Isolation of Lactic Acid Bacteria from Intestine of Freshwater Fishes and Elucidation of Probiotic Potential for Aquaculture Application.

Probiotics play significant roles in enhancing systemic immunity, improving intestinal balance and feed value, enhancing enzymatic digestion, and inhibiting pathogenic microorganisms of freshwater fish. Probiotics from an identical organism's gastrointestinal system promote effective colonization and provide greater benefits than other sources. This study aimed to evaluate the usefulness of probiotic bacteria isolated from the intestines of freshwater fishes for a dietary supplement of freshwater aquaculture. A total of 120 isolates were collected from freshwater fishes of Channa striata, Puntius filamentosus, Oreochromis mossambicus, Cirrhinus mrigala, and Rasbora daniconius. Seven of these isolates exhibited antagonistic activity against fish pathogens: Aeromonas hydrophila, Staphylococcus epidermidis, Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Pseudomonas aeruginosa. Using 16S rRNA gene sequencing analysis, the isolates were identified as Enterococcus sp., Lactococcus lactis, Weissella cibaria, and Limosilactobacillus fermentum. Of these tolerates, L. fermentum URLP18 isolated from C. mrigala exhibited high tolerance to low acidic (pH 2.0) and high bile salt (2%) concentrations, exhibiting a significant hydrophobicity and extracellular enzyme secretions like amylase, protease, and lipase. In vitro evaluations on intestinal mucus indicate that L. fermentum URLP18 have strong adherence capacity, and its survival rate increased after being administered to Artemia nauplii. The results suggest that L. fermentum URLP18 has high probiotic potential and is an effective dietary supplement for freshwater aquaculture.

Identification and characterization of virulent Aeromonas hydrophila Ah17 from infected Channa striata in river Cauvery and in vitro evaluation of shrimp chitosan.

Aeromonas hydrophila, an inhabitant in the aquatic ecosystem is considered as an important foodborne bacterial zoonotic pathogen in aquaculture. The present study aimed to identify virulent A. hydrophila from naturally infected Channa striata in river Cauvery and in vitro evaluation of shrimp chitosan. Rimler Shotts (RS) and blood agar medium identified the presence of pathogenic Aeromonas sp. from the infected C. striata. A. hydrophila Ah17 was identified using 16S rRNA nucleotide sequence. Extracellular enzymes such as amylase, lipase, and protease were screened in A. hydrophila Ah17. Antibiotic susceptibility tests showed A. hydrophila Ah17 was highly resistant against ß-lactam, glycopeptide, macrolides, phosphonic, fucidin, and oxazolidinone classes of antibiotics. Virulent genes such as hemolysin (aer and hly), heat-labile enterotoxin (act), cytotonic heat-stable enterotoxin (ast), elastase (ahyB), and lipase (lip) were identified. Growth and the viable cell population of virulent A. hydrophila Ah17 were significantly reduced in a dose-dependent manner against shrimp chitosan (CHS) from Penaeus indicus (P. indicus). Thus, the present study isolated virulent A. hydrophila Ah17 from the environmental source and characterized in vitro with shrimp chitosan.

Determination of lethal dose of Aeromonas hydrophila Ah17 strain in snake head fish Channa striata.

Aeromonas hydrophila is a major bacterial fish pathogen which causes economic losses in the aquaculture. The study determined the Letha Dose (LD50-96h) of A. hydrophila Ah17 strain (isolated from EUS infected Channa striata) in C. striata. C. striata were challenged with three different concentration of A. hydrophila Ah17 strain 1.0 × 107, 1.0 × 108, 1.0 × 109 CFU/mL. The LD50-96h values were found to be 4.1 × 108 CFU/mL. Percentage of mortality was observed as 10%, 40% and 70% in 1.0 × 107, 1.0 × 108 and 1.0 × 109 CFU/mL respectively in challenged fish. Microbial load was calculated on muscle, kidney, liver and spleen with highest load was observed in muscle and lowest in kidney. Level of liver enzymes such as Aspartate aminotransferase (AST), Alanine Aminotransferase (ALT), and Alkaline Phosphatase (ALP) were increased compared to control fish. Level of mRNA expression of antioxidant genes such as Catalase (cat), Manganese Superoxide Dismutase (MnSOD), Glutathione Peroxidase (GPx) were high in liver tissue of all treated groups than control. Clinical signs were observed after intraperitoneal treatment of Ah17 in C. striata. Clinical signs such as lesions on the site of injection, imbalanced state, changes in the movement of pectoral fins, depigmentation on the tail of caudal fin and irregular lesions on the muscle region were observed. Thus the study concluded that, the LD50-96h value of A. hydrophila Ah17 strain was 4.1 × 108 CFU/mL and exhibited potential pathogenic effect upon experimental infection in snakehead murrel C. striata.