Biogenic Amines Producers Research Articles

Screening of Yeasts Other than Saccharomyces for Amino acid Decarboxylation

This study is aimed at screening non-Saccharomyces for amino acids decarboxylation potentials. The yeasts were isolated from banana fruit and honey purchased from markets in Rivers State. The isolation and molecular identification of yeast isolates were according to standard microbiological procedures. A plate assay method for amino acid decarboxylation (biogenic amine production) screening was used. Wild Non-Saccharomyces yeast (NSY) were identified as Candida tropicalis Pe 1 (B7), Candida tropicalis WC65-1 (B10), Candida tropicalis WC57 (H4), Clavispora lusitaniae WM03 (H7), and a Commercial Wine yeast (CY) identified as Candida tropicalis zhuan4 (CY). The NSYs and CY were biogenic amine producers, from L-histidine and glutamic acid; strain variability from glycine, proline, glutamine, and asparagine decarboxylation; while L-arginine, lysine, tyrosine, cysteine, leucine, and phenylalanine were not decarboxylated at a concentration of 0.1 %. The increase in amino acid concentration influenced the number of amino acids decarboxylated - phenylalanine and leucine; L-histidine, glycine, asparagine and glutamic acid were decarboxylated by wild NSY and CY, while the strain variability of phenylalanine, proline, leucine and glutamic acid decarboxylation. The amino acids L-arginine, lysine, tyrosine, and cysteine were not decarboxylated. In terms of the concentration of amino acids, L-histidine and glutamic acid were decarboxylated and arginine, lysine, tyrosine, and cysteine were not decarboxylated by wild NSY isolates and CY. The Chi-square Test and Kendall’s Test of concordance suggest that there is no association between the amino acid concentrations (0.1 and 1 %) and biogenic amine production (P-value > 0.05). The wild NSY and CY are biogenic amine-producers, and the increase in amino acid concentration influences biogenic amine production concerning some amino acids.

Occurrence of Biogenic Amines Producers in the Wastewater of the Dairy Industry.

Out of six samples of wastewater produced in the dairy industry, taken in 2017 at various places of dairy operations, 86 bacterial strains showing decarboxylase activity were isolated. From the wastewater samples, the species of genera Staphylococcus, Lactococcus, Enterococcus, Microbacterium, Kocuria, Acinetobacter, Pseudomonas, Aeromonas, Klebsiella and Enterobacter were identified by the MALDI-TOF MS and biochemical methods. The in vitro produced quantity of eight biogenic amines (BAs) was detected by the HPLC/UV–Vis method. All the isolated bacteria were able to produce four to eight BAs. Tyramine, putrescine and cadaverine belonged to the most frequently produced BAs. Of the isolated bacteria, 41% were able to produce BAs in amounts >100 mg L−1. Therefore, wastewater embodies a potential vector of transmission of decarboxylase positive microorganisms, which should be taken into consideration in hazard analyses within foodstuff safety control. The parameters of this wastewater (contents of nitrites, nitrates, phosphates, and proteins) were also monitored.

Biogenic amines formation and their importance in fermented foods

Biogenic amines (BAs) are low molecular weight organic bases with an aliphatic, aromatic, or heterocyclic structure which have been found in many foods. biogenic amines have been related with several outbreaks of food-borne intoxication and are very important in public health concern because of their potential toxic effects. The accumulation of biogenic amines in foods is mainly due to the presence of bacteria able to decarboxylate certain amino acids. Biogenic amines are formed when the alpha carboxvl group breaks away from free amino acid precursors. They are colled after the amino acid they originated from. The main biogenic amines producers in foods are Gram positive bacteria and cheese is among the most commonly implicated foods associated with biogenic amines poisoning. The consumption of foods containing high concentrations of biogenic amines has been associated with health hazards and they are used as a quality indicator that shows the degree of spoilage, use of non-hygienic raw material and poor manufacturing practice. Biogenic amines may also be considered as carcinogens because they are able to react with nitrites to form potentially carcinogenic nitrosamines. Generally, biogenic amines in foods can be controlled by strict use of good hygiene in both raw material and manufacturing environments with corresponding inhibition of spoiling microorganisms. The aim of this review was to give some information about biogenic amines in foods.

Selection and technological potential of Lactobacillus plantarum bacteria suitable for wine malolactic fermentation and grape aroma release

Lactobacillus plantarum strains have resistance mechanisms that enable them to survive and proliferate in wine, which makes them potential malolactic fermentation (MLF) starter cultures. This work focused on the technological characterization of 11 L. plantarum strains isolated from Southern Italian wines that undergo spontaneous MLF, and proposes a selection of new L. plantarum malolactic starters. These strains were characterized according to their oenological characteristics, their ability to produce biogenic amines and bacteriocins, their response to the presence of phenolic compounds, their enzymatic activities and their ability to produce wine odorant aglycones from odourless grape glycosidic aroma precursors. Finally, the malolactic activity of one selected strain was assessed in Cabernet Sauvignon wine, using two inoculation methods. L. plantarum strains tested were not producers of biogenic amines. In particular, the M10 strain showed a good resistance to high levels of ethanol and low pH, it has a good malolactic performance and β-glucosidase activity, this last one demonstrated both directly through the measurement of this enzymatic activity and indirectly by following the release of volatile aglycones from commercial and natural grape glycosidic odourless precursors. These results demonstrated the potential applicability of M10 as a new MLF starter culture, especially for high-ethanol wines.

Identification of biogenic amines-producing lactic acid bacteria isolated from spontaneous malolactic fermentation of chilean red wines

Abstract Most Chilean wineries perform malolactic fermentation by means of non-typified autochthonous lactic acid bacteria already present in grapes and oak barrels or fermenters. The objective of this research was to investigate the principal lactic acid bacteria present during spontaneous malolactic fermentation of Chilean Cabernet Sauvignon wines and to study its role in BA formation. To the best of our knowledge this is the first time that this relation has been reported for Chilean wines. Lactic acid bacteria were isolated from five wineries located in three Chilean geographical regions. Genotypic differentiation of each bacterial isolated was performed via a restriction fragment length polymorphism method using rpoB and 16S rRNA genes and HinfI, AciI and MseI enzymes. Sixty-five colonies were isolated and identified as lactic acid bacteria, identifying two species, Lactobacillus rhamnosus and Oenococcus oeni. The predominant species was L. rhamnosus, which, to the best of our knowledge, we are describing for the first time in the vinification process. Considering that L. rhamnosus was detected in wineries from different geographical viticultural regions, it could be preliminarily considered as an endemic species. Both species were biogenic amines producers, L. rhamnosus being mainly responsible for biogenic amines present in the Cabernet Sauvignon wines studied.

Production of Biogenic Amines by Lactic Acid Bacteria Isolated from Uzicka Sausages

The aim of this study was to monitor production of seven biogenic amines (Cadaverine – CAD, Putrescine – PUT, Spermine – SPE, Spermidine – SPD, Histamine – HIS, Tyramine – TYR and Tryptamine – TRY) in selected 24 lactic acid bacteria (LAB) strains. The decarboxylase activity of the microorganisms was studied in growth medium after 24h cultivation. The ability of 24 LAB isolates cultivated in MRS broth and M17 broth supplement with 0.5% glucose to produce biogenic amines was assessed using liquid chromatography tandem mass spectrometry (LC-MS/MS). The investigation showed that LAB isolated from Uzicka sausage are not significant producers of biogenic amines in vitro.

SKRÍNING PROBIOTICKÝCH KULTUR URČENÝCH PRO VÝROBU FERMENTOVANÝCH POTRAVIN NA SCHOPNOST TVORBY BIOGENNÍCH AMINŮ

The contemporary trend is using probiotic cultures in fermented food production. They can be used as starter cultures and for their positive effect on human health. Probiotics are defined as living microorganisms present in food which consumed in adequate amounts affects positively the intestinal microflora’s composition and balance and thus human health itself. Cultures of these bacteria have to be of human origin and be able to survive the passage through the gastrointestinal tract. They also have to be able to multiply on the site of action (in intestine) and must not be toxic or pathogenic. Unfortunately, even some probiotic cultures can be counted among potential producers of biogenic amines, so their testing for the presence of biogenic amines is necessary (BURDYCHOVÁ, 2007).The aim of this study was screening of 26 types of bacterial cultures (SACCO, Italy) as probiotic cultures for their ability to produce biogenic amines tyramine and histamine. Cultivation in decarboxy­lating medium (BOVER-CID and HOLZAPFEL, 1999), HPLC descibed by BURDYCHOVÁ and DOHNAL (2007), and PCR detection of genes coding enzymes tyrosindecarboxylase and histidindecarboxylase, participating in formation of biogenic amines (COTON et al., 2004), were used as the screening methods. 19 strains of Lactobacillus spp., 3 strains of Bifidobacterium spp., 2 strains of Pediococcus spp. and 2 strains of Enterococcus spp. were examined by the methods mentioned above. The tyramine production was detected at 8 strains of Lactobacillus spp., 3 strains of Bifidobacterium spp. and 2 strains of Enterococcus spp., whereas no tested cultures were found to be able to produce histamine.The strains at which production of biogenic amines tyramine and histamine wasn’t detected are suitable for fermented food processing. When the strains at which production of tyramine was demonstrated were used in food processing, a control of concentration of this biogenic amine in final product is highly recommended.

Strains of Staphylococcus and Bacillus isolated from traditional sausages as producers of biogenic amines.

Histidine, lysine, ornithine, and tyrosine decarboxylase activities were tested in 38 strains of Staphylococcus (15 of S. equorum, 11 of S. epidermidis, 7 of S. saprophyticus, and 5 of S. pasteuri) and 19 strains of Bacillus (13 of B. subtilis and 6 of B. amyloliquefaciens) isolated from two Spanish traditional sausage varieties. The four decarboxylase activities were present in most of the strains studied, but some variability was observed between strains within each microbial species. Accumulation of putrescine and cadaverine was assessed in the culture media of the strains that displayed ornithine and lysine decarboxylase activities. The aminogenic potential of the strains was low, with amounts accumulated lower than 25 mg/L for the putrescine and than 5 mg/L for the cadaverine, with the exception of a strain of S. equorum that produced 1415 mg/L of putrescine, and of a strain of S. epidermidis that accumulated 977 mg/L of putrescine and 36 mg/L of cadaverine.

Diversity and assessment of potential risk factors of Gram-negative isolates associated with French cheeses

The goal of this study was to identify at the species level a large collection of Gram-negative dairy bacteria isolated from milks or semi-hard and soft, smear-ripened cheeses (cheese core or surface samples) from different regions of France. The isolates were then assessed for two risk factors, antibiotic resistance and volatile and non-volatile biogenic amine production in vitro. In total, 173 Gram-negative isolates were identified by rrs and/or rpoB gene sequencing. A large biodiversity was observed with nearly half of all Gram-negative isolates belonging to the Enterobacteriaceae family. Overall, 26 different genera represented by 68 species including potential new species were identified among the studied Gram-negative isolates for both surface and milk or cheese core samples. The most frequently isolated genera corresponded to Pseudomonas, Proteus, Psychrobacter, Halomonas and Serratia and represented almost 54% of the dairy collection. After Pseudomonas, Chryseobacterium, Enterobacter and Stenotrophomonas were the most frequently isolated genera found in cheese core and milk samples while Proteus, Psychrobacter, Halomonas and Serratia were the most frequently isolated genera among surface samples. Antibiotic resistance profiles indicated that resistances to the aminosid, imipemen and quinolon were relatively low while more than half of all tested isolates were resistant to antibiotics belonging to the monobactam, cephem, fosfomycin, colistin, phenicol, sulfamid and some from the penam families. Thirty-six% of isolates were negative for in vitro biogenic amine production. Among biogenic amine-producers, cadaverine was the most frequently produced followed by isoamylamine, histamine and putrescine. Only low levels (<75 mg/l) of tyramine were detected in vitro.

Detection of Biogenic Amine Producer Bacteria in a Typical Italian Goat Cheese

The aim of this study was to research decarboxylating bacterial strains and biogenic amine content in a typical Italian goat cheese (Robiola di Roccaverano). The study was performed on fresh and ripened samples of goat cheese manufactured from industrial and artisanal producers. Sixty-seven bacterial strains isolated showed decarboxylating activity, and Enterococcus faecalis was the most widespread decarboxylating species in all artisanal and industrial products. Pediococcus acidilactici and Enterococcus malodoratus were also identified as biogenic amine producers in Robiola di Roccaverano cheese. All the E. faecalis strains isolated in this study were able to decarboxylate tyrosine. Tyramine was the most abundant biogenic amine in cheese samples, while histamine was the most widespread. High amounts of these two biogenic amines were found in ripened samples (up to 2,067 mg/kg for tyramine and 1,786 mg/kg for histamine), whereas 2-phenylethylamine and tryptamine were present in almost all ripened cheeses at low concentrations. The detection of strains producing biogenic amines and the high concentrations of tyramine and histamine found in ripened Robiola di Roccaverano could represent a potential risk to the consumer.

Biogenic amine production by lactic acid bacteria isolated from cider

To study the occurrence of histidine, tyrosine and ornithine decarboxylase activity in lactic acid bacteria (LAB) isolated from natural ciders and to examine their potential to produce detrimental levels of biogenic amines. The presence of biogenic amines in a decarboxylase synthetic broth and in cider was determined by reversed-phase high-performance liquid chromatography (RP-HPLC). Among the 54 LAB strains tested, six (five lactobacilli and one oenococci) were biogenic amine producers in both media. Histamine and tyramine were the amines formed by the LAB strains investigated. Lactobacillus diolivorans were the most intensive histamine producers. This species together with Lactobacillus collinoides and Oenococcus oeni also seemed to produce tyramine. No ability to form histamine, tyramine or putrescine by Pediococus parvulus was observed, although it is a known biogenic amine producer in wines and beers. This study demonstrated that LAB microbiota growing in ciders had the ability to produce biogenic amines, particularly histamine and tyramine, and suggests that this capability might be strain-dependent rather than being related to a particular bacterial species. Production of biogenic amines by food micro-organisms has continued to be the focus of intensive study because of their potential toxicity. The main goal was to identify the microbial species capable of producing these compounds in order to control their presence and metabolic activity in foods.

Biogenic amine production by wild lactococcal and leuconostoc strains.

Two qualitative and one quantitative HPLC methods were evaluated for the detection of biogenic amine producers among wild dairy lactococcal and leuconostoc strains. High tyramine producers ranging from 370 to 807 mg l-1 were detected by qualitative methods and confirmed by HPLC analysis. Tyramine levels detected throughout the incubation time depended on the concentration of the amino acid precursor available and no tyramine production was observed when strains were grown in milk. However, increasing amounts of tyramine were detected in cultures grown in milk supplemented with different concentrations of tyrosine. Qualitative methods failed to detect weak producers so that tryptamine production (< 7 mg l-1) could only be determined by HPLC. None of the tested strains was able to produce histamine. Simultaneous production of different amines was observed by HPLC although no colour change was observed in the specific decarboxylase media. Thus, it was concluded that the amine forming ability should be taken into account when selecting starters for milk fermentations. Qualitative methods could be used as a first screening step to eliminate the highest amine producers while the quantitative methods would detect any producing strain.