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.     

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.     

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。     

Fast identification of wine related lactic acid bacteria by multiplex PCR

The microflora of must and wine consists of yeasts, acetic acid bacteria and lactic acid bacteria (LAB). The latter group plays an important role for wine quality. The malolactic fermentation carried out by LAB leads to deacidification and stabilisation of wines. Nevertheless, LAB are often associated with wine spoilage. They are mainly responsible for the formation of biogenic amines. Furthermore, some strains produce exopolysaccharide slimes, acetic acid, diacetyl and other off-flavours. In this context a better monitoring of the vinification process is crucial to improve wine quality. Moreover, a lot of biodiversity studies would also profit from a fast and reliable identification method.     

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.     

Biogenic amines content in Spanish and French natural ciders: application of qPCR for quantitative detection of biogenic amine-producers

Biogenic amines (BA) are low molecular weight nitrogenous bases commonly found in fermented foods and beverages and their consumption can induce undesirable reactions. In this work, the BA content in natural cider from Spain and France was determined. Samples from commercially available cider or obtained during the elaboration process were analyzed. A different profile and BA concentration was observed depending on cider origin. qPCR tools developed for the quantitative detection of BA producers from cheese and wine were tested in the cider samples. A good connection between the BA content and the presence of BA-producing microorganisms was observed. Based on these tools, BA-producing bacteria were isolated from the analyzed cider samples, including new potential histamine- and putrescine-producing Lactobacillus paracollinoides strains.     

Amino acid decarboxylase activities and biogenic amine formation abilities of lactic acid bacteria isolated from shalgam

56 lactic acid bacteria were isolated during shalgam fermentation and identified as Lactobacillus spp. (51 isolates), Lactococcus spp. (3 isolates), Streptococcus sp. (one isolate), and Leuconostoc sp. (one isolate). 53 of all isolates decarboxylated both arginine and tyrosine, while others decarboxylated one of arginine or tyrosine. None of the isolates could decarboxylate histidine, ornithine, lysine, phenylalanine, or tryptophan. All isolates produced both agmatine (105.8–867.5 mg L^–1) and tyramine (24.5–649.7 mg L^–1). Although none of the isolates displayed ornithine decarboxylase activity, putrescine was produced (2.1–33.3 mg L^–1) by all isolates, except one Lactobacillus strain. Therefore, lactic acid bacteria seem to be responsible mainly for tyramine and agmatine formation during shalgam fermentation, as well as a small amount of putrescine.     

Reduced biogenic amine contents in sauerkraut via addition of selected lactic acid bacteria

The contents of free amino acids and biogenic amines in spontaneously fermented sauerkraut, inoculated or not with specific lactic acid bacterium strains, were monitored throughout 45 d of storage. The strains tested were Lactobacillus plantarum 2142, Lactobacillus casei subsp. casei 2763 and Lactobacillus curvatus 2771. In both the control and the experiments, the total amino acid contents increased with time – and the predominant ones were aspartic acid, arginine and glutamic acid. However, upon inoculation with either strain, the total biogenic amine contents remained considerably lower than those of the control (especially in the cases of L. casei subsp. casei and L. curvatus); every single biogenic amine was actually below the 100 ppm-threshold. The dominant biogenic amines in the control were putrescine – and tyramine and histamine, to a lesser extent; the putrescine content was 10-fold lower if inoculation had taken place with either lactobacillus tested; and histamine and tyramine were essentially absent during storage, whereas they ranked above 200 ppm in the control by 45 d. Hence, an efficient food-grade biological tool was made available that constrains buildup of dangerous biogenic amines in fermented vegetables during storage.     

葡萄酒中的生物胺的生产与工艺控制

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酸脱羧产生生物胺。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。     

Functional properties and safety assessment of lactic acid bacteria isolated from goat colostrum for application in food fermentations

Thirty representative isolates from genotypes obtained after RAPD‐PCR analysis of 191 presumptive LAB isolates from goat colostrum were assessed. They were identified as Leu. mesenteroides, Leu. lactis and Lac. garvieae. Only Lac. garvieae P27M15 showed a high resistance to low pH, while 93.3% of the isolates retained their viability after exposure to bile salts. Leu. lactis V22M11, V42M18 and V23M15 displayed the best technological properties. Lac. garvieae P27M15 and Leu. lactis V22M11, V42M18 and V23M15 were susceptible to 8 of the 13 antibiotics assayed. Therefore, it can be concluded that the isolates P27M15, V22M11, V42M18 and V23M15, showing potentially important properties, could be valuable for practical applications in biotechnological processes.     

Biogenic amines in cold-smoked fish fermented with lactic acid bacteria

 It has been proved in previous studies that fish can be fermented by lactic acid bacteria (LAB). Added glucose must be at concentrations of almost 1% to reduce the pH to <5.3, a level which corresponds to the safe pH values for meat products. Because some LAB and some contaminating microbes form biogenic amines, this study was conducted to examine the possible formation of biogenic amines during the fish fermentation process. Other parameters under study were sensory quality, pH value, titrated acid content, weight loss, a _w value, redox potential and microbiological counts. Three groups of fish (rainbow trout) with three different LAB inocula and a group without any inoculum were made. The fermentation of the products made with LAB succeeded. The products were sensorially accepted, the inoculated LAB grew to >8 log cfu/g, the pH reduced to 5.0–5.3 and a _w to 0.927 and the pseudomonads, the predominant flora of fish raw material, disappeared. The fish raw material and the products contained low amounts of biogenic amines with one exception: cadaverine, histamine and tyramine increased in all product groups in one experimental series (II) out of three. The highest concentrations of these amines were in the control products without any LAB inoculation. The LAB used and the contaminants isolated from the products were unable to produce cadaverine, histamine or tyramine. The appearance of these amines could therefore be caused by the non-isolated contaminants of fish raw material. Received: 18 June 1999     

The inhibitory effects of wine phenolics on lysozyme activity against lactic acid bacteria

The lysozyme of hen's egg white is used in winemaking to control spontaneous lactic acid bacteria (LAB). A total of eight LAB strains, isolated from grape must and wine, were used to assess the inhibitory effects of wine phenolics on lysozyme activity. The presence of phenolics, extracted from grape pomace, in growth medium reduced the mortality rate due to the lysozyme activity. This effect was especially clear in the case of strains belonging to Lactobacillus uvarum, Pediococcus parvulus and Oenococccus oeni, which are more sensitive to lysozyme than L. plantarum and L. hilgardii strains. Cell lysis assays carried out on four strains sensitive to lysozyme and Micrococcus lysodeikticus ATCC 4698, used as a reference strain, confirmed the inhibition of grape pomace phenolics on the muramidase. There was no interference from non-flavonoids, flavanols and flavonol compounds, when they were tested individually, on the lysozyme activity against the strains. Anthocyanins extracted from grape skins slightly inhibited the activity only against M. lysodeikticus. However, proanthocyanidins extracted from seed berries, strongly inhibited the lysozyme. In this extract, dimers were the predominant oligomers of flavan-3-ol. The study demonstrated that the effectiveness of lysozyme against LAB in red winemaking is related to the amount of low molecular weight proanthocyanidins that are released when the grapes are macerating.     

Lactic acid bacteria profiles and tyramine and tryptamine contents of Turkish tulum cheeses

Tulum cheese, commonly produced in Turkey, is a traditional Turkish dairy product made from raw milk. In this research, 20 samples of tulum cheese were purchased from different markets in Isparta and Ankara (Turkey) and the populations of lactic acid bacteria (LAB) and contents of biogenic amines of Turkish tulum cheeses were investigated. The mean counts of LAB (as log10 cfu/g) were 7.22±0.67 on MRS agar, 6.93±1.77 on M17 agar, 7.28±0.72 on APT agar and 5.98±2.20 on ADA agar. Isolates of LAB obtained from 20 tulum cheeses were characterised as lactobacilli (48.5%), enterococci (32.7%), lactococci (15.8%) and streptococci (3.0%). Lactobacillus paracasei ssp. paracasei, Lactobacillus bifermentans and Enterococcus faecium predominated among the LAB in the samples of tulum cheeses; these organisms represented 30.2% of the isolates. As amino acids, tyrosine and tryptophan were determined in tulum cheeses. Tyramine and tryptamine, biogenic amines, were found at 0.00–329.00 and 0.32–40.44 mg/kg, respectively.     

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.     

Genetic screening of lactic acid bacteria of oenological origin for bacteriocin-encoding genes

A total of 330 lactic acid bacteria isolated from South African red wines during alcoholic and malolactic fermentations and 9 commercial malolactic bacteria starter cultures were screened for antimicrobial activity. Of the entire screened isolates, 26 strains, belonging to the species Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus hilgardii and Oenococcus oeni, showed activity towards various wine-related and non-wine-related indicator strains. A PCR-based screening revealed the presence of the plantaricin encoding genes plnA, plnEF, plnJ and plnK in five selected Lb. plantarum strains. Furthermore, a co-culture experiment with Lb. plantarum and Enterococcus faecalis was performed. A complete inhibition of cell growth of Ent. faecalis was observed within 72 h. Four putative bacteriocin-encoding genes in the genome of O. oeni were identified and sequenced.     

Synergistic and antagonistic effect of lactic acid bacteria on tyramine production by food-borne pathogenic bacteria in tyrosine decarboxylase broth

The effect of lactic acid bacteria (LAB) strains on tyramine (TYR) and also other biogenic amines (BA) production by eight common food-borne pathogen (FBP) in tyrosine decarboxylase broth (TDB) was investigated by using a rapid HPLC method. Significant differences were observed among the FBP strains in ammonia (AMN) and BA production apart from tryptamine, histamine (HIS) and spermine formation (p < 0.05). Salmonella paratyphi A was characterised as the main amine producer. LAB had an important synergetic role in some BA production by food-borne pathogenic bacteria, although the effect of some LAB strains on BA production was strain-dependent. Lactococcus spp. and Streptococcus spp. resulted in significantly higher TYR accumulation by Aeromonas hydrophila and Enterococcus faecalis in TDB. The presence of Lactococcus and/or Lactobacillus in TDB significantly increased HIS production by A. hydrophila, Escherichia coli, Ent. faecalis, Klebsiella pneumoniae and Pseudomonas aeruginosa, whereas HIS accumulation was significantly reduced by Staphylococcus aureus, S. paratyphi A and Listeria monocytogenes.     

Technological and safety properties of lactic acid bacteria isolated from Spanish dry-cured sausages

Technological and safety-related properties were analyzed in lactic acid bacteria isolated from Spanish dry-cured sausages in order to select them as starter cultures. In relation to technological properties, all the strains showed significative nitrate reductase activity; Lactobacillus plantarum, Lactobacillus paracasei and 52% of the Enterococcus faecium strains showed lipolytic activity and only Lactobacillus sakei strains (43%) were able to form biofilms. Related to safety aspects, E. faecium strains were the most resistant to antibiotics, whereas, L. sakei strains were the most sensitive. In relation to virulence factors, in the E. faecium strains analyzed, only the presence of efaA gene was detected. The analysis of biogenic amine production showed that most E. faecium strains and L. sakei Al-142 produced tyramine. In conclusion, L. paracasei Al-128 and L. sakei Al-143 strains possess the best properties to be selected as adequate and safe meat starter cultures.     

The function of lactic acid bacteria and brine solutions on biogenic amine formation by foodborne pathogens in trout fillets

The influences of lactic acid bacteria and brine solutions on the biogenic amine formation by Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterococus faecalis, Pseudomonas aeruginosa, Listeria monocytogenes, Aeromonas hydrophila and Salmonella paratyphi A in fermented trout fillets were investigated. Fish fillets were divided into four groups, group 1 without any lactic acid bacteria inoculation, group 2 and group 3 with different salt concentration inoculated with lactic acid bacteria and food-borne pathogens, and group 4 inoculated with lactic acid bacteria and food-borne pathogens without a salt solution. The histamine content in trout fillets in group 4 was found to be more than 10 mg/100 g, while the other groups contained less than 7.5 mg/100 g. The highest tyramine production was found for group 1 and group 3, ranging from 3 to 18 mg/100 g. Lactic acid bacteria did not seem to play an important role on biogenic amine production by food borne pathogens, while adding brine solution on fillets has inhibitory effects on some of the biogenic amines.     

Phenotypic and genetic evaluations of biogenic amine production by lactic acid bacteria isolated from fish and fish products

In this work, biogenic amine production (histamine, tyramine and putrescine) by a collection of 74 lactic acid bacteria of aquatic origin has been investigated by means of amino acid decarboxylation by growth on decarboxylase differential medium, biogenic amine detection by thin-layer chromatography (TLC) and decarboxylase gene detection by PCR. None of the evaluated strains showed neither production of histamine and putrescine, nor presence of the genetic determinants encoding the corresponding decarboxylase activities. However, the tyrosine decarboxylase gene (tdc) was present in all the enterococcal strains, and tyramine production was detected by TLC in all of them but Enterococcus faecium BCS59 and MV5. Analysis of the tyrosine decarboxylase operon of these strains revealed the presence of an insertion sequence upstream tdc that could be responsible for their lack of tyrosine decarboxylase activity.     

Occurrence and role of lactic acid bacteria in seafood products

Lactic acid bacteria (LAB) in fish flesh has long been disregarded because the high post-mortem pH, the low percentage of sugars, the high content of low molecular weight nitrogenous molecules and the low temperature of temperate waters favor the rapid growth of pH-sensitive psychrotolerant marine Gram-negative bacteria like Pseudomonas, Shewanella and Photobacterium. In seafood packed in both vacuum (VP) and modified atmosphere (MAP) packaging commonly CO₂ enriched, the growth of the Gram-negative aerobic bacteria group (predominantly pseudomonads) is effectively inhibited and the number reached by LAB during storage is higher than that achieved in air but always several log units lower than the trimethylamine oxide (TMA-O) reducing and CO₂-resistant organisms (Shewanella putrefaciens and Photobacterium phosphoreum). Accordingly, LAB are not of much concern in seafood neither aerobically stored nor VP and MAP. However, they may acquire great relevance in lightly preserved fish products (LPFP), including those VP or MAP. Fresh fish presents a very high water activity (aw) value (0.99). However, aw is reduced to about 0.96 when salt (typically 6% WP) is added to the product. As a result, aerobic Gram-negative bacteria are inhibited, which allows the growth of other organisms more resistant to reduced aw, i.e. LAB, and then they may acquire a central role in the microbial events occurring in the product. Changes in consumers’ habits have led to an increase of convenient LPFP with a relative long shelf-life (at least 3 weeks) which, on the other hand, may constitute a serious problem from a safety perspective since Listeria monocytogenes and sometimes Clostridium botulinum (mainly type E) may able to grow. In any case the LAB function in marine products is complex, depending on species, strains, interaction with other bacteria and the food matrix. They may have no particular effect or they may be responsible for spoilage and, in certain cases, they may even exert a bioprotective effect in relation to undesirable bacteria. The bioprotective potential of endogenous LAB in relation to pathogens and spoiling bacteria has often been highlighted. However, the technology is still in its infancy compared with foods dairy and meat products in which either the carbohydrate content (dairy products) or sugar and salt added (meat products) favor the acidification by LAB that enable a natural preservation of the product. Successful studies on LAB as probiotic for fish intensify, but this potential is still to be explored for human. Although not usual, some applications of LAB for fermentation of marine products and by-products are described.