Isolation and characterization of competitive exclusion microorganisms from animal wastes-based composts against Listeria monocytogenes.

Abstract

To isolate the slow-growing or viable but non-culturable competitive exclusion (CE) microorganisms from composts and then verify the anti-Listeria monocytogenes activities of those CE isolates in compost. CE strains were isolated from composts using double- or triple-layer agar methods, purified, and then characterized. Both compost extracts and solid compost samples were spiked with a cocktail of 3 L. monocytogenes strains which were co-inoculated with or without CE strain cocktail and incubated at both 22 and 35°C for 168h. Results indicated that the addition of resuscitation promoting factor (Rpf) promoted the growth of slow-growing species from composts. About 50% of the isolated CE strains (n=40) were identified as Bacillus spp., 17 strains can inhibit more than 10 tested L. monocytogenes strains, and nine strains were motile and competitive biofilm formers. In compost extracts, the growth potentials of L.monocytogenes were reduced up to 2.2 logs when co-culturing with CE strains. In compost samples, the addition of CE strains reduced L. monocytogenes population by ca. 1.3 log CFU/g at 22°C after 24-168h incubation. Our modified double/triple-layer agar procedure with Rpf as growth supplement coupled with spot-on-lawn testing can be a quick and efficient method for isolating CE candidates from composts. The efficacy of CE strains against L. monocytogenes in compost extracts and compost samples was affected by compost type, nutrient level, and incubation temperature. Compost is a rich source of CE microorganisms, and compost-adapted CE microorganisms have the potential as a biological agent to control L. monocytogenes in agricultural environments.