Screening and characterization of Sahiwal cattle calves-origin lactic acid bacteria based on desired probiotic attributes for potential application.

Probiotics are living microorganisms that confer health benefits to host when administered in adequate amounts. To develop novel host-specific probiotic for their application as feed additive, the present study was undertaken to isolate and characterize probiotic strains of indigenous cattle-calves origin. A total of 55 colonies were isolated from 12 healthy calves, with 34 of the isolates being Gram-positive, catalase-negative and vancomycin-resistant. Furthermore, eleven isolates showed tolerance to acid (pH 2.0) and thirteen isolates tolerated bile salts (0.3%). Seven common acid and bile tolerance strains were further investigated for other probiotic attributes and displayed higher (p< 0.05) auto-aggregation and cell surface hydrophobicity values. Moreover, all seven isolates had potent antibacterial activity against pathobiont E. coli as well as significant co-aggregation capacity and enzyme activity. In vitro biosafety assessment revealed that all seven isolates were non-hemolytic, negative for mucin degradation and susceptible to most of the antibiotics. Based on the obtained findings, heatmap and principal component analysis identified four highly effective probiotic candidates confirmed by 16S rDNA sequencing as Limosilactobacillus reuteri SW23, Limosilactobacillus reuteri SW26, Limosilactobacillus reuteri SW27 and Enterococcus faecium SW28, respectively. Further studies on biosafety aspect are warranted for the application of these strains in animal as potential probiotics.HIGHLIGHTSL. reuteri SW23, L. reuteri SW26, L. reuteri SW28 and Enterococcus faecium SW28 were successfully isolated and identified from indigenous calves' feces.These microbes were characterized for potential probiotics attributes.Heatmap analysis and principal component analysis (PCA) was used along with probiotic attributes to select highly effective probiotic candidates.

Techno-functional characterization of fecal lactobacilli isolates of Bos indicus calves for probiotic properties.

In this study, 105 bacterial colonies were isolated from the feces of newborn healthy Bos indicus calves and 37 isolates were confirmed using morphological, biochemical tests, and genus-specific PCR as lactobacilli. 11 isolates were then short-listed for in vitro probiotic testing based on their ability to dwell under acid and bile stress. Species-level identification using 16S rRNA gene sequencing revealed that they were Ligilactobacillus salivarius. These isolates flourished in 0.4% phenol, depicting resistance in adverse conditions encountered in the gastrointestinal tract. The results of cell surface hydrophobicity were found to be 74.50% for RBL12 and 62.62% for RBL09 in hexadecane and xylene, respectively, and that of auto-aggregation was highest in RBL26 (58.92%). These isolates also produced digestive enzymes like amylase, protease, and ß-galactosidase. Further assays reiterated their antimicrobial and coaggregation potential against diarrhea-causing pathogens like Escherichia coli ATCC-25922 and Salmonella arizonae ATCC-13314. Biosafety assessment revealed that none of the tested isolates were hemolytic and mucinolytic in nature. Furthermore, the antioxidant potential of the isolates was also confirmed using 1,1­diphenyl­2­picrylhydrazyl (DPPH) and ferric ion-reducing antioxidant power (FRAP) assay. Along with efficient utilization of inulin, isolates showed promising adhesion ability to the HT-29 cell line. The current findings hence conclude that these Lactobacillus isolates can be exploited as animal probiotics for potential application in young calves to foster gut health and immunity.