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. 2017 Jun 20;4(2):31.
doi: 10.3390/vetsci4020031.

Detection and Characterization of Histamine-Producing Strains of Photobacterium damselae subsp. damselae Isolated from Mullets

Marcello Trevisani  1 , Rocco Mancusi  2 , Matilde Cecchini  3 , Claudia Costanza  4 , Marino Prearo  5
Affiliations

Detection and Characterization of Histamine-Producing Strains of Photobacterium damselae subsp. damselae Isolated from Mullets

Marcello Trevisani et al. Vet Sci. .

Abstract

Photobacterium damselae subsp. damselae (Pdd) is considered to be an emerging pathogen of marine fish and has also been implicated in cases of histamine food poisoning. In this study, eight strains isolated from mullets of the genera Mugil and Liza captured in the Ligurian Sea were characterized, and a method to detect histamine-producing Pdd from fish samples was developed. The histamine-producing potential of the strains was evaluated in culture media (TSB+) using a histamine biosensor. Subsequently, two strains were used to contaminate mackerel fillets (4 or 40 CFU/g), simulating a cross-contamination on the selling fish stalls. Sample homogenates were enriched in TSB+. The cultures were then inoculated on thiosulfate-citrate-bile salts-sucrose agar (TCBS) and the dark green colonies were cultured on Niven agar. The violet isolates were characterized using specific biochemical and PCR based tests. All Pdd strains were histamine producers, yielding concentration varying from 167 and 8977 µg/mL in TSB+ cultures incubated at 30 °C for 24 h. Pdd colonies were detected from the inoculated mackerel samples and their histidine decarboxylase gene was amplified using species-specific primer pairs designed for this study. The results indicate that mullets can be source of Pdd and the fish retailers needs to evaluate the risk posed by cross-contamination on the selling fish stalls.

Keywords: Photobacterium damselae subsp. damselae; cross contamination; histamine biosensor; histidine decarboxylase; mullets.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Amplification plot (A) and melt curves (B) from rt-PCR of histidine decarboxylase genes of Gram-negative bacteria (primers hdc-dp). Legend: annealing temperature for gradient PCR amplification ranged from 49 (dark lines) to 53 °C (grey lines); lines with symbols Δ and ○ correspond to Pdd (3 strains)and M. morgani, respectively.
Figure 2
Figure 2
Amplification plot (A) and melt curves (B) from rt-PCR of histidine decarboxylase gene of Photobacterium damselae subsp. damselae.
See this image and copyright information in PMC

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