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Journal of Food Protection, Vol. 48, No. 2, Pages 166-168 (February 1985) Copyright0 International Association of Milk, Food, and Environmental Sanitarians

Presence of in the Bluefisii, Pomatomus saltatrix

JOHN J. KAROLUS*, DONALD H. LEBLANC, A. J. MARSH, ROGER MSHAR and TOM H. FURGALACK Downloaded from http://meridian.allenpress.com/jfp/article-pdf/48/2/166/1656614/0362-028x-48_2_166.pdf by guest on 02 October 2021 Connecticut State Department of Health Services, Laboratory and Environmental Health Services Divisions, 150 Washington Street, Hartford, Connec­ ticut 06106

(Received for publication June 16, 1984)

ABSTRACT their bodies. At a hospital emergency room, the individu­ Consumption of bluefish {Pomatomus saltatrix) has been als also exhibited a rapid pulse and elevated blood pres­ epidemiologically implicated and confirmed by laboratory sure. They were administered , which gave analyses as a cause of scombroid food . An examina­ an immediate favorable response. Two patients were also tion of marketable bluefish filets in the State of Connecticut treated with epinephrine. Within 24 h, all symptoms had found over 6.5% of the filets had histamine levels indicative ceased. Laboratory analyses of comparison samples from of . No direct correlation of the presence of his- the same catch of bluefish revealed a range of 36 to 138 tidine decarboxylating and levels of histamine was mg of histamine/100 g of . found. The bluefish should be recognized as a food capable In 1982, five additional scombroid poisoning outbreaks of causing scombroid poisoning. occurred involving 18 people. Once again the bluefish was implicated. Because of these outbreaks, it was de­ cided that a survey of bluefish would be conducted to Illnesses with symptoms similar to scombroid fish determine what levels of histamine were present in raw poisoning have occurred occasionally following con­ fish marketed for consumption. Since detection of his­ sumption since before 1800 (16). The cause of the illness tamine is a procedure involving materials and equipment is thought to be histamine. Histamine is formed in the not common in many labortories, the prevalence of his­ flesh of certain fish by bacterial decarboxylation of his- tidine decarboxylating bacteria (HDB) was also deter­ tidine to histamine. Fish in the family Scombridae con­ mined. This was done to determine if a direct correlation tain levels of as high as 1 g/100 g of fish (72). existed between levels of histamine and the levels of The histamine may act synergistically with other heat- HDB. stable substances to produce the allergic-like reaction. The incubation period is a few minutes to an hour and MATERIALS AND METHODS the major symptoms are peppery taste, intense headaches, dizziness, vomiting, facial swelling and , and oral Samples burning sensations. Recovery is within 12 h (13). A total of 44 bluefish filets was collected from large retail markets The literature is well-documented in describing scom­ and restaurants throughout the State. The dates of collection ranged from mid-August through the beginning of October when the bluefish broid food poisoning from members of the family Scom­ is plentiful in Long Island Sound. About half the samples were fileted bridae (1-5,11,13,14,17). The U.S. outbreaks implicating at the establishment; the remaining were fileted before reception at the tuna, , and bonito as the vehicles of transmis­ establishment. Most samples were collected within 48 h after the facility sion, each involved a few to over 200 people. Also re­ received the fish. Nine samples were held 4 d or greater at the estab­ ported has been the confirmation of scombroid food lishment before collection. At the time of collection all samples were stored below 4°C and delivered to the laboratory under the same condi­ poisoning involving mahi mahi (Coryphaena hippurus), tion. If the analyses could not be done immediately, the samples were also called blue dolphin, a nonscombroid fish (6). frozen. In August of 1981, an outbreak of scombroid food poisoning involving five individuals was confirmed in Histamine detection Connecticut by epidemiological investigation and labora­ The AOAC - Fluorometric Method was used (/). The photofluorome- tory findings. Eight people had consumed bluefish while ter used in the determinations was the Perkin-Elmer Model MPF-2A. dining at a restaurant. Within 1 h, 5 people developed Histidine-decarboxyiating bacteria The histamine differential and confirmatory media and methods used nausea and headache. Immediately thereafter, a flushing have been described elsewhere (15). Briefly, formulation of the diffe­ sensation was experienced and a had spread across rential medium was as follows (g/L of distilled water): L-histidine -

JOURNAL OF FOOD PROTECTION, VOL. 48, FEBRUARY 1985 HISTAMINE IN BLUEFISH 167

2HC1, 27; tryptone, 5; yeast extract, 5; sodium chloride, 5; calcium mg of histamine/100 g of fish and a corresponding HDB carbonate, 1, agar, 30; and 10 ml of brom cresol purple solution (0.6 g count of <10/g. It would appear as if there is good corre­ per 100 ml of 95% ethanol). Before sterilization (10 min at 121°C), lation between the HDB and histamine levels. However, the pH of the medium was adjusted to 5.25. The confirmatory medium the table does show samples with high histamine levels contained the following (g/L of distilled water): L-histidine - 2HC1, 27; ammonium chloride, 0.1; yeast extract, 1; D-glucose, 0.5; sodium and low HDB counts. It was thought the freezing of sam­ chloride, 2; dextran T-500, 10; agar, 30; and 3.3 ml of brom cresol ples may be partially responsible for this observation. purple solution (0.6 g per 100 ml of 95% ethanol). Before sterilization Only 3 samples received at 4°C were frozen at the labora­ (15 min at 121°C), the pH was adjusted to 5.0. Three-ml volumes of tory until the following day. All of these samples had this medium were dispensed into cotton stoppered tubes. <5.0 mg of histamine/100 g of fish and HDB values of Fifty g of bluefish were homogenized with 450 ml of sterile phos­ phate buffer 0.4 M, pH 7.2, by a Waring blender. From this serial <10/g. Only 5 samples were collected in a frozen state. dilutions were made. One ml of the dilution was placed into a sterile Two of these, with 28.0 and 11.0 mg of histamine/100 g petri dish. Differential agar was poured into the dish, followed by gen­ of fish, had HDB counts of 2000 and 100/g, respectively. tle mixing. After solidification, 10 additional ml of the agar was poured Non-frozen samples with 50.0 and 11.0 mg of histamine/ as an overlay. Incubation was at 35°C for 24 h. HDB were character­ 100 g of fish had respective HDB counts of 30 and 100/ ized by a purple colored zone surrounding the colony. Suspicious col­ g. One non-frozen sample with 0.5 mg of histamine/ Downloaded from http://meridian.allenpress.com/jfp/article-pdf/48/2/166/1656614/0362-028x-48_2_166.pdf by guest on 02 October 2021 onies were subcultured to the confirmatory medium by means of a stab into the agar. Incubation was at 35°C for 24 h. An alkaline reaction 100 g of fish had a corresponding HDB count of 1000/g. characterized by a purple color change was a positive reaction. The These differences may be a reflection not only of freezing isolates were identified by biochemical - carbohydrate reactions. the samples, but also the sensitivity of the selective medium used, death of bacteria for reasons other than RESULTS AND DISCUSSION freezing, and species differences in the capability to de- carboxylate the . All bacterial isolates were The results of the histamine analyses and HDB counts members of the family Enterobacteriaceae. It appeared are listed in Table 1. The FDA states that the defect ac­ that the level of HDB alone is not an adequate indicator tion levels for tuna are 10 mg of histamine/100 g of fish for determining the histamine quality of bluefish. when accompanied by odors of decomposition or hon­ From 1977 to 1981, scombroid poisoning ranked fourth eycombing and 20 mg of histamine/100 g of fish based in confirmed foodborne disease outbreaks reported in the on histamine alone. A level which exceeds 50 mg of his­ United States, accounting for 7.1% of the total number tamine/100 g of fish may be considered a danger to (8). Traveling in large schools as warm season migrants, health (9). No sample exhibited decomposition odors or the bluefish has a worldwide but irregular distribution in honeycombing. The table indicates that over 6.5% (3 of the warmer seas. Documentation has shown that bluefish 44) of the marketable fish had reached levels of decom­ migration includes the Gulf of Mexico and the entire At­ position. Almost 10% (4 of 44) approached the defect lantic Coast. Bluefishing is a commercial and recreational action level of 20 mg of histamine/100 g of fish. The Centers for Disease Control report that disease usually re­ industry in the United States. In 1980, the total bluefish sults when the level exceeds 20 mg of histamine/100 g caught commercially was over 14.5 million lb. The recre­ offish (5). ational industry showed a total catch of 25 million lb. Both industries may expand if foreign markets were to Concerning levels of histamine vs. levels of HDB, our develop (7). results showed 21 samples with a concentration of <5.0 Given the seasonal abundance and commerical mar­ ketability of this widely distributed food fish, its capabil­ TABLE 1. Histamine levels and HDB counts in the samples. ity to serve as a medium for histamine production under­ No. of HDB No. of Sa scores its public health significance. Of major concern (per g) is the apparent lack of knowledge by fisherman regarding Not done 3 the critical importance of prompt and continous refrigera­ <10 21 tion of landed bluefish. The survey samples found to co- 10-100 6 tain levels of histamine >10 mg/100 g of fish were all >100 3 fileted before reception by the establishment. Even filets Not done 1 which were received by the facility in an iced state con­ <10 1 tained unsafe levels. Yoshinga and Frank (19) reported 10-100 2 most spoilage organisms are found on the skin and gills >100 0 of fish. Therefore, it is easy to spread these organisms onto the muscle surface during the fileting process (78). Not done 2 It is concluded that some form of temperature-abuse oc­ <10 0 10-100 2 curred on the boat, at the dock during the fileting proce­ >100 0 dure, or during delivery to market. This is of great con­ cern especially with the recreational fishermen who most Not done 1 often sell their excess catch to whomever it is that desires <10 0 the product. Frequently the recreational fisherman be­ 10-100 1 come the "back door" supplier for many foodservice and >100 1 retail stores. Unfortunately, these fishermen may lack the

JOURNAL OF FOOD PROTECTION, VOL. 48, FEBRUARY 1985 168 KAROLUS ET AL. knowledge and facilities to adequately handle and store 7. Anonymous. 1982. Bluefish fishery management plan - a draft. their catch. Frank et al. (70) experimenting with skipjack Mid-Atlantic Fishery Management Council. tuna found very little histamine formed in 24 h at 21.1 °C. 8. Anonymous. 1983. Centers for Disease Control foodborne disease surveillance annual summary 1981. HHS Publication No. (CDC) The optimal temperature was 37.8°C. At this tempera­ 83-8185. ture, the histamine levels (mg/100 g of tuna) ranged 9. Food and Drug Administration. 1982. Defect action levels for his­ from 472 to 673 at 24 h along various sections of the tamine in tuna; availability of guide. Fed. Reg. 470:40487-40488. filet. Within 12 h, sections of the filet exceeded the de­ 10. Frank, H. A., D. H. Yoshinaga, and W. Nip. 1981. Histamine composition level. This temperature/histamine level re­ formation and honeycombing during decomposition of , Katsuwonus pelamis at elevated temperatures. Marine lationship points out the importance of adequate refrigera­ Fisheries Rev. 43:9-14. tion in retarding production in tuna. This may also 11. Lerke, P. A., S. B. Weiner, S. L. Taylor, and L. S. Guthertz. be true for other species of fish, including the bluefish. 1978. Scombroid poisoning. Report of an outbreak. West. J. Med. There was no correlation between the length of storage 129:381-386. below 4°C of the filet product and levels of histamine 12. Lukton, A., and H. S. Olcott. 1958. Content of free imidazole found in the sampled filets. compounds in the muscle tissue of aquatic animals. Food Res. 23:611-618. Downloaded from http://meridian.allenpress.com/jfp/article-pdf/48/2/166/1656614/0362-028x-48_2_166.pdf by guest on 02 October 2021 Fish is a delicate food which rapidly decomposes when 13. Merson, M. H., W. B. Baine, E. J. Gangarosa, and R. C. Swan- improperly stored and handled. The bluefish (Pomatomus son. 1974. Scombroid fish poisoning. Outbreak traced to commer­ saltatrix) should be noted by public health officials as cially canned tuna fish. J. Am. Med. Assoc. 228:1268-1269. a potential bacterial growth medium for histamine pro­ 14. Omura, Y., R. J. Price, and H. S. Olcott. 1978. Histamine-form- ing bacteria isolated from spoiled skipjack tuna and jack mackerel. duction . J. Food Sci. 43:1779-1781. 15. Smith, A. M., M. A. Hayden, S. G. McCay, F. A. Zapatka, and M. K. Hamdy. 1982. Detection and confirmation of histamine - producing bacteria. Bull. Environm. Contam. Toxicol. 29:618-623. REFERENCES 16. Staruszkiewicz, W. F., E. M. Waldron, and J. F. Bond. 1977. 1. A.O.A.C. 1980. Official methods of analysis, 13th ed. Association Fluorometric determination of histamine in tuna: development of of Official Analytical Chemists, Washington, DC. p. 296. method. J. Assoc. Off. Anal. Chem. 60:1125-1130. 2. Anonymous. 1964. Scombroid fish poisoning - California. 17. Taylor, S. L., L. S. Guthertz, M. Letherwood, and E. R. Lieber. M.M.W.R. 13:166-167. 1979. Histamine production of Klebsiella pneumoniae and an inci­ 3. Anonymous. 1968. Scombroid fish poisoning - New York City. dent of scombroid fish poisoning. Appl. Environ. Microbiol. M.M.W.R. 17:452. 37:274-278. 4. Anonymous. 1971. Scombroid fish poisoning - Florida. 18. Yamani, M. I., D. Dickertmann, and F. Untermann. 1981. His­ M.M.W.R. 20:364. tamine formation by Proteus species from tunafish. Zbl. Bakteriol. 5. Anonymous. 1980. Scombroid fish poisoning - New Jersey. Hyg., I. Abt. Orig. B. 173:478-487. M.M.W.R. 29:106-107. 19. Yoshinaga, D. H., and H. A. Frank. 1982. Histamine-producing 6. Anonymous. 1980. Scombroid fish poisoning - Illinois, Michigan. bacteria in decomposing skipjack tuna (Katsuwonus pelamis). Appl. M.M.W.R. 29:167-168. Environ. Microbiol. 44:447-452.

Bullerman, con't. from p. 165 5. Dahlberg, K.R., and J.L. Van Etten, 1982. Physiology and However, A. ochraceus at 25°C produced more ochrato- biochemistry of fungal sporulation. Ann. Rev. Phytopathol. xin in the presence of low levels of sorbate. Thus the 20:281-301. effects of sorbate on mold growth and mycotoxin produc­ 6. Hesseltine, C.W. 1976. Conditions leading to mycotoxin contami­ nation of foods and feeds. In J.V. Rodricks (ed.), Mycotoxins and tion differ depending upon temperature and the mold other fungal related food problems. Advances in Chemistry Series, strain. It is likely that other environmental factors also Number 149. American Society, Washington, D.C. modify the inhibitory effects of sorbate and other preser­ 7. Jarvis, B. 1971. Factors affecting the production of mycotoxins. vatives. J. Appl. Bacteriol. 34:199-213. ACKNOWLEDGMENT 8. Mislivec, P.B. 1983. Personal communication. 9. Mislivec, P.B., C.T. Dieter, and V.R. Bruce. 1975. Effect of tem­ The technical assistance of Ms. Cindy Hall is gratefully acknowl­ perature and relative humidity on spore germination of mycotoxin edged. species of Aspergillus and Penicillium. Mycologia 67:1187-1189. 101 . Pohlmeier, M.M. 1978. Ochratoxin and citrinin production by REFERENCES Penicillium species isolated from . M.S. Thesis, University of Nebraska, Lincoln, NE. 1. Bauer, J., A.V. Montgelas, and B. Gedek. 1983. Aflatoxin B, 111 . Smith, J.E., and D.R. Berry. 1974. An introduction to biochemis­ production in presence of preservatives/antimicrobial agent. Proc. try of fungal development. Academic Press. London and New Int. Symp. Mycotoxins, Cairo, pp. 249-225. York. 326 pp. 2. Bullerman, L.B. 1983. Effects of potassium sorbate on growth and 112 . Sofos, J.N., and F.F. Busta. 1981. Antimicrobial activity of sor­ aflatoxin production by Aspergillus parasiticus and Aspergillus bate. J. Food Prot. 44:614-622. flavus. J. Food Prot. 46:940-942. 113 . Yousef, A.E., and E.H. Marth. 1981. Growth and synthesis of af­ 3. Bullerman, L.B. 1984. Effects of potassium sorbate on growth and latoxin by Aspergillus parasiticus in the presence of sorbic acid. patulin production by Penicillium patulum and Penicillium J. Food Prot. 44:736-741. I4 roqueforti. J. Food Prot. 47:312-316. 14' . Yousef, A.E., and E.H. Marth. 1983. Incorporation of [ C]acetate 4. Collins, E.B. 1971. Preservatives in dairy foods. J. Dairy Sci. by Aspergillus parasiticus in the presence of antifungal agents. Eur. 54:148-152. J. Appl. Microbiol. Biotechnol. 18:103-108.

JOURNAL OF FOOD PROTECTION, VOL. 48, FEBRUARY 1985