The aim of this study was to assess the germination and growth of two strains of Bacillus cereus following the artificial inoculation of six selected hot dishes with spores which were then stored at temperatures of 40, 50, and 60 °C for 0.5, 1.0, 2.0, 2.5, 3.0, and 4.0 h. The water activity of the prepared meals varied between 0.967 and 0.973 and the salt content between 0.74 and 1.40%. The pH value of four dishes exceeded 6.0, but for two (tomato sauce and ratatouille) it was 4.6. The tested strain DSM 4312 showed good growth abilities and attained a population exceeding 6.0 log CFU/g within 4 h at 40 °C in foods with pH values > 6.0. The study demonstrated that a drop in food temperatures to 40 °C is risky, while no growth of B. cereus was detected within 4 h at 50 and 60 °C. The growth rate of B. cereus is conditioned not merely by environmental conditions (temperature, pH values, food composition), but also by the bacterial strain.

Department of Animal Origin Food and Gastronomic Sciences Faculty of Veterinary Hygiene and Ecology University of Veterinary Sciences Brno Palackého tř 1946 1 612 42 Brno Czech Republic

See more in PubMed

Bağcıoğlu M., Fricker M., Johler S., Ehling-Schulz M. Detection and Identification of Bacillus cereus, Bacillus cytotoxicus, Bacillus thuringiensis, Bacillus mycoides and Bacillus weihenstephanensis via Machine Learning Based FTIR Spectroscopy. Front. Microbiol. 2019;10:902. doi: 10.3389/fmicb.2019.00902. PubMed DOI PMC

Jeßberger N., Krey V.M., Rademacher C., Böhm M.E., Mohr A.K., Ehling-Schulz M., Scherer S., Märtlbauer E. From genome to toxicity: A combinatory approach highlights the complexity of enterotoxin production in Bacillus cereus. Front. Microbiol. 2015;6:560. doi: 10.3389/fmicb.2015.00560. PubMed DOI PMC

Bogdanovičová K., Kameník J., Dorotíková K., Strejček J., Křepelová S., Dušková M., Haruštiaková D. Occurrence of Foodborne Agents at Food Service Facilities in the Czech Republic. J. Food Prot. 2019;82:1096–1103. doi: 10.4315/0362-028X.JFP-18-338. PubMed DOI

Ramarao N., Tran S.L., Marin M., Vidic J. Advanced Methods for Detection of Bacillus cereus and Its Pathogenic Factors. Sensors. 2020;20:2667. doi: 10.3390/s20092667. PubMed DOI PMC

Beattie S.H., Williams A.G. Detection of Toxins. In: Robinson R.K., Batt C.A., Patel P.D., editors. Encyclopedia of Food Microbiology. 2nd ed. Academic Press; Elsevier, Ltd.; Amsterdam, The Netherlands: 2014. pp. 144–150.

Granum P.E., Lindbäck T. Bacillus cereus. In: Doyle M.P., Buchanan R.L., editors. Food Microbiology: Fundamentals and Frontiers. 4th ed. ASM Press; Washington, DC, USA: 2012. pp. 491–502.

EFSA The European Union one health 2022 zoonoses report. EFSA J. 2023;21:8442. doi: 10.2903/j.efsa.2023.8442. PubMed DOI PMC

Matthews K.R., Kniel K.E., Montville T.J. Food Microbiology: An Introduction. 4th ed. ASM Press; Washington, DC, USA: 2017. p. 597.

Food and Agriculture Organization of the United Nations . Act No. 258/2000 Coll. Act on the Protection of Public Health (Czech Republic) Food and Agriculture Organization of the United Nations; Rome, Italy: 2000. No. 258.

Czech Republic Ministry of Health . Decree No. 137/2004 Coll. On hygienic requirements for catering services and on the principles of personal and operational hygiene in epidemiologically important activities (Czech Republic) Czech Republic Ministry of Health; Prague, Czech Republic: 2004. No. 137.

FDA Food Facts. Serving Up Safe Buffets. U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition’s Food. [(accessed on 1 September 2024)];2017 :3. Available online: https://www.fda.gov/media/84739/download.

Food and Agriculture Organization of the United Nations . Decree No. 121/2023 Coll. On Food Requirements (Czech Republic) Food and Agriculture Organization of the United Nations; Rome, Italy: 2023. No. 121.

Webb M.D., Barker G.C., Goodburn K.E., Peck M.W. Risk presented to minimally processed chilled foods by psychrotrophic Bacillus cereus. Trends Food Sci. Technol. 2019;93:94–105. doi: 10.1016/j.tifs.2019.08.024. PubMed DOI PMC

Juneja V.K., Golden C.E., Mishra A., Harrison M.A., Mohr T., Silverman M. Predictive model for growth of Bacillus cereus during cooling of cooked rice. Int. J. Food Microbiol. 2019;290:49–58. doi: 10.1016/j.ijfoodmicro.2018.09.023. PubMed DOI

Kim S.H., Chung B.D. Integrated food delivery problem considering both single-order and multiple order deliveries. Comput. Ind. Eng. 2024;196:110458. doi: 10.1016/j.cie.2024.110458. DOI

Simoni M.D., Winkenbach M. Crowdsourced on-demand food delivery: An order batching and assignment algorithm. Transp. Res. Part C Emerg. Technol. 2023;149:104055. doi: 10.1016/j.trc.2023.104055. DOI

Steever Z., Karwan M., Murray C. Dynamic courier routing for a food delivery service. Comput. Oper. Res. 2019;107:173–188. doi: 10.1016/j.cor.2019.03.008. DOI

Runštuk J., Syrový F., Rusnaková S. Receptury Teplých Pokrmů®. 7th ed. Radek Runštuk-R Plus; Divec, Czech Republic: 2015. 580p. (In Czech)

Bardi C. Italian Cooking. 1st ed. McRae Publishing Ltd.; London, UK: 2008. 976p

Roux M. The Essence of French Cooking. 1st ed. Quadrille Publishing Ltd.; London, UK: 2015. 272p

Kilcullen K., Teunis A., Popova T.G., Popov S.G. Cytotoxic Potential of Bacillus cereus Strains ATCC 11778 and 14579 Against Human Lung Epithelial Cells Under Microaerobic Growth Conditions. Front. Microbiol. 2016;7:69. doi: 10.3389/fmicb.2016.00069. PubMed DOI PMC

Frentzel H., Kraemer M., Kelner-Burgos Y., Uelze L., Bodi D. Cereulide production capacities and genetic properties of 31 emetic Bacillus cereus group strains. Int. J. Food Microbiol. 2024;417:110694. doi: 10.1016/j.ijfoodmicro.2024.110694. PubMed DOI

Bursová Š., Haruštiaková D., Necidová L., Krobotová E., Mlejnková Z., Tkáč M., Stojanová K., Golian J. Evaluation of Bacillus cereus growth in cooked rice. Microbiol. Biotech. Food Sci. 2024;14:10985. doi: 10.55251/jmbfs.10985. DOI

Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Presumptive Bacillus cereus—Colony-Count Technique at 30 Degrees C—Amendment 1: Inclusion of Optional Tests. International Organization for Standardization; Geneva, Switzerland: 2004. 27p

Microbiology of the Food Chain—Horizontal Method for the enumeration of Microorganisms—Part 1: Colony Count at 30 °C by the Pour Plate Technique. International Organization for Standardization; Geneva, Switzerland: 2013. 9p

Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Beta-Glucuronidase-Positive Escherichia coli—Part 2: Colony-Count Technique at 44 Degrees C Using 5-bromo-4-chloro-3-indolyl beta-D-Glucuronide. International Organization for Standardization; Geneva, Switzerland: 2001. 8p

Yamada S., Ohashi E., Agata N., Venkateswaran K. Cloning and nucleotide sequence analysis of gyrB of Bacillus cereus, B. thuringiensis, B. mycoides, and B. anthracis and their application to the detection of B. cereus in rice. Appl. Environ. Microbiol. 1999;65:1483–1490. doi: 10.1128/AEM.65.4.1483-1490.1999. PubMed DOI PMC

Regulation Regulation (EU) no 1169/2011 of the European Parliament and of the Council of 25 October 2011 on the provision of food information to consumers, amending Regulations (EC) No 1924/2006 and (EC) No 1925/2006 of the European Parliament and of the Council, and repealing Commission Directive 87/250/EEC, Council Directive 90/496/EEC, Commission Directive 1999/10/EC, Directive 2000/13/EC of the European Parliament and of the Council, Commission Directives 2002/67/EC and 2008/5/EC and Commission Regulation (EC) No 608/2004. Off. J. Eur. Union. 2011;L 304:18.

Heo S.K., Lee J.Y., Baek S.B., Ha S.D. A Response Surface Model to Describe the Effect of Temperature and pH on the Growth of Bacillus cereus in Cooked Rice. J. Food Prot. 2009;72:1296–1300. doi: 10.4315/0362-028X-72.6.1296. PubMed DOI

Jessberger N., Dietrich R., Granum P.E., Märtlbauer E. The Bacillus cereus food infection as multifactorial process. Toxins. 2020;12:701. doi: 10.3390/toxins12110701. PubMed DOI PMC

Stenfors Arnesen L.P., Fagerlund A., Granum P.E. From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiol. Rev. 2008;32:579–606. doi: 10.1111/j.1574-6976.2008.00112.x. PubMed DOI

Rahnama H., Azari R., Yousefi M.H., Berizi E., Mazloomi S.M., Hosseinzadeh S., Derakhshan Z., Ferrante M., Conti G.O. A systematic review and meta-analysis of the prevalence of Bacillus cereus in foods. Food Control. 2023;143:109250. doi: 10.1016/j.foodcont.2022.109250. DOI

Yu S., Yu P., Wang J., Li C., Guo H., Liu C., Kong L., Yu L., Wu S., Lei T., et al. A Study on Prevalence and Characterization of Bacillus cereus in Ready-to-Eat Foods in China. Front. Microbiol. 2020;10:3043. doi: 10.3389/fmicb.2019.03043. PubMed DOI PMC

Navaneethan Y., Effarizah M.E. Prevalence, toxigenic profiles, multidrug resistance, and biofilm formation of Bacillus cereus isolated from ready-to eat cooked rice in Penang, Malaysia. Food Control. 2021;121:107553. doi: 10.1016/j.foodcont.2020.107553. DOI

Navaneethan Y., Effarizah M.E. Post-Cooking Growth and Survival of Bacillus cereus Spores in Rice and Their Enzymatic Activities Leading to Food Spoilage Potential. Foods. 2023;12:626. doi: 10.3390/foods12030626. PubMed DOI PMC

Turner N.J., Whyte R., Hudson J.A., Kaltovei S.L. Presence and growth of Bacillus cereus in dehydrated potato flakes and hot-held, ready-to-eat potato products purchased in New Zealand. J. Food Prot. 2006;69:1173–1177. doi: 10.4315/0362-028X-69.5.1173. PubMed DOI

Bozanta A., Cevik M., Kavaklioglu C., Kavuk E.M., Tosun A., Sonuc S.B., Duranel A., Basar A. Courier routing and assignment for food delivery service using reinforcement learning. Comput. Ind. Eng. 2022;164:107871. doi: 10.1016/j.cie.2021.107871. DOI

Huang Y., Flint S.H., Yu S., Ding Y., Palmer J.S. Phenotypic properties and genotyping analysis of Bacillus cereus group isolates from dairy and potato products. LWT Food Sci. Technol. 2021;140:110853. doi: 10.1016/j.lwt.2021.110853. DOI

Tirloni E., Stella S., Celandroni F., Mazzantini D., Bernardi C., Ghelardi E. Bacillus cereus in Dairy Products and Production Plants. Foods. 2022;11:2572. doi: 10.3390/foods11172572. PubMed DOI PMC

Mohammadi B., Reyes M.E.P., Smith S.A. Survival, Growth, and Toxin Production of Bacillus cereus During Cooking and Storage of Fresh Rice Noodles. J. Food Prot. 2024;87:100239. doi: 10.1016/j.jfp.2024.100239. PubMed DOI

Rouzeau-Szynalski K., Stollewerk K., Messelhaeusser U., Ehling-Schulz M. Why be serious about emetic Bacillus cereus: Cereulide production and industrial challenges. Food Microbiol. 2020;85:103279. doi: 10.1016/j.fm.2019.103279. PubMed DOI

Kranzler M., Stollewerk K., Rouzeau-Szynalski K., Blayo L., Sulyok M., Ehling-Schulz M. Temperature exerts control of Bacillus cereus emetic toxin production on post-transcriptional levels. Front. Microbiol. 2016;7:1640. doi: 10.3389/fmicb.2016.01640. PubMed DOI PMC

Schmid P.J., Maitz S., Kittinger C. Bacillus cereus in Packaging Material: Molecular and Phenotypical Diversity Revealed. Front. Microbiol. 2021;12:698974. doi: 10.3389/fmicb.2021.698974. PubMed DOI PMC

Heini N., Stephan R., Filter M., Plaza-Rodriguez C., Frentzel H., Ehling-Schulz M., Johler S. Temperature-Dependent Growth Characteristics of Bacillus thuringiensis in a Ratatouille Food Model. J. Food Prot. 2020;83:816–820. doi: 10.4315/0362-028X.JFP-19-358. PubMed DOI

Mahler H., Pasi A., Kramer J.M., Schulte P., Scoging A.C., Bär W., Krähenbühl S. Fulminant liver failure in association with the emetic toxin of Bacillus cereus. N. Engl. J. Med. 1997;336:1142–1148. doi: 10.1056/NEJM199704173361604. PubMed DOI

Naranjo M., Denayer S., Botteldoorn N., Delbrassinne L., Veys J., Waegenaere J., Sirtaine N., Driesen R.B., Sipido K.R., Mahillon J., et al. Sudden death of a young adult associated with Bacillus cereus food poisoning. J. Clin. Microbiol. 2011;49:4379–4381. doi: 10.1128/JCM.05129-11. PubMed DOI PMC

Colaco C.M., Basile K., Draper J., Ferguson P.E. Fulminant Bacillus cereus food poisoning with fatal multi-organ failure. BMJ Case Rep. 2021;14:e238716. doi: 10.1136/bcr-2020-238716. PubMed DOI PMC

Delbrassinne L., Andjelkovic M., Dierick K., Denayer S., Mahillon J., Van Loco J. Prevalence and levels of Bacillus cereus emetic toxin in rice dishes randomly collected from restaurants and comparison with the levels measured in a recent foodborne outbreak. Foodborne Pathog. Dis. 2012;9:809–814. doi: 10.1089/fpd.2012.1168. PubMed DOI

Ehling-Schulz M., Messelhäusser U. One pathogen but two different types of foodborne outbreak: Bacillus cereus in catering facilities in Germany. In: Hoorfar J., editor. Case Studies in Food Safety and Authenticity. Woodhead Publishing; Sawston, UK: 2012. pp. 63–70.

Thery M., Cousin V.L., Tissieres P., Enault M., Morin L. Multi-organ failure caused by lasagnas: A case report of Bacillus cereus food poisoning. Front. Pediatr. 2022;10:978250. doi: 10.3389/fped.2022.978250. PubMed DOI PMC

Pirhonen T.I., Andersson M.A., Jääskeläinen E.L., Salkinoja-Salonen M.S., Honkanen-Buzalski T., Johansson T.L. Biochemical and toxic diversity of Bacillus cereus in a pasta and meat dish associated with a food-poisoning case. Food Microbiol. 2005;22:87–91. doi: 10.1016/j.fm.2004.04.002. DOI

Dietrich R., Jessberger N., Ehling-Schulz M., Märtlbauer E., Granum P.E. The food poisoning toxins of Bacillus cereus. Toxins. 2021;13:98. doi: 10.3390/toxins13020098. PubMed DOI PMC

Griffiths M.W., Schraft H. Bacillus cereus Food Poisoning. In: Dodd C.E.R., Aldsworth T., Stein R.A., Cliver D.O., Riemann H.P., editors. Foodborne Diseases. 3rd ed. Academic Press; Cambridge, MA, USA: 2017. pp. 395–405.

EFSA BIOHAZ Opinion of the Scientific Panel on biological hazards (BIOHAZ) on Bacillus cereus and other Bacillus spp. in foodstuffs. EFSA J. 2005;175:1–48. doi: 10.2903/j.efsa.2005.175. DOI

Le Marc Y., Baert L., da Silva N.B., Postollec F., Huchet V., Baranyi J., Ellouze M. The effect of pH on the growth rate of Bacillus cereus sensu lato: Quantifying strain variability and modelling the combined effects of temperature and pH. Int. J. Food Microbiol. 2021;360:109420. doi: 10.1016/j.ijfoodmicro.2021.109420. PubMed DOI

Key Factors Influencing Bacillus cereus Contamination in Hot Ready-to-Eat Meal Delivery