Biogenic amines in meat inoculated with Lactobacillus sake starter strains and an amine-positive lactic acid bacterium

 Formation of biogenic amines in minced meat inoculated with two Lactobacillus sake starter culture strains and an amine-positive lactic acid bacterium (G-106) was studied. The effects of these starter cultures against the formation of biogenic amines were dependent on the kind of decarboxylating microorganisms present in the raw material and the effects were different for each amine. Starter strains maintained the microbiological quality of the meat kept at 20°C for 7 days, and inhibited the formation of putrescine and cadaverine. They also inhibited the formation of phenylethylamine caused by G-106 when it was present at an initial level of 10² colony-forming units (CFU)/g. However, the formation of tyramine was not affected and the formation of histamine was increased when starters were used in samples inoculated with G-106. Received: 4 December 1996 / Revised form: 27 January 1997     

Biogenic Amines in Dry Sausages as Affected by Starter Culture and Contaminant Amine-Positive Lactobacillus

The effect of an amine-negative starter culture, containing Pediococcus pentosaceus and Staphylococcus carnosus, on the growth and amine formation of an amine-positive contaminant lactic acid bacterium (G 106) was studied in dry sausages. Levels of biogenic amines, precursor amino acids, pH, water activity and microbial counts were measured. Levels of phenylethylamine and tyramine increased in the sausages inoculated with the amine-positive strain. The starter culture did not prevent growth of G 106 or its amine formation capability. However, levels of histamine remained low although G 106 could produce histamine in vitro.     

Effect of fermentation with mixed starter cultures on biogenic amines in bighead carp surimi

The effects of one group mixed starter cultures, combined with Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen, and Monascus anka, and a batch without starter as control on biogenic amines accumulation in the bighead carp surimi during fermentation were investigated. Determination of five different biogenic amines was carried out by reverse‐phase high‐performance liquid chromatography with UV detection. Results showed that mixed starter cultures decreased the pH quickly from initial pH value of 6.4 to 5.2, inhibited the growth of contaminant microorganisms, such as Enterobacteriaceae and Pseudomonas, present in the raw materials, and suppressed the accumulation of histamine, tyramines, spermine and spermidine. Fermentation inoculated with combination of Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen, and Monascus anka with negative‐decarboxylase activity may prevent biogenic amine formation in the fermented ssurimi and improve hygienic quality of fermented surimi.     

Biogenic amine formation in Nham,a Thai fermented sausage,and the reduction by commercial starter culture, Lactobacillus plantarum BCC 9546

Biogenic amines are of concern for sausage due to their toxicological effects on nervous, blood pressure, gastric and intestinal systems. In this study, the influence of raw pork meat quality and starter culture inoculation on biogenic amines accumulation in Nham, a Thai traditional fermented pork, were studied. Before Nham processing, pork meat was stored at 30 °C for 6 h, and at 4 and −20 °C for 2 days. Formation of biogenic amines (cadaverine, putrescine, histamine and tyramine) was significantly higher in Nham processed from stored meat. Accumulation of these biogenic amines in Nham reduced significantly by the addition of Lactobacillus plantarum BCC 9546, a commercial Nham starter culture. The results highlight the importance of using fresh meat products and the inclusion of an appropriate starter culture to minimise the formation of biogenic amines during the process of Nham fermentation.     

Bacterial community succession and biogenic amine changes during fermentation of fish-chili paste inoculated with different commercial starter cultures

The microbial dynamics, diversity and succession characterisation during the fermentation of fish-chili paste, with and without starter cultures by high-throughput sequencing, were investigated to identify the relationship between microbial composition and the accumulation of biogenic amines. Results showed that Firmicutes was the predominant phylum, and Lactobacillus and Weissella were the main genera during fermentation process, regardless of the inclusion of starter cultures. Compared with naturally fermented samples (NS; no starter cultures), Pediococcus and Staphylococcus flourished in samples inoculated with SBM-52 starter culture (Staphylococcus xylosus + Staphylococcus carnosus + Pediococcus pentosaceus + Pediococcus acidilactici). Staphylococcus also accounted for a larger proportion in samples inoculated with WBX-43 starter culture (Staphylococcus xylosus + Staphylococcus carnosus). Furthermore, Enterobacter was inhibited in inoculated samples. Unweighted pair-group method with arithmetic means and principal component analyses revealed that microbiota structures were different among NS, SBM and WBX samples. In case of biogenic amines, the concentration of putrescine, cadaverine and tyramine increased to higher levels in NS samples compared with inoculated samples. Our work also investigated the relationship between core bacterial communities and biogenic amines. This study provided an improved understanding for the effects of starter cultures on the microbial community and the quality of fish-chili paste.     

Novel starter cultures to inhibit biogenic amines accumulation during fish sauce fermentation

Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in food products such as meat and fish sausages. However, little information is available regarding the application of these bacteria in fish sauce. Hence, our study was aimed to investigate the effect of such starter cultures in reducing biogenic amines accumulation during fish sauce fermentation. Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05 isolated from fish sauce which possess amine oxidase activity were used as starter cultures in this study. Fermentation was held for 120 days at 35 °C. The pH value increased in all samples, while salt concentration remained constant throughout fermentation. Aerobic bacteria count was significantly lower (p < 0.05) in the control than in inoculated samples as a result of starter cultures addition. However, it decreased during fermentation due to the growth inhibition by high salt concentration. Proteolytic bacterial count decreased during fermentation with no significant difference (p > 0.05) among samples. These bacteria hydrolyzed protein in anchovy to produce free amino acid precursors for amines formation by decarboxylase bacteria. The presence of biogenic amines producing bacteria in this study was considered to be indigenous from raw material or contamination during fermentation, since our cultures were negative histamine producers. Amino acid histidine, arginine, lysine and tyrosine concentration decreased at different rates during fermentation as they were converted into their respective amines. In general, biogenic amines concentration namely histamine, putrescine, cadaverine and tyramine increased throughout fermentation. However, their concentrations were markedly higher (p < 0.05) in the control (without starter cultures) as compared to the samples treated with starter cultures. Histamine concentration was reduced by 27.7% and 15.4% by Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively. Both cultures could also reduce other amines during fermentation. After 120 days of fermentation, the overall biogenic amines concentration was 15.9% and 12.5% less in samples inoculated with Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively, as compared to control samples. These findings emphasized that application of starter cultures with amines oxidase activity in fish sauce fermentation was found to be effective in reducing biogenic amines accumulation.     

Malolactic fermentation of Tempranillo wine: contribution of the lactic acid bacteria inoculation to sensory quality and chemical composition

Malolactic fermentation (MLF) of Tempranillo Rioja wines (Spain) inoculated with two lactic acid bacteria (LAB) strains were studied and compared with spontaneous MLF. Inoculation with selected Oenococcus oeni lyophila shortened MLF duration up to 19 days and lead to wines with more fresh and fruity characters, especially when implantation was 100%. We demonstrated modifications in the concentration of volatile and nitrogenous compounds and a good correlation between analytical and sensory attributes was also noted. In addition, the low initial amino acid concentration and the consumption of these compounds by the inoculated yeast strain during alcoholic fermentation resulted in wines with very low biogenic amines levels (under 3.75 mg L^?1) after MLF and 3 month storage period in all cases. The results showed the significance of choose the most suitable starter to elaborate quality wines and suggest the control of amino acid content in must and wine to prevent the formation of biogenic amines.     

Biogenic amines formation in Streptococcus thermophilus isolated from home-made natural yogurt

Twelve different biogenic amines formation in 58 isolates of Streptococcus thermophilus from home-made natural yogurt were investigated in histidine (HDB) and lysine decarboxylase broth (LDB). All S. thermophilus isolates had an ability to produce twelve different biogenic amines in HDB and LDB. Most of the S. thermophilus isolates formed low amounts of histamine (1–50 mg/L) from histidine. Apart from one isolate, S. thermophilus produced tyramine at low (47 isolates) and medium (10 isolates) levels. The amount of each specific biogenic amine produced by S. thermophilus was generally lower than 100 mg L⁻¹. Also, the presence of hdcA gene was investigated using PCR technique and relation between gene and histamine production was conducted in S. thermophilus isolates. This study showed that most of the S. thermophilus isolates have the ability to form biogenic amines, especially histamine, and tyramine, which is an important consideration when selecting strains as starter cultures.     

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     

Improvement of the quality and abatement of the biogenic amines of grass carp muscles by fermentation using mixed cultures

BACKGROUND: To improve the quality of processed grass carp (Ctenopharyngodon idellus) products and control the accumulation of hazardous substances therein, minced grass carp slices were salted for 6 h at room temperature and then inoculated with mixed starter cultures of Lactobacillus casei, Streptococcus lactis, Saccharomyces cerevisiae Hansen and Monascus anka and fermented for 12 h at 30 °C. The changes in some characteristics and biogenic amine contents of the fermented muscles were investigated. RESULTS: During the 12 h fermentation at 30 °C, muscles inoculated with mixed starter cultures showed a rapid decrease in pH from 6.0 to 5.1 and suppression of the growth of enterobacteria and pseudomonads. The fermented muscles exhibited better colour, appearance, flavour and overall acceptability than the control (P < 0.05). The changes in non‐protein nitrogen and free amino acid contents of the fermented muscles and in their sodium dodecyl sulfate polyacrylamide gel electrophoresis profiles indicated that severe hydrolysis of muscle proteins occurred during fermentation. The accumulation of biogenic amines in the muscles was efficiently reduced by fermentation with mixed starter cultures. CONCLUSION: Fermentation with mixed starter cultures of L. casei, S. lactis, S. cerevisiae Hansen and M. anka significantly improved the characteristics of grass carp muscles and controlled the accumulation of biogenic amines. Copyright © 2009 Society of Chemical Industry     

The effects of a starter culture on the formation of biogenic amines in Turkish soudjoucks

The effects of starter culture containing Lactobacillus sake, Pediococcus pentosaceus, Staphylococcus carnosus plus Staphylococcus xylosus on the formation of biogenic amines during ripening of Turkish soudjoucks were investigated. Determination of eight different biogenic amines was carried out by reverse-phase high performance liquid chromatography (HPLC) using diode array detection. It was found that aerobic plate counts (APC) decreased in the samples with added starter culture. Lactic acid bacteria counts increased in both the controls and samples with added starter during ripening. Total Enterobacteriaceae (ENT) counts decreased on the 30th day for vacuum packed and starter added samples. Yeast counts decreased only in vacuum packed and starter added samples on the 30th day. No biogenic amines were found in ground meat samples. Putrescine (PU) and tyramine (TYR) were determined in all control samples during the storage at 4°C. Starter addition inhibited formation of PU but not TYR.     

Microbiological Profile of Goat Meat Inoculated with Lactic Acid Bacteria Cultures and Stored at 30��C for 7?days

The ability of lactic acid bacteria (LAB) cultures to preserve goat meat at 30°C was evaluated in the present study. Strains of Pediococcus pentosaceus GOAT 01 and Lactobacillus plantarum GOAT 012, individually and in combination, were applied as starters on sliced meat samples at 6 log cfu/g and stored for 7 days at 30°C to simulate ambient temperature in Nigeria. They were evaluated for microbiological profile during storage. Reduction in bacterial counts was recorded for enterobacteriaceae, Staphylococcus, yeasts and moulds in starter culture inoculated samples (SCIS), whereas an increase occurred in uninoculated control samples (UCS). In challenge experimental trials, two different sets of meat were inoculated individually with 6 log cfu/g each of pathogenic organisms, Listeria monocytogenes and Salmonella Typhimurium. The inoculated pathogens were monitored during storage to assess the influence of starter cultures on them. Approximately 1 log reduction was recorded in the viable count of L. monocytogenes on day 1, while counts were below detection limit (<2 log) on day 2 in meat samples inoculated with P. pentosaceus alone and in combination with L. plantarum. Counts of Salmonella Typhimurium showed about 2 log reduction in SCIS, inoculated with P. pentosaceus alone and in combination with L. plantarum, on day 2 while an increase by 4 logs was observed in UCS. Our findings suggest that the protective effect of the LAB strains could be exploited in shelf life extension and control of foodborne pathogens in goat meat. If the starter strains could be improved upon, their potential as biopreservatives may be engaged in the preservation of the meat in Nigeria, where storage systems have been very inadequate.     

Impedance measurements to assess microbial contamination of ready-to-use vegetables

 An impedometric method for estimating the microbial load of ready-to-use vegetables, either inoculated or not inoculated with antimicrobial-producing lactic acid bacteria, was investigated. The correlation (R ²) between total mesophilic bacteria (log cfu/g) and time to detection was 0.889. A discrepancy was observed for the impedometric measurement of low coliforms counts, particularly in samples inoculated with lactic acid bacteria and stored at 8°C for 6 days. Time to detection for the two groups of microorganisms was less than 11 h. Received: 25 November 1996 / Revised version: 23 January 1997     

Impedance measurements to assess microbial contamination of ready-to-use vegetables

 An impedometric method for estimating the microbial load of ready-to-use vegetables, either inoculated or not inoculated with antimicrobial-producing lactic acid bacteria, was investigated. The correlation (R ²) between total mesophilic bacteria (log cfu/g) and time to detection was 0.889. A discrepancy was observed for the impedometric measurement of low coliforms counts, particularly in samples inoculated with lactic acid bacteria and stored at 8°C for 6 days. Time to detection for the two groups of microorganisms was less than 11 h. Received: 25 November 1996 / Revised version: 23 January 1997     

Reduction of biogenic amine formation using a negative amino acid-decarboxylase starter culture for fermentation of Fuet sausages

The ability of Lactobacillus sakei CTC494, a negative amino acid-decarboxylase starter culture, to reduce biogenic amine accumulation during sausage fermentation and storage at 4 and 19 degrees C was studied. The effect on the amine formation of the tyramine producer Lactobacillus curvatus CTC371, as a positive strain, was also examined in comparison to a spontaneous fermentation process without starter culture (control batch). The polyamines spermine, spermidine, and diaminopropane were not influenced by the ripening, and their levels slightly decreased in all the batches throughout the storage. Tyramine, cadaverine, and putrescine were the main amines formed during the ripening. The addition of starter culture resulted in a decrease on the biogenic amine formation, depending on the strain inoculated. A great reduction in tyramine content was achieved when L. sakei CTC494 was inoculated, whereas L. curvatus CTC371 only attenuated tyramine accumulation compared with the control batch. Both starters were able to significantly limit the production of putrescine and cadaverine, and they inhibited tryptamine and phenylethylamine formation by the wild microbial flora. Tyramine levels of the control sausages rose during the storage at both temperatures, whereas those of cadaverine only increased at 19 degrees C. On the contrary, sausages manufactured through the starter controlled fermentation did not show changes of amine contents during the storage. The addition of a proper selected starter culture is advisable to produce safer sausages with low contents of biogenic amines.     

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.     

传统中式香肠中产生物胺氧化酶菌的分离鉴定

结合培养基显色法和氧化酶试验,对发酵香肠中产生物胺氧化酶优势菌进行了初步分离。进而利用肉品发酵剂标准剔除不符合要求的菌株。利用高效液相色谱检测符合要求的产生物胺氧化酶菌株对生物胺的氧化减少能力。最后采用16S rDNA分子鉴定方法,通过同源性比对,得到4株符合发酵剂标准的产生物胺氧化酶菌株,即1株表皮葡萄球菌,3株模仿葡萄球菌。     

不同条件对两株乳酸菌降解生物胺的影响

为探究从新疆熏马肠中分离出的植物乳杆菌和唾液乳杆菌在不同环境条件下降解生物胺的效果,通过高效液相色谱(High Performance Liquid Chromatography,HPLC)检测两株菌在不同条件下(生物胺浓度、初始pH、温度、NaCl浓度和乙醇浓度)降解生物胺的情况。结果表明:当生物胺浓度在200~300 μg/mL时两株菌生物胺降解能力基本达到最大,而当生物胺浓度大于300 μg/mL时两株菌降解生物胺的能力逐渐下降,但唾液乳杆菌(Lactobacillus salivarius)对生物胺的降解能力显著高于植物乳杆菌(Lactobacillus plantarum)(p<0.05),分别在pH为6.5和7.0时对大多数生物胺具有较高的降解能力;两株菌均在32~37 ℃温度范围内对生物胺有较好的降解效果;当NaCl浓度达到8%,乙醇浓度达到4.5%时,两株菌对生物胺的降解活性被强烈抑制。可见本研究中的唾液乳杆菌相对于植物乳杆菌更适合作为商业发酵剂,具有更好的降解生物胺的特性。     

The fate of recombinant DNA in thermally treated fermented sausages

 The fate of recombinant DNA (rDNA) from genetically modified starter organisms in thermally treated fermented sausages (Kochsalami) was monitored by a polymerase chain reaction (PCR)-based estimation of the concentration of the rDNA. We defined an rDNA titre as the smallest volume of the DNA solution which still results in a detectable amplicon on PCR. In sausages a degradation of the rDNA was observed during storage. However, this did not proceed so far that rDNA was no longer detectable after 9 weeks of incubation. It was observed that the rDNA is strongly protected against the activity of DNase I in the meat matrix. To include the aspect of release of rDNA from genetically modified starter organisms into the meat matrix, Kochsalami-type sausage was produced with the aid of recombinant strains of Lactobacillus curvatus as starter organisms. It was shown that only a minute part of the rDNA contained in the starter organisms can be recovered from the meat matrix by extraction as "free" DNA. To recover the total rDNA, cell wall lytic enzymes were necessary. No significant effect on the detectability of the rDNA was observed on variation of the ecological factors prevailing during the production and storage of the sausages, such as the temperature of thermal treatment, pH and fat content. The concomitant presence of Staphylococcus aureus (>10⁶ cfu/g) producing heat-stable nuclease did not affect the detection of the rDNA. Received: 29 March 1999     

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.