Inactivation of Morganella morganii by high hydrostatic pressure combined with lemon essential oil.

The inactivation and damage of histamine-forming bacterium, Morganella morganii, in phosphate buffer and tuna meat slurry by high hydrostatic pressure (HHP) alone or in combination with 0.2% lemon essential oil (LEO) treatments were studied using viability measurement and scanning electron microscopy (SEM). HHP alone or in combination with LEO treatments showed first-order destruction kinetics to M. morganii during pressure holding period. The D values of M. morganii (200 to 600 MPa) in phosphate buffer ranged from 16.4 to 0.08 min, whereas those in tuna meat slurry ranged from 51.0 to 0.10 min, respectively. M. morganii in tuna meat slurry had higher D values and were more resistant to HHP treatments than in phosphate buffer. In addition, the D values of HHP in combination with LEO treatment were lower than those of HHP treatment alone at <400 MPa of pressure, indicating that it is more effective to inactivate M. morganii under the same pressure. The results showed the M. morganii at HHP in combination with LEO treatment was more susceptible to pressure treatment alone. HHP with or without LEO treatments can be used to inactivate M. morganii by causing disruption to bacterial cell membrane and cell wall as demonstrated by SEM micrographs.

Effect of Brine Concentrations on the Bacteriological and Chemical Quality and Histamine Content of Brined and Dried Milkfish.

In this research, the occurrence of hygienic quality and histamine in commercial brined and dried milkfish products, and the effects of brine concentrations on the quality of brined and dried milkfish, were studied. Brined and dried milkfish products (n = 20) collected from four retail stores in Taiwan were tested to investigate their histamine-related quality. Among them, five tested samples (25%, 5/20) had histamine contents of more than 5 mg/100 g, the United States Food and Drug Administration guidelines for scombroid fish, while two (10%, 2/20) contained 69 and 301 mg/100 g of histamine, exceeding the 50 mg/100 g potential hazard level. In addition, the effects of brine concentrations (0%, 3%, 6%, 9%, and 15%) on the chemical and bacteriological quality of brined and dried milkfish during sun-drying were evaluated. The results showed that the aerobic plate count (APC), coliform, water activity, total volatile basic nitrogen (TVBN), and histamine content values of the brined and dried milkfish samples decreased with increased brine concentrations, whereas those of salt content and thiobarbituric acid (TBA) increased with increasing brine concentrations. The milkfish samples prepared with 6% NaCl brine had better quality with respect to lower APC, TVBN, TBA, and histamine levels.

Inactivation and Damage of Histamine-Forming Bacteria by Treatment with High Hydrostatic Pressure.

The inactivation and damage of histamine-forming bacteria (HFB), Enterobacter aerogenes and Staphylococcus capitis, in a 0.1 M potassium phosphate buffer (pH 6.8) and marlin meat slurry by high hydrostatic pressure (HHP) treatments were studied using viability measurement and scanning electron microscopy (SEM). HHP treatments showed first order destruction kinetics to E. aerogenes and S. capitis during the pressure holding period. HFB in marlin meat slurry had higher D values and were more resistant to HHP treatments than in phosphate buffer. In phosphate buffer, E. aerogenes had higher D values than S. capitis at >380 MPa of pressure, whereas the reverse trend was noticed at lower pressures (<380 MPa). In marlin meat slurry, S. capitis had a higher D value than E. aerogenes at the same treatment pressure, indicating that S. capitis was more resistant to HHP treatment. To our knowledge, this is the first report to demonstrate that HHP can be used to inactivate HFB, E. aerogenes, and S. capitis, by causing disruption to bacterial cell membrane and cell wall as demonstrated by SEM micrographs.

Histamine Fish Poisoning and Histamine Production by Raoultella ornithinolytica in Milkfish Surimi.

ABSTRACT: In April 2017, an outbreak of histamine fish poisoning causing illness in nine victims associated with consumption of milkfish surimi products (fish ball) occurred in Kaohsiung City, southern Taiwan. Of the two suspected frozen milkfish surimi samples, one sample contained 91.06 mg/100 g of histamine, levels that are greater than the potential hazard action level (50 mg/100 g) in most illness cases. Moreover, 28 frozen milkfish surimi samples from retail stores were collected and tested to determine the occurrence of histamine. One (3.6%) of 28 commercial surimi samples had histamine levels greater than the U.S. Food and Drug Administration (FDA) guideline for decomposition of 5 mg/100 g for scombroid fish and/or products. Thirteen histamine-producing bacterial strains isolated from suspected and commercial surimi samples were identified as prolific histamine formers, able to produce 98.4 to 121.8 mg/100 mL of histamine in Trypticase soy broth supplemented with 1.0% l-histidine. In addition, milkfish surimi was inoculated with Raoultella ornithinolytica at 5.0 log CFU/g and stored at 4, 15, 25, and 37°C to investigate bacterial growth and formation of histamine. The histamine contents quickly increased to more than 50 mg/100 g in samples stored at 37 and 25°C within 12 and 24 h, respectively, as well those stored at 15°C within 96 h. To our knowledge, this is the first report in Taiwan to demonstrate that milkfish surimi products could cause histamine intoxication.

Determination of Histamine in Japanese Spanish Mackerel (Scomberomorus niphonius) Meat Implicated in a Foodborne Poisoning.

An incident of foodborne poisoning causing illness in seven victims due to ingestion of fried Japanese Spanish mackerel (JS mackerel; Scomberomorus niphonius) meat occurred in September 2014 in Hualien County, eastern Taiwan. Of the two suspected fish meats, one raw sample contained 3,318 ppm of histamine and one fried sample contained 1,906 ppm of histamine, levels which are greater than the potential hazard action level (500 ppm) in most illness cases. Given the allergy-like symptoms of the victims and the high histamine content in the suspected fish samples, this foodborne poisoning was strongly suspected to be caused by histamine intoxication. In addition, five histamine-producing bacterial strains isolated from suspected raw fish samples, capable of producing 152 to 1,020 ppm of histamine in Trypticase soy broth supplemented with 1.0% l-histidine, were identified as Hafnia alvei (one strain), Enterobacter aerogenes (two strains), Raoultella ornithinolytica (one strain), and Morganella morganii (one strain) by 16S rDNA sequencing with PCR amplification. Moreover, 12 raw fish samples and 39 fried fish samples from retail stores were collected and tested to determine the occurrence of histamine. Two of 12 commercial raw fish samples (16.7%) had histamine levels greater than the U.S. Food and Drug Administration guideline for decomposition of 50 ppm for scombroid fish or product or a combination of both. To our knowledge, this is the first report in Taiwan to demonstrate that the JS mackerel meat products could cause histamine intoxication.

Effect of Vacuum Packaging on Histamine Production in Japanese Spanish Mackerel (Scomberomorus niphonius) Stored at Various Temperatures.

The effect of polyethylene packaging (PEP) in air cushion and vacuum packaging (VP) on histamine related to the quality of Japanese Spanish mackerel (JS mackerel) was studied with samples stored at -20, 4, 15, and 25°C. The aerobic plate count (APC), total volatile basic nitrogen (TVBN), and histamine concentrations of the PEP and VP samples stored at 25°C increased as the storage time continued. The PEP and VP samples stored at temperatures below 15°C showed lower levels of APC, TVBN, and histamine, with VP samples having considerably lower levels of APC, TVBN, and histamine than PEP samples. For the frozen JS mackerel stored at -20°C for 2 months and then thawed and stored at 25°C, the VP treatment delayed the increases of TVBN and histamine longer than did the PEP treatment. Thus, the storage of VP JS mackerel at temperatures below 4°C could prevent quality deterioration and extend shelf life.

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 69.6mg/100g and <1.0 to 7.1logCFU/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 4mg/100g, and only one mullet roe sample had the histamine content (8.18mg/100g) greater than the 5.0mg/100g allowable limit suggested by the US Food and Drug Administration. Two histamine-producing bacterial strains capable of producing 10.7ppm and 9.6ppm 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 (8.18mg/100g).

Degradation of Histamine by Lactobacillus plantarum Isolated from Miso Products.

Histamine is a toxic chemical and is the causative agent of food poisoning. This foodborne toxin may be degraded by the oxidative deamination activity of certain microorganisms. In this study, we isolated four histamine-degrading Lactobacillus plantarum bacteria from miso products. Among them, L. plantarum D-103 exhibited 100% degradation of histamine in de Man Rogosa Sharpe (MRS) broth containing 50 ppm of histamine after 24 h of incubation at 30°C. The optimal growth, histamine oxidase, and histamine-degrading activity of L. plantarum D-103 were observed in histamine MRS broth at pH 7.0, 3% NaCl, and 30°C. It also exhibited tolerance to broad ranges of pH (4 to 10) and salt concentrations (0 to 12%) in histamine MRS broth. Therefore, the histamine-degrading L. plantarum D-103 might be used as an additive culture to prevent histamine accumulation in miso products during fermentation.

The effect of vacuum packaging on histamine changes of milkfish sticks at various storage temperatures.

The effects of polyethylene packaging (PEP) (in air) and vacuum packaging (VP) on the histamine related quality of milkfish sticks stored at different temperatures (-20°C, 4°C, 15°C, and 25°C) were studied. The results showed that the aerobic plate count (APC), pH, total volatile basic nitrogen (TVBN), and histamine contents increased as storage time increased when the PEP and VP samples were stored at 25°C. At below 15°C, the APC, TVBN, pH, and histamine levels in PEP and VP samples were retarded, but the VP samples had considerably lower levels of APC, TVBN, and histamine than PEP samples. Once the frozen fish samples stored at -20°C for 2 months were thawed and stored at 25°C, VP retarded the increase of histamine in milkfish sticks as compared to PEP. In summary, this result suggested the milkfish sticks packed with VP and stored below 4°C could prevent deterioration of product quality and extend shelf-life.

Reduction of histamine and biogenic amines during salted fish fermentation by Bacillus polymyxa as a starter culture.

Bacillus polymyxa D05-1, isolated from salted fish product and possessing amine degrading activity, was used as a starter culture in salted fish fermentation in this study. Fermentation was held at 35°C for 120 days. The water activity in control samples (without starter culture) and inoculated samples (inoculated with B. polymyxa D05-1) remained constant throughout fermentation, whereas the pH value rose slightly during fermentation. Salt contents in both samples were constant in the range of 17.5-17.8% during the first 60 days of fermentation and thereafter increased slowly. The inoculated samples had considerably lower levels of total volatile basic nitrogen (p < 0.05) than control samples at each sampling time during 120 days of fermentation. Aerobic bacterial counts in inoculated samples were retarded during the first 60 days of fermentation and thereafter increased slowly, whereas those of control samples increased rapidly with increased fermentation time. However, the aerobic bacterial counts of control samples were significantly higher (p < 0.05) than those of inoculated samples after 40 days of fermentation. In general, overall biogenic amine contents (including histamine, putrescine, cadaverine, and tyramine) in the control samples were markedly higher (p < 0.05) than those of the inoculated samples throughout fermentation. After 120 days of fermentation, the histamine and overall biogenic amine contents in the inoculated samples were reduced by 34.0% and 30.0%, respectively, compared to control samples. These results emphasize that the application of starter culture with amines degrading activity in salted fish products was effective in reducing biogenic amine accumulation.

Histamine production by Raoultella ornithinolytica in mahi-mahi meat at various storage temperatures.

Mahi-mahi meat was inoculated with Raoultella ornithinolytica at 5.0 log CFU/g and stored at -20°C, 4°C, 15°C, 25°C, or 37°C to investigate bacterial growth and formation of total volatile base nitrogen and histamine in mahi-mahi meat. R. ornithinolytica grew rapidly in samples stored at temperature above 15°C. The histamine contents quickly increased to higher than 50 mg/100 g in samples stored at 25°C and 37°C within 12 hours as well as those stored at 15°C within 48 hours. The total volatile base nitrogen contents increased to higher than the index level (30 mg/100 g) for fish decomposition at 25°C within 48 hours and 37°C within 24 hours. However, bacterial growth and histamine formation were controlled by cold storage of the samples at 4°C or below. Once the frozen mahi-mahi samples stored at -20°C for 2 months were thawed and stored at 25°C after 24 hours, histamine started to accumulate rapidly (>50 mg/100 g of fish).

Reduction of Biogenic Amines during Miso Fermentation by Lactobacillus plantarum as a Starter Culture.

Lactobacillus plantarum D-103 isolated from a miso product that possesses amine-degrading activity was used as a starter culture in miso fermentation (25°C for 120 days) in this study. The salt content in control samples (without starter culture) and inoculated samples (inoculated with L. plantarum D-103) remained constant at 10.4% of the original salt concentration throughout fermentation, whereas the pH value decreased from 6.2 to 4.6 during fermentation. The inoculated samples had significantly lower (P < 0.05) levels of total volatile basic nitrogen than control samples after 40 days of fermentation. After 120 days of fermentation, the histamine and overall biogenic amine contents in inoculated samples were reduced by 58 and 27%, respectively, compared with control samples. To our knowledge, this is the first report to demonstrate that application of a starter culture with amine-degrading activity in miso products was effective in reducing the accumulation of biogenic amines.

Determination of histamine in milkfish stick implicated in food-borne poisoning.

An incident of food-borne poisoning causing illness in 37 victims due to ingestion of fried fish sticks occurred in September 2014, in Tainan city, southern Taiwan. Leftovers of the victims' fried fish sticks and 16 other raw fish stick samples from retail stores were collected and tested to determine the occurrence of histamine and histamine-forming bacteria. Two suspected fried fish samples contained 86.6 mg/100 g and 235.0 mg/100 g histamine; levels that are greater than the potential hazard action level (50 mg/100 g) in most illness cases. Given the allergy-like symptoms of the victims and the high histamine content in the suspected fried fish samples, this food-borne poisoning was strongly suspected to be caused by histamine intoxication. Moreover, the fish species of suspected samples was identified as milkfish (Chanos chanos), using polymerase chain reaction direct sequence analysis. In addition, four of the 16 commercial raw milkfish stick samples (25%) had histamine levels greater than the US Food & Drug Administration guideline of 5.0 mg/100 g for scombroid fish and/or products. Ten histamine-producing bacterial strains, capable of producing 373-1261 ppm of histamine in trypticase soy broth supplemented with 1.0% L-histidine, were identified as Enterobacter aerogenes (4 strains), Enterobacter cloacae (1 strain), Morganella morganii (2 strains), Serratia marcescens (1 strain), Hafnia alvei (1 strain), and Raoultella orithinolytica (1 strain), by 16S ribosomal DNA sequencing with polymerase chain reaction amplification.

Hygienic quality, adulteration of pork and histamine production by Raoultella ornithinolytica in milkfish dumpling.

Ten milkfish dumpling products purchased from retail stores in southern Taiwan were collected to determine the occurrence of biogenic amines, histamine-forming bacteria, and adulteration of pork. This study showed the high contents of aerobic plate count (APC), total coliforms (TC) and Escherichia coli in tested milkfish dumpling samples, whereas the average content of various biogenic amines in all tested samples was < 1.6 mg/100 g (< 0.05 to 1.54 mg/100 g). Three histamine-producing bacterial strains (2 isolates of Raoultella ornithinolytica and 1 isolate of Enterobacter aerogenes) isolated from tested samples produced 276.6 ppm to 561.8 ppm of histamine in trypticase soy broth supplemented with 1.0% L-histidine (TSBH). Assay of multiplex polymerase chain reaction (PCR) revealed that the adulteration rates were 50% (5/10) for pork in milkfish dumplings. In addition, milkfish dumpling stuffing was inoculated with R. ornithinolytica at 5.0 log colony forming units (CFU)/g and stored at various temperatures from 4°C to 37°C to investigate bacterial growth and formation of histamine. The histamine contents quickly increased to higher than 50 mg/100 g in samples stored at 37°C and 25°C within 24 hours and 36 hours, respectively, as well as stored at 15°C within 48 hours. Therefore, bacterial growth and histamine formation were controlled by cold storage of the samples at 4°C.

Histamine production by Enterobacter aerogenes in sailfish and milkfish at various storage temperatures.

Enterobacter aerogenes was studied for its growth and ability to promote the formation of total volatile base nitrogen (TVBN) and histamine in sailfish (Istiophorus platypterus) and milkfish (Chanos chanos) stored at various temperatures from -20 to 37 degrees C. The optimal temperature for bacterial growth in both fish species was 25 degrees C, whereas the optimal temperature for histamine formation was 37 degrees C. The two fish species inoculated with E. aerogenes, when not properly stored at low temperatures such as 15 degrees C for 36 h, formed histamine at above the U.S. Food and Drug Administration hazardous guideline level of 50 mg/100 g. Milkfish was a better substrate than sailfish for histamine formation by bacterial histidine decarboxylation at elevated temperatures (> 15 degrees C). Although higher contents of TVBN were detected in the spiked sailfish than milkfish during the same storage time at temperatures above 15 degrees C, the use of the 30-mg/100 g level of TVBN as a determination index for fish quality and decomposition was not a good criterion for assessing potential histamine hazard for both fish species. Bacterial growth was controlled by cold storage of the fish at 4 degrees C or below, but histamine formation was stopped only by frozen storage. Once the frozen fish samples were thawed and stored at 25 degrees C, histamine started to accumulate rapidly and reached levels greater than the hazardous action level in 36 h.

The histamine content of dried flying fish products in Taiwan and the isolation of halotolerant histamine-forming bacteria.

Thirty dried flying fish products were purchased from fishing village stores in Taiwan and tested to detect the presence of histamine and histamine-forming bacteria. Except for histamine and cadaverine, the average content of various biogenic amines in the tested samples was less than 3.5 mg/100 g. Eight (26.6%) dried flying fish samples had histamine levels greater than the United States Food and Drug Administration guideline of 5 mg/100 g for scombroid fish and/or scombroid products, whereas four (13.3%) samples contained more than the hazard action level of 50 mg/100 g. One histamine-producing bacterial isolate was identified as Staphylococcus xylosus by 16S rDNA sequencing with polymerase chain reaction amplification. This isolate was capable of producing 507.8 ppm of histamine in trypticase soy broth supplemented with 1.0% l-histidine (TSBH). The S. xylosus isolate was a halotolerant bacterium that had a consistent ability to produce more than 300 ppm of histamine at 3% sodium chloride concentration in TSBH medium after 72 hours.

Histamine-related hygienic qualities and bacteria found in popular commercial scombroid fish fillets in Taiwan.

To determine the histamine-related hygienic qualities and bacteria of scombroid fish fillets sold in traditional retail markets, 61 samples were collected from northern and southern Taiwan. It was found that the content of volatile base nitrogen in most samples was below 25 mg/100 g, which is the regulatory level in Taiwan. The ratio of unacceptable samples/total samples for aerobic plate count and Escherichia coli was 100% and 15% in northern samples and 100% and 20% in southern samples, respectively, compared with the requirements of hygienic standards. The average content of various biogenic amines in all samples were lower than 3 mg/100 g, except for histamine average content (4.6 mg/100 g) in southern samples. Among southern samples, four samples contained 12.8 to 28.8 mg/100 g histamine, which is more than 5 mg/100 g that is the allowable limit suggested by the U.S. Food and Drug Administration. Furthermore, 14 bacterial strains were isolated from sailfish fillets on a selective medium for histamine-forming bacteria. These presumptive histamine-forming strains, such as Proteus, Enterobacter, Klebsiella, Rahnella, and Acinetobacter, have been identified and found to produce 20 to 2,000 ppm histamine after incubating at 37 degrees C for 24 h.

Antimicrobial effect of extracts of cruciferous vegetables.

The cruciferous vegetables cauliflower, broccoli, cabbage, Chinese radish, Chinese kale, and Chinese kitam were used in this study to prepare water-soluble and methanol-water extracts. Crude protein extracts were also obtained by diethylaminoethyl (DEAE) anion exchange chromatography. Water-soluble polysaccharides were prepared by ethanol precipitation followed by ultrafiltration. The antimicrobial effects of all these extracts were evaluated against Gram-positive bacteria, Gram-negative bacteria, and yeast. Crude protein extracts exhibited the greatest antimicrobial activity in monoculture experiments. The antimicrobial effects of cruciferous vegetables were also studied by steeping beef, carrot, and celery in chlorine (10 ppm) or citric acid solution (1%) containing the crude protein extract (500 ppm) for different time periods. Total aerobic plate counts and coliform counts on these foods decreased significantly after 10 minutes in all steeping solutions (p < 0.05).

Biogenic amine content, histamine-forming bacteria, and adulteration of pork in tuna sausage products.

Twenty-five tuna sausage products were purchased from retail markets in Taiwan. The rates of occurrence of biogenic amines, histamine-forming bacteria, and adulteration by pork and poultry were determined. The average content of various biogenic amines in all tested samples was less than 2.0 mg/100 g (<0.05 to 1.85 mg/100 g). Thirteen histamine-producing bacterial strains isolated from tested samples produced 12.1 to 1,261 ppm of histamine in Trypticase soy broth supplemented with 1.0% L-histidine. Among them, Raoultella ornithinolytica (one strain), Enterobacter aerogenes (one strain), and Staphylococcus pasteuri (two strains) were identified as prolific histamine formers. PCR assay revealed that the adulteration rates were 80% (20 of 25) and 4% (1 of 25) for pork and poultry, respectively, in tuna sausage. The fish species in the tuna sausage samples were identified as Thunnus albacares for 22 samples (88%), Thunnus alalunga for 1 sample (4%), and Thunnus thynnus for 1 sample (4%), whereas the remaining sample was identified as Makaira nigricans (blue marlin).

Effect of salt concentrations and drying methods on the quality and formation of histamine in dried milkfish (Chanos chanos).

The effects of salt concentrations (0-15.0%) and drying methods on the quality of dried milkfish were studied. The results showed that the levels of aerobic plate counts, total coliform, water activity, moisture contents, total volatile basic nitrogen (TVBN) and thiobarbituric acid (TBA) of the dried milkfish samples prepared with the same drying method decreased with increased salt concentrations. The samples prepared with the cold-air drying method had better quality in term of lower TVBN and TBA values than those of samples prepared with other drying methods. The histamine contents in all samples, except two, prepared with various salt concentrations by different drying methods were less than 1.9 mg/100 g. Two unsalted samples prepared with hot-air drying at 35 °C and sun drying methods were found to contain histamine at levels of 249.7 and 67.4 mg/100 g, respectively, which were higher than the potential hazard level of 50 mg/100 g.