medigraphic.com
ISSN 2500-7106 (Electronic)
ISSN 0123-2576 (Print)
Medicina & Laboratorio

2014, Number 09-10

<< Back Next >>

Medicina & Laboratorio 2014; 20 (09-10)

Direct detection of toxigenic genes of Bacillus cereus in corn starch and wheat flour using multiplex polymerase chain reaction (mPCR)

Sánchez-Chica JA, Correa-Ochoa MM, Aceves-Díez ÁE, Castañeda-Sandoval LM

Language: Spanish
References: 26
Page: 441-451
PDF size: 322.71 Kb.


ABSTRACT

Introduction: Bacillus cereus is a human pathogen that causes two kinds of foodborne diseases, the emetic syndrome caused by emetic toxin or cereulide (Ces), and the diarrheal syndrome caused by three different enterotoxins, the hemolytic enterotoxin or hemolysin BL (Hbl), the nonhemolytic enterotoxin (Nhe) and the cytotoxin K (CytK). Objective: To determine the presence of toxigenic genes of Bacillus cereus in DNA samples directly obtained from corn starch and wheat flour using multiplex polymerase chain reaction. Material and methods: The presence of toxigenic genes of Bacillus cereus were determined in DNA samples directly extracted from corn starch and wheat flour, using a multiplex polymerase chain reaction technique specific for cesB, hblC, nheA and cytK genes. Results: From a total of 76 corn starch samples, 60.5% had toxigenic genes of Bacillus cereus and were grouped in six consortia: I: hblC, cytK (30.4%), II: nheA, hblC, cytK (21.7%), III: hblC (19.6%), IV: nheA (15.2%), V: nheA, hblC (10.9%) and VI: nheA, hblC, cytK, cesB (2.2%). From 79 wheat flour samples tested, 65.8% had toxigenic genes of Bacillus cereus and were grouped into four consortia: I: nheA, hblC, cytK (80.8%), II: hblC, cytK (11.5%), III: hblC (5.8%) and IV: nheA, hblC (1.9%). Conclusions: It was found that in corn starch the enterotoxigenic consortia predominated over the emetic while in wheat flour only enterotoxigenic consortia were detected. The study describes a multiplex polymerase chain reaction that allowed direct, rapid, and simultaneous detection of toxigenic genes of Bacillus cereus in foods.


REFERENCES

  1. Stenfors Arnesen LP, Fagerlund A, Granum PE. From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiol Rev 2008; 32: 579-606.

  2. Bhunia AK. Bacillus cereus and Bacillus an-thracis. In: Bhunia A, ed. Foodborne Microbial Pathogens: Mechanisms and Pathogenesis. Nueva York, Estados Unidos: Springer Sci-ence+Business Media; 2008: 135-148.

  3. Dommel MK, Lucking G, Scherer S, Eh-ling-Schulz M. Transcriptional kinetic analyses of cereulide synthetase genes with respect to growth, sporulation and emetic toxin produc-tion in Bacillus cereus. Food Microbiol 2011; 28: 284-290.

  4. Granum P. Bacillus cereus. In: Fratamico PM, Bhunia AK, Smith JL, eds. Foodborne Patho-gens: Microbiology and Molecular Biology. Norfolk, Inglaterra: Caister Academic Press; 2005: 409-419.

  5. Økstad OA, Kolstø AB. Genomics of Bacil-lus Species. In: Wiedmann M, Zhang W, eds. Genomics of Foodborne Bacterial Pathogens. Nueva York, Estados Unidos: Springer Sci-ence+Business Media; 2011: 29-53.

  6. Kotiranta A, Lounatmaa K, Haapasalo M. Epi-demiology and pathogenesis of Bacillus cereus infections. Microbes Infect 2000; 2: 189-198.

  7. Logan NA. Bacillus and relatives in foodborne illness. J Appl Microbiol 2012; 112: 417-429.

  8. Instituto Colombiano de Normas Técnicas y Certificación. Norma Técnica Colombiana NTC: 467

  9. Método horizontal para el recuento de Bacillus cereus. Técnica de recuento de colo-nias. Bogotá D.C., Colombia; 2006.9.Ehling-Schulz M, Fricker M, Scherer S. Iden-tification of emetic toxin producing Bacillus ce-reus strains by a novel molecular assay. FEMS Microbiol Lett 2004; 232: 189-195.

  10. Ombui J, Gitahi J, Gicheru M. Direct detection of Bacillus cereus enterotoxin genes in food by multiplex Polymerase Chain Reaction. Int J In-tegr Biol 2008; 2: 172-181.

  11. Rather MA, Aulakh RS, Gill JPS, Rao TS, Hassan MN. Direct Detection of Bacillus cereus and its Enterotoxigenic Genes in Meat and Meat Products by Polymerase Chain Reaction. J Adv Vet Res 2011; 1: 99-104.

  12. D’Alessandro B, Antúnez K, Piccini C, Zunino P. DNA extraction and PCR detection of Paeni-bacillus larvae spores from naturally contami-nated honey and bees using spore-decoating and freeze-thawing techniques. World J Micro-biol Biotechnol 2007; 23: 593–597.

  13. Moravek M, Wegscheider M, Schulz A, Diet-rich R, Burk C, Martlbauer E. Colony immu-noblot assay for the detection of hemolysin BL enterotoxin producing Bacillus cereus. FEMS Microbiol Lett 2004; 238: 107-113.

  14. Guinebretière MH, Broussolle V, Nguy-en-The C. Enterotoxigenic profiles of food-poi-soning and food-borne Bacillus cereus strains. J Clin Microbiol 2002; 40: 3053-3056.

  15. Ehling-Schulz M, Vukov N, Schulz A, Sha-heen R, Andersson M, Martlbauer E, et al. Identification and partial characterization of the nonribosomal peptide synthetase gene respon-sible for cereulide production in emetic Bacillus cereus. Appl Environ Microbiol 2005; 71: 105-113.

  16. Ngamwongsatit P, Buasri W, Pianariyanon P, Pulsrikarn C, Ohba M, Assavanig A, et al. Broad distribution of enterotoxin genes (hblC-DA, nheABC, cytK, and entFM) among Bacillus thuringiensis and Bacillus cereus as shown by novel primers. Int J Food Microbiol 2008; 121: 352-356.

  17. Ehling-Schulz M, Svensson B, Guinebretiere MH, Lindback T, Andersson M, Schulz A, et al. Emetic toxin formation of Bacillus cereus is restricted to a single evolutionary lineage of closely related strains. Microbiology 2005; 151: 183-197.

  18. Park YB, Kim JB, Shin SW, Kim JC, Cho SH, Lee BK, et al. Prevalence, genetic diversity, and antibiotic susceptibility of Bacillus cereus strains isolated from rice and cereals collected in Korea. J Food Prot 2009; 72: 612-617.

  19. Samapundo S, Heyndrickx M, Xhaferi R, Devlieghere F. Incidence, diversity and toxin gene characteristics of Bacillus cereus group strains isolated from food products marketed in Belgium. Int J Food Microbiol 2011; 150: 34-41.

  20. Ankolekar C, Rahmati T, Labbe RG. Detec-tion of toxigenic Bacillus cereus and Bacillus thuringiensis spores in U.S. rice. Int J Food Mi-crobiol 2009; 128: 460-466.

  21. López AC, Alippi AM. Enterotoxigenic gene profiles of Bacillus cereus and Bacillus megate-rium isolates recovered from honey. Rev Argent Microbiol 2010; 42: 216-225.

  22. Chaves JQ, Pires ES, Vivoni AM. Genetic di-versity, antimicrobial resistance and toxigenic profiles of Bacillus cereus isolated from food in Brazil over three decades. Int J Food Microbiol 2011; 147: 12-16.

  23. Altayar M, Sutherland AD. Bacillus cereus is common in the environment but emetic toxin producing isolates are rare. J Appl Microbiol 2006; 100: 7-14.

  24. Kim JB, Park JS, Kim MS, Hong SC, Park JH, Oh DH. Genetic diversity of emetic toxin producing Bacillus cereus Korean strains. Int J Food Microbiol 2011; 150: 66-72.

  25. Lee N, Sun JM, Kwon KY, Kim HJ, Koo M, Chun HS. Genetic diversity, antimicrobial resis-tance, and toxigenic profiles of Bacillus cereus strains isolated from Sunsik. J Food Prot 2012; 75: 225-230.

  26. López AC, Minnaard J, Pérez PF, Alippi AM. A case of intoxication due to a highly cytotox-ic Bacillus cereus strain isolated from cooked chicken. Food Microbiol 2014; Ago 27 [Epub ahead of print].




2020     |     www.medigraphic.com