Isolation of Shewanella putrefaciens GRD 03 from Fish and Explication of Biofilm Adherence Potency on Different Substrates

Abstract

Foodborne pathogens are the main threat and cause of food poisoning. The majority of food infections have been related to the biofilm formation of foodborne pathogens in the food industry. Shewanella putrefaciens (KX355803, GRD 03), a Gram-negative pathogen isolated from mackerel fish, was identified and recognized as a food spoilage bacterium and a strong biofilm producer. The adhesion or attachment ability of Shewanella putrefaciens was determined on steel, plastic, glass, PVC and wood. NB (Nutrient broth), LB (Luria-Bertani broth), TSB (Tryptic soy broth) and BHI (Brain heart infusion broth) were enriched with glucose and shows optimum for bacterial adhesion. In the microtiter plate method (MTP), the strong attachment was observed at 48 and 72 hours of incubation and significant differences were obtained at p < 0.05. As the incubation period increases, the OD value (Optical density) of samples also increase. Biofilm formation is the major cause cross-contamination, and shows resistance to certain disinfectants, which leads to environmental stress tolerance. This study suggested with optimum biofilm production of isolate from fish by using glucose enriched media on different substrates, also comparing different growth media provide a detailed idea about biofilm-forming ability at different incubation time intervals.

Shewanella putrefaciens, a rare human pathogen: A review from a clinical perspective

“…S. putrefaciens is a gramnegative, facultatively anaerobic, rod shaped, oxidase and catalase positive, motile bacterium with a single polar flagellum ( Nozue et al., 1992 ; Héritier et al., 2004 ; Holt et al., 2005 ; Sharma and Kalawat, 2010 ; Yu et al., 2022 ). The bacterium has a G+C content between 45-48 mol% and grows in 1-2mm large, yellowish-brown colonies ( Nozue et al., 1992 ; Holt et al., 2005 ; Jayalekshmi et al., 2022 ). Hydrogen sulfide generation is the main phenotypic feature ( Holt et al., 2005 ).…”

“…They have also been isolated from a variety of foods including milk, cream, butter, eggs, poultry, (raw) fish or seafood and beef products ( Chen et al., 1997 ; Khashe and Janda, 1998 ; Bulut et al., 2004 ; Oh et al., 2008 ). S. putrefaciens is a biofilm former and able to reduce TMAO to trimethylamine ( Vogel et al., 1997 ; Bagge et al., 2001 ; Holt et al., 2005 ; Jayalekshmi et al., 2022 ). This seems to be a relevant reason why this bacterium is an important spoilage agent of protein-rich refrigerated foods, especially for frozen white-fleshed fish from temperate waters ( Brink et al., 1995 ; Vogel et al., 1997 ; Jayalekshmi et al., 2022 ).…”

“…S. putrefaciens is a biofilm former and able to reduce TMAO to trimethylamine ( Vogel et al., 1997 ; Bagge et al., 2001 ; Holt et al., 2005 ; Jayalekshmi et al., 2022 ). This seems to be a relevant reason why this bacterium is an important spoilage agent of protein-rich refrigerated foods, especially for frozen white-fleshed fish from temperate waters ( Brink et al., 1995 ; Vogel et al., 1997 ; Jayalekshmi et al., 2022 ).…”

“…grow after an incubation period of 18-24 hours on conventional solid culture media, S. putrefaciens e.g. on Luria-Bertani (LB) broth, and on media commonly used in microbiological diagnostics, such as Mac-Conkey agar ( Holt et al., 2005 ; Jayalekshmi et al., 2022 ).…”

Fish as a source of foodborne bacteria

“…Bacterial colonies or communities directly attached to a specific surface initiate the formation of biofilm, which is a major issue in food processing industries. Cross-contamination is the main cause of biofilm formation, and it occurs when food moves through some unhygienic surfaces, leading to biofilm production and contamination (38).…”