Histamine contents of salted seafood products in Taiwan and isolation of halotolerant histamine-forming bacteria

Fifty-seven salted seafood products sold in the fishing village stores in Taiwan, including salted fish product, salted mollusc product and salted shrimp product, were tested to determine the occurrence of histamine and histamine-forming bacteria. Although the average content of each of nine different biogenic amines in all samples was less than 5.0 mg/100 g, 10.5% (6/57) of tested samples had the histamine content greater than the 5.0 mg/100 g allowable limit suggested by the US Food and Drug Administration. One histamine-producing bacterial strain, capable of producing 78.5 ppm of histamine in trypticase soy broth supplemented with 1.0% l-histidine (TSBH), was identified as Bacillus megaterium. The B. megaterium isolate was a halotolerant bacterium which grew well to an elevated NaCl concentration of 15% in TSBH medium. Besides, it had a consistent ability to produce >300 ppm of histamine at 10% NaCl concentration in TSBH medium after 72 h.     

Histamine formation by histamine-forming bacteria in douchi,a Chinese traditional fermented soybean product

Seven soybean and 19 black bean douchi products sold in the supermarkets in southern Taiwan were purchased and tested to determine the occurrence of histamine and histamine-forming bacteria. The levels of pH, salt content, water content, yeast and mold, and aerobic plate count (APC) in all samples ranged from 4.7 to 5.9, 4.4% to 14.0%, 6.8% to 51.6%, 3.0 to 5.1 log CFU/g, and 5.2 to 9.2 log CFU/g, respectively. None of these samples contained total coliform and Escherichia coli. Although black bean douchi products had an average histamine content of 29.0 mg/100 g, 18 of them had histamine contents greater than 5 mg/100 g, the allowable level set by the US Food and Drug Administration (FDA) for scombroid fish and/or products. In contrast, only four soybean douchi products had histamine levels greater than 5 mg/100 g. Among the black bean samples, four contained histamine at 56.3, 62.1, 80.2 and 80.8 mg/100 g, that are above the 50 mg/100 g hazard action level. Eight histamine-forming bacterial strains, capable of producing 11.7–601 ppm of histamine in trypticase soy broth (TSB) supplemented with 1% l-histidine (TSBH), were identified as Bacillus subtilis (four strains) Staphylococcus pasteuri (one strain) and Staphylocuccus capitis (three strains) by 16S rDNA sequencing with PCR amplification. S. capitis, which was previously reported to be halotolerant, was a potent histamine-former capable of producing more than 500 ppm of histamine in TSBH in the presence of 0.5–10% NaCl.     

Chemical characterisation and histamine-forming bacteria in salted mullet roe products

Sixteen salted mullet roe products sold in the retail markets in Taiwan were purchased and tested to determine the occurrence of histamine and histamine-forming bacteria. The levels of pH, salt content, water content, total volatile basic nitrogen (TVBN) and aerobic plate count (APC) in all samples ranged from 5.4 to 5.8, 5.1% to 7.2%, 15.4% to 27.3%, 32.0 to 69.6 mg/100 g and <1.0 to 7.1 log CFU/g, respectively. None of these samples contained total coliform and Escherichia coli. The average content of each of the nine biogenic amines in all samples was less than 4 mg/100 g, and only one mullet roe sample had the histamine content (8.18 mg/100 g) greater than the 5.0 mg/100 g allowable limit suggested by the US Food and Drug Administration. Two histamine-producing bacterial strains capable of producing 10.7 ppm and 9.6 ppm of histamine in trypticase soy broth (TSB) supplemented with 1.0% l-histidine (TSBH) were identified as Staphylococcus carnosus by 16S rDNA sequencing with PCR amplification, and they were isolated from the sample with higher histamine content (8.18 mg/100 g).     

Histamine content and histamine-forming bacteria in dried milkfish (Chanos chanos) products

Thirty-two dried milkfish products sold in five retail markets in southern Taiwan were purchased and tested to determine the occurrence of histamine and histamine-forming bacteria. Except for histamine and cadaverine, the average content of various biogenic amines in tested samples was less than 8.5 mg/100 g. Most of the tested dried milkfish products (78.1%) had histamine levels greater than the FDA guideline of 5 mg/100 g for scombroid fish and/or product, while fourteen of them (43.7%) contained > 50 mg/100 g hazard action level. Thirty histamine-producing bacterial strains, capable of producing 5.4 ppm to 562 ppm of histamine in trypticase soy broth supplemented with 1.0% L-histidine (TSBH), were identified as Enterobacter aerogenes (seven strains), Citrobacter sp. (one strain), Staphylococcusxylosus (ten strains), S. sciuri (one strain), Bacillus thuringiensis (two strains), Citrobacter freundii (five strains), Klebsiella pneumoniae (one strain) and E. cloacae (three strains), by 16S rDNA sequencing with PCR amplification.     

Histamine and other biogenic amines and histamine-forming bacteria in miso products

Twenty-seven miso products sold in supermarkets and 13 products sold in retail markets were purchased from southern Taiwan, and tested to determine the occurrence of histamine and histamine-forming bacteria. The levels of pH, salt content, and aerobic plate count (APC) in all samples ranged from 5.1 to 5.8, 6.1% to 13.8%, and 2.1 to 9.1 log CFU/g, respectively. Only one of the supermarket miso products contained 100 MPN/g total coliform. None of these samples contained Escherichia coli. Although the average content for each of the nine biogenic amines in all samples was less than 5 mg/100 g, two supermarket samples (22.1 and 11.9 mg/100 g) and one retail market sample had histamine content (10.2 mg/100 g) greater than the 5.0 mg/100 g allowable limit suggested by the US Food and Drug Administration. Eight histamine-producing bacterial strains, capable of producing 10.4–39.4 ppm of histamine in trypticase soy broth (TSB) supplemented with 1.0% l-histidine (TSBH), were identified as Staphylococcus pasteuri (one strain), Bacillus sp. (one strain), B. amyloliquefaciens (two strains), B. subtilis (two strains) and B. megaterium (two strains), by 16S rDNA sequencing with PCR amplification.     

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.     

Histamine formation by histamine-forming bacteria and yeast in mustard pickle products in Taiwan

The occurrence of histamine, histamine-forming bacteria and yeast were tested in 37 mustard pickle products sold in both retail markets and supermarkets in southern Taiwan. Aerobic plate count (APC), total coliform, and Escherichia coli were also tested for microbiological quality. Salt content, pH value, titratable acidity and sulphite content were determined for quality of mustard pickle products. Only one retail market sample and one supermarket sample had 8.9 and 7.4 mg histamine per 100 g products, although the average content for each of the nine biogenic amines was less than 2 mg/100 g. Ten histamine-forming bacterial strains and 6 histamine-producing yeast strains capable of producing 8.7 to 1260 ppm of histamine in trypticase soy broth (TSB) supplemented with 1% l-histidine (TSBH) were identified as Staphylococcus capitis (four strains),Staphylococcus pasteuri (two strains), Enterobacter cloacae (four strains), Candida glabrata (two strains) and Candida rugosa (four strains). S. capitis, which was previously reported to be halotolerant, was a potent histamine-former, capable of producing more than 1000 ppm of histamine in TSBH in the presence of 0.5–10% NaCl. The numbers of the aerobic plate count (APC) in all samples were below the Taiwanese regulatory level of 5 log CFU/g. None of the samples contained total coliform or E. coli. The values of pH, salt content, titratable acidity and sulphite content in all samples ranged from 3.8% to 5.0%, 2.0% to 10.0%, 0.21% to 1.18% and <2.0–1876 ppm, respectively.     

Proteinase-producing halophilic lactic acid bacteria isolated from fish sauce fermentation and their ability to produce volatile compounds

Halophilic lactic acid bacteria were isolated from fish sauce mashes fermented at 1 to 12 months. Seven out of sixty-four isolates were selected according to their proteolytic activity and growth at 25% NaCl for characterization and investigation of volatile compound production. All selected isolates were Gram-positive cocci with pairs/tetrads and grew at 0-25% NaCl, pH 4.5-9.0. Results of 16S rRNA gene sequence analysis showed 99% homology to Tetragenococcus halophilus ATCC 33315. The restriction fragment length polymorphism (RFLP) patterns of all isolates were also similar to those of T. halophilus ATCC 33315. These isolates were, thus, identified as T. halophilus. All isolates hydrolyzed fish protein in the medium containing 25% NaCl. Intracellular aminopeptidase of 7 isolates exhibited the highest activity of 2.85-3.67 U/ml toward Ala-p-nitroanilide (Ala-pNA). T.halophilus strains MS33 and M11 showed the highest alanyl aminopeptidase activity (P < 0.05), and produced histamine in mGYP broth containing 5 and 25% NaCl in the level of 6.62-22.55 and 13.14-20.39 mg/100 ml, respectively. Predominant volatile compounds of fish broth containing 25% NaCl inoculated with T. halophilus MS33 and MRC5-5-2 were 1-propanol, 2-methylpropanal, and benzaldehyde, corresponding to major volatile compounds in fish sauce. T.halophilus appeared to play an important role in volatile compound formation during fish sauce fermentation.     

Determination of histamine and histamine-forming bacteria in tuna dumpling implicated in a food-borne poisoning

An incident of food-borne poisoning causing illness in seven victims, due to ingestion of tuna dumpling, occurred in March 2006, in Chiayi Prefecture, southern Taiwan. The leftovers of the victims’ tuna dumpling and the five other tuna dumpling samples from five other retail stores were collected and tested to determine the occurrence of histamine and histamine-forming bacteria. The levels of pH, salt content, aerobic plate count (APC), total volatile basic nitrogen (TVBN), total coliform (TC) and Escherichia coli in all samples ranged from 6.08 to 6.43, 0.46% to 0.81%, 5.90 to 8.95 log CFU/g, 6.38 to 21.29 mg/100 g, 750 to 8000 most probable number (MPN)/g, and <3 to 1000 MPN/g, respectively. The suspected tuna dumpling contained 160.8 mg/100 g of histamine greater than the hazard action level of 50 mg/100 g set by the US Food and Drug Administration (FDA) for tuna fish. Given the allergy-like symptoms of the victims and the high histamine content in the suspected tuna dumpling, this food-borne poisoning was strongly suspected to be due to histamine intoxication. In addition, although thirteen histamine-producing bacteria strains capable of producing 8.1–19.7 ppm of histamine in trypticase soy broth (TSB) supplemented with 1.0% l-histidine (TSBH), were identified as Enterobacter sp. (three strains), Pantoea agglomerans (two strains), Klebsiella variicola (four strains) and Serratia marcescens (four strains), by 16S rDNA sequencing with PCR amplification, they were not determined to be the main contributors to histamine accumulation in suspected tuna dumpling.     

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).     

蔗糖对复合菌快速发酵虾头酱产品品质的影响

研究蔗糖添加量对复合菌种发酵产品品质的影响.选取凡纳滨对虾虾头为原料,发酵前期加入不同浓度的蔗糖,追踪虾头酱发酵期间的理化及安全指标(pH、组胺、TVB-N和AA-N等),对成品酱营养成分测定分析,并进行感官评定.结果表明,蔗糖组发酵后期pH稳定在8.6左右,菌落总数减少.添加蔗糖后虾头酱粗脂肪、蛋白质含量无显著变化(...     

Diversity of plasmids encoding histidine decarboxylase gene in Tetragenococcus spp. isolated from Japanese fish sauce

Nineteen isolates of histamine producing halophilic bacteria were isolated from four fish sauce mashes, each mash accumulating over 1000 ppm of histamine. The complete sequences of the plasmids encoding the pyruvoyl dependent histidine decarboxylase gene (hdcA), which is harbored in histamine producing bacteria, were determined. In conjunction, the sequence regions adjacent to hdcA were analyzed to provide information regarding its genetic origin. As reference strains, Tetragenococcus halophilus H and T. muriaticus JCM10006^T were also studied. Phenotypic and 16S rRNA gene sequence analyses identified all isolates as T. halophilus, a predominant histamine producing bacteria present during fish sauce fermentation. Genetic analyses (PCR, Southern blot, and complete plasmid sequencing) of the histamine producing isolates confirmed that all the isolates harbored approximately 21-37 kbp plasmids encoding a single copy of the hdc cluster consisting of four genes related to histamine production. Analysis of hdc clusters, including spacer regions, indicated > 99% sequence similarity among the isolates. All of the plasmids sequenced encoded traA, however genes related to plasmid conjugation, namely mob genes and oriT, were not identified. Two putative mobile genetic elements, ISLP1-like and IS200-like, respectively, were identified in the up- and downstream region of the hdc cluster of all plasmids. Most of the sequences, except hdc cluster and two adjacent IS elements, were diverse among plasmids, suggesting that each histamine producers harbored a different histamine-related plasmid. These results suggested that the hdc cluster was not spread by clonal dissemination depending on the specific plasmid and that the hdc cluster in tetragenococcal plasmid was likely encoded on transformable elements.     

An amperometric biosensor for the rapid assessment of histamine level in tiger prawn (Penaeus monodon) spoilage

Histamine could accumulate in seafood when bacteria spoilage commenced and caused histamine poisoning without altering the fish normal appearance and odor. Therefore, a histamine biosensor using immobilized enzyme diamine oxidase (DAO) has been developed for the rapid monitoring of the histamine levels in tiger prawn (Penaeus monodon). The histamine biosensor had a response time of <1 min and optimum pH of operation was 7.4 with reproducibility and repeatability (n = 5) of 4.87% and 5.26% relative standard deviations (RSD) respectively. Recoveries ranging from 93.11% to 100.58% were obtained for histamine spiked at levels from 5 to 20 ppm. The variation in histamine levels of some tiger prawn samples after a 5-h exposure at temperature of 30 °C ± 2 were studied using the histamine biosensor and the results were comparable to histamine levels determined by an HPLC method. The two methods showed a linear correlation with R² = 0.9612 (Y = 0.9164x + 5.58). The limit of detection was 0.65 ppm of histamine, which is below the indicator level of 50 ppm established by USA FDA. The reusable biosensor is simple and can be used for direct histamine determination without further sample pretreatment, and is suitable for the routine analysis of histamine in tiger prawns to monitor spoilage.     

Analysis of biogenic amines in fermented fish products consumed in Korea

The aim of this study was to investigate the biogenic amine (BA) contents of fermented fish foods in Korea. A total of 47 samples of anchovy and sand lance sauces and squid, clam, and shrimp pastes were used for high performance liquid chromatography (HPLC) analysis. The histamine and tyramine contents of the anchovy and sand lance sauces were significantly high. The average histamine contents of all tested samples were >300 mg/kg and the histamine contents were higher in the sauce (aekjeot) samples than in the paste (jeotgal) samples. This result strongly suggests careful monitoring of BA contents in these products is necessary to ensure consumer health.     

Quality characteristics of Dutch-style fermented sausages manufactured with partial replacement of pork back-fat with pure, pre-emulsified or encapsulated fish oil

Dutch-style fermented sausages were manufactured with 15% and 30% pork back-fat substitution by pure or commercial encapsulated fish oil, either added as such or as pre-emulsified mixture with soy protein isolate. Adding commercial encapsulated fish oil was the most important factor influencing the chemical composition. The fat content was not significantly different between products (p > 0.05). The n−6/n−3 ratio decreased from 8.49 in controls to 0.90-2.47 in modified products. Lipid oxidation parameters (propanal and hexanal) showed much higher values for sausages with pure fish oil than for products with encapsulated oil. For the latter, lipid oxidation was similar to controls. Products with encapsulated or pre-emulsified oil were significantly firmer than products from other treatments in physical and sensory analysis (p < 0.05). Overall, it is technologically feasible to enrich dry fermented sausages with n−3 fatty acids from fish oil and the application of commercial encapsulated fish oil seems to be the best in retaining overall quality.     

Occurrence of histamine and histamine-forming bacteria in kimchi products in Taiwan

Twenty kimchi products sold in supermarkets and 17 products sold in retail markets were purchased from southern Taiwan and tested to determine the occurrence of histamine and histamine-forming bacteria. The levels of pH and salt content in all samples ranged from 3.6 to 5.1 and 1.5–16.0%, respectively. The supermarket kimchi products had 1–7.2 log CFU/g of APC and <3–600 MPN/g of total coliform (TC), and the retail market kimchi products had 4–8.03 log CFU/g of APC and <3 to >2400 MPN/g of TC. Only one of the retail market kimchi products contained 20 MPN/g Escherichia coli. Although, supermarket kimchi products had an average histamine content of 49.8 mg/100 g, 15 of them had histamine content greater than 5 mg/100 g, the allowable level set by the US Food and Drug Administration (FDA) for scombroid fish and/or product. In contrast, only eight retail market kimchi products had histamine levels greater than 5 mg/100 g. Among the supermarket samples, three contained histamine at 50.2, 273 and 535 mg/100 g, that are more than the 50 mg/100 g hazard action level. Four histamine-producing bacteria capable of producing 13.6–43.1 ppm of histamine in MRS broth supplemented with 0.25% l-histidine were identified as Lactobacillus para. paracasei (one strain), Lb. brevis (one strain), and Brevibacillus brevis (two strains). To our knowledge, this is the first report to demonstrate the occurrence of high contents of histamine and histamine-forming bacteria in kimchi products.     

Seasonal effects on the physicochemical characteristics of fish sauce made from capelin (Mallotus villosus)

Fresh capelin (Mallotus villosus) was harvested from the North Atlantic during both summer and winter fishing seasons. Reaction conditions for fish sauce processing were optimized with respect to temperature, salt concentration and reaction time, using a response surface methodology (RSM) experimental design. Whole capelin was minced and samples were ground with increasing salt concentrations. RSM optimizations were conducted, ranging from 5% to 30% salt, and incubating at 5° intervals from 0 to 65 °C. Autolytic activity was estimated by extracting the liquid formed by the mixture with trichloroacetic acid and estimating protein content by the Lowry method. Samples for fish sauce production were then prepared under optimized conditions by mixing ground capelin with 10% salt and incubating at 50 °C for up to 270 days for the summer capelin and up to 360 days for the winter capelin. Samples were collected at regular intervals and analyzed for liquid yield, moisture, protein, soluble solids, specific gravity, pH, colour and amino acid content. Kjeldahl protein content in the fish sauce from summer capelin was 2.03% after 250 days of fermentation and twice as high as that in winter capelin fish sauce. Moisture content and pH were lower in the summer capelin fish sauce, but Brix and density were higher than those in fish sauce from winter capelin. Brown colour formation was very rapid in the summer capelin fish sauce but slow in the winter capelin fish sauce. Summer capelin may be successfully utilized for the production of fish sauce without added enzymes.     

Effects of dietary replacement of fish oil by conjugated linoleic acid on some meat quality traits of Pacific white shrimp Litopenaeus vannamei

This experiment was conducted to evaluate the effects of fish oil replacement by conjugated linoleic acid (CLA) on body proximate analysis and thiobarbituric acid-reactive substances, fat content, shear force and fatty acid composition in musculature of Pacific white shrimp (Litopenaeus vannamei). Graded levels of CLA (0%, 0.5%, 1% and 2%) were added to the basic diet of shrimp at the expense of fish oil. Results showed that fat content (p = 0.036) and shear force (p = 0.001) in shrimp musculature were enhanced with increasing dietary CLA inclusion. Fish oil replacement by CLA significantly promoted the incorporation of cis-9, trans-11 CLA (p = 0.0001) and trans-10, cis-12 CLA (p < 0.0001) into shrimp musculature; moreover, the polyunsaturated fatty acid was elevated (p = 0.020) and monounsaturated fatty acid was reduced by CLA inclusion (p = 0.024). It was concluded that replacement of fish oil by CLA could improve some meat quality traits of shrimp and 1% CLA was an appropriate amount.     

Characterisation of histamine-producing bacteria from farmed blackspot seabream (Pagellus bogaraveo) and turbot (Psetta maxima)

Turbot (Psetta maxima) and blackspot seabream (Pagellus bogaraveo) represent two of the most important emerging farmed fish species in European countries. However, no information of the presence and development of histamine-producing bacteria on them has been reported so far. Accordingly, the aim of this study was to isolate and identify the main histamine-producing bacteria in farmed turbot and blackspot seabream. For this study, 24 isolates (12 from turbot and 12 from blackspot seabream) were preliminarily selected on Niven medium. Two of these isolates were confirmed as prolific histamine producers by HPLC. Thus, Pseudomonas fragi (isolated from turbot) and Pseudomonas syringae (isolated from blackspot seabream) were able to produce 272 ± 69 ppm and 173 ± 45 ppm of histamine in vitro, respectively, after incubation at 30 °C/24 h. While turbot fillets proved to be quite resistant to histamine formation at temperatures below 10 °C, blackspot seabream fillets inoculated with P. syringae and the prolific histamine former Morganella morganii accumulated 696 ± 84 and 760 ± 59 ppm histamine, respectively, under such conditions. Genetic identification based on 16S rRNA sequencing was performed in parallel with the investigation of characteristic mass spectral profiles of the isolates by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). The MALDI-TOF MS analyses provided species-specific fingerprints, which allow rapid identification and classification of the isolates. Six genus-specific mass peaks in the range of 2218-4434 m/z were shared by both strains. Bacterial identification was achieved by the identification of six species-specific mass peaks in the ranges of 2534-7183 m/z and 2536-9113 m/z for P. fragi and P. syringae, respectively.     

Histamine levels in seventeen species of fresh and processed South African seafood

Histamine levels were determined in fresh and processed seafood from a representative range of 10 outlets after several incidents of scombroid seafood poisoning occurred. Species included seventeen fresh and processed scombroid- and non-scombroid fish, marine mollusks and crustaceans. Histamine levels in fresh seafood were generally low (0–9 ppm) with the exception of one sample of snoek (scombroid fish; >50 ppm) and one sample of yellowtail (non-scombroid fish; >50 ppm). Both species are rich in free histidine (1.5–5.3 ppb), a precursor of histamine. Processed seafood had, in general, low histamine concentrations (0–3 ppm) with the exception of fish meal (76 ppm), salted herring (47 ppm), one sample of smoked snoek (>50 ppm) and dried tuna (8000 ppm). In total, 5 of 80 examined samples (6%) contained histamine concentrations above the legal limit of 50 ppm. Experimental formation of histamine was demonstrated to be strongly temperature- and time-dependent. Samples were not contaminated with Vibrio spp., Pseudomonas spp., Klebsiellas spp. or Enterobacteria.