Screening and activity of potential gastrointestinal probiotic lactic acid bacteria against Yersinia ruckeri O1b.

Yersiniosis of cultured Atlantic salmon is a recurrent fish health management challenge in many continents. The causative organism, Yersinia ruckeri, can reside latently in the gut and lead to acute infection and disease during hatchery and sea-transfer stages. One potential prevention approach is the administration of probiotic bacteria to suppress gut colonization of Y. ruckeri. Our study aimed to isolate and identify anti-Yersinia activity among lactic acid bacteria (LAB) isolated from the gastrointestinal tract (GIT) of aquatic animals. Of the 186 aquatic GIT isolates examined, three strains showed diffusible antimicrobial activity towards Y. ruckeri O1b. Analysis of 16 s rRNA gene sequences indicated the three bacterial strains were Enterococci, related to Enterococcus sp. (99%), Enterococcus thailandicus (99%), and Enterococcus durans (99%). Anti-Yersinia activity was maintained at neutral pH (~6.5-7.0), and in-vitro environmental tolerance assays showed the three strains could withstand simulated salmonids gastrointestinal tract conditions of: low pH (3.4) and 3% bile salt content. All three Enterococci strains showed higher adhesion to the intestinal mucus of Atlantic salmon than Y. ruckeri O1b (E. durans 24%, E. enterococcus sp. 25% and E. thailandicus 98%, compared to Y. ruckeri O1b 5%). However, only Enterococcus sp. and E. thailandicus were able to grow in the salmon intestinal mucus broth while E. durans showed no growth. Anti-Yersinia activity was completely inactivated by proteinase-K treatment, suggesting that the active compound/s are proteinaceous and may be bacteriocin-like inhibitory substances (BLIS). Our data indicate that Enterococcus sp. MA176 and E. thailandicus MA122 are potential probionts for the prevention of yersiniosis in salmonids. Further in-vivo studies are required to determine whether these bacteria reduce the incidence of yersiniosis in Atlantic salmon.

Spoilage microbial community profiling by 16S rRNA amplicon sequencing of modified atmosphere packaged live mussels stored at 4oC.

There is little information on the microbial communities associated with modified atmosphere (MA)-packaged live mussels. There is also a dearth of information on how pre-packaging depuration modifies the microbial communities and spoilage of live mussels. Amplicon sequencing was used to describe spoilage microbial succession in MA-packaged live mussels during storage at 4 °C. Proteobacteria, Cyanobacteria and Firmicutes were the three major phyla observed in the mussel meat and pouch water of undepurated and depurated mussels. Among these phyla, Cyanobacteria was more predominant on day 0 in mussel meat of undepurated and depurated mussels while Proteobacteria was predominant in commercially-depurated mussels. Synechococcus was apparently dominant on days 0-7 in the meat of undepurated mussels and days 0-10 in depurated mussels. Shewanella was dominant on day 0 in commercially-depurated mussels and dominant on day 15 in undepurated while Acidaminococcus was dominant in depurated mussels on day 15. Psychromonas was observed to be dominant in commercially-depurated mussels on day 7 and further shifted to Acinetobacter by day 10 and 15. In the pouch water, Acinetobacter was dominant throughout the storage days in undepurated mussels while Psychrobacter was predominant in both depurated and commercially-depurated mussels. This study demonstrated the impact of depuration on the microbiota and the spoilage mechanism of MA-packaged live mussels. Shewanella was easily removed through depuration. However, spoilage bacteria such as Acidaminococcus could not be easily removed although they are not important at the beginning but grew at the end. Pouch water contributed suitable biological medium for the growth of Acinetobacter and Psychrobacter and both enhanced the growth of spoilage bacteria such as Shewanella and Acidaminococcus.

Evaluation of spoilage potential and volatile metabolites production by Shewanella baltica isolated from modified atmosphere packaged live mussels.

Under the current commercial practice, live mussels only have 10days' shelf-life. Observed spoilage indices reduce consumers' acceptance, palatability and shelf-life of modified atmosphere packaged (MAP) live mussels. The aims of this study are to isolate specific spoilage bacteria from modified atmosphere packaged live mussels, evaluate isolates for microbial spoilage indices using qualitative methods and volatile metabolites production. Forty-six hydrogen sulphide producing bacteria were isolated and evaluated for trimethylamine n-oxide (TMAO) reduction, proteolytic and lipolytic activities and hydrogen sulphide production. Twenty-eight isolates were obtained from pouch water and 18 from mussel meat. All the isolates could produce H2S on Iron agar at 25°C while 30/46 produced H2S at 4°C and tolerate 0-6% NaCl. Four (4/46) isolates could not hydrolyse mussel protein. Over 80% isolates reduced TMAO to TMA in 3days with the production of H2S. Results of this study shows hydrogen sulphide producing bacteria isolated from MAP live mussels produce microbial spoilage indices. Isolate with highest enzymatic activities and hydrogen sulphide production was identified as Shewanella baltica using 16S rRNA gene. Axenic culture of the isolate was inoculated into sterile mussel broth. Inoculated sample was further stored at 4°C for 10days for spoilage study. Volatile metabolites produced during storage were evaluated using headspace solid phase micro-extraction gas chromatography mass spectrometry (HS-SPME GC/MS). A total of 44 compounds were identified in the sample after 10days while 27 compounds were identified in inoculated mussel broth. Group of compounds identified are alcohols, aldehydes, phenol, furans, ketone, esters, organic acid, aromatic hydrocarbons, alkanes, nitrogen and sulphur containing compounds. Dimethyl trisulphide, methyl-phenol, 3,5-octadiene and thiohexene were unique to inoculated mussel broth. Understanding spoilage mechanism and attendant spoilage indices will help in designing effective mussel quality protocols and shelf-life extension.

Seafood spoilage microbiota and associated volatile organic compounds at different storage temperatures and packaging conditions.

Seafood comprising of both vertebrate and invertebrate aquatic organisms are nutritious, rich in omega-3 fatty acids, essential vitamins, proteins, minerals and form part of healthy diet. However, despite the health and nutritional benefits, seafood is highly perishable. Spoilage of seafood could be as a result of microbial activity, autolysis or chemical oxidation. Microbial activity constitutes more spoilage than others. Spoilage bacteria are commonly Gram negative and produce off odours and flavours in seafood as a result of their metabolic activities. Storage temperature, handling and packaging conditions affect microbial growth and thus the shelf-life of seafood. Due to the complexity of the microbial communities in seafood, culture dependent methods of detection may not be useful, hence the need for culture independent methods are necessary to understand the diversity of microbiota and spoilage process. Similarly, the volatile organic compounds released by spoilage bacteria are not fully understood in some seafood. This review therefore highlights current knowledge and understanding of seafood spoilage microbiota, volatile organic compounds, effects of storage temperature and packaging conditions on quality of seafood.

Application of electrolysed oxidising water as a sanitiser to extend the shelf-life of seafood products: a review.

Electrolysed oxidising water (E.O. water) is produced by electrolysis of sodium chloride to yield primarily chlorine based oxidising products. At neutral pH this results in hypochlorous acid in the un-protonated form which has the greatest oxidising potential and ability to penetrate microbial cell walls to disrupt the cell membranes. E.O. water has been shown to be an effective method to reduce microbial contamination on food processing surfaces. The efficacy of E.O. water against pathogenic bacteria such as Listeria monocytogenes, Escherichia coli and Vibrio parahaemolyticus has also been extensively confirmed in growth studies of bacteria in culture where the sanitising agent can have direct contact with the bacteria. However it can only lower, but not eliminate, bacteria on processed seafoods. More research is required to understand and optimise the impacts of E.O. pre-treatment sanitation processes on subsequent microbial growth, shelf life, sensory and safety outcomes for packaged seafood products.