Safety assessment and characterisation of Ligilactobacillus salivarius PS21603 as potential feed additive for swine

Abstract

The present study aimed to characterise in vitro properties of the strain Ligilactobacillus salivarius PS21603 and evaluate in vivo piglets' tolerance for its use as feed additive in swine. The ability of L. salivarius PS21603 of inhibiting enteropathogens' growth in vitro was evaluated using a co-culture assay. Low pH tolerance, bile tolerance, and resistance to osmotic changes were evaluated. The antibiotic susceptibility profile of L. salivarius PS21603 was assessed through broth microdilution method. Whole genome sequencing (WGS) was performed to exclude the presence of antibiotic resistance genes. L. salivarius PS21603 showed a high antimicrobial activity in vitro, reducing in a mean of 6.16 log cfu/ml eight different enterotoxigenic Escherichia coli strains. Moreover, L. salivarius PS21603 showed resistance to osmotic changes and was able to survive to a pH above 3.5 during 24 h and up to pH 2 at least during 2 h. In addition, WGS revealed that L. salivarius PS21603 did not harbour any resistance genes and thus there was no risk of transmissibility. Finally, an in vivo 28-days safety and tolerance study was performed. For that, 384 healthy piglets (28±2 days old and 7.5±1.5 kg, at weaning) were divided into three treatment groups receiving a different dose of L. salivarius PS21603: T1, 10⁹ cfu/day; T2, 10⁷ cfu/day; T3, control. Piglet's health status was daily controlled. Individual bodyweight and feed intake per pen were weekly recorded to determine performance parameters. Blood samples were collected in 16 piglets from each treatment group on days 0 and 28 for determination of cytokine profiles. L. salivarius PS21603 was safe and well tolerated by piglets, there were no differences in performance nor cytokine profile between treatment groups. In conclusion, L. salivarius PS21603 is a potential candidate for a probiotic prevention strategy against pig diarrhoea.

Keywords: Escherichia coli; Ligilactobacillus salivarius; diarrhoea; feed additive; pig.