Isolation and Characterization of Isolates of Bacillus cereus Group and their Comparative Antibiotic Susceptibility Testing against Oxytetracycline
-
Kirti Saini
,
Anil Kumar Teotia
,
Prasad Thota
,
Ajay Chandrakant Lagashetti
,
Manoj Kumar Pandey
,
Ajayendra Kumar Keshari
,
Sanjay Mendiratta
,
Meenakshi Dahiya
,
Vivekanandan Kalaiselvan
Abstract
Background: Bacillus cereus is a very important organism widely used for microbiological assays of pharmaceutical products especially in antibiotic susceptibility testing. The increasing emergence of antibiotic-resistant bacterial strains has emphasized the need for effective antibiotic susceptibility testing of pathogens for the management of patient’s health.
Methods: The present study isolated of 5 strains of B. cereus (R-01, R-02, C-01, C-03 & SS-01) from different samples collected from Ghaziabad, Uttar Pradesh, India. Pure cultures were then characterized based on microscopy, cultural and biochemical characteristics and also compared with standard reference strain B. cereus MTCC 430. Further, all the 5 isolates were subjected to MALDI-TOF analysis to confirm their identity. Finally, antimicrobial susceptibility of all the indigenous strains was tested in comparison with B. cereus MTCC 430 using the agar well diffusion method at various concentrations of Oxytetracycline (1.6, 2.0, 2.5, 3.12, 3.91, 4.0, 8.0, 12.0, & 16.0 µg/mL).
Results: Morphological, cultural and biochemical characteristics confirmed the identity of all the bacterial isolates as B. cereus which was further authenticated by MALDI-TOF analysis. The antibiotic susceptibility tests revealed that out of 5 indigenous isolates, R-01 and R-02 were found to be susceptible at all tested concentrations of Oxytetracycline; whereas C-01, C-03 and SS-01 were found to be resistant at a concentration 2.5 µg/mL and below.
Conclusion: The antibiotics susceptibility test results displayed sensitivity profile of all 05 indigenous isolates of B. cereus against the Oxytetracycline and revealed that 02 isolates (R-01 and R-02) were more sensitive to Oxytetracycline compared to others.
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24. Bhutia MO, Thapa N, Tamang JP. Molecular characterization of bacteria, detection of enterotoxin genes, and screening of antibiotic susceptibility patterns in traditionally processed meat products of Sikkim, India. Front Microbiol 2021; 11:599606.
25. MacWilliams MP. Citrate test protocol. Am Soc Microbiol 2009; 1-7.
26. Harish K, Jagadeesh Babu A, Madhava Rao T, et al. Isolation and identification of cellulose producing Bacillus cereus from soil samples from Tirupati, India. Pharma Innov J 2021; SP10(11):1993-97.
27. Kumar A, Goura MRS. 2025. Experiment no. 4: biochemical tests for bacterial identification. In: Shakya, P., Garg, A., Bisen, A., Singh, G. (Eds.), A Manual on Veterinary Bacteriology, Virology and Mycology. Salabad, Danpur, Buland Shahar, pp. 71-95.
28. Garcia LS. 2007. Clinical Microbiology Procedures Handbook, 2nd Edition Updates, Volume 1. American Society for Microbiology,
ASM Press, Washington, D.C.
29. Salam MA, Al-Amin MY, Salam MT, et al. Antimicrobial resistance: a growing serious threat for global public health. Healthcare 2023; 11(13):1946.
30. Lopez AC, De Ortuzar RV, Alippi AM. Tetracycline and Oxytetracycline resistance determinants detected in Bacillus cereus strains isolated from honey samples. Rev Argent Microbiol 2008; 40(4):231-7.
31. Mohammadi B, Gorkina N, Pérez-Reyes ME, et al. Profiling toxin genes and antibiotic resistance in Bacillus cereus isolated from pre-launch spacecraft. Front Microbiol 2023; 14:1231726.
32. Khan ZA, Siddiqui MF, Park S. Current and emerging methods of antibiotic susceptibility testing. Diagnostics 2019; 9(2):49.
33. Fuchs E, Raab C, Brugger K, et al. Performance testing of Bacillus cereus chromogenic agar media for improved detection in milk and other food samples. Foods 2022; 11(3):288.
34. Solanki KS, Parmar BC, Brahmbhatt MN, et al. Cultural and biochemical characterization of B. cereus isolates and multidrug resistant detection of B. cereus isolates collected from various chicken shops of market in and around Anand, Gujarat, India. Int J Curr Microbiol App Sci 2019; 8(3):910-5.
35. Kalhoro DH, Rahu N, Kalhoro MS, et al. Antimicrobial sensitivity of bacterial species isolated from sub-clinical mastitis in sheep. Science International (Lahore) 2016; 28(5):4793-8.
2. Nemati M, Hamidi A, Dizaj SM, et al. An overview on novel microbial determination methods in pharmaceutical and food quality control. Advanced Pharmaceutical Bulletin 2016; 6(3):301-08.
3. Sandle T., 2015. Pharmaceutical Microbiology: Essentials for Quality Assurance and Quality Control. Woodhead Publishing, Amsterdam.
4. Al-Bayati FA, Al-Mola HF. Antibacterial and antifungal activities of different parts of Tribulus terrestris L. growing in Iraq. J Zhejiang Univ Sci B 2008; 9(2):154-9.
5. Indian Pharmacopoeia Commission, Ministry of Health and Family Welfare, Government of India, 2022. Indian Pharmacopoeia 2022, Volume 1, p. 1399, Indian Pharmacopoeia Commission, Ghaziabad, Uttar Pradesh, India.
6. Kaya PH, Kaya B, Tuncel NB, et al. Fermentation of sainfoin seed flour with Saccharomyces boulardii: effects on total dietary fiber, anti-nutrients, antimicrobial activity, and bioaccessibility of bioactive compounds. Microorganisms 2025; 13(6):1421.
7. Ang ST, Kim TH, Cheesman MJ, et al. Antibacterial and synergistic effects of Terminalia citrina leaf extracts against gastrointestinal pathogens: insights from metabolomic analysis. Antibiotics 2025; 14(6):593.
8. Drobniewski FA. Bacillus cereus and related species. Clinl Microbiol Reviews 1993; 6(4):324-38.
9. Li C, Yuan X, Li N, et al. Isolation and characterization of Bacillus cereus phage vB_BceP-DLc1 reveals the largest member of the Φ29-like phages. Microorganisms 2020; 8(11): 1750.
10. Stenfors Arnesen LP, Fagerlund A, Granum PE. From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiol Rev 2008; 32(4):579-606
11. Chaves JQ, Pires ES, Vivoni AM. Genetic diversity, antimicrobial resistance and toxigenic profiles of Bacillus cereus isolated from food in Brazil over three decades. Int J Food Microbiol 2011; 147(1):12-6.
12. Tirloni E, Stella S, Celandroni F, et al. Bacillus cereus in dairy products and production plants. Foods 2022; 11(17):2572.
13. Celandroni F, Vecchione A, Cara A, et al. Identification of Bacillusspecies: Implication on the quality of probiotic formulations. PlosOne 2019; 14(5):e0217021.
14. Duc LH, Hong HA, Barbosa TM, et al. Characterization of Bacillus probiotics available for human use. Appl Environ Microbiol 2004; 70(4):2161-71.
15. Cutting SM. Bacillus probiotics. Food Microbiol 2011; 28(2):214-20.
16. Kulkova I, Dobrzyński J, Kowalczyk P, et al. Plant growth promotion using Bacillus cereus. Int J Mol Sci 2023; 24(11):9759.
17. Fiedler G, Schneider C, Igbinosa EO, et al. Antibiotics resistance and toxin profiles of Bacillus cereus-group isolates from fresh vegetables from German retail markets. BMC Microbiol 2019; 19(1):250.
18. Pena-Miller R, Laehnemann D, Jansen G, et al. When the most potent combination of antibiotics selects for the greatest bacterial load: the smilefrown transition. PLoS Biology 2013; 11(4):e1001540.
19. Onyankouang IS, Nieko NP, Morabandza CJ, et al. Antibiotics resistance profile of Bacillus cereus strains isolated from soil and pepper in Brazzaville. J Biosci Med 2022; 10(9):30-40.
20. Abdelaziz MN, Zayda MG, Maung AT, et al. Genetic characterization, antibiotic resistance, and virulence genes profiling of Bacillus cereus strains from various foods in Japan. Antibiotics 2024; 13(8):774.
21. Mahbubani S, Sangeetha Vani G. Isolation and
identification of Bacillus cereus from cooked rice at refrigerated & unrefrigerated conditions. J Emerg Tech Innov Res 2024; 11(10):C752-C762.
22. Adetitun DO, Tomilayo RB, Oguntoye MB, et al. Biodegradation of hydrocarbons by Bacillus cereus isolated from indoor and outdoor air of selected hospitals in Ilorin, Kwara State, Nigeria. J Appl Sci Env Manage 2020; 7: 24(6):985-9.
23. Rasool U, Ahmad A, Badroo GA, et al. Isolation and identification of Bacillus cereus from fish and their handlers from Jammu, India. Int J Curr Microbiol App Sci 2017; 6(8):441-7.
24. Bhutia MO, Thapa N, Tamang JP. Molecular characterization of bacteria, detection of enterotoxin genes, and screening of antibiotic susceptibility patterns in traditionally processed meat products of Sikkim, India. Front Microbiol 2021; 11:599606.
25. MacWilliams MP. Citrate test protocol. Am Soc Microbiol 2009; 1-7.
26. Harish K, Jagadeesh Babu A, Madhava Rao T, et al. Isolation and identification of cellulose producing Bacillus cereus from soil samples from Tirupati, India. Pharma Innov J 2021; SP10(11):1993-97.
27. Kumar A, Goura MRS. 2025. Experiment no. 4: biochemical tests for bacterial identification. In: Shakya, P., Garg, A., Bisen, A., Singh, G. (Eds.), A Manual on Veterinary Bacteriology, Virology and Mycology. Salabad, Danpur, Buland Shahar, pp. 71-95.
28. Garcia LS. 2007. Clinical Microbiology Procedures Handbook, 2nd Edition Updates, Volume 1. American Society for Microbiology,
ASM Press, Washington, D.C.
29. Salam MA, Al-Amin MY, Salam MT, et al. Antimicrobial resistance: a growing serious threat for global public health. Healthcare 2023; 11(13):1946.
30. Lopez AC, De Ortuzar RV, Alippi AM. Tetracycline and Oxytetracycline resistance determinants detected in Bacillus cereus strains isolated from honey samples. Rev Argent Microbiol 2008; 40(4):231-7.
31. Mohammadi B, Gorkina N, Pérez-Reyes ME, et al. Profiling toxin genes and antibiotic resistance in Bacillus cereus isolated from pre-launch spacecraft. Front Microbiol 2023; 14:1231726.
32. Khan ZA, Siddiqui MF, Park S. Current and emerging methods of antibiotic susceptibility testing. Diagnostics 2019; 9(2):49.
33. Fuchs E, Raab C, Brugger K, et al. Performance testing of Bacillus cereus chromogenic agar media for improved detection in milk and other food samples. Foods 2022; 11(3):288.
34. Solanki KS, Parmar BC, Brahmbhatt MN, et al. Cultural and biochemical characterization of B. cereus isolates and multidrug resistant detection of B. cereus isolates collected from various chicken shops of market in and around Anand, Gujarat, India. Int J Curr Microbiol App Sci 2019; 8(3):910-5.
35. Kalhoro DH, Rahu N, Kalhoro MS, et al. Antimicrobial sensitivity of bacterial species isolated from sub-clinical mastitis in sheep. Science International (Lahore) 2016; 28(5):4793-8.
| Files | ||
| Issue | Vol 14 No 1 (2026) | |
| Section | Original Articles | |
| Keywords | ||
| Antibiotic resistance Bacillus cereus Characterization Oxytetracycline Susceptibility. | ||
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How to Cite
1.
Saini K, Teotia A, Thota P, Lagashetti A, Pandey M, Keshari A, Mendiratta S, Dahiya M, Kalaiselvan V. Isolation and Characterization of Isolates of Bacillus cereus Group and their Comparative Antibiotic Susceptibility Testing against Oxytetracycline. J Med Bacteriol. 2026;14(1):11-23.
