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Effect of oxygen and temperature on the dynamic of the dominant bacterial populations of pig manure and on the persistence of pig-associated genetic markers, assessed in river water microcosms
Authors:Marti R  Mieszkin S  Solecki O  Pourcher A-M  Hervio-Heath D  Gourmelon M
Institution:Cemagref, UR GERE, Rennes, France.
Abstract:Aims: The aim is to evaluate the dynamic of Bacteroides–Prevotella and Bacillus–Streptococcus–Lactobacillus populations originating from pig manure and the persistence of pig‐associated markers belonging to these groups according to temperature and oxygen. Methods and Results:  River water was inoculated with pig manure and incubated under microaerophilic and aerobic conditions, at 4 and 20°C over 43 days. The diversity of bacterial populations was analysed by capillary electrophoresis‐single‐strand conformation polymorphism. The persistence of the pig‐associated markers was measured by real‐time PCR and compared with the survival of Escherichia coli and enterococci. Decay was characterized by the estimation of the time needed to produce a 1‐log reduction (T90). The greatest changes were observed at 20°C under aerobic conditions, leading to a reduction in the diversity of the bacterial populations and in the concentrations of the Pig‐1‐Bac, Pig‐2‐Bac and Lactobacillus amylovorus markers with a T90 of 10·5, 8·1 and 17·2 days, respectively. Conclusions: Oxygen and temperature were found to have a combined effect on the persistence of the pig‐associated markers in river waters. Significance and Impact of the Study: The persistence profiles of the Pig‐1‐Bac, Pig‐2‐Bac and Lact. amylovorus markers in addition to their high specificity and sensitivity support their use as relevant markers to identify pig faecal contamination in river waters.
Keywords:Lactobacillus amylovorus  microbial source tracking  oxygen  persistence  pig‐associated Bacteroidales markers  quantitative real‐time PCR  river water  temperature
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