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.     

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.     

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.     

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.     

The effect of ripening and storage conditions on the distribution of tyramine,putrescine and cadaverine in Edam-cheese

The aim of the work was to describe the development of selected biogenic amines (histamine, tyramine, putrescine and cadaverine) in 4 layers of Dutch-type cheese (Edam-cheese) depending on 3 ripening/storage regimes during a 98-day period. Biogenic amines were analysed by means of ion-exchange chromatography. A further goal was to identify microbial sources of biogenic amines in the material analysed. Phenotype characterization and repetitive sequence-based PCR fingerprinting were used to identify the isolated bacteria. The highest content of tyramine, putrescine and cadaverine was determined in cheeses stored in a ripening cellar at a temperature of 10 °C during the whole observation period. Lower biogenic amines content was determined in samples which were moved into a cold storage device (5 °C) after 38 days of storage in a ripening cellar (10 °C). The lowest concentrations of biogenic amines were detected in cheeses which were moved into a cold storage device (5 °C) after 23 days of storage in a ripening cellar (10 °C). During the 98-day period, histamine was not detected in any of the regimes. Within the cheeses analysed, non-starter lactic acid bacteria Lactobacillus curvatus, Lactobacillus casei/paracasei and Lactobacillus plantarum were detected as the main producers of the biogenic amines tested. In starter bacteria Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris the decarboxylase activity tested was not detected.     

Microbiological attributes and biogenic amine content of probiotic Turkish fermented sausage

Microbiological attributes and biogenic amine content of Turkish fermented sausage manufactured by using probiotic starter cultures (Lactobacillus casei, L. acidophilus or their combination) were investigated before and after fermentation-drying period and during refrigerated storage at 4?±?1?°C for 8?months at 2?month intervals. As results of the study, during fermentation and storage biogenic amine content (histamine, putrescine, cadaverine and tyramine) of the samples were increased significantly. Salmonella, Staphylococcus aureus, coliform and fecal coliform microorganisms were not detected during fermentation and storage. Probiotic microorganism counts of all samples were higher than the lower limit of 6.0 log cfu/g which is requested for probiotic foods.     

Effects of food additives on biogenic amine formation in Myeolchi-jeot, a salted and fermented anchovy (Engraulis japonicus)

This study was carried out to reduce biogenic amine contents in Myeolchi-jeot, a salted and fermented anchovy (Engraulis japonicus). The effects of various food additives on biogenic amine formation were determined by HPLC. The greatest inhibitory effect on biogenic amine production was observed in the culture treated with glycine. In the culture, the contents of putrescine, cadaverine, histamine, tyramine and spermidine were reduced by 32.6%, 78.4%, 93.2%, 100.0% and 100.0%, respectively, compared to control. The other additives tested had less effect in inhibiting biogenic amine production. Out of food additives tested, glycine was finally applied to the ripening of Myeolchi-jeotin situ, and then overall production of biogenic amines was reduced by up to 63.0% and 73.4%, compared to controls prepared with no and 20% NaCl, respectively. Therefore, it is expected that the findings of this study enhance the safety of not only Myeolchi-jeot but other salted and/or fermented seafood.     

Microbiological attributes and biogenic amine content of probiotic Turkish fermented sausage

Microbiological attributes and biogenic amine content of Turkish fermented sausage manufactured by using probiotic starter cultures (Lactobacillus casei, L. acidophilus or their combination) were investigated before and after fermentation-drying period and during refrigerated storage at 4 ± 1 °C for 8 months at 2 month intervals. As results of the study, during fermentation and storage biogenic amine content (histamine, putrescine, cadaverine and tyramine) of the samples were increased significantly. Salmonella, Staphylococcus aureus, coliform and fecal coliform microorganisms were not detected during fermentation and storage. Probiotic microorganism counts of all samples were higher than the lower limit of 6.0 log cfu/g which is requested for probiotic foods.     

Limosilactobacillus fermentum FSCBAD033控制带鱼加工下脚料发酵鱼露中生物胺含量研究

目的 控制带鱼加工下脚料低盐发酵鱼露中的生物胺含量。方法 以耐盐性高效生物胺降解乳酸菌Limosilactobacillus fermentum FSCBAD033为功能发酵剂控制带鱼加工下脚料发酵鱼露中的生物胺含量,并研究其对鱼露发酵过程中的pH值、NaCl含量、总可溶性氮(Total Soluble Nitrogen,TSN)含量、氨基酸态氮(Amino Acid Nitrogen,AAN)含量、挥发性盐基氮(Total Volatile Basic Nitrogen,TVB-N)含量以及发酵结束时感官风味的影响。结果 与Control组相比,接种L. fermentum FSCBAD033发酵剂对鱼露发酵过程中的NaCl含量影响不大(p>0.05),但可促进pH值下降以及TSN和AAN含量上升,并显著抑制TVB-N的产生(p<0.05)；显著降低发酵结束时鱼露中52.38%尸胺、40.21%组胺、45.44%酪胺、23.74%腐胺、52.67%苯乙胺和43.26%总生物胺(p<0.05),对含量较低的色胺、精胺和亚精胺影响不大；此外,接种L. fermentum FSCBAD033发酵剂可增强发酵结束时鱼露的鲜味和肉味,减弱氨味、腥味和臭味,整体提升鱼露感官风味。结论 L. fermentum FSCBAD033可作为带鱼加工下脚料低盐发酵的功能发酵剂,用于生产低盐、营养丰富、感官风味好且生物胺含量低的高品质鱼露产品。     

Effect of different starter cultures on the biogenic amines production as a critical control point in fresh fermented sausages

The selection of critical control points is one of the most important steps in the design of a hazard analysis and critical control points system. In this study, the influence of starter culture inoculation on biogenic amine accumulation in fresh fermented sausage as critical control point was studied. The effect of three starter culture Lactobacillus plantarum plus Bifidobacterium lactis, L. plantarum plus Bifidobacterium bifidum or L. plantarum plus B. lactis and B. bifidum on amines production were investigated. Tyramine (TY), Cadaverine (CA), Putrescine (PU), Histamine (HIS) and Tryptamine (TR) contents increased during the fermentation and storage stage, and TY was the prevailing amine in the final product. Sausages produced by fermentation with starters, as compared to natural fermentation (control), had a lower amount of biogenic amine. A great reduction in biogenic amine content was achieved when Bifidobacterium sp. were inoculated, also levels of species of Enterobacteriaceae and Pseudomonas decreased, while lactic acid bacteria counts increased during fermentation and storage.     

The biogenic amines and bacterial changes of farmed rainbow trout (Oncorhynchus mykiss) stored in ice

The biogenic amine content and related bacterial changes (Pseudomonas spp., psychrotrophic and mesophilic counts) in whole farmed rainbow trout (Oncorhynchus mykiss) were monitored during ice storage for 18 days (at 0, 3, 6, 9, 12, 15 and 18 days). Levels of putrescine, cadaverine and histamine, and bacterial loads, increased (P < 0.05) during storage, but tyramine was not detected in any of the tested samples. Concentration of putrescine ranged from 0.4 initially to 8.97 μg/g, and psychrotrophic microorganisms were dominant. The best linear regressions (correlations) were for putrescine and Pseudomonas spp. and psychrotrophs (r = 0.98), and for cadaverine with Pseudomonas spp. (r = 0.82). Putrescine content was a good quality marker. Histamine was detected only at later stages of storage and was therefore less suitable than the other biogenic amines as freshness indicator.     

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.     

Changes of yolk biogenic amine concentrations during storage of shell hen eggs

The RP-HPLC/UV method, using dabsyl derivatization, optimised for the determination of biogenic amines in egg yolk, was appropriate for quantification of putrescine, cadaverine, histamine, ethylamine, propylamine, ethanolamine, tyramine, tryptamine, spermine, spermidine, phenylethylamine. Detection limits ranged between 0.05 and 0.06 mg of biogenic amine/kg of egg yolk. Two experiments using, respectively, farm and avian eggs were conducted to evaluate yolk biogenic amine concentrations of fresh and stored eggs, and to explain the effect of temperature and time of storage in the levels of biogenic amines during egg shelf-life. Only five of the 11 biogenic amines under study were detected: putrescine, cadaverine, propylamine, ethylamine and ethanolamine. Storage time during shelf-life presented a significant effect on the levels of the five amines (p < 0.01). On the contrary, storage temperature did not presented a significant effect on the levels of the mentioned amines, p > 0.01. The significant reduction of biogenic amine concentration during the shelf-life justified the application of a multiple linear regression using stepwise method to estimate the storage time. The regression equation was applied with success to confirm the storage time of farm eggs and avian eggs that were stored at two different temperatures.     

Tyrosine- and histidine-decarboxylase positive lactic acid bacteria and enterococci in dry fermented sausages

Lactic acid bacteria (LAB) and enterococci were isolated immediately after stuffing (day 0), at the end of ripening (28th day) and at the end of storage (112th day) from dry fermented sausages produced by two different producers (K; R) in two diameters (4.5 and 7 cm) using either of two spice mixtures (P; H) and either of two starter cultures (Pediococcus pentosaceus, C; Lactobacillus curvatus + Staphylococcus carnosus, F), resulting in a total of 16 different combinations. Tyrosine-decarboxylase DNA sequence (tyrdc) was identified on average in 88% and 44% of enterococci and LAB isolates, respectively at the end of ripening, the corresponding figures regarding histidine-decarboxylase gene sequence (hisdc) was 71% and 16%, respectively. Lactobacillus plantarum, L. brevis and L. casei/paracasei, and Enterococcus faecium and Enterococcus faecalis were identified as tyramine/histamine producers in the sausages.     

Biogenic amines and potential histamine – Forming bacteria in Rihaakuru (a cooked fish paste)

The biogenic amine concentration in Rihaakuru (a fish paste) (n = 28), obtained from different parts of the Maldives (North, South, and Central), was determined by HPLC. Ten biogenic amines were detected; agmatine, not detected (ND) – 161 ppm; cadaverine, ND – 387 ppm; histamine, ND – 5487 ppm; putrescine, ND – 290 ppm; phenylethylamine, ND – 23 ppm; serotonin, ND – 91 ppm; spermine, ND – 329 ppm; spermidine, ND – 79 ppm; tryptamine, ND – <5 ppm; and tyramine, ND – 50 ppm. Nine biogenic amines were found in 3 samples, 8 in 10 samples, 7 in 6 samples, 6 in 3 samples, 4 in 5 samples, and 1 was found in 1 sample. Histamine was detected at levels that are regarded as a risk to human health. Fourteen isolates were selected from two randomly selected samples out of the 28 samples of Rihaakuru and screened for histamine production. Twelve of the 14 isolates produced histamine, with the highest histamine producers being Bacillus massiliensis Nai5 (6.65 ppm) and Bacillus polyfermenticus (5.58 ppm); while Bacillus malacitensis produced the least (<0.5 ppm).     

发酵香肠中生物胺生成量的菌株效应分析

本实验对乳杆菌(米酒乳杆菌和戊糖乳杆菌)和片球菌(戊糖片球菌和小片球菌)在发酵香肠中生物胺的生成量进行了分析。结果表明,供试乳杆菌和片球菌均能产生不等量的组胺、尸胺、腐胺、酪胺、亚精胺和精胺,但都不产生色胺,产品中的生物胺生成量表现出明显的菌株效应。采用复配菌种(片球菌+葡萄球菌)发酵可以明显降低香肠中组胺、尸胺、腐胺和酪胺以及总生物胺的含量。     

Use of a starter culture of lactic acid bacteria in plaa-som, a Thai fermented fish

Plaa-som is a Thai fermented fish prepared from freshwater fish and various ingredients. In this study, two strains of lactic acid bacteria (LAB) isolated from natural plaa-som fermentation were used as starter cultures: Lactobacillus plantarum IFRPD P15 and Lactobacillus reuteri IFRPD P17. These strains were used as a mixed starter culture for plaa-som using an air-drying method (laminar airflow) with sterilized rice grains as the filler. This method produced a suitable starter culture, which was maintained at 4 °C for more than 20 weeks. LAB were the dominant bacteria in the starter culture and produced high acidity from 24 h until the end of fermentation. This resulted in decreased pH in plaa-som. L. plantarum IFRPD P15 was dominant as an acidity producer, whereas L. reuteri IFRPD P17 showed an ability to suppress and eliminate pathogenic bacteria such as Escherichia coli within 24 h. The use of a single starter culture for plaa-som resulted in incomplete suppression of pathogenic bacteria and elimination of E. coli. Thus, L. plantarum IFRPD P15 and L. reuteri IFRPD P17 have great potential for use as a mixed starter culture in plaa-som fermentation and may possibly help to reduce fermentation time.     

Technological conditions influence aminogenesis during spontaneous sausage fermentation

The influence of two manufacturing processes on biogenic amine formation during the manufacture of Spanish dry fermented sausages of different diameters (fuet and llonganissa) was evaluated to elucidate which conditions allow better control of the aminogenic activity of spontaneous microbiota. Technological conditions affected both the amounts and the qualitative profile of biogenic amine accumulated. The higher processing temperature and relative humidity in process A (simulating those applied in industrial manufacture) favoured aminogenesis, since biogenic amine accumulation was faster and higher than in sausages manufactured under the process B (close to those used in traditional practices). The major amine differed depending on the diameter of the sausages, tyramine being the major amine in fuet (2.5 cm diameter sausage), and putrescine in llonganissa (4.5 cm). Moreover, sausages of higher diameter (llonganissa) had higher biogenic amine contents compared with the thinnest sausages (fuet). Conditions would modulate biogenic amine accumulation not only due to its influence on development of the bacterial population but also on its aminogenic activity. From the biogenic amine point of view, when sausages are spontaneously fermented, traditional lower temperatures and relative humidities are more appropriate than those usually applied in industrial processes.     

Mixed starter cultures to control biogenic amine production in dry fermented sausages

Several combinations of an amine-negative Lactobacillus sakei strain, along with proteolytic Staphylococcus carnosus or Staphylococcus xylosus strains, were used to study the influence of mixed starter cultures on biogenic amine production during the manufacture of dry fermented sausages. Changes in pH, water content, proteolysis, microbial counts, and biogenic amine contents were simultaneously examined in a spontaneously fermented batch and in three mixed starter-mediated batches. A double-controlled microbial charge initially inoculated as mixed starter culture of L. sakei and Staphylococcus spp. (all amine-negative strains) drastically reduced tyramine, cadaverine, and putrescine accumulation. No production of other aromatic amines such as histamine, phenylethylamine, or tryptamine was observed in any batch. The polyamines, spermine and spermidine, were found in raw materials and their levels decreased slightly in the spontaneously fermented batch. No correlation between proteolysis and biogenic amine production was observed. The use of proper technological conditions favoring starter development and the use of the raw materials with good hygienic quality make it possible to produce fermented sausages nearly free of biogenic amines.     

The effect of different starter cultures and ripening temperatures on formation of biogenic amine in Turkish fermented sausages

Different starter culture added groups (Group I: Lactobacillus sake, Staphylococcus xylosus; Group II: Lactobacillus plantarum, Staphylococcus carnosus; Group III: Lactobacillus curvatus, Staphylococcus carnosus, and Staphylococcus xylosus) and control group sausage samples were produced experimentally at two different temperatures (22 and 26 °C). In raw materials and all groups, by the 0th, 2nd, 5th and 7th days of the ripening period, the analysis of HPLC and biogenic amine (tyramine, histamine, cadaverine, putrescine, phenylethylamine, tryptamine, spermine, and spermidine) amounts, microbiological (Lactic acid bacteria, Micrococcus/Staphylococcus, Enterobacteriaceae, Enterococcus spp., and moulds–yeasts) and chemical (pH, dry matter, and salt) have been done. During the ripening period, there is no statistical discrepancy (P > 0.05) in terms of biogenic amine amounts, microbiological, and chemical values detected from the sausage samples produced at 26 and 22 °C. However, in both levels of temperature, there is statistical discrepancy (P < 0.001) detected in terms of tyramine, putrescine values and the count of Enterococcus spp. between the starter culture added samples and control group samples. By this study, it has been stated that the ripening temperature does not make any statistical discrepancy (P > 0.05) for all values but the use of starter culture prevents the formation of biogenic amine in Turkish fermented sausages.