Dataset Information

Preliminary in vitro assessment of probiotic properties of Bacillus subtilis GM1, a spore forming bacteria isolated from goat milk.

ABSTRACT:

Background

Species of the Bacillus genus have a long history of use in biotechnology. Some Bacillus strains have recently been identified for food applications and industrial as safe bacteria, which mostly have been recognized as probiotic strains.

Aims

The primary purpose of the current study was to evaluate the probiotic characteristics of Bacillus subtilis strains isolated and identified from the goat milk samples.

Methods

After sampling from 40 goat milk and cultivation, suspected colonies were subjected to biochemical and molecular identification. Then, the confirmed isolate was assessed for in vitro probiotic tests, including hemolysis and lecithinase properties, bile salt, acid, and artificial gastric juice resistance, antioxidant activity, antibiotics susceptibility, enterotoxin genes detection, and attachment capacity to the HT-29 cells.

Results

Among 11 suspected isolates evaluated, only one isolate was identified as B. subtilis. In vitro tests for this strain showed similar results to other probiotic strains. The B. subtilis strain was susceptible to various antibiotics. The enterotoxin genes were not detected based on PCR assay. Concerning its probiotic characteristics assessment, especially tolerance to bile salts and acidic conditions, the Bacillus strain could have the potential to consider as a probiotic.

Conclusion

Goat milk can be recommended as a source of Bacillus isolates. Also, the isolated strain showed high adaptability to the gastrointestinal environment, relatively equal percentages of adhesion properties, and some safety aspects, having the potential to be considered as an appropriate probiotic.

SUBMITTER: Daneshazari R

PROVIDER: S-EPMC10291520 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Preliminary <i>in vitro</i> assessment of probiotic properties of <i>Bacillus subtilis</i> GM1, a spore forming bacteria isolated from goat milk.

Daneshazari R R Rabbani Khorasgani M M Hosseini-Abari A A

Iranian journal of veterinary research 20230101 1

<h4>Background</h4>Species of the <i>Bacillus</i> genus have a long history of use in biotechnology. Some <i>Bacillus</i> strains have recently been identified for food applications and industrial as safe bacteria, which mostly have been recognized as probiotic strains.<h4>Aims</h4>The primary purpose of the current study was to evaluate the probiotic characteristics of <i>Bacillus subtilis</i> strains isolated and identified from the goat milk samples.<h4>Methods</h4>After sampling from 40 goat ...[more]

PMID: 37378382

Similar Datasets

Project description:BackgroundClonorchiasis, a food-borne zoonosis, is caused by Clonorchis sinensis. The intestinal tract and bile ducts are crucial places for C. sinensis metacercariae to develop into adult worms. The endospore of Bacillus subtilis is an ideal oral immunization vehicle for delivery of heterologous antigens to intestine. Cysteine protease of C. sinensis (CsCP) is an endogenous key component in the excystment of metacercariae and other physiological or pathological processes.MethodsWe constructed a fusion gene of CotC (a coat protein)-CsCP and obtained B. subtilis spores with recombinant plasmid of pEB03-CotC-CsCP (B.s-CotC-CsCP). CotC-CsCP expressed on spores' surface was detected by Western blotting and immunofluorescence. Immunological characteristics of recombinant spore coat protein were evaluated in a mouse model. The levels of CsCP-specific antibodies were detected by ELISA. Effects of recombinant spores on mouse intestine were evaluated by histological staining. The activities of biochemical enzymes in serum were assayed by microplate. Liver sections of infected mice were evaluated by Ishak score after Masson's trichrome.ResultsThe B.s-CotC-CsCP spores displayed CsCP on their coat. Specific IgG and isotypes were significantly induced by coat proteins of B.s-CotC-CsCP spores after subcutaneous immunization. IgA levels in intestinal mucus and bile of B.s-CotC-CsCP orally treated mice significantly increased. Additionally, more IgA-secreting cells were observed in enteraden and lamina propria regions of the mouse jejunum, and an increased amount of acidic mucins in intestines were also observed. There were no significant differences in enzyme levels of serum among groups. No inflammatory injury was observed in the intestinal tissues of each group. The degree of liver fibrosis was significantly reduced after oral immunization with B.s-CotC-CsCP spores.ConclusionsBacillus subtilis spores maintained the original excellent immunogenicity of CsCP expressed on their surface. Both local and systemic specific immune responses were elicited by oral administration of B.s-CotC-CsCP spores. The spores effectively promoted intestinal health by inducing secretion of acidic mucins, with no other side effects to the liver or intestine. Oral administration of spores expressing CsCP could provide effective protection against C. sinensis. This study may be a cornerstone for development of antiparasitic agents or vaccines against clonorchiasis based on B. subtilis spore expressing CsCP on the surface.

Project description:In the current study, we screened 46 isolates of lactic acid bacteria (LAB) derived from goat milk for bacteriocin producers that can inhibit common foodborne pathogens (Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus). The following three strains that showed antimicrobial activity against all indicators were identified: Enterococcus faecalis DH9003 and DH9012, and Lactococcus lactis DH9011. Their antimicrobial products exhibited typical bacteriocin characteristics, such as heat stability and proteinase nature. The bacteriostatic activity of concentrated bacteriocins produced by these LAB was observed at low concentrations (half-minimum inhibitory concentration [MIC50] and 4MIC50), whereas complete inhibition activity against Listeria monocytogenes was detected at high concentrations (16MIC50) of the two Enterococcus faecalis strains (DH9003 and DH9012). Furthermore, the probiotic potentials of the three strains were investigated and described. The results revealed that none of the strains had hemolytic activity, whereas all: were sensitive to ampicillin (50 mg/mL) and streptomycin sulfate (100 mg/mL); were resistant to bile, artificial simulated intestinal tract, and gastric juice at different pH levels (2.5, 3.0, 3.5); and had β-galactosidase activity. Furthermore, all strains exhibited an auto-aggregating phenotype, with self-aggregation ranging from 30% to 55%. DH9003 and DH9012 co-aggregated well with Listeria monocytogenes and Escherichia coli (52.6% and 63.2%, 68.5% and 57.6%, respectively), whereas DH9011 co-aggregated poorly with Listeria monocytogenes (15.6%) and did not co-aggregate with Escherichia coli. Furthermore, our results showed that all three isolates exhibited strong antibacterial activity, tolerance to bile and simulated gastrointestinal environments, adhesion capability, and safety. Finally, DH9003 was selected and used for gavage in rats. By observing the pathological characteristics of rat intestinal and liver tissue sections, DH9003 showed no harmful effects on the intestine and liver of rats, but rather resulted in a denser and longer intestinal mucosa, as well as improving the intestinal mucosa of rats. Considering their substantial prospective applications, we concluded that these three isolates are potential probiotic candidates.