Concentrations of biogenic amines in fish,squid and octopus and their changes during storage

The concentrations of seven biogenic amines (BA) were simultaneously determined in 74 samples of fish, squid and octopus, by the method of HPLC coupled with pre-column derivatisation. The relationship between the formation of BA in aquatic products and the growth of microbial flora during storage was also investigated. Results showed that putrescine, cadaverine, histamine and tyramine were the dominant BA in the studied samples, but the concentrations of histamine and tyramine were mostly less than 50 and 100 mg kg⁻¹, respectively. Freezing can effectively prevent the formation of BA, but the levels of putrescine, cadaverine, histamine and tyramine significantly increased (p < 0.05) during storage at 4 and 25 °C. The growth of mesophilic or psychrophilic bacteria in blue scad and octopus strongly and positively correlated with the formation of amines (such as putrescine, cadaverine, histamine and tyramine) during storage, except for histamine in octopus.     

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.     

Manufacture of Turkish Beyaz cheese added with probiotic strains

The survival of the probiotic strains Lactobacillus fermentum (AB5-18 and AK4-120) and Lactobacillus plantarum (AB16-65 and AC18-82), all derived from human faces, was investigated in Turkish Beyaz cheese production. Three batches of Turkish Beyaz cheese were produced: one with the test probiotic culture mix (P), another with a commercial starter culture mix including Lactoccocus lactis subsp. cremoris, Lactococcus lactis subsp. lactis (C) and the third with equal parts of the commercial starter culture mix and test probiotic culture mix (CP). The cheeses were ripened at 4 °C for 120 days and the viability of cultures was determined monthly. Cheese samples were analyzed for total solids, fat in solids, titratable acidity, pH, salt in total solids, proteolysis, sensory evaluation, aroma compounds and biogenic amines. While initial lactic acid bacteria load in P cheese was 2.7 × 10⁹ at the beginning, it was 7.42 × 10⁷ cfu/g at the end of 120 days of ripening. The results showed that test probiotic culture mix was successfully used in cheese production without adversely affecting the cheese quality during ripening. The chemical composition and sensory quality of P cheeses were also comparable with C cheeses. The present study indicates that probiotic cultures of human origin are feasible for Turkish Beyaz cheese production.     

Monitoring of biogenic amines in cheeses manufactured at small-scale farms and in fermented dairy products in the Czech Republic

The aim of the study was the monitoring of six biogenic amines (histamine, tyramine, phenylethylamine, tryptamine, putrescine, and cadaverine) and two polyamines (spermidine and spermine) in 112 samples of dairy products purchased in the Czech Republic, namely in 55 cheeses made in small-scale farms and in 57 fermented dairy products. The products were tested at the end of their shelf-life period. Neither tryptamine nor phenylethylamine was detected in the monitored samples; histamine was found only in four cheese samples containing up to 25 mg/kg. The contents of spermine and spermidine were low and did not exceed the values of 35 mg/kg. Significant amounts of tyramine, putrescine, and cadaverine occurred especially in cheeses produced from ewe’s milk or in long-term ripened cheeses. In about 10% of the tested cheeses, the total concentration of all the monitored biogenic amines and polyamines exceeded the level of 200 mg/kg, which can be considered toxicologically significant. In fermented dairy products, the tested biogenic amines occurred in relatively low amounts (generally up to 30 mg/kg) that are regarded safe for the consumer’s health.     

Impact of fermentation conditions and refrigerated storage on microbial quality and biogenic amine content of sauerkraut

White cabbage (Brassica oleracea L. var. capitata cv. Bronco) was fermented, at 0.5% and 1.5% NaCl, using Lactobacillus plantarum or Leuconostoc mesenteroides as starter cultures and, subsequently, sauerkraut was stored at 4 °C for 3 months. Microbial populations and six biogenic amines (putrescine, cadaverine, histamine, tyramine, spermine and spermidine) were investigated. Fermentation and storage increased aerobic mesophilic bacteria and LAB populations in sauerkrauts, and this was accompanied by a rise in biogenic amine content. L. plantarum sauerkrauts produced with 0.5% NaCl had the highest microbial counts, whilst no differences between salt contents were found with L. mesenteroides. Total biogenic amine amount was lower at 0.5% NaCl than at 1.5% in both induced fermentations and L. mesenteroides produced a lower content than did L. plantarum. Spermidine was the major contributor to the total biogenic amine content, followed by putrescine, whilst histamine was present at the lowest level. The individual and total biogenic amine levels in the experimental sauerkrauts stored at 4 °C for 3 months were below the upper limits reported in the literature for fermented products, indicating good quality and safety of the sauerkrauts. L. mesenteroides starter and 0.5% NaCl were the optimal fermentation conditions for producing sauerkrauts with the lowest biogenic amine contents.     

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.     

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.     

Effect of low-dose irradiation on biogenic amines formation in vacuum-packed trout flesh (Oncorhynchus mykiss)

The effects of vacuum packaging followed by high-energy electron beam irradiation on the shelf-life of fillets of rainbow trout (Oncorhynchus mykiss) were examined. Samples were irradiated at doses of 0.25, 0.50, 0.75, 1.0 and 2.0 kGy. The control and treated packs were stored at 3.5 °C for up to 28, 42, 70 and 98 days (control - 0, 0.25, 0.50 and >0.50 kGy respectively). The content of seven biogenic amines (putrescine, cadaverine, spermidine, spermine, histamine, tyramine and tryptamine) were determined. Putrescine, cadaverine and tyramine showed very good correspondence with the irradiation dose and organoleptic properties. Samples of good quality contained less than 10 mg/kg of each of these amines. The polyamines spermidine and spermine did not show statistically significant changes with the irradiation dose and the time of storage. With few exceptions, histamine was not detected in the samples treated with radiation. Tryptamine was not detected in any of the samples.     

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.     

Changes in biogenic amine and polyamine contents in smear-ripened cheeses during storage

The content of eight biologically active amines in two groups of smear-ripened cheeses (unwashed-rind and washed-rind) was analysed. The biogenic amine levels were determined after storage at 5 °C in three periods of shelf lifetime: at the minimum durability date; 1 and 2 weeks after this date. The analytical procedure consisted of amine extraction from sample matrix, derivatisation with dansyl chloride and ultra-performance liquid chromatography quantification. The content of biogenic amines and polyamines significantly differed according to the technology of ripening. The cheeses unwashed during ripening had much higher contents of all observed amines and polyamines in comparison with the washed-rind cheeses. The mean content of putrescine, cadaverine and tyramine exceeded 100 mg/kg in unwashed-rind cheeses, while the other amines occurred at lower levels. The content of all detected amines was very low in washed-rind cheeses; no tryptamine, phenylethylamine and histamine were found. The effect of storage on the amine formation was not confirmed.     

Formation of biogenic amines by Gram-negative bacteria isolated from poultry skin

The aim of this study is to explore production of biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) by 88 Gram-negative bacteria isolated from poultry skin (41 isolates of family Enterobacteriaceae, 21 isolates of genus Aeromonas, 16 isolates of genus Pseudomonas, and 10 isolates of other Gram-negative rods). Ion-exchange chromatography was employed to analyse the above mentioned amines. Enterobacteria were found to be the largest producers of amines with proved presence of tyramine, agmatine, putrescine, and cadaverine in cultivation broth after incubation of bacteria. Putrescine and cadaverine were the most abundant products. Presence of at least two biogenic amines, i.e. mainly concurrent presence of putrescine and cadaverine, was revealed in 19 enterobacteria strains. Eleven isolates classified into Aeromonas genus produced putrescine and five of them also formed cadaverine. The other observed biogenic amines (histamine, spermine and spermidine) were not found among tested isolates. Production of biogenic amines by any Pseudomonas family isolates was not proved.     

不同NaCl添加量对牦牛乳硬质干酪成熟过程中生物胺产生的影响

以不同盐添加量（1.0%、1.3%、1.8%和2.3%）牦牛乳硬质干酪为研究对象,分析牦牛乳硬质干酪0～6 个月成熟过程中生物胺的动态变化,并对干酪中产胺微生物进行筛选。结果显示：不同盐添加量牦牛乳硬质干酪中生物胺主要为色胺、β-苯乙胺、尸胺、酪胺和腐胺,未检测到组胺,生物胺积累阶段主要集中在成熟后期。不同盐添加量干酪中色胺含量最低,且在成熟后期含量差异较小。当盐添加量从2.3%减少到1.0%时,干酪中β-苯乙胺含量减少。盐添加量分别为1.0%和1.3%时,干酪中尸胺含量较低,且未检测出腐胺。当盐添加量在1.8%～2.3%时,随着盐添加量的增加,干酪中尸胺和腐胺含量整体呈现增加趋势。不同盐添加量干酪成熟过程中,其酪胺含量范围为3.13～49.81 mg/kg,且盐添加量为1.3%干酪中酪胺含量较高。不同盐添加量干酪中生物胺总量最高为304.18 mg/kg。采用显色培养基筛选出一株产胺微生物,经分子生物学鉴定为Enterococcus durans。     

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 amines and HL60 citotoxicity of alfalfa and fenugreek sprouts

The use of germinated seeds as food originated in far east countries and has recently spread to the western world where they are seen as fresh and healthy ingredients. While sprouted alfalfa is widely consumed, sprouted fenugreek seeds are not commonly produced, yet could be active ingredients for blood glucose and cholesterol control. As part of a safety evaluation of sprouted alfalfa and fenugreek flours, as novel ingredients for use in functional foods, their contents of biogenic amines and HL60 cytotoxicity were studied. Alfalfa (Medicago sativa) and fenugreek (Trigonella foenum-graecum) were germinated or 4 days at 20 °C and 30 °C, with and without light. Ungerminated seeds contained putrescine, cadaverine, histamine, tyramine, spermidine and spermine. Bioactive amine levels found in alfalfa sprouts were twice higher than those found in raw seeds and germination at 20 °C without light provided the lowest levels of total biogenic amines. In sprouted fenugreek, only putrescine and cadaverine increased during germination and temperature and light exposure brought about little change. The amount of biogenic amines in sprout seeds was always below acceptable healthy levels. Results obtained in HL60 leukemic cells showed apoptosis, cell proliferation and cell viability values similar to those found for distilled water and no toxic effects were found. The results provide support for the use of germinated alfalfa and fenugreek seeds as ingredients in functional foods.     

Biogenic amine content and biogenic amine quality indices of sardines (Sardina pilchardus) stored in modified atmosphere packaging and vacuum packaging

A comparative study of the effects of packaging on the formation of biogenic amines during storage of sardines (Sardina pilchardus) at 4 °C in air, modified atmosphere pack (MAP) and vacuum pack (VP) was carried out. Sardines were organoleptically acceptable for up to 3 days in air, 12 days in MAP and 9 days in VP. The biogenic amine content generally increased in all treatments with increasing storage time. The concentrations of putrescine and/cadaverine in fish stored in air reached maximum levels of 12.2 mg/100g at 12 days and 10.0 mg/l00 g at 15 days. Significant differences were found (P < 0.05) in the levels of cadaverine and putrescine among the three treatments. Spermidine and spermine levels increased slightly and did not change much throughout the storage period for all experimental conditions. The amine contents of sardine were highest in sardine stored in air, followed by VP and MAP. Quality indices related to the contents of the major biogenic amines were calculated and they correlated well with organoleptic qualities.     

Content and distribution of biogenic amines in Dutch-type hard cheese

Content of biogenic amines (BA; histamine, tyramine, tryptamine, putrescine, 2-phenylethylamine, cadaverine, spermidine, spermine) and counts of microorganisms (total aerobic and facultative anaerobic mesophilic microorganisms, TAC; lactic acid bacteria, LAB; coliforms; total anaerobic microorganisms, TAN; yeasts + moulds), were evaluated within the ripening interval of 1–6 months in Dutch-type hard cheese, produced by two different producers (the cheeses designated as R30 and R45) using pasteurized milk and different starter cultures (designated as R30-FD and R30-CH in the case of the R30 cheeses). The cheeses from the two producers differed in their fat content (30% and 45% of fat in dry matter, respectively).     

The effect of storage temperatures and packaging methods on properties of Motal cheese

The effects of storage temperature (+4°C and -18°C) and packaging method (nonvacuum and vacuum) on biogenic amines in Motal cheese during storage periods were investigated. In addition, dry matter, titratable acidity, total nitrogen, water-soluble nitrogen, trichloroacetic acid-soluble nitrogen, phosphotungstic acid-soluble nitrogen, free amino group (proteolysis), electrophoretic patterns of casein, and amounts of lactic acid bacteria and coliforms were determined. Storage period had a significant effect on all of the biogenic amines. When compared with vacuum packaging, normal packaging had higher amounts of putrescine, cadaverine, histamine, and tyramine. Coliforms were not found at detectable levels (<100 cfu/g) in all cheese samples. Results of urea-PAGE analysis of cheese samples were in good agreement with biogenic amine results and other proteolysis parameters.     

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.     

The effect of different packaging methods on the formation of biogenic amines and organic acids in Kashar cheese

The effects of packaging methods (nonvacuum and vacuum) on biogenic amines (cadaverine, putrescine, tyramine, tryptamine, phenylethylamine, and histamine) and organic acids (citric, lactic, formic, acetic, propionic, and butyric) during storage for 180 d at 4°C were investigated in Kashar cheese. Dry matter, titratable acidity, total nitrogen, water-soluble nitrogen, trichloroacetic acid-soluble nitrogen, phosphotungstic acid-soluble nitrogen, free amino group (proteolysis), pH, fat, and acid degree value were also determined. Storage period had a significant effect on all of the biogenic amines. When compared with vacuum packaging, nonvacuum packaging resulted in no large differences among the amounts of biogenic amines. Vacuum-packaged cheeses had more lactic, formic, acetic, and butyric acids than did cheeses packaged without vacuum. Water-soluble nitrogen, trichloroacetic acid-soluble nitrogen, phosphotungstic acid-soluble nitrogen, proteolysis, pH, and acid degree values of the cheese samples increased continuously until the end of the ripening in all the samples. No significant change was observed in total nitrogen, dry matter, or fat content within the ripening period, whereas titratable acidity values changed significantly in vacuum-packaged cheese and decreased slightly in the non-vacuum-packaged cheeses. The results of this study showed that storage period and packaging method had significant effects on the quality of Kashar cheese.     

Biogenic amines and their producers in Akawi white cheese

The aim of this work was to study the biogenic amine content of brine‐ripened cheeses after one year of storage and then to investigate possible contaminating micro‐organisms with decarboxylase activity. The biogenic amine production of isolates was tested in vitro. The most frequent biogenic amines were putrescine, histamine and tyramine. The biogenic amine content detected in one cheese sample was above 120 mg/kg; this can be considered toxicologically relevant. Decarboxylase activity was found for 33 contaminating micro‐organisms. Isolates belonging to Bacillus licheniformis, Debaryomyces hansenii, Staphylococcus equorum and Serratia marcescens produced significant amounts of putrescine and cadaverine.