Inhibition of Zoonotic Pathogens Naturally Found in Pig Manure by Black Soldier Fly Larvae and Their Intestine Bacteria

SIMPLE SUMMARY: With the rapid development of the economy and the improvement of people’s living standards, people need to rear a lot of livestock to meet demand for proteins. This also involves an increase in the production of livestock manure. The expanding rate of livestock manure has become a thorny issue, owing to characteristics such as plentiful nitrogen and abundant zoonotic pathogens. The saprophagous larvae of the black soldier fly (BSF) are often associated with animal manure and can significantly reduce the populations of different zoonotic pathogens in livestock manure. However, reports about the mechanisms of this phenomenon are scarce. In this study, we investigated the potential mechanisms of BSF larvae in reducing the zoonotic pathogens naturally found in pig manure. The results clearly showed that zoonotic pathogens in pig manure were significantly decreased after being treated with BSF larvae, and also suggested that the antimicrobial peptides produced by the BSF larvae and gut-associated bacteria are able to antagonize the zoonotic pathogens. This study will contribute to reveal the potential antagonistic mechanisms of BSF larvae against zoonotic pathogens and improve the safety of organic waste conversion by BSF larvae. ABSTRACT: Black soldier fly (BSF) larvae are often exposed to organic waste which harbors abundant zoonotic pathogens. We investigated the ability of BSF larvae to inhibit the zoonotic pathogens naturally found in pig manure. The zoonotic pathogens populations were detected by using selective medium during the conversion. Results showed that the viability of the zoonotic pathogens in pig manure was significantly affected. After eight days of conversion, the Coliform populations were undetected, and Staphylococcus aureus and Salmonella spp. decreased significantly on the eighth day. Antimicrobial assays of the purified recombinant defensin-like peptide 4 (DLP4) showed that this peptide exhibits inhibitory activity against S. aureus, Salmonella enterica serovar typhimurium, and Escherichia coli in vitro. Bacteria BSF-CL and BSF-F were isolated from the larvae gut, and both inhibited the growth of S. aureus and E. coli, but Salmonella spp. was sensitive to the BSF-CL strain (but not to the BSF-F strain). The results from our experiments indicate that BSF larvae are capable of functionally inhibiting potential zoonotic pathogens in pig manure through a variety of mechanisms including antimicrobial peptides expression and the gut associate microorganisms. This study provides a theoretical basis for further study on the combined mechanism of BSF larvae immunity and its gut microbes against the zoonotic pathogens in pig manure.