Isolation and characterization of bacillus cereus strains isolated from a beef pizza food poisoning incident in Hanoi

UDC: 
614.446
DOI: 
10.21668/health.risk/2025.2.08.eng
Authors: 

Pham Ngoc Ha1, Ninh Thi Hanh1, Vu Khanh Van1, Tran Le Minh2, Nguyen Thanh Trung1

Organization: 

1National Institute for Food Control, 65 Pham Than Duat St., Hanoi, Vietnam
2Hanoi Medical University, 1 Ton That Tung St., Hanoi, Vietnam

Abstract: 

Bacillus cereus is one of the global causes of food poisoning. In this study, we isolated 10 strains of B. cereus from beef pizza samples identified as the cause of food poisoning among students at two kindergartens A and B in Vietnam in 2024.

Species identification was carried out using biochemical tests and MALDI-TOF technology; antibiotic resistance profile was constructed according to the M45 CLSI guidelines, and examined the presence of cytK, bceT, hbl (hblA, hblC, and hblD) and nhe (nheA, nheB, and nheC) toxin genes among these isolated strains. The antibiotic resistance testing results showed that isolated B. cereus strains were significantly resistant to several strong antibiotics, including penicillin (100 %), vancomycin (100 %), strepto-mycin (90 %), tetracycline (80 %), ampicillin (70 %), and erythomycin (70 %). In addition, 100 % of B. cereus strains (10/10) in the beef pizza sample were positive for the bceT toxin gene, 80 % of strains (8/10) were positive for the cytK toxin gene, and 60% of strains (6/10) were positive for the nheA and nheC toxin genes, and negative for the NRPS emetic toxin gene. Our study contributes to the antibiotic resistance database for B. cereus associated with food poisoning in Vietnam and provides a valuable resource for developing reference materials aimed at the rapid diagnosis of food poisoning caused by diarrhea B. cereus type.

Keywords: 
Bacillus cereus, food poisoning, antibiotic resistance, M45 CLSI, bceT, cytK, hbl, nhe
Pham Ngoc Ha, Ninh Thi Hanh, Vu Khanh Van, Tran Le Minh, Nguyen Thanh Trung. Isolation and characterization of bacillus cereus strains isolated from a beef pizza food poisoning incident in Hanoi. Health Risk Analysis, 2025, no. 2, pp. 98–106. DOI: 10.21668/health.risk/2025.2.08.eng
References: 
  1. McDowell R.H., Sands E.M., Friedman H. Bacillus Cereus. StatPearls. Treasure Island (FL), StatPearls Publ., 2024. Available at: http://www.ncbi.nlm.nih.gov/books/NBK459121/ (September 19, 2024).
  2. Duan S., Yu Y., Guo Y., Lu D., Li N., Liu Z., Liang J., Jiang Y. [et al.]. Epidemiological Evaluation of Bacillus cereus-Induced Foodborne Outbreaks – China, 2010–2020. China CDC Wkly, 2023, vol. 5, no. 33, pp. 737–741. DOI: 10.46234/ccdcw2023.140
  3. Bottone E.J. Bacillus cereus, a Volatile Human Pathogen. Clin. Microbiol. Rev., 2010, vol. 23, no. 2, pp. 382–398. DOI: 10.1128/CMR.00073-09
  4. Etikala A., Thamburaj S., Johnson A.M., Sarma C., Mummaleti G., Kalakandan S.K. Incidence, toxin gene profile, antibiotic resistance and antibacterial activity of Allium parvum and Allium cepa extracts on Bacillus cereus isolated from fermented millet-based food. LWT, 2022, vol. 160, no. 11, pp. 113314. DOI: 10.1016/j.lwt.2022.113314
  5. Jensen L.B., Baloda S., Boye M., Aarestrup F.M. Antimicrobial resistance among Pseudomonas spp. and the Bacillus cereus group isolated from Danish agricultural soil. Environ. Int., 2001, vol. 26, no. 7–8, pp. 581–587. DOI: 10.1016/s0160-4120(01)00045-9
  6. Dietrich R., Jessberger N., Ehling-Schulz M., Märtlbauer E., Granum P.E. The Food Poisoning Toxins of Bacillus ce-reus. Toxins (Basel), 2021, vol. 13, no. 2, pp. 98. DOI: 10.3390/toxins13020098
  7. Ramm F., Stech M., Zemella A., Frentzel H., Kubick S. The Pore-Forming Hemolysin BL Enterotoxin from Bacillus cereus: Subunit Interactions in Cell-Free Systems. Toxins (Basel), 2021, vol. 13, no. 11, pp. 807. DOI: 10.3390/toxins13110807
  8. Zhao Y., Sun L. Bacillus cereus cytotoxin K triggers gasdermin D-dependent pyroptosis. Cell Death Discov., 2022, vol. 8, no. 1, pp. 305. DOI: 10.1038/s41420-022-01091-5
  9. Choma C., Granum P.E. The enterotoxin T (BcET) from Bacillus cereus can probably not contribute to food poisoning. FEMS Microbiol. Lett., 2002, vol. 217, no. 1, pp. 115–119. DOI: 10.1111/j.1574-6968.2002.tb11464.x
  10. Hansen B.M., Høiby P.E., Jensen G.B., Hendriksen N.B. The Bacillus cereus bceT enterotoxin sequence reappraised. FEMS Microbiol. Lett., 2003, vol. 223, no. 1, pp. 21–24. DOI: 10.1016/S0378-1097(03)00249-0
  11. Marxen S., Stark T.D., Rütschle A., Lücking G., Frenzel E., Scherer S., Ehling-Schulz M., Hofmann T. Depsipeptide Intermediates Interrogate Proposed Biosynthesis of Cereulide, the Emetic Toxin of Bacillus cereus. Sci. Rep., 2015, vol. 5, pp. 10637. DOI: 10.1038/srep10637
  12. Ehling-Schulz M., Fricker M., Scherer S. Identification of emetic toxin producing Bacillus cereus strains by a novel molecular assay. FEMS Microbiol. Lett., 2004, vol. 232, no. 2, pp. 189–195. DOI: 10.1016/S0378-1097(04)00066-7
  13. Cở sở dữ liệu nhiệm vụ KHCN – Đánh giá khả năng phát hiện trực tiếp gen độc tố của Bacillus Cereus trong một số thực phẩm có nguồn gốc từ gạo bằng kỹ thuật Multiplex PCR. BỘ KHOA HỌC VÀ CÔNG NGHỆ HỆ THỐNG THÔNG TIN KHOA HỌC VÀ CÔNG NGHỆ. Available at: https://sti.vista.gov.vn/tw/Pages/tai-lieu-khcn.aspx?ItemID=199450&Type_...... (October 08, 2024) (in Vietnamese).
  14. Saeed B.M.S., Abbas B.A., Al-Jadaan S.A.N. Detection of Bacillus cereus genes responsible for diarrheal and emetic toxins. J. Phys. Conf. Ser., 2021, vol. 1879, no. 2, pp. 022034. DOI: 10.1088/1742-6596/1879/2/022034
  15. National Advisory Committee On Microbiological Criteria For Foods. Response to Questions Posed by the Department of Defense Regarding Microbiological Criteria as Indicators of Process Control or Insanitary Conditions. J. Food Prot., 2018, vol. 81, no. 1, pp. 115–141. DOI: 10.4315/0362-028X.JFP-17-294
  16. Abdelaziz M.N.S., Zayda M.G., Maung A.T., El-Telbany M., Mohammadi T.N., Lwin S.Z.C., Linn K.Z., Wang C. [et al.]. Genetic Characterization, Antibiotic Resistance, and Virulence Genes Profiling of Bacillus cereus Strains from Various Foods in Japan. Antibiotics (Basel), 2024, vol. 13, no. 8, pp. 774. DOI: 10.3390/antibiotics13080774
  17. Nakayama T., Yamaguchi T., Jinnai M., Yamamoto S., Li H.T., Ngo P.T., Tran D.N.M., Nguyen O.T.H. [et al.]. Un-targeted Phylogenetic Group III of Multi-drug-Resistant Bacillus cereus Isolated Using Fraser Medium from Retail Chickens in Ho Chi Minh City. Curr. Microbiol., 2021, vol. 78, no. 8, pp. 3115–3123. DOI: 10.1007/s00284-021-02562-1
  18. Algammal A.M., Alfifi K.J., Mabrok M., Alatawy M., Abdel-Moneam D.A., Alghamdi S., Azab M.M., Ibrahim R.A. [et al.]. Newly Emerging MDR B. cereus in Mugil seheli as the First Report Commonly Harbor nhe, hbl, cytK, and pc-plc Viru-lence Genes and bla1, bla2, tetA, and ermA Resistance Genes. Infect. Drug Resist., 2022, vol. 15, pp. 2167–2185. DOI: 10.2147/IDR.S365254
  19. Jessberger N., Dietrich R., Granum P.E., Märtlbauer E. The Bacillus cereus Food Infection as Multifactorial Process. Toxins, 2020, vol. 12, no. 11, pp. 701. DOI: 10.3390/toxins12110701
  20. Bağcıoğlu M., Fricker M., Johler S., Ehling-Schulz M. Detection and Identification of Bacillus cereus, Bacillus cyto-toxicus, Bacillus thuringiensis, Bacillus mycoides and Bacillus weihenstephanensis via Machine Learning Based FTIR Spectros-copy. Front. Microbiol., 2019, vol. 10. DOI: 10.3389/fmicb.2019.00902
Received: 
13.12.2024
Approved: 
25.04.2025
Accepted for publication: 
14.06.2025

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