Evaluating the probiotic potential and adhesion characteristics of Bacillus spp. isolated from the intestine of Rhynchocypris lagowskii Dybowski

Probiotics have been widely used in aquaculture worldwide especially in China as antibiotics have been banned in aquaculture. In the present study, 27 potential strains were isolated from Rhynchocypris lagowskii Dybowski intestinal tract towards the assessment of their probiotic potential. Two potential probiotics were finally screened from the 27 candidate strains according to the results of enzyme-producing ability, in vitro against pathogens ability and antibiotic sensitivity test. The 2 potential probiotics were identified and confirmed on the basis of their colony morphology, biochemical characteristics, and 16S rRNA sequencing. The probiotic strains, LSG2-3–2 and LSG3-6, were identified to be Bacillus methylotrophicus and B. tequilensis, respectively. Further studies showed that LSG2-3–2 and LSG3-6 had excellent tolerance to high temperature (80℃), low pH (3–5), bile salts (0.3%), intestinal juice (10%), and gastric juice (0.5%). The adhesion rates of LSG2-3–2 and LSG3-6 in the in vitro intestinal mucosal adhesion model were 17.74% and 24.04%, respectively. Analysis of their bacteria surface adhesive proteins revealed that the lectins on the bacterial surface of LSG2-3–2 and LSG3-6 were mainly protein and glycoprotein properties, respectively. The adhesion receptor components in the mucus proteins of the two strains were all protein properties. The results of the inhibitory adhesion test indicated that LSG3-6 had a higher inhibitory effect on Aeromonas hydrophila and LSG2-3–2 had a better inhibitory effect on A. veronii. The biosafety assay confirmed that the isolates were not pathogenic to the host fish. Based on the presently observed characteristic features, it can be concluded that LSG2-3–2 (B. methylotrophicus) and LSG3-6 (B. tequilensis) retrieved from the digestive tract of R. lagowskii can be used for the healthy breeding and disease prevention of R. lagowskii.