Characterization of ready-to-eat fish surface as a potential source of contamination of Vibrio parahaemolyticus biofilms

Abstract

The worldwide consumption of ready-to-eat seafood products has steadily increased due to a range of health benefits. However, depending on the handling or cutting process of raw fish, ready-to-eat sashimi can be exposed to microbiological risks that can lead to foodborne infection by marine pathogens. Since surface characteristics are key factors for microbial adhesion and biofilm formation, the present study aims to determine the correlation between raw fish skin properties and Vibrio parahaemolyticus biofilm formation. We analyzed V. parahaemolyticus biofilms (ATCC 17802; initially inoculated ca. 2 or 4 log CFU/cm²) on fish skin (gizzard shad, pomfret, red snapper, and mackerel; fish species served as sashimi without peeling the skin) formed under simulated marine environments (incubating in artificial seawater with rocking motion at 30 °C, the maximum temperature of seasonal seawater) for 24 h. The characteristics of fish skin were determined using confocal laser scanning microscopy/scanning electron microscopy. V. parahaemolyticus showed higher biofilm counts on fish skins than on stainless steel, which was used as a control (P < 0.05). In particular, V. parahaemolyticus formed biofilms with significantly higher levels of bacterial populations on gizzard shad and pomfret (ca. 5 log CFU/cm²; P < 0.05), highlighting the relationship between the biofilm formation level and the characteristics of gizzard shad and pomfret skins. The surface roughness of fish skins, including the main roughness parameters (Ra, Rq, and Rz), influenced the attachment of V. parahaemolyticus (P < 0.05). Additionally, images of V. parahaemolyticus biofilms suggested that different topographical profiles of fish species (e.g., mucus, unique structural features, etc.) could cause V. parahaemolyticus to exhibit different biofilm phenotypes, such as sticking to or entangling on the fish skin surface. The major findings of this study provide various phenotypic adhesions of V. parahaemolyticus to fish skin in considerations of the potential hazard for the consumption of ready-to-eat sashimi served with its skin.

Keywords: Biofilm; Cross-contamination; Raw fish; Surface roughness; Surface topography; Vibrio parahaemolyticus.