Detection of Gram-Negative Histamine-Producing Bacteria in Fish: A Comparative Study

• Published in: Journal of food protection 2009-09, Vol.72 (9), p.1987-1991
• Main Authors: Bjornsdottir, Kristin, Bolton, Gregory E, McClellan-Green, Patricia D, Jaykus, Lee-Ann, Green, David P
• Format: Article
• Language: English
• Subjects: Agar; Amines; Animals; Bacteria; bacterial contamination; biogenic amines; Biological and medical sciences; carboxy-lyases; Chromatography; Chromatography, High Pressure Liquid; Colony Count, Microbial - methods; Comparative studies; Consumer Product Safety; Fish; Fishes - microbiology; food contamination; Food contamination & poisoning; Food Contamination - analysis; Food industries; Food microbiology; food pathogens; Food safety; Foodborne Diseases - microbiology; Fundamental and applied biological sciences. Psychology; General aspects; genes; Genetic testing; Gram-negative bacteria; Gram-Negative Bacteria - isolation & purification; Gram-Negative Bacteria - metabolism; Histamine; Histamine - biosynthesis; Histamine - toxicity; histide decarboxylyase; Humans; Ingestion; isolation; Liquid chromatography; Methods; Methods of analysis, processing and quality control, regulation, standards; microbial detection; Poisoning; Polymerase Chain Reaction; potentiometers; Predictive Value of Tests; Public health; rapid methods; reversed-phase high performance liquid chromatography; scombroid poisoning; Seafood; Seafood - microbiology; secondary metabolites; selective media; Sensitivity and Specificity; virulence
• ISSN: 0362-028X; 1944-9097
• DOI: 10.4315/0362-028x-72.9.1987

Poisoning due to ingestion of foods with elevated levels of biogenic amines (histamine, putrescine, cadaverine, and tyramine) is well documented. Histamine fish poisoning largely is due to growth of naturally occurring bacteria associated with scombroid fish species. A rapid and reliable method is needed to screen for the presence of histamine-forming bacteria in fish. This study included a comparison of three methods for the detection of histamine-producing bacteria. A total of 152 histamine-producing and non-histamine-producing bacteria from multiple sources were screened using a modified Niven's agar method, a potentiometric method, and a PCR-based assay targeting a 709-bp fragment of the histidine decarboxylase gene. Histamine production by bacterial isolates was confirmed by high-performance liquid chromatography (HPLC). Bacterial strains were categorized as producing high amounts of histamine, low amounts of histamine, or no histamine. Of the 152 strains tested, 128 (84%) were positive with the Niven's agar method, 73 (48%) were positive with the potentiometric technique, and 74 (49%) were positive with the PCR assay. Overall, a 38% false-positive rate was observed with the modified Niven's agar method, although this method detected both low-histamine and high-histamine strains. There was a high degree of concordance (>99%) between results of the potentiometric and PCR methods, but neither of these methods detected low-histamine bacteria. These observations support the need for a simple and straightforward yet sensitive method for detecting histamine-producing bacteria in seafood and environmental samples.