Prevalence and Molecular Detection of Toxigenic Genes of Bacillus cereus in Rice-Based Street Foods in Tasikmalaya, Indonesia
Abstract
Bacillus cereus is a spore-forming foodborne pathogen frequently associated with rice-based foods and capable of causing emetic and diarrheal illnesses through the production of heat-stable and heat-labile toxins. This study investigated the prevalence, contamination levels, and toxigenic gene profiles of B. cereus in rice-based street foods. A total of 12 samples, including fried rice, chicken porridge, and nasi kuning, were collected from street vendors in Kahuripan Sub-district, Tasikmalaya, Indonesia. Presumptive B. cereus was isolated using mannitol yolk polymyxin agar (MYPA) as a selective medium and confirmed biochemically and molecularly by PCR targeting the gyrB gene. Toxigenic potential isolate was evaluated by detecting the nheA, hblD, cytK, entFM, and ces genes. Presumptive B. cereus was detected in 91.67% (11/12) of samples, while molecular confirmation identified gyrB-positive isolates in 42% (5/12) of samples, originating from fried rice and chicken porridge. No significant association was found between food type and gyrB detection (p > 0.05). Contamination levels ranged from < 1.5 × 10¹ to 2.95 × 10⁶ CFU g-1. All confirmed isolates harbored at least one toxigenic gene, with nheA being the most prevalent (80%), followed by cytK (60%), entFM (40%), ces (40%), and hblD (20%). Notably, one of the fried rice isolates carried all 5 toxin-related genes, indicating a high virulence potential. These findings demonstrate that rice-based street foods in Tasikmalaya may pose a significant public health risk due to contamination with toxigenic B. cereus. Enhanced hygiene practices, strict temperature control, and molecular surveillance are essential to reduce the risk of foodborne illness associated with street-vended rice products.
Keywords
Full Text:
PDFReferences
Albaridi, N. (2022). Risk of Bacillus cereus contamination in cooked rice. Food Science and Technology, 42, e108221. https://doi.org/10.1590/fst.108221
Apriliansyah, M., Zuhrotun, A., & Astrini, D. (2022). Bakteri utama penyebab kejadian luar biasa keracunan pangan. Jurnal Farmasi Klinik Indonesia, 11(3), 239–255. https://doi.org/10.15416/ijcp.2022.11.3.239
Arisanti, R. R., Indriani, C., & Wilopo, S. A. (2018). Kontribusi agen dan faktor penyebab kejadian luar biasa keracunan pangan di Indonesia: Kajian sistematis. Berita Kedokteran Masyarakat, 34(3), 99–106. Retrieved from https://pdfs.semanticscholar.org/3eff/b3eb0947f23b18f079eb5fb1857dc62eb75e.pdf
Arta, M. J. (2016). Identifikasi bakteri patogen dan kualitas mikrobiologi nasi kuning berbahan beras organik dan non organik dengan metode pemasakan tradisional dan modern. Universitas Katolik Soegijapranata. Retrieved from https://repository.unika.ac.id/11642/1/12.70.0037%20Maria%20Jessica%20Arta%20COVER.pdf
Asano, S.-I., Nukumizu, Y., Bando, H., Iizuka, T., & Yamamoto, T. (1997). Cloning of novel enterotoxin genes from Bacillus cereus and Bacillus thuringiensis. Applied and Environmental Microbiology, 63(3), 1054–1057. Retrieved from https://journals.asm.org/doi/pdf/10.1128/aem.63.3.1054-1057.1997
BAM. (2020). BAM Chapter 14 : Bacillus cereus (Issue October). Bacteriological Analytical Manual. U.S Food & Drug Administration. Retrieved from https://www.fda.gov/food/laboratory-methods-food/bam-chapter-14-bacillus-cereus
Bursová, Š., Haruštiaková, D., Necidová, L., Krobotová, E., Mlejnková, Z., Tkáč, M., ... & Golian, J. (2024). Evaluation of Bacillus cereus growth in cooked rice. Journal of Microbiology, Biotechnology and Food Sciences, 14(1), e10985–e10985. https://doi.org/10.55251/jmbfs.10985
Chen, J., Zhang, J., Zhan, L., Chen, H., Zhang, Z., Huang, C., & Yue, M. (2022). Prevalence and antimicrobial-resistant characterization of Bacillus cereus isolated from ready-to-eat rice products in Eastern China. Frontiers in Microbiology, 13, 964823. https://doi.org/10.3389/fmicb.2022.964823
Cruz-Facundo, I. M., Adame-Gómez, R., Castro-Alarcón, N., Toribio-Jiménez, J., Castro-Coronel, Y., Santiago-Dionisio, M. C., ... & Ramírez-Peralta, A. (2023). Enterotoxigenic profiles and submerged and interface biofilms in Bacillus cereus group isolates from foods. Revista Argentina de Microbiología, 55(3), 262–271. https://doi.org/10.1016/j.ram.2023.01.007
da Silva, N. B., Ellouze, M., Rouzeau-Szynalski, K., Johnson, N., Zwietering, M. H., & den Besten, H. M. W. (2022). Predicting B. cereus growth and cereulide production in dairy mix. International Journal of Food Microbiology, 364, 109519. https://doi.org/10.1016/j.ijfoodmicro.2021.109519
Didier, A., Dietrich, R., & Märtlbauer, E. (2016). Antibody binding studies reveal conformational flexibility of the Bacillus cereus non-hemolytic enterotoxin (Nhe) A-component. PLoS One, 11(10), e0165135. https://doi.org/10.1371/journal.pone.0165135
Dietrich, R., Jessberger, N., Ehling-Schulz, M., Märtlbauer, E., & Granum, P. E. (2021). The food poisoning toxins of Bacillus cereus. Toxins, 13(2), 98. https://doi.org/10.3390/toxins13020098
Ellouze, M., Buss Da Silva, N., Rouzeau-Szynalski, K., Coisne, L., Cantergiani, F., & Baranyi, J. (2021). Modeling Bacillus cereus growth and cereulide formation in cereal-, dairy-, meat-, vegetable-based food and culture medium. Frontiers in Microbiology, 12, 639546. https://doi.org/10.3389/fmicb.2021.639546
Fagerlund, A., Ween, O., Lund, T., Hardy, S. P., & Granum, P. E. (2004). Genetic and functional analysis of the cytK family of genes in Bacillus cereus. Microbiology, 150(8), 2689–2697. https://doi.org/10.1099/mic.0.26975-0
Fitrian, A. M. (2023). 51 Warga Leuwisari Tasikmalaya keracunan termasuk bayi 2,5 tahun, setelah makan nasi kuning acara syukuran. Kabar Priangan. Retrieved from https://kabarpriangan.pikiran-rakyat.com/kabar-priangan/pr-1486825496/51-warga-leuwisari-tasikmalaya-keracunan-termasuk-bayi-25-tahun-setelah-makan-nasi-kuning-acara-syukuran?page=all
Fuchs, E., Raab, C., Brugger, K., Ehling-Schulz, M., Wagner, M., & Stessl, B. (2022). Performance testing of Bacillus cereus chromogenic agar media for improved detection in milk and other food samples. Foods, 11(3), 288. https://doi.org/10.3390/foods11030288
Gdoura-Ben Amor, M., Siala, M., Zayani, M., Grosset, N., Smaoui, S., Messadi-Akrout, F., ... & Gdoura, R. (2018). Isolation, identification, prevalence, and genetic diversity of Bacillus cereus group bacteria from different foodstuffs in Tunisia. Frontiers in Microbiology, 9, 447. https://doi.org/10.3389/fmicb.2018.00447
Granum, P. E., O’sullivan, K., & Lund, T. (1999). The sequence of the non-haemolytic enterotoxin operon from Bacillus cereus. FEMS Microbiology Letters, 177(2), 225–229. https://doi.org/10.1111/j.1574-6968.1999.tb13736.x
Izumi, S., & Suzuki, K. (2016). A novel PCR detection method for major fish pathogenic bacteria of Vibrio anguillarum. Turkish Journal of Fisheries and Aquatic Sciences, 16(3), 571–578. https://doi.org/10.4194/1303-2712-v16_3_09
Jovanovic, J., Tretiak, S., Begyn, K., & Rajkovic, A. (2022). Detection of enterotoxigenic psychrotrophic presumptive Bacillus cereus and cereulide producers in food products and ingredients. Toxins, 14(4), 289. https://doi.org/10.3390/toxins14040289
Kabir, M. S., Hsieh, Y. H., Simpson, S., Kerdahi, K., & Sulaiman, I. M. (2017). Evaluation of two standard and two chromogenic selective media for optimal growth and enumeration of isolates of 16 unique Bacillus species. Journal of Food Protection, 80(6), 952–962. https://doi.org/10.4315/0362-028X.JFP-16-441
Kim, J. M., Forghani, F., Kim, J. B., Park, Y. B., Park, M. S., Wang, J., ... & Oh, D. H. (2012). Improved multiplex PCR assay for simultaneous detection of Bacillus cereus emetic and enterotoxic strains. Food Science and Biotechnology, 21(5), 1439–1444. https://doi.org/10.1007/s10068-012-0189-8
Komprda, T., Cwiková, O., Kumbár, V., Franke, G., Kouřil, P., Patloka, O., ... & Zouharová, A. (2025). Key factors influencing Bacillus cereus contamination in hot ready-to-eat meal delivery. Foods, 14(15), 2605. https://doi.org/10.3390/foods14152605
Kristiadi. (2020). Bertambah, korban keracunan makanan di Tasikmalaya jadi 146 orang. Media Indonesia. Retrieved from https://mediaindonesia.com/nusantara/351363/bertambah-korban-keracunan-makanan-di-tasikmalaya-jadi-146-orang
Marrollo, R. (2016). Bacillus cereus food-borne disease. The diverse faces of Bacillus cereus (pp. 61–72). Elsevier. https://doi.org/10.1016/B978-0-12-801474-5.00005-0
Martín-Miguélez, J. M., Delgado, J., Ruiz-Carrascal, J., Olegario, L. S., & González-Mohino, A. (2025). Physicochemical and sensory properties of dry-fermented sausages as influenced by the surface inoculation with non-traditional molds. Food Bioscience, 71, 107144. https://doi.org/10.1016/j.fbio.2025.107144
McDowell, R. H., Sands, E. M., & Friedman, H. (2023). Bacillus cereus. StatPearls [Internet]. StatPearls Publishing. Retrieved from https://www.ncbi.nlm.nih.gov/sites/books/NBK459121/
Meng, Q., Song, L., Chi, S., Wang, H., Li, J., Chen, Y., ... & Wang, X. (2025). Effect of environmental stress factors on the expression of virulence genes and pathogenicity of lethal Bacillus cereus of bovine origin. Frontiers in Microbiology, 16, 1519202. https://doi.org/10.3389/fmicb.2025.1519202
Mohammadi, B., Reyes, M. E. P., & Smith, S. A. (2024). Survival, growth, and toxin production of Bacillus cereus during cooking and storage of fresh rice noodles. Journal of Food Protection, 87(3), 100239. https://doi.org/10.1016/j.jfp.2024.100239
Navaneethan, Y., & Effarizah, M. E. (2023). Post-cooking growth and survival of Bacillus cereus spores in rice and their enzymatic activities leading to food spoilage potential. Foods, 12(3), 626. https://doi.org/10.3390/foods12030626
Oliveira, M., Carvalho, M., & Teixeira, P. (2023). Characterization of the toxigenic potential of Bacillus cereus sensu lato isolated from raw berries and their products. Foods, 12(21), 4021. https://doi.org/10.3390/foods12214021
Osimani, A., Aquilanti, L., & Clementi, F. (2018). Bacillus cereus foodborne outbreaks in mass catering. International Journal of Hospitality Management, 72, 145–153. https://doi.org/10.1016/j.ijhm.2018.01.013
Oyetibo, O. B., Hutman, A., & Oni, E. O. (2022). Prevalence and characterisation of Bacillus cereus in cooked rice retailed in Ilara-mokin, Nigeria. Journal of Food Safety and Hygiene, 8(3), 156–164. https://doi.org/10.18502/jfsh.v8i3.11017
Perera, M. L., & Ranasinghe, G. R. (2012). Prevalence of Bacillus cereus and associated risk factors in chinese-style fried rice available in the City of Colombo, Sri Lanka. Foodborne Pathogens and Disease, 9(2), 125–131. https://doi.org/10.1089/fpd.2011.0969
Permatasari, N. C. (2016). Identifikasi bakteri penyebab kerusakan pada nasi uduk berbahan baku beras organik dan non organik varietas IR-64. Universitas Katolik Soegijapranata. Retrieved from https://repository.unika.ac.id/11710/
Rajkovic, A., Uyttendaele, M., Vermeulen, A., Andjelkovic, M., Fitz‐James, I., In ‘t Veld, P., ... & Debevere, J. (2008). Heat resistance of Bacillus cereus emetic toxin, cereulide. Letters in Applied Microbiology, 46(5), 536–541. https://doi.org/10.1111/j.1472-765X.2008.02350.x
Reyes-Roldán, J., Cano-Ponce, M., Gaspar-Nava, L. F., Pérez-Olais, J. H., Muñoz-Barrios, S., Rodríguez-Ruíz, H. A., & Ramírez-Peralta, A. (2024). Enterotoxigenic profile, biofilm production, and antimicrobial resistance of Bacillus cereus isolated from rice-based food marketed in southern Mexico. Biotecnia, 26, 1–9. https://doi.org/10.18633/biotecnia.v26.2429
Rodrigo, D., Rosell, C. M., & Martinez, A. (2021). Risk of Bacillus cereus in relation to rice and derivatives. Foods, 10(2), 302. https://doi.org/10.3390/foods10020302
Saba, C. K. S., Antwi, M. V., & Adzitey, F. (2019). Prevalence of Bacillus cereus in ready-to-eat boiled and fried rice in the Tamale Metropolis of Ghana. Journal of Food Safety and Hygiene, 5(1), 19–23. https://doi.org/10.18502/jfsh.v5i1.3880
Sánchez-Chica, J., Correa, M. M., Aceves-Diez, A. E., & Castañeda-Sandoval, L. M. (2021). Enterotoxin gene distribution and genotypes of Bacillus cereus sensu lato isolated from cassava starch. Toxins, 13(2), 131. https://doi.org/10.3390/toxins13020131
Sandra, A., Afsah-Hejri, L., Tunung, R., Zainazor, T. T., Tang, J. Y. H., Ghazali, F. M., ... & Son, R. (2012). Bacillus cereus and Bacillus thuringiensis in ready-to-eat cooked rice in Malaysia. International Food Research Journal, 19(3), 829. Retrieved from https://www.academia.edu/79480205/Bacillus_cereus_and_Bacillus_thuringiensis_in_ready_to_eat_cooked_rice_in_Malaysia
Sastalla, I., Fattah, R., Coppage, N., Nandy, P., Crown, D., Pomerantsev, A. P., & Leppla, S. H. (2013). The Bacillus cereus Hbl and Nhe tripartite enterotoxin components assemble sequentially on the surface of target cells and are not interchangeable. PLOS ONE, 8(10), e76955. https://doi.org/10.1371/journal.pone.0076955
Statistics Indonesia. (2025). Rata-rata konsumsi perkapita seminggu menurut kelompok makanan dan minuman jadi per kabupaten/kota (satuan komoditas), 2024. Retrieved from https://www.bps.go.id/id/statistics-table/2/MjEwNyMy/rata-rata-konsumsi-perkapita-seminggu-menurut-kelompok-makanan-dan-minuman-jadi-per-kabupaten-kota.html
Talahmeh, N., Abu-Rumeileh, S., & Al-Razem, F. (2020). Development of a selective and differential media for the isolation and enumeration of Bacillus cereus from food samples. Journal of Applied Microbiology, 128(5), 1440–1447. https://doi.org/10.1111/jam.14561
Tirloni, E., Bernardi, C., Ghelardi, E., Celandroni, F., Cattaneo, P., & Stella, S. (2019). Bacillus cereus in fried rice meals: Natural occurrence, strain dependent growth and haemolysin (HBL) production. Lwt, 114, 108393. https://doi.org/10.1016/j.lwt.2019.108393
Tuipulotu, D. E., Mathur, A., Ngo, C., & Man, S. M. (2021). Bacillus cereus: Epidemiology, virulence factors, and host–pathogen interactions. Trends in Microbiology, 29(5), 458–471. https://doi.org/10.1016/j.tim.2020.09.003
Wei, S., Chelliah, R., Park, B. J., Park, J. H., Forghani, F., Park, Y. S., ... & Oh, D. H. (2018). Molecular discrimination of Bacillus cereus group species in foods (lettuce, spinach, and kimbap) using quantitative real-time PCR targeting groEL and gyrB. Microbial Pathogenesis, 115, 312–320. https://doi.org/10.1016/j.micpath.2017.12.079
Woh, P. Y., & Ng, C. (2024). Bacillus cereus in rice: A review on food poisoning, antimicrobial resistance, and control measures. Tropical Biomedicine, 41(3), 298–309. http://dx.doi.org/10.47665%2Ftb.41.3.010
Yennie, Y., Dewanti-Hariyadi, R., Kusumaningrum, H. D., & Poernomo, A. (2022). Kontaminasi Staphylococcus aureus dan Bacillus cereus pada sushi di tingkat ritel di wilayah Jabodetabek. Jurnal Pengolahan Hasil Perikanan Indonesia, 25(2), 331–344. http://dx.doi.org/10.17844/jphpi.v25i2.42066
Zhang, Z., Feng, L., Xu, H., Liu, C., Shah, N. P., & Wei, H. (2016). Detection of viable enterotoxin-producing Bacillus cereus and analysis of toxigenicity from ready-to-eat foods and infant formula milk powder by multiplex PCR. Journal of Dairy Science, 99(2), 1047–1055. https://doi.org/10.3168/jds.2015-10147
Refbacks
- There are currently no refbacks.










