Selection and characterization of probiotic lactic acid bacteria and its impact on growth, nutrient digestibility, health and antioxidant status in weaned piglets

• Published in: PloS one 2018-03, Vol.13 (3), p.e0192978-e0192978
• Main Authors: Dowarah, Runjun, Verma, Ashok Kumar, Agarwal, Neeta, Singh, Putan, Singh, Bhoj Raj
• Format: Article
• Language: English
• Subjects: Acids; Animal Nutritional Physiological Phenomena; Animals; Anti-Bacterial Agents - pharmacology; Antibiotics; Antimicrobial agents; Antioxidants; Antioxidants (Nutrients); Antioxidants - metabolism; Azithromycin; Bacilli; Bacteria; Bacterial infections; Bile salts; Biology and Life Sciences; Body weight; Body Weight - physiology; Carbohydrates; Catalase; Cell culture; Cell surface; Cephalothin; Chloramphenicol; Ciprofloxacin; Citric acid; Clindamycin; Cotrimoxazole; Diet; Dietary supplements; Digestion - physiology; Drug resistance; E coli; Ecology and Environmental Sciences; Epithelium; Erythrocytes; Erythromycin; Feces; Feces - microbiology; Feeds; Fermented food; Food and nutrition; Glycerol; Health aspects; Hogs; Host specificity; Hydrophobicity; Identification and classification; Intestine; Lactic acid; Lactobacillales - classification; Lactobacillales - genetics; Lactobacillales - physiology; Lactobacillus; Lactobacillus acidophilus; Lipase; Livestock; Medicine and Health Sciences; Microbial Sensitivity Tests; Nutrient status; Nutrient utilization; Nutrients; Nutrition research; Pediococcus acidilactici; Penicillin; pH effects; Phylogeny; Physiological aspects; Phytase; Probiotics; Probiotics - administration & dosage; RNA, Ribosomal, 16S - genetics; Salmonella; Salts; Species Specificity; Superoxide dismutase; Swine; Toluene; Vancomycin; Weaning
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0192978

The present study was aimed to develop an effective probiotic lactic acid bacteria (LAB) from piglet feces and in vitro characterization of probiotic properties. To confirm host-species specificity of probiotics, the efficacy of isolated LAB on growth, nutrient utilization, health and antioxidant status was observed in early weaned piglets. A total of 30 LAB were isolated from feces of five healthy piglets (28d old). All isolates were Gram positive, cocco-bacilli and catalase negative. Out of thirty LAB isolates, twenty were shortlisted on the basis of their tolerance to pH (3.0, 4.0, 7.0 and 8.0) and bile salts (0.075, 0.15, 0.3 and 1.0%). Whereas, fourteen isolates were selected for further in vitro probiotic characterization due higher (P45 percent). These isolates fermented twenty-seven different carbohydrates but were negative for ONPG, citrate and malonate. Also enabled to synthesize amylase, protease, lipase and phytase. They were sensitive to penicillin, azithromycin, lincomycin, clindamycin, erythromycin, cephalothin and chloramphenicol and resistant to ciprofloxacin, ofloxacin, gatifloxacin, vancomycin and co-trimoxazole. Except three isolates, all showed antagonistic activity (>60% co-culture activity) against Escherichia coli, Salmonella Enteritidis, Salmonella serotype (ser.) Typhimurium, Staphylococcus intermedius, Staph. chromogenes, Proteus mirabillis, Areomonas veonii, Bordetella bronchioseptica and Klebsialla oxytoca. The isolate Lacp28 exhibited highest tolerance to acidic pH and bile salts (up to 0.3%), phytase activity, cell surface hydrophobicity, antagonistic activity and co-culture assay (>80% growth inhibition). Host specificity of Lacp28 was further confirmed by heavy in vitro adhesion to pig intestinal epithelium cells compared to chicken. Hence, Lacp28 was selected and identified by phylogenetic analysis of 16S rRNA as Pediococcus acidilactici strain FT28 with 100% similarity (GenBank accession nos. KU837245, KU837246 and KU837247). The Pediococcus acidilactici FT28 was selected as potential probiotic candidature for in vivo efficacy in weaned pigs. Thirty-six crossbred piglets (28d) were randomly distributed into three groups (four replicates of three each) namely, basal diet without probiotics (T0) or with Lactobacillus acidophilus NCDC15 (conventional dairy-specific probiotic; T1) or Pediococcus acidilactici FT28 (swine-specific probiotic; T2). At end of the experiment, six pigl