Identification and Characterisation of Potential Probiotic Lactic Acid Bacteria Extracted from Pig Faeces

Abstract

Given that probiotics always have host-homologous and strain-specific effects on the hosts, lactic acid bacteria extracted and identified from porcine specimens can be potentially developed as probiotics for pig production. We aimed to identify lactic acid bacteria that are potentially probiotic, have good capacity of inhibiting pathogenic bacteria in intestine and are promising to be used as substitutes for antibiotics in pig production. Potential probiotic strains were extracted from 15 fecal specimens collected from 15 apparently healthy pigs, and were identified via 16S rDNA sequencing. The antimicrobial activity, tolerance to acid and bile salts, Caco-2 cell adhesiveness and susceptibility to antibiotics of the isolates were evaluated in vitro, and oral toxicity of the isolates were evaluated in mice. One Lactiplantibacillus plantarum (BJR2), two Lacticaseibacillus casei (HJD and TH2), one Lacticaseibacillus rhamnosus (MRS1), and two Enterococcus faecium (S-3 and S-4-H) were extracted from healthy pigs and underwent 16S rDNA sequencing identification. L. plantarum BJR2 and L. casei HJD exhibited broad-spectrum and higher antimicrobial activity against indicator enteric pathogens, including Salmonella choleraesuis CVCC 2139, Escherichia coli (O147:K89) CVCC 199, Escherichia coli (O141:K99) CVCC 223 and Escherichia coli (O139) CVCC 1496, among 6 tested strains. In addition, both L. plantarum BJR2 and L. casei HJD exhibited good tolerance to low pH (pH 2.5 and pH 3.5) and 0.30% bile salts, had relatively strong Caco-2 adhesiveness and carried no transferable resistant genes against antibiotics encoded by plasmid. In safety trials, these two isolates had no α or β-hemolysis activity, and were proved safe through oral toxicity tests in mice. It is concluded that L. plantarum BJR2 and L. casei HJD are potential probiotic candidate strains and their probiotic effects need to be further studied in pigs.