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<p>An indigenous autochthonous bacterium, <i>Bacillus cereus</i> PX472023 isolated from the gut of <i>Lates calcarifer</i> was evaluated for its potential probiotic property. The isolated bacteria produced hydrolytic enzyme activities such as starch, esculin and casein hydrolysis along with positive result for biofilm formation. The isolate possessed desirable features like tolerance to low pH (3), high bile salt (up to 4%), pepsin (0.3%), high saline conditions (up to 3%), along with optimum hydrophobicity, auto and co aggregation activity. In addition, the isolate exhibited strong antagonistic activity against <i>L. calcarifer</i> pathogens, including <i>Photobacterium damselae</i> subsp. <i>damselae</i>, <i>Lactococcus garvieae</i>, <i>Aeromonas veronii</i> and <i>Pseudomonas putida</i>. In addition, PCR amplification revealed the presence of two probiotic marker genes (<i>LuxS</i> gene and <i>E1 β-subunit</i> of the pyruvate dehydrogenase complex gene) as well. Draft genome assembly analyses revealed genome size to be 56,16,837&#xa0;bp, with GC content of 34.97% and 5,605 protein coding sequences. Downstream analyses following the functional annotation of the genome demonstrated the presence of variety of bacteriocin gene clusters and presence of several genes in favour of probiotic potential of the isolate. Safety evaluation demonstrated that the isolate showed no haemolysis on sheep blood agar, was devoid of virulence-associated genes in its genome and did not induce mortality in the in vivo challenge study. The genomic investigation of antimicrobial resistance determinants revealed the absence of acquired antimicrobial resistance genes in the isolate, thereby alleviating primary concerns regarding horizontal transfer of resistance genes. The integration of in vitro and genomic analyses provided preliminary evidence supporting the probiotic potential of <i>Bacillus cereus</i> PX472023 in <i>L. calcarifer</i> and highlighted its prospective suitability for in vivo application in the same host under local environmental conditions.</p>

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Genomic and in Vitro Evaluation of Probiotic Traits in a Gut-Derived Bacillus cereus PX472023 from Farmed Lates Calcarifer, Asian Seabass

  • 期刊论文

  • 发表日期：

    2026年6月2日

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  • Amulya S G,
  • Gayatri Tripathi,
  • Samad Sheikh,
  • R Bharathi Rathinam

摘要

An indigenous autochthonous bacterium, Bacillus cereus PX472023 isolated from the gut of Lates calcarifer was evaluated for its potential probiotic property. The isolated bacteria produced hydrolytic enzyme activities such as starch, esculin and casein hydrolysis along with positive result for biofilm formation. The isolate possessed desirable features like tolerance to low pH (3), high bile salt (up to 4%), pepsin (0.3%), high saline conditions (up to 3%), along with optimum hydrophobicity, auto and co aggregation activity. In addition, the isolate exhibited strong antagonistic activity against L. calcarifer pathogens, including Photobacterium damselae subsp. damselae, Lactococcus garvieae, Aeromonas veronii and Pseudomonas putida. In addition, PCR amplification revealed the presence of two probiotic marker genes (LuxS gene and E1 β-subunit of the pyruvate dehydrogenase complex gene) as well. Draft genome assembly analyses revealed genome size to be 56,16,837 bp, with GC content of 34.97% and 5,605 protein coding sequences. Downstream analyses following the functional annotation of the genome demonstrated the presence of variety of bacteriocin gene clusters and presence of several genes in favour of probiotic potential of the isolate. Safety evaluation demonstrated that the isolate showed no haemolysis on sheep blood agar, was devoid of virulence-associated genes in its genome and did not induce mortality in the in vivo challenge study. The genomic investigation of antimicrobial resistance determinants revealed the absence of acquired antimicrobial resistance genes in the isolate, thereby alleviating primary concerns regarding horizontal transfer of resistance genes. The integration of in vitro and genomic analyses provided preliminary evidence supporting the probiotic potential of Bacillus cereus PX472023 in L. calcarifer and highlighted its prospective suitability for in vivo application in the same host under local environmental conditions.