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Bacillus cereus: public health burden associated with ready-to-eat foods in Himachal Pradesh, India

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Abstract

The study determined incidence, enterotoxigenecity and antimicrobial susceptibility profiles of Bacillus cereus isolated from ready-to-eat (RTE) milk products (n = 80), RTE meat products (n = 40), beverages (n = 40) and water samples (n = 60, from food preparing and serving outlets/restaurants) collected from eight different tourist places of Himachal Pradesh. 11.4% (25/220) samples were contaminated with Bacillus and isolates were identified as B. cereus (76.0%, n = 19), B. alvei (12.0%, n = 3), B. polymyxa (8.0%, n = 2) and B. firmus (4.0%, n = 1) by conventional and molecular methods. B. cereus incidence was highest in cheese based foods (25.0%) followed by vegetable soups (16.7%), khoa based foods (14.0%), milk based beverages (10.5%), paneer based foods (8.6%), cream based foods (8.3%) and water (8.3%) samples. Multiplex polymerase chain reaction detected enterotoxigenic genes only in B. cereus isolates. nhe complex (encoding non-haemolytic enterotoxins, ABC) genes were detected only in B. cereus isolates. 57.6% (11/19), 36.8% (7/19) and 5.3% (1/19) harboured all three (nheA, nheB, nheC), two (nheB, nheC) and one (nheC) nhe gene, respectively. Among hbl complex genes (encoding haemolytic enterotoxins CAD), only hblC (36.8%, 7/19) was detected. Incidence B. cereus cytK (encoding cytotoxin enterotoxin) was 52.6% (10/19). Each B. cereus isolate harboured two or more enterotoxigenic genes. Seven isolates had at least one gene from haemolytic and non-haemolytic complexes along with cytK. High levels (> 50%) of antimicrobial resistance were recorded for penicillin, amoxicillin, ampicillin cefixime and ceftazidine in tested B. cereus isolates. Two isolates were identified as multidrug resistant isolates with resistance to ≥ 3 antibiotic classes.

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References

  • Abbas BA, Khudor MH, Saeed BMS (2014) Detection of hbl, nhe and bceT toxin genes in Bacillus cereus isolates by multiplex PCR. Int J Curr Microbiol Appl Sci 3:1009–1016

    Google Scholar 

  • Agwa OK, Uzoigwe CI, Wokoma EC (2012) Incidence and antibiotic sensitivity of Bacillus cereus isolated from ready to eat foods sold in some markets in Portharcourt, Rivers state, Nigeria. Asian J Microbiol Biotechnol Environ Sci 14:13–18

    CAS  Google Scholar 

  • Ahmadi SA, Panda AK (2015) Prevalence of Escherichia coli and Salmonella spp. in ready-to-eat milk and milk products in Himachal Pradesh. J Vet Public Health 13:25–29

    Google Scholar 

  • Aksu H, Bostan K, Ergün Ö (2000) Presence of Bacillus cereus in packaged some spices and herbs sold in Istanbul. Pak J Biol Sci 3:710–712

    Google Scholar 

  • Al-Khatib MS, Khyami-Horani H, Badran E, Shehabi AA (2007) Incidence and characterization of diarrheal enterotoxins of fecal Bacillus cereus isolates associated with diarrhea. Diag Microbiol Infect Dis 59:383–387

    CAS  Google Scholar 

  • Andersson A, Ronner U, Granum PE (1995) What problems does the food industry have with the spore-forming pathogens Bacillus cereus and Clostridium perfringens? Int J Food Microbiol 28:145–155

    CAS  PubMed  Google Scholar 

  • Aruwa CE, Akinyosoye FA (2015) Microbiological assessment of ready-to-eat foods (RTEs) for the presence Bacillus species. J Adv Biol Biotechnol 3:145–152

    Google Scholar 

  • Bashir M, Malik MA, Moien J, Badroo GA, Mohd Bhat A, Singh M (2017) Prevalence and characterization of Bacillus cereus in meat and meat products in and around Jammu region of Jammu and Kashmir, India. Int J Curr Microbiol Appl Sci 6:1094–1106

    Google Scholar 

  • Bauer AW, Kirby WM, Sherris JC, Turck M (1966) Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 45:493–496

    CAS  PubMed  Google Scholar 

  • Beecher DJ, Schoeni JL, Wong AC (1995) Enterotoxic activity of hemolysin BL from Bacillus cereus. Infect Immun 63:4423–4428

    CAS  PubMed  PubMed Central  Google Scholar 

  • Berthold-Pluta A, Puta A, Debevere J (2015) The effect of selected factors on the survival of Bacillus cereus in the human gastro-intestinal tract. Microb Pathog 82:7–14

    CAS  PubMed  Google Scholar 

  • Bottone EJ (2010) B. cereus, a volatile human pathogen. Clin Microbiol Rev 23:382–398

    PubMed  PubMed Central  Google Scholar 

  • Cadirci O, Gucukoglu A, Goknur T, Kevenk O, Mustafa A (2013) Determination of enterotoxigenic gene profiles of Bacillus cereus isolates isolated from dairy desserts by multiplex PCR. Kafkas Univ Vet Fak Derg 19:869–874

    Google Scholar 

  • Chettri R, Tamang JP (2015) Bacillus species isolated from Tungrymbai and Bekang, naturally fermented soybean foods of India. Int J Food Microbiol 197:72–76

    PubMed  Google Scholar 

  • CLSI (2013) Performance standards for antimicrobial susceptibility testing. Twenty-third informational supplement. CLSI document M100-S23. Clinical and Laboratory Standards Institute, Wayne

    Google Scholar 

  • Fayaz S, Badroo GA, Ahmad A, Rasool U, Mustafa R, Mudasir M (2017) Molecular characterization of enterotoxigenic Bacillus cereus species isolated from meat using conventional PCR and multiplex PCR. Int J Curr Microbiol Appl Sci 6:324–328

    Google Scholar 

  • Fernandes MDS, Fujimoto G, Schneid I, Kabuki DV, Kuaye AY (2014) Enterotoxigenic profile, antimicrobial susceptibility and biofilm formation of Bacillus cereus isolated from ricotta processing. Int Dairy J 38:16–23

    CAS  Google Scholar 

  • Fernández-No IC, Guarddon M, Böhme K, Cepeda A, Calo-Mata P, Barros-Velázquez J (2011) Detection and quantification of spoilage and pathogenic Bacillus cereus, Bacillus subtilis and Bacillus licheniformis by real-time PCR. Food Microbiol 28:605–610

    PubMed  Google Scholar 

  • Gomaa OM, Momtaz OA (2006) 16S rRNA characterization of a Bacillus isolate and its tolerance profile after subsequent subculturing. Arab J Biotechnol 10:107–116

    Google Scholar 

  • Gopal N, Hill C, Ross PR, Beresford TP, Fenelon MA, Cotter PD (2015) The prevalence and control of Bacillus and related spore-forming bacteria in the dairy industry. Front Microbiol 6:1418

    PubMed  PubMed Central  Google Scholar 

  • Hadithi AL, Hadeel T, Entesar AK (2016) Incidence of Bacillus cereus in food and environmental sources in Basrah/Iraq. Antibiotic resistance profiles of recovered isolates. Int J Res Acad Rev 4:117–127

    Google Scholar 

  • Hanlin JH (1998) Spoilage of acidic products by Bacillus species. Dairy Food Environ Sanit 18:655–659

    Google Scholar 

  • Iurlina MO, Saiz AI, Fuselli SR, Fritz R (2006) Prevalence of Bacillus spp. in different food products collected in Argentina. LWT 39:105–110

    CAS  Google Scholar 

  • Keisam S, Tuikhar N, Ahmed G, Jeyaram K (2019) Toxigenic and pathogenic potential of enteric bacterial pathogens prevalent in the traditional fermented foods marketed in the Northeast region of India. Int J Food Microbiol 296:21–30

    CAS  PubMed  Google Scholar 

  • Kim JM, Forghani F, Jung-Beom K, Yong-Bae P, Myoung-Su P, Wang J, Park JH, Deog-Hwan O (2011) Improved multiplex PCR assay for simultaneous detection of Bacillus cereus emetic and enterotoxic isolates. Food Sci Biotechnol 21:1439–1444

    Google Scholar 

  • Kirk MD, Pires SM, Black RE, Caipo M, Crump JA, Devleesschauwer B, Dopfer D, Fazil A, Fischer-Walker CL, Hald T, Hall AJ, Keddy KH, Lake RJ, Lanata CF, Torgerson PR, Havelaar AH, Angulo FJ (2015) World Health Organization estimates of the global and regional disease burden of 22 foodborne bacterial, protozoal, and viral diseases, 2010: a data synthesis. PLoS Med 12:e1001921

    PubMed  PubMed Central  Google Scholar 

  • Kotiranta A, Lounatmaa K, Haapasalo M (2000) Epidemiology and pathogenesis of B. cereus infections. Microbes Infect 2:189–198

    CAS  PubMed  Google Scholar 

  • Kramer JM, Gilbert RJ (1989) B. cereus and other Bacillus species. In: Doyle MP (ed) Foodborne bacterial pathogens. Marcel Dekker, New York, pp 21–77

    Google Scholar 

  • Lakhanpal P, Panda AK, Rajesh Chahota, Shivani Choudhary, Thakur SD (2019) Incidence and antimicrobial susceptibility of Staphylococcus aureus isolated from ready-to-eat animal foods from tourist destinations of north western Himalayas, Himachal Pradesh, India. J Food Sci Technol 56:1078–1083

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lindbäck T, Fagerlund A, Rødland MS, Granum PE (2004) Characterization of the Bacillus cereus nhe enterotoxin. Microbiology 150:3959–3967

    PubMed  Google Scholar 

  • Logan NA (2011) Bacillus and relatives in food borne illness. J Appl Microbiol 112:417–429

    PubMed  Google Scholar 

  • Ngamwongsatit P, Buasri W, Pianariyanon P, Pulsrikarn C, Ohba M, Assavanig A, Panbangred W (2008) Broad distribution of enterotoxin genes (hblCDA, nheABC, cytK and entFm) among Bacillus thuringiensis and Bacillus cereus as shown by novel primers. Int J Food Microbiol 121:352–356

    CAS  PubMed  Google Scholar 

  • Organji SR, Abulreesh HH, Elbanna K, Ebrahim G, Osman H, Khider M (2015) Occurrence and characterization of toxigenic Bacillus cereus in food and infant feces. Asian Pac J Trop Biomed 5:515–520

    Google Scholar 

  • Owusu-Kwarteng J, Wuni A, Akabanda F, Tano-Debrah K, Jespersen L (2017) Prevalence, virulence factor genes and antibiotic resistance of Bacillus cereus sensu lato isolated from dairy farms and traditional dairy products. BMC Microbiol 17:65

    PubMed  PubMed Central  Google Scholar 

  • Perera ML, Ranasinghe GR (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

    CAS  PubMed  Google Scholar 

  • Public Health England (2018) UK standards for microbiology investigations. Identification of Bacillus species. Standards Unit, Microbiology Services, Public Health England. Bacteriology-identification 3.1, pp 1–27. https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/697260/ID_9i3.1.pdf. Accessed on 12 May 2018

  • Rajkovic A (2014) Microbial toxins and low level of food borne exposure. Trends Food Sci Technol 38:149–157

    CAS  Google Scholar 

  • Rather MA, Aulak RS, Gill JPS, Rao TS, Hassan MN (2011) Direct detection of Bacillus cereus and its enterotoxigenic genes in meat and meat products by polymerase chain reaction. J Adv Vet Res 1:99–104

    Google Scholar 

  • Roy A, Moktan B, Sarkar PK (2007) Characteristics of Bacillus cereus isolates from legume-based Indian fermented foods. Food Control 18:1555–1564

    CAS  Google Scholar 

  • Sadashiv SO, Kaliwal BB (2014) Isolation, characterization and antibiotic resistance of Bacillus spp. from bovine mastitis in the region of north Karnataka, India. Int J Curr Microbiol Appl Sci 3:360–373

    Google Scholar 

  • Salem NA, Jakee JE, Nasef SA, Badr H (2015) Prevalence of Bacillus cereus in milk and milk products. Anim Health Res J 3:168–172

    Google Scholar 

  • Schneider KR, Parish M E, Goodrich RM, Cookingham T (2004) Preventing food borne illness: B. cereus and Bacillus anthracis. Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida. FSHN04-05

  • Singh VK, Shukla S, Chaturvedi A (2015) Study the incidence of Bacillus cereus isolates from dairy foods. Pharm Innov J 3:41–43

    CAS  Google Scholar 

  • Sood B, Sahota PP, Hunjan M (2017) Multidrug resistant Bacillus cereus in fresh vegetables: a serious burden to public health. Int J Curr Microbiol Appl Sci 6:649–661

    CAS  Google Scholar 

  • Tewari A, Abdullah S (2015) Bacillus cereus food poisoning: international and Indian perspective. J Food Sci Technol 52:2500–2511

    CAS  PubMed  Google Scholar 

  • Tewari A, Singh SP, Singh R (2015) Incidence and enterotoxigenic profile of Bacillus cereus in meat and meat products of Uttarakhand, India. J Food Sci Technol 52:1796–1801

    CAS  PubMed  Google Scholar 

  • Tirloni E, Ghelardi E, Celandroni F, Bernardi C, Stella S (2017) Effect of dairy product environment on the growth of Bacillus cereus. J Dairy Sci 100:7026–7034

    CAS  PubMed  Google Scholar 

  • Vos PD, Garrity GM, Jones D, Krieg NR, Ludwig W, Rainey FA, Schleifer KH, William BW (2009) Bergey’s manual of systematic bacteriology, vol 3, 2nd edn. The Firmicutes. Springer, Dordrecht

    Google Scholar 

  • Waters AE, Contente-Cuomo T, Buchhagen J, Liu CM, Watson L, Pearce K, Foster JT, Bowers J, Driebe EM, Engelthaler DM, Keim PS, Price LB (2011) Multidrug-resistant Staphylococcus aureus in US meat and poultry. Clin Infect Dis 52:1227–1230

    PubMed  PubMed Central  Google Scholar 

  • Yang IC, Shih DY, Huang TP, Huang YP, Wang JY, Pan TM (2005) Establishment of a novel multiplex PCR assay and detection of toxigenic strains of the species in the Bacillus cereus group. J Food Prot 68:2123–2130

    CAS  PubMed  Google Scholar 

  • Yıbar A, Çetinkaya F, Soyutemiz E, Yaman G (2017) Prevalence, enterotoxin production and antibiotic resistance of Bacillus cereus isolated from milk and cheese. Kafkas Univ Vet Fak Derg 23(4):635–642

    Google Scholar 

  • Yusuf U, Kotwal SK, Gupta S, Ahmed T (2018) Identification and antibiogram pattern of Bacillus cereus from the milk and milk products in and around Jammu region. Vet World 11:186–191

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We are thankful to CSK Himachal Pradesh Agricultural University for financial and infrastructure support for this research. The necessary help and technical support provided by Head, Department of Veterinary Microbiology, Dr. GC Negi College of Veterinary and Animal Sciences, CSK Himachal Pradesh Agricultural is duly acknowledged.

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Correspondence to Sidharath Dev Thakur.

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Rana, N., Panda, A.K., Pathak, N. et al. Bacillus cereus: public health burden associated with ready-to-eat foods in Himachal Pradesh, India. J Food Sci Technol 57, 2293–2302 (2020). https://doi.org/10.1007/s13197-020-04267-y

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