Enterotoxigenic profile, biofilm production, and antimicrobial resistance of Bacillus cereus isolated from rice-based food marketed in southern Mexico

Authors

  • Joel Reyes-Roldan Laboratorio de Investigación en Patometabolismo Microbiano, Universidad Autónoma de Guerrero;
  • Mariela Cano-Ponce Laboratorio de Investigación en Patometabolismo Microbiano, Universidad Autónoma de Guerrero
  • Luis-Fernando Gaspar-Nava Laboratorio de Investigación en Patometabolismo Microbiano, Universidad Autónoma de Guerrero
  • José-Humberto Perez-Olais Unidad de Investigación en Virología y Cancer, Hospital Infantil de México Federico Gómez
  • Salvador Muñloz-Barrios Laboratorio de Investigación en Inmunología y Microbiología, Universidad Autónoma de Guerrero
  • Hugo-Alberto Rodriguez-Ruiz Laboratorio de Investigación en Biomedicina Molecular, Universidad Autónoma de Guerrero
  • Arturo Ramirez- Peralta Universidad Autonoma de Guerrero

DOI:

https://doi.org/10.18633/biotecnia.v26.2429

Keywords:

Bacillus cereus, rice, virulence

Abstract

Bacillus cereus is responsible for food poisoning worldwide; thus, the characterization of strains isolated from food, in this case rice, is essential.

The objective of this study was to identify the toxigenic profile, lytic enzymes, antimicrobial resistance, and biofilm production of B. cereus strains isolated from rice. The genetic profile of toxins and biofilm-related genes in the strains was determined by endpoint PCR. Biofilm formation was visualized using safranin staining, and the evaluation of lytic enzymes was conducted in culture media.

Psychrophilic characteristics were monitored by assessing the growth of the strains at refrigeration temperature. The GTG5 technique was employed to determine the genetic diversity of the strains, and their antimicrobial resistance was validated through minimum inhibitory concentration testing.

The strains of B. cereus s.l. isolated from rice contained genes for enterotoxins and genes associated with biofilm production. However, the strains did not possess the cereulide gene. The strain isolated from fried rice was the only one that contained the hbl toxin gene and the eps2 operon.

Interestingly, this strain was the only one that did not produce biofilm. It exhibited intermediate sensitivity to erythromycin, was positive for amylase activity, showed high lecithinase activity, and was capable of growing at refrigeration temperature.

Downloads

Download data is not yet available.

Author Biography

Arturo Ramirez- Peralta, Universidad Autonoma de Guerrero

Dr. Arturo Ramírez Peralta

Laboratorio de Investigación en Patometabolismo Microbiano

Facultad de Ciencias Químico Biologicas

References

Adame-Gomez, R., Castro Alarcón, N., Vences-Velázquez, A., Rodríguez-Bataz, E., Santia-go-Dionisio, M.C., Ramírez-Peralta, A., 2019. Prevalencia de cepas del grupo de Bacillus cereus productoras de biopelicula en helados comercializados en México. https://doi.org/10.5281/ZENODO.3520530

Adame-Gómez, R., Cruz-Facundo, I.-M., García-Díaz, L.-L., Ramírez-Sandoval, Y., Pérez-Valdespino, A., Ortuño-Pineda, C., Santiago-Dionisio, M.-C., Ramírez-Peralta, A., 2020a. Biofilm Produc-tion by Enterotoxigenic Strains of Bacillus cereus in Different Materials and under Different Environmental Conditions. Microorganisms 8, 1071. https://doi.org/10.3390/microorganisms8071071

Adame-Gómez, R., Muñoz-Barrios, S., Castro-Alarcón, N., Leyva-Vázquez, M.-A., Toribio-Jiménez, J., Ramírez-Peralta, A., 2020b. Prevalence of the Strains of Bacillus cereus Group in Artisanal Mexican Cheese. Foodborne Pathog. Dis. 17, 8–14. https://doi.org/10.1089/fpd.2019.2673

Ankolekar, C., Rahmati, T., Labbé, R.G., 2009. Detection of toxigenic Bacillus cereus and Bacillus thu-ringiensis spores in U.S. rice. Int. J. Food Microbiol. 128, 460–466. https://doi.org/10.1016/j.ijfoodmicro.2008.10.006

Arslan, S., Eyi, A., Küçüksarı, R., 2014. Toxigenic genes, spoilage potential, and antimicrobial re-sistance of Bacillus cereus group strains from ice cream. Anaerobe 25, 42–46. https://doi.org/10.1016/j.anaerobe.2013.11.006

Berthold-Pluta, Pluta, Garbowska, Stefańska, 2019. Prevalence and toxicity characterization of Bacillus cereus in food products from Poland. Foods 8, 269. https://doi.org/10.3390/foods8070269

Candela, T., Fagerlund, A., Buisson, C., Gilois, N., Kolstø, A.-B., Økstad, O.A., Aymerich, S., Niel-sen-Leroux, C., Lereclus, D., Gohar, M., 2019. CalY is a major virulence factor and a biofilm matrix protein. Mol. Microbiol. 111, 1416–1429. https://doi.org/10.1111/mmi.14184

Cano- Ponce, M., Ramirez- Peralta, A., 2023. Aislamiento de B. Cereus en arroz comercializado en el sur de México. Tlamati Press.

Caro-Astorga, J., Álvarez-Mena, A., Hierrezuelo, J., Guadix, J.A., Heredia-Ponce, Z., Arbole-da-Estudillo, Y., González-Munoz, E., de Vicente, A., Romero, D., 2020. Two genomic regions encoding exopolysaccharide production systems have complementary functions in B. cereus multicellularity and host interaction. Sci. Rep. 10, 1000. https://doi.org/10.1038/s41598-020-57970-3

Caro-Astorga, J., Pérez-García, A., de Vicente, A., Romero, D., 2014. A genomic region involved in the formation of adhesin fibers in Bacillus cereus biofilms. Front. Microbiol. 5, 745. https://doi.org/10.3389/fmicb.2014.00745

Castulo-Arcos, D.A., Adame-Gómez, R., Castro-Alarcón, N., Galán-Luciano, A., Santiago Dionisio, M.C., Leyva-Vázquez, M.A., Perez-Olais, J.-H., Toribio-Jiménez, J., Ramirez-Peralta, A., 2022. Genetic diversity of enterotoxigenic Bacillus cereus strains in coriander in southwestern Mexico. PeerJ 10, e13667. https://doi.org/10.7717/peerj.13667

Chen, J., Zhang, J., Zhan, L., Chen, H., Zhang, Z., Huang, C., Yue, M., 2022. Prevalence and antimicro-bial-resistant characterization of Bacillus cereus isolated from ready-to-eat rice products in Eastern China. Front. Microbiol. 13, 964823. https://doi.org/10.3389/fmicb.2022.964823

Chen, L., Daniel, R.M., Coolbear, T., 2003. Detection and impact of protease and lipase activities in milk and milk powders. Int. Dairy J. 13, 255–275. https://doi.org/10.1016/S0958-6946(02)00171-1

Claus, D., Berkeley, C., 1986. The genus Bacillus, in: Bergey’s Manual of Systematic Bacteriology. The Williams & Wilkins Co, Baltimore, pp. 1105–39.

Cruz-Facundo, I., Adame-Gómez, R., Vences-Velázquez, A., Rodríguez-Bataz, E., Muñoz-Barrios, S., Pérez-Oláis, J., Ramírez-Peralta, A., 2022. Bacillus Cereus in Eggshell: Enterotoxigenic Profiles and Biofilm Production. Braz. J. Poult. Sci. 24, eRBCA-2021-1535. https://doi.org/10.1590/1806-9061-2021-1535

Cruz-Facundo, I.-M., Toribio-Jiménez, J., Castro-Alarcón, N., Leyva-Vázquez, M.-A., Rodríguez-Ruíz, H.-A., Pérez-Olais, J.-H., Adame-Gómez, R., Rodríguez-Bataz, E., Reyes-Roldán, J., Mu-ñoz-Barrios, S., Ramírez-Peralta, A., 2023. Bacillus cereus in the Artisanal Cheese Production Chain in Southwestern Mexico. Microorganisms 11, 1290. https://doi.org/10.3390/microorganisms11051290

De Jonghe, V., Coorevits, A., Vandroemme, J., Heyrman, J., Herman, L., De Vos, P., Heyndrickx, M., 2008. Intraspecific genotypic diversity of Bacillus species from raw milk. Int. Dairy J. 18, 496–505. https://doi.org/10.1016/j.idairyj.2007.11.007

De-la-Pinta, I., Cobos, M., Ibarretxe, J., Montoya, E., Eraso, E., Guraya, T., Quindós, G., 2019. Effect of biomaterials hydrophobicity and roughness on biofilm development. J. Mater. Sci. Mater. Med. 30, 77. https://doi.org/10.1007/s10856-019-6281-3

Dietrich, R., Jessberger, N., Ehling-Schulz, M., Märtlbauer, E., Granum, P.E., 2021. The Food Poisoning Toxins of Bacillus cereus. Toxins 13, 98. https://doi.org/10.3390/toxins13020098

Dogsa, I., Brloznik, M., Stopar, D., Mandic-Mulec, I., 2013. Exopolymer diversity and the role of levan in Bacillus subtilis biofilms. PloS One 8, e62044. https://doi.org/10.1371/journal.pone.0062044

Duport, C., Jobin, M., Schmitt, P., 2016. Adaptation in Bacillus cereus: From Stress to Disease. Front. Microbiol. 7. https://doi.org/10.3389/fmicb.2016.01550

Ehling-Schulz, M., Fricker, M., Scherer, S., 2004. Bacillus cereus, the causative agent of an emetic type of food-borne illness. Mol. Nutr. Food Res. 48, 479–487. https://doi.org/10.1002/mnfr.200400055

Ehling-Schulz, M., Guinebretiere, M.-H., Monthán, A., Berge, O., Fricker, M., Svensson, B., 2006. Toxin gene profiling of enterotoxic and emetic Bacillus cereus. FEMS Microbiol. Lett. 260, 232–240. https://doi.org/10.1111/j.1574-6968.2006.00320.x

Ehling-Schulz, M., Lereclus, D., Koehler, T.M., 2019. The Bacillus cereus Group: Bacillus Species with Pathogenic Potential. Microbiol. Spectr. 7, 7.3.6. https://doi.org/10.1128/microbiolspec.GPP3-0032-2018

Enosi Tuipulotu, D., Mathur, A., Ngo, C., Man, S.M., 2021. Bacillus cereus: Epidemiology, Virulence Factors, and Host-Pathogen Interactions. Trends Microbiol. 29, 458–471. https://doi.org/10.1016/j.tim.2020.09.003

Flores-Urbán, K.A., Natividad-Bonifacio, I., Vázquez-Quiñones, C.R., Vázquez-Salinas, C., Quiño-nes-Ramírez, E.I., 2014. Detection of toxigenic Bacillus cereus strains isolated from vegetables in Mexico City. J. Food Prot. 77, 2144–2147. https://doi.org/10.4315/0362-028X.JFP-13-479

Fraccalvieri, R., Bianco, A., Difato, L.M., Capozzi, L., Del Sambro, L., Simone, D., Catanzariti, R., Ca-ruso, M., Galante, D., Normanno, G., Palazzo, L., Tempesta, M., Parisi, A., 2022. Toxigenic Genes, Pathogenic Potential and Antimicrobial Resistance of Bacillus cereus Group Isolated from Ice Cream and Characterized by Whole Genome Sequencing. Foods Basel Switz. 11, 2480. https://doi.org/10.3390/foods11162480

Grutsch, A.A., Nimmer, P.S., Pittsley, R.H., Kornilow, K.G., McKillip, J.L., 2018. Molecular Pathogen-esis of Bacillus spp., with Emphasis on the Dairy Industry. Fine Focus 4, 203–222. https://doi.org/10.33043/FF.4.2.203-222

Hayrapetyan, H., Muller, L., Tempelaars, M., Abee, T., Nierop Groot, M., 2015. Comparative analysis of biofilm formation by Bacillus cereus reference strains and undomesticated food isolates and the effect of free iron. Int. J. Food Microbiol. 200, 72–79. https://doi.org/10.1016/j.ijfoodmicro.2015.02.005

Hayrapetyan, H., Siezen, R., Abee, T., Nierop Groot, M., 2016. Comparative Genomics of Iron-Transporting Systems in Bacillus cereus Strains and Impact of Iron Sources on Growth and Biofilm Formation. Front. Microbiol. 7, 842. https://doi.org/10.3389/fmicb.2016.00842

Hindler, J.A., Richter, S.S., 2016. Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria: M45, 3rd edition. ed, Documents / Clinical and La-boratory Standards Institute. Clinical and Laboratory Standards Institute, Wayne, PA.

Ivanova, N., Sorokin, A., Anderson, I., Galleron, N., Candelon, B., Kapatral, V., Bhattacharyya, A., Rez-nik, G., Mikhailova, N., Lapidus, A., Chu, L., Mazur, M., Goltsman, E., Larsen, N., D’Souza, M., Walunas, T., Grechkin, Y., Pusch, G., Haselkorn, R., Fonstein, M., Ehrlich, S.D., Overbeek, R., Kyrpides, N., 2003. Genome sequence of Bacillus cereus and comparative analysis with Bacillus anthracis. Nature 423, 87–91. https://doi.org/10.1038/nature01582

Jessberger, N., Dietrich, R., Granum, P.E., Märtlbauer, E., 2020. The Bacillus cereus Food Infection as Multifactorial Process. Toxins 12, 701. https://doi.org/10.3390/toxins12110701

Kim, Booyoung, Bang, J., Kim, H., Kim, Y., Kim, Byeong-sam, Beuchat, L.R., Ryu, J.-H., 2014. Bacil-lus cereus and Bacillus thuringiensis spores in Korean rice: Prevalence and toxin production as affected by production area and degree of milling. Food Microbiol. 42, 89–94. https://doi.org/10.1016/j.fm.2014.02.021

Kim, S.K., Kim, K.-P., Jang, S.S., Shin, E.M., Kim, M.-J., Oh, S., Ryu, S., 2009. Prevalence and Toxi-genic Profiles of Bacillus cereus Isolated from Dried Red Peppers, Rice, and Sunsik in Korea. J. Food Prot. 72, 578–582. https://doi.org/10.4315/0362-028X-72.3.578

Kotiranta, A., Lounatmaa, K., Haapasalo, M., 2000. Epidemiology and pathogenesis of Bacillus cereus infections. Microbes Infect. 2, 189–198. https://doi.org/10.1016/s1286-4579(00)00269-0

Liu, Y., Lai, Q., Göker, M., Meier-Kolthoff, J.P., Wang, M., Sun, Y., Wang, L., Shao, Z., 2015. Genomic insights into the taxonomic status of the Bacillus cereus group. Sci. Rep. 5, 14082. https://doi.org/10.1038/srep14082

Majed, R., Faille, C., Kallassy, M., Gohar, M., 2016. Bacillus cereus Biofilms-Same, Only Different. Front. Microbiol. 7, 1054. https://doi.org/10.3389/fmicb.2016.01054

Ołtuszak-Walczak, E., Walczak, P., Modrak, R., 2006. Detection of enterotoxic Bacillus cereus produc-ing hemolytic and non hemolytic enterotoxins by PCR test. Pol. J. Microbiol. 55, 113–118.

Park, Y.-B., Kim, J.-B., Shin, S.-W., Kim, J.-C., Cho, S.-H., Lee, B.-K., Ahn, J., Kim, J.-M., Oh, D.-H., 2009. Prevalence, Genetic Diversity, and Antibiotic Susceptibility of Bacillus cereus Strains Isolated from Rice and Cereals Collected in Korea. J. Food Prot. 72, 612–617. https://doi.org/10.4315/0362-028X-72.3.612

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 Pathog. Dis. 9, 125–131. https://doi.org/10.1089/fpd.2011.0969

SAGARPA, 2017. Arroz Mexicano. (Federal), Planeacion agricola nacional 2017. 2030. SAGARPA, Ciudad de Mexico.

Schoeni, J.L., Wong, A.C.L., 2005. Bacillus cereus food poisoning and its toxins. J. Food Prot. 68, 636–648. https://doi.org/10.4315/0362-028x-68.3.636

Shemesh, M., Ostrov, I., 2020. Role of Bacillus species in biofilm persistence and emerging antibiofilm strategies in the dairy industry. J. Sci. Food Agric. 100, 2327–2336. https://doi.org/10.1002/jsfa.10285

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

Wei, S., Chelliah, R., Park, B.-J., Park, J.-H., Forghani, F., Park, Y.-S., Cho, M.-S., Park, D.-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. Microb. Pathog. 115, 312–320. https://doi.org/10.1016/j.micpath.2017.12.079

Wijman, J.G.E., de Leeuw, P.P.L.A., Moezelaar, R., Zwietering, M.H., Abee, T., 2007. Air-liquid inter-face biofilms of Bacillus cereus: formation, sporulation, and dispersion. Appl. Environ. Micro-biol. 73,

Graphical abstract

Downloads

Published

2024-12-17

How to Cite

Reyes-Roldan, J., Cano-Ponce, M., Gaspar-Nava, L.-F., Perez-Olais, J.-H., Muñoz-Barrios, S., Rodriguez-Ruiz, H.-A., & Ramirez- Peralta, A. (2024). Enterotoxigenic profile, biofilm production, and antimicrobial resistance of Bacillus cereus isolated from rice-based food marketed in southern Mexico. Biotecnia, 26, e2429. https://doi.org/10.18633/biotecnia.v26.2429

Issue

Vol. 26 (2024): January - December

Section

Research Articles

License

Copyright (c) 2023

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Creative Commons License

The journal Biotecnia is licensed under the Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) license.

Metrics