Effects of Storage Time and Temperature on the Microflora and Amine-Development in Spanish Mackerel (Scomberomorus maculatus)

Microbial content was characterized and levels of three amines (histamine, cadaverine, and putrescine) were determined in Spanish mackerel (Scomberomorus maculatus) decomposed at PC, WC, and 30°C for varying lengths of time. Correlations were shown (1) between the levels of the histamine, cadaverine, and putrescine and the time and temperature of decomposition, (2) between the ratios of cadaverine/histamine and putrescine/histamine levels and the temperature of decomposition, and (3) between increasing total microbial counts and rising amine levels. A total of 14 bacterial species with histidine decarboxylase activity were isolated from decomposing fish, including three species (Acinetobacter lwoffi, Pseudomonas putrefaciens, and Aeromonas hydrophila) not previously reported to have the potential to produce histamine.     

Histamine Production in Soy Extended Tuna Salads

Histamine production in tuna salads extended with textured soy protein (TSP) was evaluated. Salads were inoculated with five known histamine-producing bacteria and held at 8°C, 24°C, and 37°C for up to 48 hr. Addition of 30% TSP to tuna salads resulted in higher initial pH and favorable growth conditions for microorganisms and histidine decarboxylase activity. Addition of 15% TSP provided an initial pH for maximal enzyme and histamine production but somewhat slower microbial growth. Tuna salad extended with either 15% or 30% TSP developed toxic levels of histamine (>50 mg/l00 g) when held at either 24° or 37°C for 6 hr. Nonextended tuna salads did not develop toxic levels of histamine even when inoculated with known histamine-producing bacteria and held at 24° or 37°C for 48 hr.     

Histamine and cadaverine production by bacteria isolated from fresh and frozen albacore (Thunnus alalunga).

Two hundred twenty-seven bacterial strains were isolated from fresh and frozen albacore stored either at -18 or -25 degrees C and investigated for their abilities to produce biogenic amines. As a preliminary screening, all 227 strains were tested in either Niven or Niven modified medium, which allowed the selection of 25 presumptive histamine-producing strains. High-pressure liquid chromatography revealed that only 10 of the 25 strains selected were able to produce low histamine concentrations (<25 ppm) in tryptic soy broth medium supplemented with 2% histidine. None of the 25 strains tested produced putrescine or spermine, whereas 6 strains produced spermidine. Histamine production by Stenotrophomonas maltophilia strain 25MC6 was not prevented at 4 degrees C, and the levels of this amine reached concentrations of 25.8 ppm after 6 days. Three S. maltophilia strains showed strong lysine-decarboxylating activity. Their cadaverine formation capacity was determined by high-pressure liquid chromatography in tryptic soy broth supplemented with 1% lysine; this revealed that the three S. maltophilia strains tested produced more than 700 ppm of cadaverine during the first 24 h of incubation at 37 degrees C. S. maltophilia strain 15MF, initially obtained from fresh albacore tuna, produced up to 2,399 ppm and 4,820 ppm of cadaverine after 24 and 48 h of incubation at 37 degrees C, respectively. To our knowledge, this is the first report on histamine and cadaverine production by strains of the species S. maltophilia, previously known as Pseudomonas and Xanthomonas maltophilia, isolated from fresh and frozen albacore tuna.     

Formation of biogenic amines by Gram-negative rods isolated from fresh,spoiled, VP-packed and MAP-packed herring (<Emphasis Type="Italic">Clupea harengus</Emphasis>)

Bacterial strains (120) were isolated from fresh, spoiled, VP-packed and MAP-packed herring. Identified bacterial strains were investigated for their abilities to produce biogenic amines in histidine, lysine and ornithine decarboxylase broth by a rapid high-performance liquid chromatography (HPLC) method. The microflora of fresh herring was dominantly Pseudomonas (30%), Enterobacteriaceae (23.2%), Vibrio (13.3%) and Moraxella (13.3%) but, the microflora of herring stored in VP and MAP was dominated by species belonging to Vibrio (23.3%) and Moraxella (20%), which indicates that these packaging systems prevented the growth of Pseudomonas and Enterobacteriaceae. In a laboratory medium containing amino acid (histidine, ornithine and lysine), most of bacterial strains produced histamine, putrescine and cadaverine. The highest histidine decarboxylase activities were observed in Klebsiella oxytoca, Hafnia alvei and Proteus vulgaris which produced 396, 232 and 54 mg histamine/L, respectively in histidine-enriched broth. The accumulation of cadaverine by Klebsiella oxytoca and Hafnia alvei was 325 and 208 mg/l, respectively. All strains isolated produced putrescine in an ornithine-enriched broth, ranging from 3 to 249 mg/l. The production of putrescine by Klebsiella oxytoca and Hafnia alvei was 249 and 195 mg/l, respectively. Moraxella spp and Acinetobacter spp did not produce histamine which indicates they did not have histidine decarboxylase activity.     

Hygienic status and biogenic amine content of mung bean sprouts

 Microbiological analyses of commercial mung bean sprouts showed the total, viable microbiological population to exceed 10⁸ cfu/g. Enterobacter cloacae, Klebsiella pneumoniae and Enterobacter agglomerans were found to be the dominant and most frequently isolated microbial species. Putrescine, cadaverine, spermidine and spermine were detected in all samples investigated. Formation of biogenic amines by pure culture isolates was studied in a modified decarboxylase medium at different temperatures, pH values and atmospheres. Highest activities were found under aerobic conditions at 20  °C. K. pneumoniae 861 produced 1.2 mg cadaverine/ml after an incubation period of 24 h and E. cloacae 862 produced 2 mg putrescine/ml after 48 h of incubation. For E. agglomerans 863, no biogenic amines were detected under these conditions. Production of cadaverine by E. cloacae 862 and K. pneumoniae 861 under aerobic conditions is presumably related to lysine decarboxylase activities. Although highest decarboxylase activities have usually been found at acidic pH values, amine production reached a maximum at pH 7. Under anaerobic conditions, E. cloacae 862 produced only about half the amount of putrescine as under aerobic conditions, whilst K. pneumoniae 861 produced significantly less cadaverine but was able to produce putrescine. Received: 10 October 1997 / Revised version: 21 January 1998     

Bacterial Histamine Production as a Function of Temperature and Time of Incubation

Optimal temperature, lower temperature limit, extent, and rate of histamine production in a tuna fish infusion broth (TFIB) varied for the strains of Proteus morganii, Klebsiella pneumoniae, Hafnia alvei, Citrobacter freundii, and Escherichia coli studied. P. morganii and K. pneumoniae produced large quantities of histamine in a relatively short incubation period (<24 hr) at 15°C, 30°C, and 37°C; production was fastest at 37°C. H. alvei, C. freundii, and E. coli produced toxicologically significant levels of histamine (>2500 nmoles/ml) only at 30°C and 37°C on prolonged incubation (≥48 hr). At 72 hr of incubation, optimal temperature for histamine production was 37°C for E. coli and C freundii; 30°C for P. morganii strain 110SC2, K. pneumoniae, and H. alvei; and 15°C for P. morganii strain JM. The lower temperature limits for production of toxicologically significant levels of histamine in TFIB were 7°C for K. pneumoniae; 15°C for both P. morganii strains; and 30°C for H. alvei, C. freundii, and E. coli.     

Hygienic status and biogenic amine content of mung bean sprouts

 Microbiological analyses of commercial mung bean sprouts showed the total, viable microbiological population to exceed 10⁸ cfu/g. Enterobacter cloacae, Klebsiella pneumoniae and Enterobacter agglomerans were found to be the dominant and most frequently isolated microbial species. Putrescine, cadaverine, spermidine and spermine were detected in all samples investigated. Formation of biogenic amines by pure culture isolates was studied in a modified decarboxylase medium at different temperatures, pH values and atmospheres. Highest activities were found under aerobic conditions at 20  °C. K. pneumoniae 861 produced 1.2 mg cadaverine/ml after an incubation period of 24 h and E. cloacae 862 produced 2 mg putrescine/ml after 48 h of incubation. For E. agglomerans 863, no biogenic amines were detected under these conditions. Production of cadaverine by E. cloacae 862 and K. pneumoniae 861 under aerobic conditions is presumably related to lysine decarboxylase activities. Although highest decarboxylase activities have usually been found at acidic pH values, amine production reached a maximum at pH 7. Under anaerobic conditions, E. cloacae 862 produced only about half the amount of putrescine as under aerobic conditions, whilst K. pneumoniae 861 produced significantly less cadaverine but was able to produce putrescine. Received: 10 October 1997 / Revised version: 21 January 1998     

Effects of on-board and dockside handling on the formation of biogenic amines in mahimahi (Coryphaena hippurus), skipjack tuna (Katsuwonus pelamis), and yellowfin tuna (Thunnus albacares)

Consumer illnesses by scombroid poisonings have been a continuing problem for many years. The intoxications follow the ingestion of fish such as tuna and mahimahi that have undergone bacterial decomposition, leading to the formation of biogenic amines. Research studies have concluded that histamine is one of the indicators of scombrotoxic fish and that other amines, such as cadaverine, could be involved in the illnesses. Guidance for the handling of fish on board fishing vessels to prevent the production of scombrotoxic fish has been limited by a lack of data addressing changes that occur in fish from the water to delivery at dockside. In this study, the changes in selected biogenic amines were determined in mahimahi and tuna, which were captured and held in seawater at 25 to 35 degrees C for incubation times up to 18 h. The fillets from the treated fish were sectioned by transverse cuts and analyzed for histamine, cadaverine, and putrescine. Results showed that at 26 degrees C, more than 12 h of incubation were required before a histamine concentration of 50 ppm was reached in mahimahi. At 35 degrees C, 50 ppm histamine formed within 9 h. Similar results were found for skipjack and yellowfin tuna. Histamine concentrations exceeded 500 ppm within an additional 3 h of incubation in mahimahi. At both temperatures, an increase in the concentration of cadaverine preceded an increase in histamine levels. Changes in putrescine concentrations in the fish were less pronounced. The study also demonstrated that histidine decarboxylase activity was retained in some frozen samples of fish and could result in further increases in histamine on thawing.     

Endotoxin and biogenic amine levels in Atlantic mackerel (Scomber scombrus), sardine (Sardina pilchardus) and Mediterranean hake (Merluccius merluccius) stored at 22 °C

Whole Atlantic mackerel (Scomber scombrus), sardine (Sardina pilchardus) and Mediterranean hake (Merluccius merluccius) from the Croatian Adriatic were stored at 22 °C and changes in histamine, putrescine, tyramine and cadaverine levels were monitored in relation to bacterial endotoxin. After 12 h, histamine levels in sardine were above the legal limit of 50 mg kg^?1, set by the US Food and Drug Administration, and an increase in putrescine content preceded the increase in histamine. After 24 h, histamine contents in mackerel and sardine reached 1090 ± 101 and 577 ± 275 mg kg^?1, respectively, which exceeded the toxic threshold of 500 mg kg^?1. At the same time, the putrescine content was also high in both fish (353–420 mg kg^?1). The time-course of endotoxin production was similar in all fish species stored at 22 °C. A high correlation was found between endotoxin and histamine, and between endotoxin and putrescine in mackerel and sardine. On the other hand, high endotoxin levels in hake, after 24 h, were associated with the low histamine and putrescine content (40–60 mg kg^?1).     

Effects of heat treatment and storage temperature on the biogenic amine content of straw mushroom (Volvariella volvacea)

The changes in the biogenic amines tryptamine, 2-phenylethylamine, putrescine, cadaverine, histamine and tyramine of straw mushroom Volvariella volvacea (Bull, ex Fr) Sing produced by heat treatment and during storage at different temperatures were studied. About 80% of the original content of these amines was lost during cooking. Commercial canned straw mushroom contained low amounts of amines. The amine contents of straw mushroom increased during storage at 4°C, with particularly notable increases in the concentrations of 2-phenylethylamine and tyramine after 5 days' storage. However, the levels of all amines increased more markedly during storage at 25°C, and the increases in the putrescine and cadaverine concentrations were much greater than those reached at 4°C.     

Inhibitory effect of spices on in vitro histamine production and histidine decarboxylase activity ofMorganella morganii and on the biogenic amine formation in mackerel stored at 30°C

The inhibitory effects of clove, cinnamon, cardamom, turmeric and pepper on the histamine production and histidine decarboxylase activity ofMorganella morganii (a potent histamine-producing bacteria in fish) was examined at 30°C using HPLC. Cinnamon and clove exhibited a significant (P<0.01) inhibitory effect, whereas turmeric and cardamom had a moderate effect. These spices were applied to whole mackerel at a level of 3% and their inhibitory effect on biogenic amine formation at 30°C was also examined. As in the in vitro study, clove and cinnamon showed a significant (P< 0.05) inhibitory effect on histamine, putrescine and tyramine formation but not on that of cadaverine. Cardamom and turmeric exhibited a moderate effect and pepper was ineffective. Therefore, clove and cinnamon are more helpful than cardamom and turmeric in the minimization of the formation of toxic histamine in mackerel.     

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.     

Histamine Formation in Abusively Stored Pacific Mackerel: Effect of CO2-Modified Atmosphere

Whole Pacific mackerel (Scomber japonicus) were abusively stored at 20°C in air or 80% CO₂, balance air. Samples were analyzed for amines using a modified amino acid analyzer. Following 24 hr storage, levels of histamine, tyramine, putrescine, and cadaverine increased only slightly above the low levels observed initially. During the next 24 hr, the amine content increased dramatically. Levels in the air control samples were about twice those in the modified atmosphere samples. In a separate trial, amine levels in fish stored 3 days were higher still and similar in the two atmospheres. Thus, in neither trial did CO₂-modified atmosphere storage lead to increased production of potentially toxic amines.     

Tissue degradation,amino acid liberation and bacterial decomposition of bulk stored capelin

Bulk stored capelin with high contents of feed in the gut is easily solubilised because of high proteolytic activity. The amino acids most quickly liberated are arginine, serine, histidine, leucine, lysine and tyrosine; glycine, aspartic acid and glutamic acid are liberated at a slower rate. Bacterial activity results in a rapid and considerable decrease in total amounts of tyrosine, lysine, serine, arginine and histidine. The main products formed by bacterial decomposition of lysine, histidine and arginine are cadaverine, histamine and putrescine, respectively. Storage of capelin in the presence of antibiotics increased the amounts of free amino acids and total amino acids. After 10 days' storage at 6°C, about 30% of the total amino acid content in fish treated with antibiotics were fee amino acids.     

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.     

Determination of trans‐ and cis‐Urocanic Acid in Relation to Histamine,Putrescine, and Cadaverine Contents in Tuna (Auxis Thazard) at Different Storage Temperatures

Scombroid fish poisoning is usually associated with consumption of fish containing high levels of histamine. However, reports indicate that some cases have responded to antihistamine therapy while ingested histamine levels in these cases were low. Potentiation of histamine toxicity by some biogenic amines, and release of endogenous histamine by other compounds such as cis‐urocanic acid (UCA) are some hypotheses that have been put forth to explain this anomaly. Very little is known about the effects of storage conditions on the production of both UCA isomers and biogenic amines in tuna. Thus, the production of trans‐ and cis‐UCA, histamine, putrescine, and cadaverine in tuna during 15 d of storage at 0, 3, and 10 °C and 2 d storage at ambient temperature were monitored. The initial trans‐ and cis‐UCA contents in fresh tuna were 2.90 and 1.47 mg/kg, respectively, whereas the levels of putrescine and cadaverine were less than 2 mg/kg, and histamine was not detected. The highest levels of trans‐ and cis‐UCA were obtained during 15 d storage at 3 °C (23.74 and 21.79 mg/kg, respectively) while the highest concentrations of histamine (2796 mg/kg), putrescine (220.32 mg/kg) and cadaverine (1045.20 mg/kg) were obtained during storage at room temperature, 10 and 10 °C, respectively. Histamine content increased considerably during storage at 10 °C whereas trans‐ and cis‐UCA contents changed slightly. The initial trans‐UCA content decreased during storage at ambient temperature. Thus, unlike histamine, concentrations of trans‐ and cis‐UCA did not result in elevated levels during storage of tuna.     

Histamine, Putrescine and Cadaverine Formation in Spanish Semipreserved Anchovies as Affected by Time/Temperature

We studied the changes in counts of mesophilic, psychrotrophic, entera-bacteria, sulphite-reducing and Vibrio microorganisms in commercial samples of Spanish semipreserved anchovies. The influence of NaCl, oxygen concentration and pH on bacterial growth and histamine, putrescine and cadaverine formation were also studied. Notable histamine formation was detected in samples incubated at 20°C and preserved in olive oil (443.68 ppm-3012.13 ppm). This accumulation was probably caused by the conditions of the product, pH 5–6, relatively low NaCl (< 15%), and permissive temperature (product not refrigerated). Correlation between the microorganisms and histamine formation was not clear. Histamine was produced by too high storage temperatures but not by the ripening process.     

Evaluation of Histamine and Other Biogenic Amines and Bacterial Isolation in Canned Anchovies Recalled by the USFDA

ABSTRACT: Histamine and other biogenic amines were evaluated in canned anchovies recalled by the U.S. Food and Drug Administration. In addition, bacteria were isolated from the products and identified to species. The recalled products were divided into 2 groups of high and low histamine, depending on the histamine contents as determined by the AOAC method. The high histamine group had the histamine contents >200 ppm, and 24 of the 30 cans analyzed belonged to this group. The most prevalent biogenic amine in this group was histamine followed by cadaverine. On the other hand, the low histamine group of 6 cans contained approximately 50 ppm histamine. The most prevalent biogenic amine found in this group of samples was cadaverine at levels >200 ppm. Other biogenic amines, such as putrescine, serotonin, and spermidine, were also detected in all the products, although at varied levels. Aerobic and anaerobic bacterial counts, if present in all of the recalled products, were below the detection limit of 10² colony-forming units (CFU)/g. Bacteria were recovered only after enrichment of the test samples. They were mostly halophilic bacteria. Bacillus spp. were most frequently identified, followed by Staphylococcus spp. However, these isolates produced negligible amounts of histamine in culture broth, indicating that they are not the contributors to histamine accumulation in the canned anchovies.     

EFFECT OF ENDOGENOUS PROTEINASES ON HISTAMINE AND HONEYCOMB FORMATION IN MACKEREL

Mackerel muscle homogenate treated with visceral trypsin-like proteinme was found to contain higher free histidine and histamine than that of an untreated sample by a difference of 120ppm histamine ajier 8 days at 4°C; 5oOppm ajier 24 h at 28°C; and 205ppm ajier 4 h at 37°C. The contents of histamine and free histidine were both found to be higher in ungutted mackerel than in gutted ones. When mackerel fillets were treated with muscle alkaline proteinase and heated, it produced maximal increase in soluble peptides at 60°C. Honeycombing appeared in the cooked fillet previously treated with muscle alkaline proteinase, while no honeycombing was observed in the control. Cooked mackerel fillets previously incubated with mackerel pybric collagenase at 25°C and 55°C resulted in increases in free hydroxyproline. The release of hydroxyproline increased with prolonged incubation at 25°C but stayed constant at 55°C which was probably related to the optimal temperature of collagenase being 25°C. Honeycombing was not noticeable in the cooked jlllet previously incubated with or without collagenase at 55°C for 6 h. Extended incubation to 20h at 55°C showed honeycombing in both control and collegenase-treated fillets. 7he possible roles of pyloric collagenase and muscle alkaline proteinase in honeycombing are discussed.