Functional and probiotic characterization of Ligilactobacillus salivarius CPN60 isolated from calf faeces and its appraisal in rats.

Emerging concern about the emergence of antimicrobial resistance has limited the use of antibiotics in calves. Hence, there is a need to find suitable alternatives to antibiotics to manage gastrointestinal infections in neonatal calves. The objective of the present study was to develop a probiotic of calf-origin for its potential application in calf nutrition. Accordingly, 69 lactic acid bacteria (LAB) strains were isolated from faeces of newborn calves, out of which 10 strains were short-listed for further in vitro testing based on the aggregation time and cell surface hydrophobicity. The results of acid-, bile- and phenol-tolerance tests indicated that out of the ten strains, the isolate CPN60 had better resistance to these adverse conditions likely to be encountered in the gastrointestinal tract. The isolate also showed an optimal ability to produce biofilm. Further assessments reiterated its superiority in terms of co-aggregation and antagonistic activity against pathogenic strains of Escherichia coli. Subsequently, the isolate was identified through 16S rRNA sequencing and sequence homology and designated as Ligilactobacillus salivarius CPN60. The candidate probiotic was evaluated in vivo using 48 male (5 weeks old) Wistar rats, divided into two equal groups viz. control (CON) and probiotic (PRO). During the 4-weeks feeding trial, the PRO group rats were gavaged with one mL culture of L. salivarius CPN60 equivalent to 10 CFU/rat. The in vivo trial results indicated better nutrient utilization efficiency and growth performance (p < 0.001) of the PRO group of rats. The probiotic supplementation improved the faecal concentration of lactate (p < 0.001) and individual as well as total short-chain fatty acids (p < 0.001) production. The cell-mediated immune response, assessed as a delayed-type hypersensitivity reaction to phytohaemagglutinin-P, was improved (p < 0.001) in PRO compared to the CON rats. It is concluded that the calf-origin probiotic L. salivarius CPN60, in addition to possessing all the in vitro functional attributes of a candidate probiotic, also has desirable potential for its future use in young calves to promote gut health and immunity.