Histamine Production by Photobacterium spp. in Tuna and Mahi-Mahi under Various Storage Temperatures.

Histamine is produced from histidine using histidine decarboxylase of histamine-producing bacteria. However, associated histamine food poisoning demands microbiological controls. Furthermore, studies reported that histamine production by histamine-producing bacteria is affected by temperature. Therefore, to prevent histamine food poisoning, it is desirable to store foods below 4℃. However, it is challenging to maintain the storage temperature of food substances in refrigerators constantly below 4℃. Thus, we investigated histamine production capacity using seven histamine-producing bacterial strains under storage at 10℃, a more reasonable cold storage condition. Subsequently, we examined the variation of histamine production in buffers, the correlation between bacterial density and histamine production quantities, and the growth rate in broths. Results showed that similar levels of histamine were produced in buffers even after 5 days of storage under certain conditions in which histamine-producing bacteria did not grow. Moreover, bacterial density was proportional to histamine production, and the coefficient of determination was more than 0.97, and the bacterial density required to produce 200 μg/mL of histamine during storage at 10℃ was calculated to be 4×107-4×108 CFU/mL. When the initial bacterial density was 102-103 CFU/mL, psychrophilic bacteria required 2 or 3 days and mesophilic bacteria required more than 4 days to grow above 107 CFU/mL. The above results suggest that understanding the capacity of histamine-producing bacteria to produce histamine and its growth rate in foods is important for the prevention of histamine food poisoning.

Histamine fish poisoning becomes highly concern not only in public health but also economic aspect. Histamine is produced from histidine in fish muscles by bacterial decarboxylase enzyme. Several techniques have been developed to determine the level of histamine in fish and their products but the effective method for detecting histamine producing bacteria is still required. This study was attempted to detect histamine producing bacteria by newly developed PCR condition. Histamine producing bacteria were isolated from scombroid fish and determined the ability to produce histamine of isolated bacteria by biochemical and TLC assays. PCR method was developed to target the histidine decarboxylase gene (hdc). The result showed that fifteen histamine producing bacterial isolates and three standard strains produced an amplicon at the expected size of 571bp after amplified by PCR using Hdc_2F/2R primers. Fifteen isolates of histamine producing bacteria were classified as M. morganii, E. aerogenes, and A. baumannii. The lowest detection levels of M. morganii and E. aerogenes were 10(2) and 10(5) Cfu/mL in culture media and 10(3) and 10(6) Cfu/mL in fish homogenates, respectively. The limit of detection by this method was clearly shown to be sensitive because the primers could detect the presence of M. morganii and E. aerogenes before the histamine level reached the regulation level at 50ppm. Therefore, this PCR method exhibited the potential efficiency for detecting the hdc gene from histamine producing bacteria and could be used to prevent the proliferation of histamine producing bacteria in fish and fish products.

Histamine and Biogenic Amine Production by Morganella morganii Isolated from Temperature-Abused Albacore

In an allergic inflammation model of air pouch type in rats, histamine level in the pouch fluid and histidine decarboxylase activity of pouch wall tissues in the postanaphylaxis phase were increased. Although treatment with dexamethasone failed to inhibit histamine release from mast cells in the anaphylaxis phase, histamine production in the postanaphylaxis phase was inhibited dose dependently. Histamine production-increasing activity in the pouch fluid collected 8 h after the Ag challenge, which was estimated by an activity to stimulate histamine production by bone marrow cells, was decreased by the administration of dexamethasone at the time of the Ag challenge. The addition of steroidal antiinflammatory drugs, dexamethasone, prednisolone, or hydrocortisone, into the incubation medium inhibited the pouch fluid-induced histamine production by bone marrow cells. Hydrocortisone mesylate antagonized the inhibitory effect of dexamethasone on histamine production by bone marrow cells. However, hydrocortisone mesylate failed to recover the decrease in histamine production-increasing activity of the pouch fluid collected from dexamethasone-treated rats. In addition, the dialyzed sample of pouch fluid obtained from dexamethasone-treated nonsensitized rats did not reduce the stimulated histamine production by the pouch fluid sample obtained from the sensitized rats. However, increase in histamine production of bone marrow cells stimulated by the pouch fluid was not inhibited by cyclosporin A that inhibited histamine production induced by Con A. This observation indicates that the pouch fluid has no effect to induce production of the histamine production-increasing factor by bone marrow cells. Consequently, it is suggested that dexamethasone inhibits not only the production of histamine production-increasing factor but also the response of histamine-producing cells to this factor.

Morganella morganii, a strong histamine-producing bacterium (HPB), has been frequently detected in seafood, such as skipjack tuna. Temperature fluctuations and improper packaging have resulted in bacterial proliferation and histamine production. This study aimed to determine the effects of different packaging and temperature conditions on M. morganii growth and to examine histamine formation in skipjack tuna. A factorial design with two factors, namely packaging type (vacuum and non-vacuum) and storage temperature (4, 15, 30, and 40°C), was used in this study. The bacterial growth model over time was analyzed using DMFit software. Histamine production was analyzed using thin-layer chromatography (TLC) combined with ImageJ program visualization. The results indicated that different temperatures significantly affected the bacterial growth rate (p < 0.05). The application of vacuum packaging at 4 °C retarded histamine formation in skipjack tuna cubes. The highest growth rate (0.2652 log CFU-1h-1) was observed in samples under non-vacuum packaging stored at 40 °C. M. morganii. At 15 °C, a 3 to 4 log increase was observed, starting from 3.2 to 7.5 (vacuum packaging) and from 5.8 to 8.3 log CFU-1 mL-1 (non-vacuum packaging) at the end. Nevertheless, the production of histamine in vacuum-packed samples stored at 15°C after days 3 and 4 of incubation were 446 ppm and 443.5 ppm, respectively. These findings highlight the importance of proper packaging of skipjack tuna using a cold chain system during storage. This study also confirmed the potential application of TLC for the detection of histidine and histamine.

Effect of Sodium Hypochlorite on Biofilm-Forming Ability of Histamine-Producing Bacteria Isolated from Fish

Histamine (scombrotoxin) food poisoning is a community health risk associated with consumption of some marine fish species and it has been proposed that some anaerobic bacteria contribute to this form of food poisoning. However, histamine levels and histamine producing bacteria in fish marketed in Tanzania have not been appraised. This study was conducted to determine histamine levels and histamine-producing bacteria in the <i>E. affinis</i> (tuna) value chain in Tanga and Mtwara regions. High-performance thin-layer chromatography (HPTLC) was used to determine histamine levels, Bacteriology and biochemical tests were employed to determine histamine-producing bacteria. A total of 64 tuna fish samples were collected from deep sea fish market and Sahare Tanga and ferry market, Mtwara coasts. The mean level of histamine was found to be 86.4±43.9 in Tanga and 64.8±47.5 in Mtwara and the overall mean was 77.1±46.4. All fish 64 samples were positive for histamine; however, the mean level was within the recommended limit for consumed fishery products according to European Union regulation of 200 mg/kg (m) to 400 mg/kg (m) (EC, 2005).<i> Klebsiella spp, pseudomonas spp, </i>and <i>proteus spp</i> were isolated and confirmed as histamine-producing bacteria based on bacteriology and biochemical characteristics. It concluded that although within the recommended levels, all fish samples were positive to histamine and three bacterial species associated with its production isolated. To prevent raising histamine to concern levels, maintenance of hygiene and low temperature along the production chain should be observed.

Histamine is the main causative agent of scombroid fish poisoning (SFP). To prevent SFP outbreaks, methods for controlling histamine accumulation have been studied. Histamine accumulation in raw fish, including frozen and semi-dried fish products, can be prevented by employing Hazard Analysis and Critical Control Point (HACCP) and Good Agricultural Practices (GAP). However, there is little information about histamine producers and the methods for preventing histamine accumulation in fermented fishery products in Japan. In 2011, CODEX adopted a limit of 400 ppm for histamine content in fish sauce, indicating that many manufacturers of fermented fish products are required to implement appropriate countermeasures. Presented here is recent information about histamine-producing bacteria in fermented fish products.

Effects of gluconodeltalactone and Lactobacillus plantarum on the production of histamine and tyramine in fermented sausages

Elevation of histamine levels in rat and mouse tissues by the deacetylation of administered N-acetylhistamine

The consumption of certain fish products containing high levels of histamine (and other biogenic amines) can result in an acute illness with allergy-like symptoms called scombroid syndrome. Fish accumulate toxic levels of histamine when their high level of histidine in muscle tissues is coupled with a proliferation of bacteria rich in the enzyme histidine decarboxylase. Other vasoactive amines—cadaverine, putrescine, etc.—may inhibit detoxification mechanisms that reduce the intestinal absorption of histamine. Moreover, histidine can be transformed by means of another metabolic pathway leading to accumulation in fish muscle of urocanic acid. Recently, interest has been extended to mesophilic and psychrotolerant bacteria. Histamine accumulation is traditionally correlated to microbially contaminated fish and poor storage conditions. In addition, the high thermal stability has to be considered. At present, different methods are available for the analytical determination of histamine ranging from the AOAC fluorometric method to HPLC, ELISA and rapid stick methods.

Growth and metabolic activity of Shewanella putrefaciens maintained under different CO 2 and O 2 concentrations

The term histamine intolerance (HIT) describes a proposed disease in which patients develop a variety of symptoms following the ingestion of foods rich in histamine.1 ,2 Due to an assumed...

Scombrotoxic fish poisoning (SFP) is associated with the consumption of contaminated fish of the Scombroid family (including tuna, mackerel, herring, marlin, bonito and jacks). SFP is a chemical intoxication and symptoms occur within 10 minute to 2 h after consumption of preformed histamine in scombroid fish and include rash on the face, neck and upper chest, flushing, sweating, nausea, vomiting, diarrhoea, abdominal cramps, headache, dizziness, palpitations, oral burning sensation, metallic taste and hypotension. Symptoms usually resolve within 24 h. Although most cases do not seek medical attention, in some instances symptoms may be of sufficient severity to prompt cases to seek urgent medical attention, where antihistamine drugs may be used. Scombroid fish naturally contain high levels of histidine which is converted to toxic histamine as a result of histidine decarboxylase-producing bacteria if storage conditions are inadequate to control bacterial growth (Fig. 1). Histamine is heat stable and survives subsequent processing, including canning, and ingestion of fish with histamine at levels in excess of 1000 ppm (100 mg/100 g) can result in illness. Bacterial spoilage and production of histamine may occur at any stage in the food chain (i.e. from landing the fish, at the processing plant or in the distribution system or in catering premises or homes) and adequate temperature control is key in preventing bacterial growth and histamine formation. For control of fish belonging to the scombroid family the permissible level set by EU legislation for each batch of fish is an average histamine concentration lower than 100 ppm (10 mg/100 g).1 Surveillance of SFP is based on results from samples submitted to the Health Protection Agency Centre for Infections (Food Safety Microbiology Laboratory). However, this data may represent an under-ascertainment because investigations may take place through other laboratories such as public analysts. However, between 1992 and 2004, 56 outbreaks of SFP were reported to the Health Protection Agency from cases in England and Wales (0–10 incidents per year) affecting 296 people (Table 1). Analysis of outbreaks associated with fish and shellfish between 1992 and 1999 identified that SFP accounted for 32 per cent of these and that SFP outbreaks occurred more frequently during the summer months.2 Examples of incidents involving spoilage at the point of manufacture in a tuna factory in Sri Lanka and at the point of production in a sandwich bar in London are summarized in Table 2. There has been a recent increase in the number of outbreaks reported. Between December, 2004, and August, 2005, 24 SFP outbreaks and incidents (46 ill, 3 hospitalized) were reported to the HPA from several regions within England and Wales. Sixteen of the outbreaks occurred between December to June and the remaining eight occurred during the summer months up to August. A seasonal increase during the summer has been observed previously; however, the increase seen in the winter and spring months was unusual since outbreaks are more likely to occur during warmer weather after consumption of fish that has been improperly stored, handled and prepared.2 The outbreaks were associated with 21 catering premises (sandwich shops, restaurants, hotels and public houses) and three domestic setting, of which 23 were caused by consumption of tuna. Six of the catering premises were supplied with tuna from the same supplier in sealed foil vacuum packs that had low levels of histamine ( 3000 ppm). This suggests poor food handling and inadequate refrigeration during storage at these premises after the tuna packs were opened. In one of the other outbreaks, toxic histamine levels (>1970 ppm) were present in both sealed packs of raw and cooked tuna indicative of poor temperature control at some stage post-harvest, storage or transportation. Maintenance of microbiological quality from postharvest until the moment of consumption is essential if SFP associated with fish is to be avoided. The EU General Food Law Regulation 178/2002 requires the removal of contaminated food from the marketplace together with efficient and complete trace-back of incriminated food to the point of origin. The EU legal limits for histamine in food is <100 ppm, and following the recent SFP outbreaks, the Food Standards Agency has circulated a letter to food enforcement authorities to make them aware of the issue and emphasized the importance of following strict hygiene during

The key factors influencing histamine accumulation in fermented sausages were investigated. Although Lactobacillus salivarius R06 with positive amino acid decarboxylase was inoculated, histamine was not detected in batch A without histidine availability. About 52.1 mg/kg of histamine was observed in batch B with the addition of 80 mg/kg histidine. Under sterilized condition, both histamine production and histamine‐forming bacteria were not detected in batch C. About 37.6 mg/kg histamine and 4.19 log10 cfu/g of histamine‐forming bacteria were presented in batch D by a spontaneous fermentation. Approximately 60% of histamine in batch F was degraded by Staphylococcus epidermidis R11 with amine oxidase activity. Additionally, histamine content in fermented sausages increased with rising temperature at storage. These findings indicate that histidine availability as a limiting factor and the presence of histamine‐forming bacteria as indispensable factor are responsible for histamine formation. The strains with amine oxidase activity contribute to histamine accumulation reduction. Storage temperature has a direct effect on further formation of histamine in fermented sausages.Practical ApplicationsHigh levels of histamine have been described in fermented sausages, which are a major problem for public health and food safety. Thus, it is worthwhile to improve knowledge of the factors involved in histamine accumulation to control histamine accumulation in fermented sausages. In this study, the key factors, including histidine availability, the presence of strains with decarboxylase activity, the presence of strains with amine oxidase activity and storage temperature influencing histamine accumulation in fermented sausages were investigated. These findings indicate that the key actions to prevent histamine accumulation must be addressed toward a reduction of the numbers of undesirable microorganisms and histamine‐forming microorganisms to control the proteolytic and decarboxylase activities and use of adjunct cultures with amine oxidase activity to degrade histamine. Additionally, fermented sausages should be stored immediately at low temperature after ripening.