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.     

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.     

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.     

Biogenic amine formation and bacterial contribution in fish,squid and shellfish

Forty-one species of fish, squid and shellfish were analyzed for biogenic amine (BA) contents. Most of the fish samples showed lower BA contents, whereas some samples showed higher contents than the allowable levels. Shellfish and squid samples had negligible BA levels. Four fish species containing high BA levels were analyzed for changes in histamine contents during storage. In the most samples, the histamine contents remarkably increased up to 36.6–2123.9 mg/kg after 24 h of storage at 25 °C, while the contents began to gradually increase after 2–3 days of storage at 4–10 °C. The dominant microbial group was enterobacteria throughout the storage period. Meanwhile, out of total 119 strains isolated from different fish species showing high BA levels, 23 strains identified as Enterobacter aerogenes produced large amounts of histamine, putrescine and cadaverine, and 33 strains identified as two different Enterobacter spp. produced less histamine but large amounts of putrescine and cadaverine.     

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.     

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.     

Safety Assessment of Crayfish (Astacus Leptodactylus ESCH., 1823) from Microbial Load and Biogenic Amines Signature: Impact of Post-Catch Icing and Frozen Storage

In prevalent conditions, fresh-caught fish were held on ice until storage at optimal temperatures. The aim of this study was to investigate and confirm biogenic amines formation and microbiological quality of crayfish during 2 days post-catch icing and 90 days frozen storage. Of the considered biogenic amines in fresh crayfish, puterscine and cadaverine were detected and initial concentrations of them were 5.33 and 50.57 μg/g of flesh, respectively. Psychrotrophs and cadaverine were the major bacteria and biogenic amines detected in crayfish at all sampling stages, respectively. At the end of ice storage, samples had higher biogenic amines and bacterial load when compared with fresh samples (P < 0.05). During the first 30 days of frozen storage, simultaneous with slight changes of biogenic amines, bacterial load significantly decreased (P < 0.05), but as frozen storage time lengthened, progressive development of biogenic amines and microbial load (except for Pseudomonas spp.) was observed. The best correlation was for psychrotrophic with histamine (r = 0.82). At the end of storage, although final values of bacterial load were very negligible, total BAs (487.03 μg/g), especially histamine (110.22 μg/g) exceeded the proposed tolerable maximum levels for total biogenic amines (300 μg/g) and histamine (50 μg/g). It could be concluded that crayfish can be hazardous after 60 days.     

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.     

Microbiological Properties and Biogenic Amines of Whole Pike‐Perch (Sander Lucioperca,Linnaeus 1758): A Perspective on Fish Safety During Postharvest Handling Practices and Frozen Storage

Abstract: The biogenic amines (tyramine, histamine, cadaverine, and puterscine) and microbiological properties (mesophilic, psychrotrophic, and Pseudomonas spp.) of whole pike‐perch (Sander lucioperca) was investigated during 2 d prestorage icing and 90 d frozen storage (−24 °C). At the end of ice storage, a noticeable increase only was found for puterscine level (P < 0.05), and microbial loads of fish increased in comparison with fresh fish (P < 0.05). During the frozen storage, as time passed, a continuous increase of biogenic amines and decrease of bacterial load (except for Pseudomonas spp. at the last 30 d) was detected (P < 0.05). The total contents of biogenic amines ranged from 6.24 to 91.76 μg/g during the investigated period. Puterscine was the major amine detected in pike‐perch and its concentration varied between 1.75 and 56.95 μg/g; due to a more step‐wise increase it was a good quality indicator. At the end of storage, all of the obtained values are below the tolerable maximum amounts based on available regulations. Based on biogenic amines content and microbial load, it could be concluded that pike‐perch can be consumed without any health risks after 2 d icing condition and 90 d frozen storage. Practical Application: Biogenic amines as one of the commonest forms of food intoxication occur in protein‐rich food such as fish. Short‐time icing during transportation is the simplest method to fish preserving for processing or long‐term storage. In this study formation of biogenic amines and bacterial changes in ungutted pike‐perch as highly demanded fish species for human consumption, during transportation and frozen storage was investigated. The results of the research can be advantageously used by fish industry. These findings suggest that the production and storage practices of fish in the retails condition could have acceptable food quality level.     

Biochemical,sensory and microbiological attributes of wild turbot (Scophthalmus maximus), from the Black Sea,during chilled storage

Freshness of wild turbot (Scophtalmus maximus) stored in ice was assessed by chemical, sensory and microbiological methods. The limit for sensory acceptability of wild turbot stored in ice was ∼12–15 days. The quality of turbot decreased on day 15 (B) and they were no longer acceptable on day 19 (C). The TVB-N level showed fluctuations during storage, indicating that TVB-N could not be a good indicator of turbot quality. The release of FFA increased from an initial value of 6.33 (expressed as % of oleic acid) to a final value of 20.6 during the storage period. The initial PV value was 5.60 meq/kg for turbot stored in ice and it started to increase to 21.6 meq/kg on day 12 and then started to decrease to 13.6 meq/kg at the end of storage period. The level of TMA in wild turbot increased sharply from an initial value of 9.36 mg/kg to a final value of 38.9 mg/kg. Linear regressions (r²) obtained from K, K_i, G, P, H and F_r were 0.92, 0.89, 0.99, 0.89, 0.96 and 0.89, respectively, for the wild turbot stored in ice. Turbot maintained high (E) and good quality (A) during the first 12 days of chilled storage when the average K, K_i and P values were ∼78–85%, and H, F_r and G values were ∼45%, 15% and 149%, respectively. Eight biogenic amines were investigated, namely, histamine, putrescine, cadaverine, spermidine, spermine, tryptamine, tyramine, and 2-phenylethylamine, three amines (histamine, tyramine, and tryptamine) were not detected in any of the fish samples during the storage period. As storage time progressed, putrescine and cadaverine became the dominant amines, reaching 22.7, and 16.9 mg/kg, respectively, at 19 days of storage in ice. Total viable counts of whole gutted turbot increased from the initial value of 3.3 log cfu g⁻¹ (day 0) to 7.87 log cfu g⁻¹ (day 19) over the period of storage. If 10⁶ microorganisms/g are considered to be the TVC limit of acceptability, the shelf life of turbot was approximately ∼13–14 days.     

Analysis of Vacuum Packed Beef Regarding Psychrotrophic Bacteria Growth and Biogenic Amines Content

It has been recognized that biogenic amines (BA) content in meat can be considered a freshness marker. Considerable amounts of some BAs can appear during food storage under certain conditions, according to the handling of the raw material, technology applied, storage temperature and time, packaging condition, mainly if amino acid - decarboxylase positive microorganisms are present. The aim of this study was to evaluate the psychrotrophic bacteria growth and metabolic production of BAs during chill storage of beef. The vacuum packed beef cuts (Longissimus dorsi muscle) were analyzed during storage at 7 °C at 0, 15, 30, 45, and 60 d, to determine the psychrotrophic bacteria growth and the BAs amount. The BAs considered were: putrescine, cadaverine, histamine, spermidine, and spermine. The BAs quantitative determination was carried out by reversed phase high-performance liquid chromatography (HPLC) with UV detection. Statistic procedures were performed using SAS statistical software. The growth parameters of psychrotrophic bacteria including lag phase, maximum specific growth rate, maximum bacterial cell density, initial population, mean square error, and coefficient of determination were determined according to Baranyi and Roberts model. The values of histamine and spermidine increased significantly (P < 0.0001) during storage time, while the levels of spermine decreased (P < 0.0001). Psychrotrophic bacteria counts increased significantly (P < 0.0001) reaching 7.6 log cfu/g over time. The counts of this group positively correlated to histamine and spermidine (r = 0.68 and 0.61, respectively), while spermine showed a negative correlation (r = -0.70). Conversely, no significant correlation was found between psychrotrophics counts and putrescine or psychrotrophics counts and cadaverine.     

Nucleotide degradation and biogenic amine formation of wild white grouper (Epinephelus aeneus) stored in ice and at chill temperature (4 °C)

Sensory (cooked and uncooked), chemical (proximate composition, TVB-N, nucleotide degradation products and biogenic amines) and microbiological quality (TVC and total coliform) changes were investigated during storage of ungutted white grouper kept in ice and at chill temperature (4 °C). According to the sensory assessment, the shelf life of white grouper was 16 days in ice and 4 days for fish stored at chill temperature. TVB-N values increased with storage time. Amines found in white grouper stored in ice were TMA, putrescine, cadaverine, 2-phenylethylamine, dopamine, agmatine, tryptamine and serotonin. Histamine, spermine, spermidine were never detected with either storage condition. The acceptability limit in terms of microbial count was exceeded at 8 days in ice and at 4 days for fish stored at chill temperature. Total coliform count was 2.8 log₁₀ cfu/ml at 1 day and reached 10⁵ cfu/ml for both storage conditions.     

Microbiological changes and its correlation with quality indices during aerobic iced storage of sea salmon (Pseudopercis semifasciata)

Pseudomonas spp. and Shewanella putrefasciens were the dominant bacteria during the ice stored period. Among the specific spoilage organisms (SSO), S. putrefasciens was identified as the most dominant spoilage bacterium, followed by Aeromonas spp. A good correlation (r=0.9829) between log₁₀ counts of SSO bacteria and total volatile bases (TVB) was observed in this study, while trimethylamine (TMA) increased more slowly along the storage. According to microbial changes, TBV (35 mg nitrogen/ 100 g sample), TMA (15.75 mg nitrogen/100 g sample), pH (7.2) and alteration of general organoleptic characteristics, the shelf life of sea salmon stored in ice at 0 °C was considered less than 10 days.     

Determination of biogenic amines in foods using ultra-performance liquid chromatography (UPLC)

A rapid ultra-performance liquid chromatographic (UPLC) method for the determination of biogenic amines (putrescine, cadaverine, spermidine, spermine, phenylethylamine, histamine, tyramine and tryptamine) in selected food samples is described. The eight biogenic amines, which are the most important to be determined in food samples, were derivatized with dansyl chloride prior to UPLC separation. The dansylated amines were separated on an Agilent Zorbax Eclipse XDB – C18 column (50 × 4.6 mm ID, 1.8 μm) using gradient elution with a binary system of acetonitrile–water, a flow rate of 1.0 ml/min and UV detection at 225 nm. The analysis is very fast, all amines are well resolved and are eluted from the column in less than 6 min. The average repeatability of the method ranged between 1.02% and 2.14%. Limits of detection (LODs) for considered amines ranged between 0.032 and 0.098 μg/l; calibration curves showed very good linearity (r = 0.9994–1.0000). The method was applied to the analysis of amines in pork, beef, chicken and fish meat, cheese and edible mushrooms.     

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.     

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).     

Effect of UV-C Radiation on Shelf life of Vacuum Package Colossoma Macropomum x Piaractus Mesopotamicus Fillets

Colossoma macropomum x Piaractus mesopotamicus (CP) is a freshwater fish with greatest commercial importance in Brazil. Fillets of CP are highly perishable food and preservation technology with UV-C could improve food safety and extend shelf life. Fillet samples were submitted at UV-C (55.83mJ/cm²) and examined for mesophilic and psychrotrophic count and biogenic amines over 6 days. UV-C reduced the bacterial growth and number of colonies in the stationary phase; also increase the levels of cadaverine, putrescine and histamine. The results suggest that UV-C enhanced the shelf-life of CP fillets by at least 50%.     

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.     

Effects of water extract of Urtica dioica L. and modified atmosphere packaging on the shelf life of ground beef

Effects of lyophilized Urtica dioica L. water extract (LUWE) and modified atmosphere packaging (MAP) on the quality and shelf life of ground beef were investigated. Ground beef was stored as aerobic control, MAP (80%O₂ + 20% CO₂), MAP + 250 ppm LUWE and MAP and 500 ppm LUWE at 2 ± 0.5 °C for 14 days. MAP and LUWE had significant effects on mesophilic, psychrotrophic and lactic acid bacteria and Pseudomonas counts. Depending on the level of LUWE, Pseudomonas and psychrotrophic counts decreased. Treatment with 500 ppm LUWE + MAP showed the lowest TBARS values compared to other groups during storage. 80% O₂-MAP increased TBARS values. Treatment had no significant effect on L* and b* values of the exterior of the ground beef, but had significant effects on the color of interior sections.     

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.