Selective enumeration and identification of mixed cultures of Streptococcus thermophilus,Lactobacillus delbrueckii subsp. bulgaricus,L. acidophilus,L. paracasei subsp. paracasei and Bifidobacterium lactis in fermented milk

This study describes selective plating methodologies for enumeration of mixed cultures of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, L. acidophilus, L. paracasei subsp. paracasei and Bifidobacterium lactis in fermented milk based on selective antibiotic-free media. Enumeration of S. thermophilus was performed using M17-lactose. MRS-fructose was suitable for enumeration of L. bulgaricus and MRS-maltose for differentiation between L. acidophilus and L. paracasei. The selective enumeration of B. lactis was obtained using MRS-raffinose containing 0.05% LiCl. The bacterial counts obtained using selective methods were equivalent to those under optimum culture conditions at a probability level of 95%. Performance of the methods was verified in fermented milk products where identification of the enumerated species was confirmed by species-specific polymerase chain reaction. This study shows that combination of species-specific polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) analysis has great detection and identification potential for verification of accurate species labelling in fermented milk without prior isolation of the bacteria.     

Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia

Spontaneous milk fermentation has a long history in Mongolia, and beneficial microorganisms have been handed down from one generation to the next for use in fermented dairy products. The objective of this study was to investigate the diversity of lactic acid bacteria (LAB) communities in fermented yak, mare, goat, and cow milk products by analyzing 189 samples collected from 13 different regions in Mongolia. The LAB counts in these samples varied from 3.41 to 9.03 log cfu/mL. Fermented yak and mare milks had almost identical mean numbers of LAB, which were significantly higher than those in fermented goat milk but slightly lower than those in fermented cow milk. In total, 668 isolates were obtained from these samples using de Man, Rogosa, and Sharpe agar and M17 agar. Each isolate was considered to be presumptive LAB based on gram-positive and catalase-negative properties, and was identified at the species level by 16S rRNA gene sequencing, multiplex PCR assay, and restriction fragment length polymorphism analysis. All isolates from Mongolian dairy products were accurately identified as Enterococcus faecalis (1 strain), Enterococcus durans (3 strains), Lactobacillus brevis (3 strains), Lactobacillus buchneri (2 strains), Lactobacillus casei (16 strains), Lactobacillus delbrueckii ssp. bulgaricus (142 strains), Lactobacillus diolivorans (17 strains), Lactobacillus fermentum (42 strains), Lactobacillus helveticus (183 strains), Lactobacillus kefiri (6 strains), Lactobacillus plantarum ssp. plantarum (7 strains), Lactococcus lactis ssp. lactis (7 strains), Leuconostoc lactis (22 strains), Leuconostoc mesenteroides (21 strains), Streptococcus thermophilus (195 strains), and Weissella cibaria (1 strain). The predominant LAB were Strep. thermophilus and Lb. helveticus, which were isolated from all sampling sites. The results demonstrate that traditional fermented dairy products from different regions of Mongolia have complex compositions of LAB species. Such diversity of LAB provides useful information for further studies of probiotic strain selection and starter culture design, with regard to the industrial production of traditional fermented milk.     

Isolation and identification of exopolysaccharide producer lactic acid bacteria from Turkish yogurt

Lactic acid bacteria (LAB) were isolated from traditional yogurt samples and genotypic characterization of these isolates revealed the presence of 21 distinct LAB strains belonging to Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Leuconostoc mesenteroides, and Lactobacillus plantarum as new LAB strains. Determination of the exopolysaccharide (EPS) production characteristics of the selected strains of each species revealed that all strains possessed at least one gene required for both homopolymeric‐ and heteropolymeric‐type EPS production. Structural analysis of the EPSs showed that L. delbrueckii subsp. bulgaricus Y39 and S. thermophilus Y102 produced heteropolymeric EPS containing glucose and galactose, whereas Leuc. mesenteroides Y35 and L. plantarum Y36 produced homopolymeric glucan‐type EPS. The level of EPS production in these strains was found to be in a similar range. These strains with EPS production characteristics are good candidates for future studies as new LAB for yogurt production.

Practical applications

Recent trends in yogurt production technology have led to an increased use of ropy starter cultures in yogurt production due to the technological roles of exopolysacharides (EPS) produced by these cultures. The main role of EPS in yogurt production is to improve the textural properties of yogurt as an in situ produced natural polymer. In addition to the yogurt starter cultures, use of adjunct cultures during production of yogurt is also of special interest to enhance the technological and nutritional characteristics of yogurt. Therefore, in this study, potential yogurt starter and adjunct cultures from traditional yogurt samples with EPS production characteristics were isolated. From these isolates, Lactobacillus delbrueckii subsp. bulgaricus Y39 and Streptococcus thermophilus Y102 produced heteropolymeric EPS containing glucose and galactose, whereas Leuconostoc mesenteroides Y35 and Lactobacillus plantarum Y36 produced homopolymeric glucan.     

Diversity of Lactic Acid Bacteria Isolated from Brazilian Water Buffalo Mozzarella Cheese

The water buffalo mozzarella cheese is a typical Italian cheese which has been introduced in the thriving Brazilian market in the last 10 y, with good acceptance by its consumers. Lactic acid bacteria (LAB) play an important role in the technological and sensory quality of mozzarella cheese. In this study, the aim was to evaluate the diversity of the autochthones viable LAB isolated from water buffalo mozzarella cheese under storage. Samples were collected in 3 independent trials in a dairy industry located in the southeast region of Brazil, on the 28th day of storage, at 4 ºC. The LAB were characterized by Gram staining, catalase test, capacity to assimilate citrate, and production of CO₂ from glucose. The diversity of LAB was evaluated by RAPD‐PCR (randomly amplified polymorphic DNA‐polymerase chain reaction), 16S rRNA gene sequencing, and by Vitek 2 system. Twenty LAB strains were isolated and clustered into 12 different clusters, and identified as Streptococcus thermophilus, Enterococcus faecium, Enterococcus durans, Leuconostoc mesenteroides subsp. mesenteroides, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus delbrueckii subsp. bulgaricus, and Lactobacillus helveticus. Enterococcus species were dominant and citrate‐positive. Only the strains of L. mesenteroides subsp. mesenteroides and L. fermentum produced CO₂ from glucose and were citrate‐positive, while L. casei was only citrate positive. This is the first report which elucidates the LAB diversity involved in Brazilian water buffalo mozzarella cheese. Furthermore, the results show that despite the absence of natural whey cultures as starters in production, the LAB species identified are the ones typically found in mozzarella cheese.     

Rheological and physical characteristics of non-fat set yogurt prepared with EPS-producing Streptococcus thermophilus and an H+-ATPase-defective mutant Lactobacillus delbrueckii subsp. bulgaricus

The rheological and physical characteristics of non-fat set yogurt produced using co-cultures of ropy EPS-producing Streptococcus thermophilus S3.3 and Lactobacillus delbrueckii subsp. bulgaricus LTM and of Streptococcus thermophilus S3.3 and Lactobacillus delbrueckii subsp. bulgaricus H+-ATPase-defective mutant L6 under different conditions were studied. Yogurts produced with mutant L6 had a higher pH and water-holding capacity and better textural properties compared to those produced with the LTM strain. The highest storage modulus (G′) and thixotropic loops were found for yogurt made with L6 at 42°C on day 21 of cold storage. This yogurt contained a network of thick, continuous protein aggregates and large void spaces.     

Productions and monomer compositions of exopolysaccharides by Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains isolated from traditional home-made yoghurts and raw milk

In this study, a total of forty‐five strains of lactobacilli and streptococci were determined exopolysaccharide (EPS) production in skim milk and Man Rogosa and Sharpe (MRS)/M17 medium, viscosity and proteolytic activity. The exopolysaccharide production by lactobacilli strains during growth in MRS medium was twenty‐one to 211 mg L^?1, while in skim milk was to thirty‐six to 315 mg L^?1. The EPS production by streptococci strains during growth in M17 medium was sixteen to 114 mg L^?1, while in skim milk was to twenty‐four to 140 mg L^?1. The EPS production of strains was lower in MRS/M17 medium than skim milk. Results showed that it was not clear correlation between the viscosity and EPS production of some strains. All strains were shown proteolytic activity. Positive correlations between exopolysaccharide production and proteolytic activity in skim milk were found some strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. These results indicated that the high exocellular protease‐producing strains can produce high EPS in skim milk. The monomer compositions of the EPSs formed by selected five strains were analysed. Mannose dominated (99–100%) on the EPS produced by L. delbrueckii subsp. bulgaricus and S. thermophilusstrains (except L. delbrueckii subsp. bulgaricus 22) in skim milk and MRS/M17 medium. Besides, the EPSs of strains in skim milk contained small amount of lactose.     

Antibacterial activity of selenium-enriched lactic acid bacteria against common food-borne pathogens in vitro

Selenium (Se) is an essential trace element for human health and animal nutrition. The aim of this study was to evaluate the inhibitory activities of Se-enriched lactic acid bacteria (LAB), Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus, against pathogenic Salmonella typhimurium, Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes in vitro. The results indicated that the accumulation amount of Se by Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus reached 12.05 ± 0.43 µg/mL and 11.56 ± 0.25 µg/mL, respectively, accompanied by the relative maximum living cells when sodium selenite was 80 µg/mL. Oxford cup double plate assay showed that bacterial culture solution and cell-free culture supernatant (CFCS) from Se-enriched LAB exerted stronger antibacterial activity than those from the non-Se strains. The growth of pathogenic bacterial culture with CFCS at any growth stages was worse than that without CFCS; moreover, the inhibiting effect of CFCS of Se-enriched LAB was more significant than that of non-Se strains. Results from a scanning electron microscope equipped with energy dispersion X-ray spectrometry showed that elemental Se nanoparticles, which characteristically energy peak around 1.42 keV, were deposited on the cell surfaces of Lactobacillus delbrueckii ssp. bulgaricus. In addition, CFCS of Se-enriched LAB induced more serious cell structure damage of pathogenic bacteria than did non-Se LAB.     

Selective and differential enumerations of Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium spp. in yoghurt--a review

Yoghurt is increasingly being used as a carrier of probiotic bacteria for their potential health benefits. To meet with a recommended level of ≥ 10⁶ viable cells/g of a product, assessment of viability of probiotic bacteria in market preparations is crucial. This requires a working method for selective enumeration of these probiotic bacteria and lactic acid bacteria in yoghurt such as Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lb. acidophilus, Lb. casei and Bifidobacterium. This chapter presents an overview of media that could be used for differential and selective enumerations of yoghurt bacteria. De Man Rogosa Sharpe agar containing fructose (MRSF), MRS agar pH 5.2 (MRS 5.2), reinforced clostridial prussian blue agar at pH 5.0 (RCPB 5.0) or reinforced clostridial agar at pH 5.3 (RCA 5.3) are suitable for enumeration of Lb. delbrueckii subsp. bulgaricus when the incubation is carried out at 45 °C for 72 h. S. thermophilus (ST) agar and M17 are recommended for selective enumeration of S. thermophilus. Selective enumeration of Lb. acidophilus in mixed culture could be made in Rogosa agar added with 5-bromo-4-chloro-3-indolyl-β-d-glucopyranoside (X-Glu) or MRS containing maltose (MRSM) and incubation in a 20% CO₂ atmosphere. Lb. casei could be selectively enumerated on specially formulated Lb. casei (LC) agar from products containing yoghurt starter bacteria (S. thermophilus and Lb. delbrueckii subsp. bulgaricus), Lb. acidophilus, Bifidobacterium spp. and Lb. casei. Bifidobacterium could be enumerated on MRS agar supplemented with nalidixic acid, paromomycin, neomycin sulphate and lithium chloride (MRS-NPNL) under anaerobic incubation at 37 °C for 72 h.     

Lysis of starters in UF cheeses: Behaviour of mesophilic lactococci and thermophilic lactobacilli

The objective of this work was to study the autolytic behaviour of strains of mesophilic (Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris) and thermophilic lactic acid bacteria (Lactobacillus helveticus, Lb. delbrueckii subsp. lactis and Streptococcus thermophilus) in UF cheese. Cheeses were made from a UF-retentate (milk concentrated by a factor of 6) of microfiltered milk (0.8 μm pore size membrane) using the following starter systems: (1) single inocula of autolytic strains of L. lactis (US3, AM2 or AM1), non-autolytic strains of L. lactis (AM2-C or CNRZ-144), (2) a co-inocula of strains of Lb. helveticus (ITG-LH1, CNRZ-32 or CNRZ-303), Lb. delbrueckii subsp. lactis (ITG-LL14 or ITG-LL51) with the same strain of S. thermophilus CNRZ-1358. Cell viability was monitored over a 28 day ripening period by enumeration on selective media. Degree of lysis was determined by the measurement of the intracellular marker lactate dehydrogenase (LDH) activity, and also by immunodetection of intracellular proteins with species specific antibodies. In UF cheeses, lysis of autolytic strains of L. lactis was significantly delayed, showing release of intracellular components after 21 days of ripening. No lysis was observed for non-autolytic L. lactis strains or for S. thermophilus. Lysis of thermophilic lactobacilli (Lb. helveticus, Lb. delbrueckii), was observed from the start of ripening, but the onset and the level of lysis observed was strain and species dependent.     

Selection of starter cultures forthe fermentation of soya milk

The growth of four cultures selected on the basis of their ability to produce lactic acid in soya milk and/or utilize oligosaccharides, namelyStreptococcus thermophilus, Lactobacillus delbrueckiisubspbulgaricus, Lactobacillus fermentiandLactobacillus fermentum, was examined in reconstituted, low fat, spray-dried soya milk powder (12% total solids). The single culture ofS. thermophilusproduced a drop in pH from 6.5 to 4.7 over a 10-h period, and reduced the level of stachyose from 8.5 mg ml^-;1in the original milk to 3.2 mg ml^-;1; after 24 h fermentation, pH fell to 4.5, and the stachyose concentration to 3.0 mg ml^-;1. The paired culture ofS. thermophilusandL. fermentumbehaved in a similar fashion, but with only a slight improvement in stachyose utilization. When yeast extract (0.3%) and glucose (1.0%) were added to the soya milk, acid production by all the cultures increased dramatically, andL. delbrueckiisubspbulgaricusalone or in combination withS. thermophiluslowered the pH of the milk to 4.3 over 10 h. This combination ofS. thermophilusandL. delbrueckiisubspbulgaricuswas considered to be a likely combination for the production of a fermented product from soya milk or modified soya milk, as neitherL. fermentinorL. fermentumwere appreciably more effective in lowering the concentrations of oligosaccharides.     

Shelf life of alginate beads containing lactobacilli and bifidobacteria: characterisation of microspheres containing Lactobacillus delbrueckii subsp. bulgaricus

Capsules, containing different microorganisms (Lactobacillus plantarum, L. casei, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. delbrueckii, L. reuteri, L. rhamnosus, Bifidobacterium breve and B. animalis subsp. lactis), were produced through a simple extrusion protocol and dipped in a CaCl₂ solution (0.5% or 8%). Beads were characterised, to assess the encapsulation yield (EY) and the shelf life at 4 °C (defined as the time to retain inside a cell count of 6 log CFU g^?1); then, L. delbrueckii subsp. bulgaricus was used as target for system optimisation, through the study of the kinetic of cell release, as a function of the concentration of CaCl₂ solution, agitation (150 rpm) and recycling. The EY was 50% or higher, and the shelf life was at least 30 days. Concerning beads containing L. delbrueckii subsp. bulgaricus, the release of cells from capsules followed a biphasic (diauxic) trend and the microsphere could be used successfully for two times, as only after third reuse, the amount of release cells decreased.     

Competition mechanisms of lactic acid bacteria and bifidobacteria: Fermentative metabolism and colonization

Enzyme activities (α- and β-glucosidases, α- and β-galactosidases and β-fructofuranosidase) and organic acid production of four strains of lactic acid bacteria (LAB; Streptococcus thermophilus STY-31, Lactobacillus delbrueckii subsp. bulgaricus LBY-27, Lactobacillus casei LC-01 and Lactobacillus acidophilus LA-5) and Bifidobacterium lactis BB-12 were tested on milk and MRS fermentation broth with glucose, lactose or fructooligosaccharides (FOS) as carbon source. The highest β-galactosidase activity was found in L. acidophilus growing on milk. As compared to milk, α-glucosidase activity was increased with FOS by B. lactis, L. acidophilus and L. casei. The analysis of organic acids and short-chain fatty acids in the medium growth showed that lactate and acetate were the major fermentation metabolites produced by LAB and bifidobacteria, respectively. However, a metabolic shift towards more acetate and formate production, at the expense of lactate production, was observed during growth of L. casei on FOS. When grown on FOS as sole carbon source, L. acidophilus showed the highest production of lactate among the species tested. In addition, L. acidophilus demonstrated resistance to colonization against the intestinal pathogens Escherichia coli and Salmonella enterica in competition assays.     

In Vivo Study of the Survival of Lactobacillus delbruecki subsp. bulgaricus CECT 4005T and Streptococcus thermophilus CECT 801 by DVC‐FISH after Consumption of Fermented Milk

Abstract: Direct Viable Count (DVC) method has been recently combined with fluorescent in situ hybridization (FISH) for the specific detection of viable cells of Lactobacillus delbrueckii subsp. bulgaricus CECT 4005T and Streptococcus thermophilus CECT 801. This method has been used to determine their in vitro viability to gastrointestinal juices, being the resistance of L. delbrueckii subsp. bulgaricus and S. thermophilus 26.2% and 9.2%, respectively. On the other hand, an in vivo study has been carried out with the application of this technique for their detection in human feces, after consuming fermented milk. Cells of L. delbrueckii subsp. bulgaricus CECT 4005T were not detected, whereas viable cells of S. thermophilus CECT 801 were detected in a number higher than 10³ cells per gram in a 30% of the samples after 4 wk of consumption. DVC‐FISH is a quick and culture‐independent useful method, which has been applied for the 1st time in an in vivo survival study of LAB.     

Proteolytic profiles of yogurt and probiotic bacteria

Nine strains of Streptococcus thermophilus, 6 strains of Lactobacillus delbrueckii ssp. bulgaricus, 14 strains of Lactobacillus acidophilus and 13 strains of Bifidobacterium spp. were screened for proteolytic, amino-, di-, tri- and endopeptidase activity by using the o-pthaldialdehyde-based spectrophotometric assay. Strains showing the highest and lowest proteolytic activity were further studied for their peptidase activities at the extracellular and intracellular levels. The amounts of free amino groups released by S. thermophilus, L. delbrueckii ssp. bulgaricus and L. acidophilus strains were higher than that by Bifidobacterium strains. Aminopeptidase activity was detected for all bacterial strains both at the extracellular and intracellular levels. The specific activity towards the six substrates studied was higher at the intracellular level for all strains. High dipeptidase activity was demonstrated by all bacterial strains for L. delbrueckii ssp. bulgaricus, L. acidophilus, and Bifidobacterium spp. whereas S. thermophilus had greater dipeptidase activity at the extracellular level. All bacterial cultures tested were able to hydrolyse large biologically active peptides, bradykinin, Ala–Ala–Ala–Ala–Ala and the tripeptide substrate Gly–Ala–Tyr at both the extracellular and intracellular levels. However, with the substrates ending with a C-terminal of phenylalanine, the hydrolysis only occurred at the intracellular level.     

Influence of compounds associated with fermented dairy products on the growth of lactic acid starter and probiotic bacteria

The growth of 24 strains of lactic acid starter bacteria (Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus and Lactococcus lactis) and 24 strains of probiotic bacteria (Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus and bifidobacteria) in liquid media containing different substances was assessed. The substances used were salts (NaCl and KCl); sugars (sucrose and lactose); sweeteners (acesulfame and aspartame); aroma compounds (diacetyl, acetaldehyde and acetoin); natural colorings for fermented milk (red, yellow and orange colorings); flavoring agents (strawberry, vanilla, peach and banana essences); flavoring–coloring agents (strawberry, vanilla and peach); nisin, natamycin and lysozyme. Bacterial growth in the presence of natural fruit juices (green apple, kiwi, pineapple, peach and strawberry) with or without neutralization and cell viability in lactic acid acidified (pH 4 and 5) milk for 4 weeks at 5°C were also studied.Some compounds (KCl, sweeteners, aroma compounds, natamycin, flavoring agents and the peach flavoring–coloring agent) did not influence the growth of the strains in the concentrations commonly used in the dairy industry. The effect of other substances (especially flavoring–coloring agents) on the growth of lactic acid starters and probiotic bacteria was strain-dependent. Natural fruit juices weakly inhibited mainly S. thermophilus strains. Cell viability during cold storage in acidified milk was satisfactory for L. delbrueckii subsp. bulgaricus and L. casei group strains. For L. acidophilus and Bifidobacterium, the decreases in cell counts at pH 5 were negligible. Nevertheless, decreases from 1.6 to 6.2 and from 0.1 to 7.6 log orders, respectively were observed at pH 4.     

Microbial and physiochemical changes in yoghurts containing different Lactobacillus delbrueckii subsp. bulgaricus strains in association with Lactobacillus plantarum as an adjunct culture

Two strains of slow and normal acid‐producing Lactobacillus delbrueckii subsp. bulgaricus, mixed separately with Streptococcus thermophilus, were used in the preparation of yoghurt with or without Lactobacillus plantarum A7. Viable cell counts of lactic bacteria, pH, lactic acid and acetic acid concentrations, soluble nitrogen content, spontaneous syneresis, viscosity and firmness were studied at selected points during the refrigerated storage period. Results showed that survival of L. plantarum A7 in yoghurt was independent of the acidification capability of L. delbrueckii subsp. bulgaricus strains which had already been characterised in their pure culture. Thus, in the preparation of probiotic yoghurt, using slow acid‐producing L. delbrueckii subsp. bulgaricus strain in combination with nondairy lactobacilli postfermentation acidification is not restricted but instability in other physicochemical properties of the yoghurt may be expected.     

Influence of casein hydrolysates on the growth and lactic acid production of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus

The growth performance of Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) and Streptococcus thermophilus (S. thermophilus) was determined in the presence of casein hydrolysates produced by the action of five proteolytic enzymes (Alcalase, Flavorzyme, Neutrase, Papain and Trypsin) with various degrees of hydrolysis (DH). In addition, these five kinds of casein hydrolysates were fractionated by ultrafiltration and the influence of the amino acid composition of the peptides on the growth and lactic acid yield of yoghurt lactic acid bacteria (LAB) was studied. The results showed that the ultrafiltered fraction (<3000 Da) was the determinant stimulator in crude hydrolysates. Furthermore, the hydrophilic amino acid residua including His, Lys, Glu and Ser were beneficial for bacterial growth. Compared with control, the cell growth and lactic acid yield of yoghurt LAB were increased with the supplementation of the peptides fraction (<3000 Da) produced with papain by 65.1% and 49.6%, respectively.     

Tyramine production of technological important strains of <Emphasis Type="Italic">Lactobacillus</Emphasis>, <Emphasis Type="Italic">Lactococcus</Emphasis> and <Emphasis Type="Italic">Streptococcus</Emphasis>

The aim of this paper was to study the biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) production of selected technological important lactic acid bacteria (strains of the genera Lactococcus, Lactobacillus and Streptococcus). Three methods (ion-exchange chromatography (IEC), PCR and cultivation method with pH indicator) were used. Within the 39 strains of lactic acid bacteria tested, the production of tyramine (formed by tyrosine decarboxylase) was detected in eight strains (3 strains of Lactococcus lactis subsp. lactis, three strains of Lactococcus lactis subsp. cremoris, 1 strain of Streptococcus thermophilus and 1 strain of Lactobacillus delbrueckii subsp. bulgaricus). The other tested biogenic amines were not detected. Cultivation in decarboxylation broth seems to be the least accurate method for the detection of biogenic amines due to enhanced risk of false-positive reactions. Therefore, in order to detect bacteria producing biogenic amines, the combination of PCR and chromatographic methods (e.g. IEC) can be recommended.     

Screening selected strains of probiotic lactic acid bacteria for their ability to produce biogenic amines (histamine and tyramine)

The aim of this study was to investigate the production of biogenic amines (BA), histamine and tyramine by some probiotic lactic acid bacteria (LAB). Fifteen strains representing six LAB species were screened qualitatively by growing them in a decarboxylase medium. Quantitative analysis was carried out by HPLC analysis with direct derivatization of acid extracts. Lactobacillus casei (TISTR 389) and Lactobacillus delbrueckii subsp. bulgaricus (TISTR 895) were found to produce BA. The highest levels of histamine (1820.9 ± 3.5 mg L^?1) and tyramine (5486.99 ± 47.6 mg L^?1) formation were observed for the TISTR 389 strain, while TISTR 895 produced only histamine (459.1 ± 0.63 mg L^?1) in the decarboxylase broth. Biogenic amine potential was not observed for the Lactobacillus acidophilus, Lactobacillus lactis subsp. lactis, Lactococcus lactis subsp. lactis, and Lactobacillus plantarum strains studied. This study confirmed that BA formation is strain dependent and not related to the species. Therefore, careful screening for amino acid decarboxylase activity is recommended before selecting LAB as appropriate starter or probiotic strains in food and dairy industry.