Evaluation of Lactobacillus sobrius/L. amylovorus as a New Microbial Marker of Pig Manure

Abstract

Based on a comparison of the dominant microbial populations in 17 pig manure samples and using a molecular typing method, we identified a species, Lactobacillus sobrius and Lactobacillus amylovorus (which now are considered a single species and are designated L. sobrius/amylovorus here), that was consistently found in manure. The aim of the present study was to confirm by real-time PCR the relevance of this species as a marker of pig fecal contamination. The specificity of L. sobrius/amylovorus was evaluated in human and animal DNA extracted from feces. The real-time PCR assay then was applied to water samples, including effluents from urban wastewater treatment plants, runoff water, and rivers. L. sobrius/amylovorus was consistently present in all samples of swine origin: 48 fecal samples, 18 from raw manure and 10 from biologically treated manure at mean concentrations of 7.2, 5.9, and 5.0 log 10 cells/g, respectively. The species was not detected in any of the other livestock feces (38 samples from cattle and 16 from sheep), in the 27 human fecal samples, or in the 13 effluent samples from urban wastewater treatment plants. Finally, L. sobrius/amylovorus was not detected in runoff water contaminated by cattle slurry, but it was quantified at concentrations ranging from 3.7 to 6.5 log 10 cells/100 ml in runoff water collected after pig manure was spread on soil. Among the stream water samples in which cultured Escherichia coli was detected, 23% tested positive for L. sobrius/amylovorus. The results of this study indicate that the quantification of L. sobrius/amylovorus using real-time PCR will be useful for identifying pig fecal contamination in surface waters.

Relevance of Bacteroidales and F-Specific RNA Bacteriophages for Efficient Fecal Contamination Tracking at the Level of a Catchment in France

“…However, pig pasturing has not been reported or observed on the Daoulas catchment. Separately, Lactobacillus amylovorus GE39/GE40, another pig-associated marker (19), was quantified at concentrations ranging from 4.6 to 6.5 log 10 copies/100 ml in 5 out of the 5 water samples collected downstream of the accidental spillage monitored in the present study (Olivia Solecki, Irstea, Rennes, France, personal communication). This marker has a greater persistence than Pig2Bac in fresh and marine waters (32) and might be considered for pig manure tracking at the catchment level.…”

Analysis of bacterial community shifts in the gastrointestinal tract of pigs fed diets supplemented with β-glucan from Laminaria digitata, Laminaria hyperborea and Saccharomyces cerevisiae

“…L. amylovorus was a stable member of the Lactobacillus community and it was not affected by any of the diets investigated. The prevalence of this species in pig faeces coincides with its use as a microbial marker of pig manure (Marti et al, 2010). Konstantinov et al (2004) demonstrated that the combination of dietary fibre and oligosaccharides present in the diet may specifically stimulate the L. amylovorus-like population along the gut of weaning pigs.…”

Microbial community analysis and identification of alternative host-specific fecal indicators in fecal and river water samples using pyrosequencing

“…As a result, many assays with host-specific primer sets do not have reliable specificities for identifying and quantifying fecal pollution sources (McLain et al, 2009), which requires alternative fecal indicators to increase the choice of tools and accurately identify the fecal pollution source in water (Marti et al, 2010).…”