Purpose: To compare the qualitative and quantitative parameters of various Bacillus clausii brands that are available in the Indian market with the International reference product, Enterogermina.

Study Centre: JSS College of Pharmacy, Ooty

Study Period: 06^th Nov 2022 to 30^th Dec 2022

Methods & Materials: Various parameters like total spore count, number of strains isolated (based on their sensitivity/ resistance pattern to antibiotics), pH & Transmittance, antibiotic resistance test, microbial purity, DNA sequencing and species identification were tested with various Bacillus Clausii brands that are available in Indian market and were compared with the International reference product, Enterogermina.

Results: Only Bifilac Clausi (Bacillus clausii - TIL 19T, TIL 21C, TIL 28S, TIL 30R) a product of Tablets (India) Limited passed all the qualitative and quantitative parameters in terms of assay, same number of strains (4 strains), taxonomical similarity, pH and suspension clarity, as well as resistance/sensitivity pattern to commonly used antibiotics.

Conclusion: Based on the results of the testing of various Bacillus clausii formulations, we conclude that only Bifilac Clausi (Bacillus clausii - TIL 19T, TIL 21C, TIL 28S, TIL 30R) showed a similarity to the International reference product, Enterogermina. Hence Bifilac Clausi was found to be probiosimilar to Enterogermina and is a viable alternative to Enterogermina to provide similar probiotic benefits.

Keywords: [Probiosimilar, Bacillus clausii formulation, Indian market, Qualitative & Quantitative parameters, 16S RNA sequence]

1. Amara AA, Shibl A. Role of Probiotics in health improvement, infection control and disease treatment and management. Saudi Pharm J SPJ. 2015 Apr;23(2):107–14.
2. Kechagia M, Basoulis D, Konstantopoulou S, Dimitriadi D, Gyftopoulou K, Skarmoutsou N, et al. Health Benefits of Probiotics: A Review. ISRN Nutr. 2013 Jan 2; 2013:481651.
3. Nagpal R, Kumar A, Kumar M, Behare PV, Jain S, Yadav H. Probiotics, their health benefits, and applications for developing healthier foods: a review. FEMS Microbiol Lett. 2012 Sep 1;334(1):1–15.
4. Kort R, Westerik N, Mariela Serrano L, Douillard FP, Gottstein W, Mukisa IM, et al. A novel consortium of Lactobacillus rhamnosus and Streptococcus thermophilus for increased access to functional fermented foods. Microb Cell Factories. 2015 Dec 8;14(1):195.
5. Merenstein D, Salminen S. Probiotics, and prebiotics. 2017. World Gastroenterology Organisation Global Guidelines.
6. Su GL, Ko CW, Bercik P, Falck-Ytter Y, Sultan S, Weizman AV, et al. AGA Clinical Practice Guidelines on the Role of Probiotics in the Management of Gastrointestinal Disorders. Gastroentero-logy. 2020 Aug 1;159(2):697–705.
7. McFarland LV, Evans CT, Goldstein EJC. Strain-Specificity and Disease-Specificity of Probiotic Efficacy: A Systematic Review and Meta-Analysis. Front Med [Internet]. 2018 [cited 2022 Dec 29];5. Available from: https://www.frontiersin.org/articles/10.3389/fmed.2018.00124
8. Szajewska H, Canani RB, Guarino A, Hojsak I, Indrio F, Kolacek S, et al. Probiotics for the prevention of antibiotic-associated diarrhea in children. J Pediatr Gastroenterol Nutr. 2016;62(3):495–506.
9. Khatri I, Tomar R, Ganesan K, Prasad G, Subramanian S. Complete genome sequence and comparative genomics of the probiotic yeast Saccharomyces boulardii. Sci Rep. 2017;7(1):1–12.
10. Cruchet S, Furnes R, Maruy A, Hebel E, Palacios J, Medina F, et al. The use of probiotics in pediatric gastroenterology: a review of the literature and recommenda-tions by Latin-American experts. Pediatr Drugs. 2015;17(3):199–216.
11. Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, et al. Expert consensus document: The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol. 2014.
12. Imperial ICVJ, Ibana JA. Addressing the Antibiotic Resistance Problem with Probiotics: Reducing the Risk of Its Double-Edged Sword Effect. Front Microbiol. 2016 Dec 15; 7:1983.
13. Ghelardi E, Celandroni F, Salvetti S, Gueye SA, Lupetti A, Senesi S. Survival, and persistence of Bacillus clausii in the human gastrointestinal tract following oral administration as spore-based probiotic formulation. J Appl Microbiol. 2015 Aug;119(2):552–9.
14. Thota P, Thota A, Medhi B, Sidhu S, Kumar P, Selvan VK, et al. Drug safety alerts of pharmacovigilance programme of India: A scope for targeted spontaneous reporting in India. Perspect Clin Res. 2018;9(1):51–5.
15. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016 Jul;33(7):1870–4.
16. Technical guidance - Update of the criteria used in the assessment of bacterial resistance to antibiotics of human or veterinary importance | EFSA [Internet]. [cited 2022 Dec 29]. Available from: https://www.efsa.europa.eu/en/efsajournal/pub/732
17. Sharma S, Arora M, Baldi A. Probiotics in India: Current status and future prospects. Pharm Aspire. 2013; 1:1–12.
18. Patrone V, Molinari P, Morelli L. Microbiological and molecular characterization of commercially available probiotics containing Bacillus clausii from India and Pakistan. Int J Food Microbiol. 2016 Nov 21; 237:92-97.
19. Ouwehand AC, Forssten S, Hibberd AA, Lyra A, Stahl B. Probiotic approach to prevent antibiotic resistance. Ann Med. 2016;48(4):246-55.

Raman Rajeshkumar

Department of Pharmaceutical Biotechnology, JSS College of Pharmacy, JSS Academy of Higher Education & Research, Ooty, The Nilgiris, Tamil Nadu, India