Description
Bacillus clausii (TBC-034) is a rod-shaped, Gram-positive, motile and spore-forming bacterium that lives in the soil. It is classified as probiotic microorganism that maintains a symbiotic relationship with the host organism. It is currently being studied in respiratory infections and some gastrointestinal disorders. Bacillus clausii, has been found to produce antimicrobial substances that are active against gram positive bacteria including Staphylococcus aureus, Enterococcus faecium, and Clostridium difficile. The alkaliphilic nature of the organism has also proved it to be useful in preventing and treating various gastrointestinal disorders as an oral bacteriotherapy.
Country Of Origin
India
Manufacturing
Bacillus clausii is manufactured through a fermentation process and freeze-dried at a DSIR (Govt. of India) certified, cGMP certified plant.
Morphology
Bacillus clausii is a rod shaped, gram-positive microbe, meaning it is surrounded by a thick cell wall. The cell wall is made up of the peptidoglycan murien. B. clausii cells tend to line up into chain-like formation, observable as a long rod cell. B. clausii is an endospore producing microbe that creates ellipsoidal spored located subterminally or paracentrally in the sporangium. Spores of B. clausii are resistant to many antibiotics including erythromycin, lincomycin, cephalosporins, and cycloserine.
Identification
By 16s rRNA sequencing method; STRAIN CODE: TBC-034
FASTA Sequence:
TFPL Bacillus clausii strain TBC-034 16S ribosomal RNA gene, partial sequence
TTAGCGGCGGACGGGTGAGTAACACGTGGGCAACCTGCCCCTTAGACTGGGATAACTCCGGGAAACCGGAGCTAATACCGGATAATCCCTTTCTCCACCTGGAGAGAGGGTGAAAGATGGCTTCGGCTATCACTAAGGGATGGGCCCGCGGCGCATTAGCTAGTTGGTAAGGTAACGGCTTACCAAGGCAACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGAGGAAGGCCTTCGGGTCGTAAAGCTCTGTTGTGAGGGAAGAAGCGGTACCGTTCGAATAGGGCGGTACCTTGACGGTACCTCACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCTTCTTAAGTCTGATGTGAAATCTCGGGGCTCAACCCCGAGCGGCCATTGGAAACTGGGGAGCTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGGA
BLAST result of TBC-034 (Sequences producing significant alignments with NCBI Genomic data)
Potency
Customized
Storage
In a sealed package store in a cool, dry place, preferably at temperature not exceeding 8°C.
Gastric Juice, Intestinal Juice, and Bile Salt Tolerance
Bacillus clausii (TBC-034) exhibited excellent probiotic properties in vitro, including artificial gastric juice, intestinal juice, and bile salt tolerance
| Table: Survival rate of Bacillus clausii at artificial gastric juice, intestinal juice, and bile (%) | |||||
|---|---|---|---|---|---|
| Strain | Gastric Juice | Intestinal Juice | Gastric Juice | Intestinal Juice | Bile Salt Tolerance |
| 1h incubation | incubation 1h | incubation 2h | incubation 2h | incubation | |
| Bacillus Subtilis | 100 | 100 | 92.5 | 97.05±0.5 | 95±0.5 |
| Values were expressed as means ± SDs of three independent repetitions. | |||||
Neutral and Alkaline Protease Production
| Table: Neutral and alkaline protease production of Bacillus subtilis. | |
|---|---|
| Neutral protease (U/ mL) | Alkaline protease (U /mL) |
| 198 | 144 |
| Values were expressed as means ± SDs of three independent repetitions. | |
Antimicrobial Activity Against Potentially Pathogenic Bacteria
Bacillus clausii (TBC-034) Showed good antimicrobial activities
Figure : Antibacterial activity of Bacillus clausii against four pathogenic bacteria (Enteropathogenic Escherichia coli, Salmonella, Staphylococcus aureus and Clostridium perfringens. Bars represented the means ± SDs of three independent repetitions.
Antibiotic Resistance
Safety consideration for probiotic microbes is the presence of DNA that provides Assessment of undesirable side-effects. Protection against the effects of antibiotics. Such antibiotic resistant genes are found naturally in bacteria, but the question is whether they are resistant to antibiotics that are used in the treatment of humans and also whether they are the type of genes that can be transferred to other bacteria. If such genes are transferrable to other bacteria, then they could be taken up by bacteria in the human gut flora and subsequently passed onto pathogens creating a new type of resistant pathogen. We tested our B. clausii strains against a range of antibiotics, and strains were sensitive (not resistant) to all the antibiotics important in medical treatment, as listed in a report of the European Food Safety Authority.
- Not resistant to tetracycline
- Not resistant to aminoglycoside
- Not resistant to chloramphenicol
- Not resistant to MLS
- Not resistant to macrolide
- Not resistant to β-lactams
- Not resistant to quinupristin–dalfopristin
Toxin Production
Pathogenic genes; specifically, the organism were tested for the presence of genes responsible for the production of various toxins, and other harmful substances such as haemolysin (blood cell disruption) and lecithinase (cell membrane disruption). No such genes were found.
Figure : Fed with the diet of 0 and 106 CFU g−1 Bacillus clausii were inoculated intraperitoneally with 1.5 mL of enterohemorrhagic Escherichia coli bacterial suspension (108 CFU mL−1) after 7 weeks of feeding. (A) The survival rate of rabbits after infection with E. coli: control and fed with the diet of 106 CFU g−1 B. clausii (n = 14). (B) E. coli content of infected rabbits at 1 and 3 dpi (log10 CFU g−1). Bars represented the means ± SDs of three independent experiments. Mann–Whitney U test was conducted to examine differences. *P < 0.05.
Product Specifications
Bluk Density : ≥0.73 g/ml
Packaging : High barrier foil laminate bags.
Purity And Legal Status : Local regulations should always be consulted concerning the status of the product as legislation regarding its intended use may vary from country to country.
Microbiological Specifications
- Total Viable Cell: Count : >100Billion CFU/G
- Foreign Yeast and Mold : <100/g
- Enterococci : <100/g
- S. aureus : Absent/g
- E. coli : Absent/Gm
- Salmonella : Absent/Gm
- Coliform : Absent/Gm
Other Specifications
| Test | Specifications |
|---|---|
| Culture Description | Creamish white, lyophilized powder |
| Moisture Content | Less than 5% |
| Morphological Characteristics | Bacillus clausii is is a gram positive, rod shaped, and can form a tough, protective endospore, grow in the mesophilic temperature range. The optimal temperature is 25-35°C. |
| Bulk Density | ≥ 0.7 g/ml |
| Particle Size | ≥ 90% < 250 μm |
| Heavy metals (Pb) | ≤ 1ppm |
| Heavy metals (As) | ≤ 2ppm |
| Total Heavy Metals (Ar, Cd, Pb, Cu and Hg) | ≤ 10 ppm |
| Smell (Olfactory) | Sweet, fermented Smell |