able to degrade histamine, 18% tyramine and 18% putrescine, whereas none of the commercial malolactic starter cultures or type strains were able to degrade any of the tested amines. The greatest biogenic amine-degrading ability was exhibited by 9 strains belonging to the Lactobacillus and Pediococcus groups, and most of them were able to simultaneously degrade at least two of the three studied biogenic amines. Further experiments with one of the strains that showed high biogenic amine-degrading ability (L. casei IFI-CA 52) revealed that cell-free extracts maintained this ability in comparison to the cell suspensions at pH 4.6, indicating that amine-degrading enzymes were effectively extracted from the cells and their action was optimal in the degradation of biogenic amines. In addition, it was confirmed that wine components such as ethanol (12%) and polyphenols (75 mg/L), and wine additives such as SO₂ (30 mg/L), reduced the histamine-degrading ability of L. casei IFI-CA 52 at pH 4.6 by 80%, 85% and 11%, respectively, in cell suspensions, whereas the reduction was 91%, 67% and 50%, respectively, in cell-free extracts. In spite of this adverse influence of the wine matrix, our results proved the potential of wine-associated LAB as a promising strategy to reduce biogenic amines in wine.     

Determination of biogenic amines by RP-HPLC of the dabsyl derivates

An automated derivatization with dabsyl chloride in combination with a high-performance liquid Chromatographic analysis is described for the determination of biogenic amines in complex matrices. The sample clean-up procedure consists of an ultrafiltration step, resulting in average recoveries for cheese in the range of 88% to 100%. A linear relation between the area of the peak and amine concentration was observed between 0.5 and 500 pmol for all amines under investigation and the detection limits ranged between 0.34 and 0.76 pmol. The average repeatability of both the performance of the Chromatographic determination and the whole method, including sample preparation, examined using cheese samples containing different amounts of biogenic amines was found to be between 2.0% and 3.7%. The method was applied to the analysis of several types of food and feed, selected examples are given by the separation of dabsyl derivates from cheese, wine, and salami-type sausage.     

Formation of biogenic amines by Gram-negative bacteria isolated from poultry skin

The aim of this study is to explore production of biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) by 88 Gram-negative bacteria isolated from poultry skin (41 isolates of family Enterobacteriaceae, 21 isolates of genus Aeromonas, 16 isolates of genus Pseudomonas, and 10 isolates of other Gram-negative rods). Ion-exchange chromatography was employed to analyse the above mentioned amines. Enterobacteria were found to be the largest producers of amines with proved presence of tyramine, agmatine, putrescine, and cadaverine in cultivation broth after incubation of bacteria. Putrescine and cadaverine were the most abundant products. Presence of at least two biogenic amines, i.e. mainly concurrent presence of putrescine and cadaverine, was revealed in 19 enterobacteria strains. Eleven isolates classified into Aeromonas genus produced putrescine and five of them also formed cadaverine. The other observed biogenic amines (histamine, spermine and spermidine) were not found among tested isolates. Production of biogenic amines by any Pseudomonas family isolates was not proved.     

生物胺对食品安全和人类健康的重要性

胺是含氧的化合物,是由一个、两个或三个氢原子与氨以烃基或芳基结合的基团。在食物中,氨基酸在脱羧基作用下形成胺类,当这些胺类在活体中与氨基酸以脱羧基的方式结合后,就成为了生物性的胺,简称生物胺。     

Estimation of biogenic amines in foods

Biogenic amines in foods have been proposed as initiators of dietary-induced migraine. Studies of levels of 2-phenylethylamine, p-tyramine and the diamines histamine, putrescine and cadaverine in a number of Australian and other foods were made by ion exchange and high pressure liquid chromatographic methods. In the latter method the amines were chromatographed as their fluorescamine derivatives. The use of such derivatives also enabled confirmation of the identity of the amine by field desorption-mass spectrometry. Extraction procedures were devised to give satisfactory recoveries of amines in foods and to overcome problems associated with the binding of amines to other food constituents.     

Effects of specific lactic acid bacteria species on biogenic amine production by foodborne pathogen

The influences of lactic acid bacteria (LAB) on biogenic amines formation by foodborne pathogens (FBP) were investigated. Biogenic amines production by single and mix cultures was tested in histidine decarboxylase broth. All of the mix cultures (LAB with FBP) inhibited significantly (P < 0.05) the ammonia accumulation except for Escherichia coli with Lactococcus lactic subsp. lactic and Lactobacillus plantarum. Although LAB with most of the pathogen showed considerably stimulation effects on putrescine, cadaverine, spermine, 2‐phenylethylamine, histamine (HIS), tyramine, trimethylamine, dopamine and agmatine (AGM), some of the LAB with pathogens showed poor inhibition effect on different amines formation. HIS production by Klebiella pneumonia was 0.16 mg L^?1, whereas HIS value in presence of Lb. plantarum increased to 30.92 mg L^?1. Consequently, LAB inhibition of the ammonium production by FBP is favourable, while the stimulation effects of LAB on biogenic amines formation by FBPs are not desirable for food industry.     

Evolution of amino acids and biogenic amines throughout storage in sausages made of horse,beef and turkey meats

The changes in concentration of free amino acids and biogenic amines, along 28 d of storage at 4 °C, were monitored in a wide range of European ripened sausages manufactured from horse, beef and turkey meats. Generally speaking, both chemical families became more concentrated with elapsing time – but rather distinct patterns were followed in each meat type: total free amino acids increased by 13-fold in the case of horse sausages, and 5-fold in the case of beef sausages, but decreased to one third in the case of turkey sausages; and total biogenic amines attained 730 mg/kg in turkey sausages, 500 mg/kg in beef sausages and 130 mg/kg in horse sausages by 28 d of refrigerated storage. For putrescine, maximum levels of 285 mg/kg were attained in turkey and 278 mg/kg in beef sausages; for cadaverine, maximum levels of 6 mg/kg in turkey and 9 mg/kg in beef; and for histamine, maximum levels of 263 mg/kg in turkey and 26 mg/kg in beef. Hence, public safety concerns may be raised in the case of turkey sausages.     

Quality and safety of irradiated food regarding biogenic amines: Ras cheese

Food safety and quality became very important, especially with the challenge to ensure safe and healthy foods in regard to chemical hazards. So, this study was conducted to evaluate the quality and safety of irradiated Ras cheese during the storage period, with respect to biogenic amines (BAs). Ras cheese was manufactured, ripened and irradiated by γ‐irradiation at 0, 5, 10 and 15 kGy. The samples were stored in refrigerator at 5 ± 1 °C from where samples were withdrawn at 0, 2, 4 and 6 months for analysis. The results revealed that most sensory scores and chemical properties showed insignificant differences (P ≤ 0.05). The microbial counts were reduced with different degrees according to both storage period and irradiation dose. Also, irradiation was effective in reducing the content of BAs without harming the chemical properties of Ras cheese. The total content of BAs reflects the safety of irradiated Ras cheese and also indicates a high‐quality product in comparison with nonirradiated samples.     

PCR detection of foodborne bacteria producing the biogenic amines histamine, tyramine, putrescine, and cadaverine

This study describes an easy PCR method for the detection of foodborne bacteria that potentially produce histamine, tyramine, putrescine, and cadaverine. Synthetic oligonucleotide pairs for the specific detection of the gene coding for each group of bacterial histidine, tyrosine, ornithine, or lysine decarboxylases were designed. Under the conditions used in this study, the assay yielded fragments of 372 and 531 bp from histidine decarboxylase-encoding genes, a 825-bp fragment from tyrosine decarboxylases, fragments of 624 and 1,440 bp from ornithine decarboxylases, and 1,098- and 1,185-bp fragments from lysine decarboxylases. This is the first PCR method for detection of cadaverine-producing bacteria. The method was successfully applied to several biogenic amine-producing bacterial strains.     

Occurrence of lactic acid bacteria and biogenic amines in biologically aged wines

Biologically aged sherry-type wines are elaborated by the so-called 'criadera and solera' system, which essentially involves development of the yeast on the wine surface forming a film velum for several years. Lactic acid bacteria can also develop and contribute to sherry-type wine quality, although their presence and role in this enological process have received very little attention. In this study, lactic acid bacteria microbiota and the presence of biogenic amines were investigated throughout the manufacture and biological aging of 36 samples of sherry wines. Malolactic fermentation was found to mainly take place during the first stage of biological ageing. The incidence and populations of lactic acid bacteria in sherry wines were low. The diversity of bacterial species isolated from the wines was greater than previously reported and included species of Lactobacillus, with prevailing Lactobacillus hilgardii, Lactobacillus plantarum and Lactobacillus zeae and Leuconostoc mesenteroides. The biogenic amine-producing capacity of the isolates was also determined. Five strains were putrescine producers, while another strain was shown to produce tyramine and phenylethylamine, simultaneously. L. zeae was one of the predominant species in wines during the biological aging and seemed to be one of the main putrescine producers. The biogenic amine composition of the wines investigated was similar to that reported for other types of wines. Putrescine was the major amine, followed by cadaverine, histamine and tyramine. The amine contents detected were lower than those usually reported in red wines.     

High content of biogenic amines in Pecorino cheeses

Pecorino refers to Italian cheeses made exclusively from raw or pasteurized ewes' milk, characterized by a high content of fat matter and it is mainly produced in the Middle and South of Italy by traditional procedures. The autochthonous microbiota plays an important role in the organoleptic traits of Pecorino cheese and it can influence biogenic amines (BA) content.     

Effect of grape storage time on biogenic amines content in must

The presence of amines in wine can produce a negative impact on the aroma of the product and, in some cases, detrimental health effects. Several factors affect the content of amines in wine, among those, the presence of biogenic amines in must.     

冰鲜鸡中主要腐败菌的分离鉴定及其产生物胺特性分析

本文用不同选择性培养基对冰鲜鸡的主要腐败菌进行分离,利用16S r RNA基因序列测定法对分离菌进行鉴定,最后利用薄层层析和HPLC法对分离菌是否代谢产生物胺进行分析。研究发现:从冰鲜鸡中共分离筛选得到11株腐败菌,分别命名为C1、C4、C7、C8、PCA1、PCA2、PCA3、PCA4、PCA5、PCA7、PCA8。PCA1与Stentrophomona(狭长平胞属)、PCA4与Kouria(考克氏菌属)、PCA3与Enterobacter asburiae(阿氏肠杆菌)、PCA7与Enterobacter cancerogenus(生癌肠杆菌)、C1与Enterobacter ludwigii(肠杆菌属)、C4与Enterobacter cloacae(阴沟肠杆菌)、C7与Enterobacter hormaechei(肠杆菌属)、C8与Enterobacter hormaechei(肠杆菌属)、PCA2与Kocuria rhizophila(克氏库克菌属)、PCA5和PCA8与Staphylococcus(葡萄球菌属)亲缘关系近。通过薄层层析初步鉴定菌是否有产生物胺能力,发现PCA2、PCA5、PCA8不产生物胺,其余菌均产生物胺。利用HPLC进一步确定产生物胺菌发酵液中生物胺组成和含量,结果 PCA7只产生腐胺,含量为844.937μg/m L;其余菌株都产含量不同的腐胺和尸胺。本文研究表明,冰鲜鸡易受肠道菌污染,且产腐胺与尸胺,为冰鲜鸡的防腐与安全食用提供一定的参考依据。      

Determination of biogenic amines in selected Malaysian food

The biogenic amines tryptamine (TRP), putrescine (PUT), histamine (HIS), tyramine (TYR) and spermidine (SPD) were determined in 62 selected food items commonly consumed in Malaysia. This include the local appetisers “budu” and “cincalok”, canned fish, salt-cured fish, meat products, fruit juice, canned vegetables/fruits and soy bean products. After the aqueous extraction, the samples were derivatised with dansyl chloride before analysing using reversed phase HPLC with UV detection. Mean levels of TRP, PUT, HIS, TYR and SPD in eight budu samples were 82.7, 38.1, 187.7, 174.7 and 5.1 mg kg⁻¹, respectively. The main biogenic amines found in cincalok were PUT, HIS and TYR where the mean values were 330.7, 126.1 and 448.8 mg kg⁻¹, respectively. With the exception of “pekasam” and “belacan”, significantly lower levels of biogenic amines were found in canned fish and salt-cured fish samples. Non detectable or low levels of biogenic amines were found in meat, fruit juice and canned vegetables/fruit samples.     

Occurrence of biogenic amines in wine: The role of grapes

The evolution of biogenic amines from must to wine has been studied in seven different grape cultivars before and after malolactic fermentation. Alcoholic and malolactic fermentations have been carried out using selected yeasts and bacteria that, in a previous study, were unable to produce biogenic amines. The study has been performed under aseptic conditions to exclude possible interferences due to uncontrolled contaminating microorganisms present in grapes and/or in the environment. The goal of this work was to investigate the influence of grape on biogenic amines content of the wine. The results obtained showed that grape variety is related to the presence of some biogenic amines in wines and that, climatic conditions also affect the accumulation of these compounds in grapes.     

Levels of biogenic amines in retail market fermented meat products

The qualitative and quantitative profile of biogenic amines (BA) in 50 samples of dry fermented sausages sold in Greek retail markets were determined by HPLC. Putrescine, cadaverine, tryptamine, β-phenylethylamine spermidine, spermine were analysed by UV detection after pre-column derivatization with benzoyl chloride, whereas tyramine and histamine were analysed by fluorescence detection after post-column derivatization with o-phthaldialdehyde (OPA). With the exception of spermidine and spermine a wide variation of BA levels was observed among the samples. Of the BA examined, tyramine, putrescine, histamine and cadaverine showed high concentrations ranging from: 0 to 510 mg/kg (median: 197.7 mg/kg), 0 to 505 mg/kg (median: 96.5 mg/kg), 0 to 515 mg/kg (median: 7.0 mg/kg) and 0 to 690 mg/kg (median: 3.6 mg/kg), respectively. The histamine content of 28% of the samples exceeded the toxicity limit of 100 mg/kg set for histamine in some fish species. Levels of tryptamine and β-phenyl-ethylamine never exceeded 50 and 29 mg/kg, respectively. Results of the present study suggest that the amounts of BA in dry fermented sausages, sold in Greek retail markets, may pose a potential health risk for sensitive individuals or for those undergoing classical monoamine oxidase inhibiting (MAOI) drug therapy.     

A survey of biogenic amines in chinese red wines

Eight biogenic amines were analyzed in 38 samples of red wine from five wine-making regions of China. The analysis was carried out by reverse phase high performance liquid chromatography with fluorescence detector and dansyl chloride precolumn derivation. Tryptamine was not found in any of the samples. Putrescine was detected in all samples (100%), followed by phenylethylamine (84.2%), spermidine (60.5%), histamine (57.8%), tyramine (57.8%), cadaverine (47.4%), and spermine (36.8%). In all the samples, the levels of aromatic and heterocyclic amines with toxicological effects were: 0–4.58 mg/l for phenylethylamine, 0–10.51 mg/l for histamine, and 0–9.13 mg/l for tyramine. The amount of histamine and tyramine in most of the samples (94.7%) was less than 8 mg/l. The amines associated with sanitary conditions were also found to be present in a very low range, between 0 and 12.98 mg/l for putrescine, 0 and 19.01 mg/l for cadaverine, respectively. In the case of other amines such as spermine and spermidine, they yielded very low levels varying between 0 and 2.64 mg/l for spermine, 0 and 3.82 mg/l for spermidine, respectively.     

葡萄酒中生物胺的产生与工艺控制

生物胺存在于多种发酵食品中，人体吸收过量的生物胺后会引起不良的生理反应。在葡萄酒苹果酸—乳酸发酵(MLF)过程中，有些乳酸菌能够对氨基波脱投产生生物胺。利用PCR与DNA探针技术能够快速检测葡萄酒中的组胺产生菌。工艺上采用接种法进行MLF，并在MLF完成后对乳酸菌进行有效清除、可以显著降低葡萄酒中生物胺的含量。     

A survey of biogenic amines in Chinese rice wines

Fourteen Chinese rice wines from four rice wine-making regions of China were analyzed for the first time, using HPLC with diode array detection after pre-column derivatization with dansyl chloride. The results showed that Chinese rice wines contained the five biogenic amines histamine, tyramine, cadaverine, spermine and spermidine. Histamine was detected in all samples (100%), followed by spermine (93%), cadaverine (87%), tyramine (79%) and spermidine (79%). The mean total level of biogenic amines in samples was 107 mg/l with a range from 39.30 to 241 mg/l. These levels are below the level that may elicit direct adverse reactions for most consumers. However, patients being treated with monoamine oxidase inhibitors should be aware of the presence of amines in rice wines and limit their consumption.     

Detection of ten biogenic amines in Chinese commercial soybean paste by HPLC

This paper aimed to develop a high-performance liquid chromatography (HPLC) method that can simultaneously detect ten biogenic amines (BAs) in commercially available soybean paste and to determine the contents of BAs in these products. Heptylamine (Hep) was used as the internal standard reagent and benzoyl chloride as the sample derivatization reagent; Inertsil ODS-3 chromatographic column was set as the stationary phase while the acetonitrile and 5 mmol/L ammonium acetate solution as the mobile phases; the samples were treated with gradient elution and detected at 254 nm (UV). Perchloric acid solution (0.4 mol/L) was employed as the sample extraction solvent to determine the contents of ten BAs in soybean paste products. All BAs were separated within 20 min. The methodological validation suggested good linearity, whereas the linear correlation coefficient of all standards was greater than 0.99 with the linear range of 0.1–80 μg/g, detection limits of 0.01–0.03 μg/g, and quantitation limits of 0.03–0.10 μg/g. For 17 commercially available soybean paste products involved in our study, different samples were distinguished from each other not only by the dramatic differences in the contents of BAs but also by the variance in BA types. Of these BAs, tyramine (Tyr) and agmatine (Agm) were found to be the highest contents in soybean paste products; the contents of histamine (His), 2-phenylethylamine (Phe), serotonin (Ser), spermidine (Spd), and putrescine (Put) were lower, followed by cadaverine (Cad), tryptamine (Try), and spermine (Spm). This method proved to be highly sensitive and effective in determining the contents of BAs in soybean paste products; there was a dramatic difference in the contents of ten BAs among different soybean paste products.