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The prevalence and molecular typing of Clostridium perfringens in ground beef and sheep meats

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Abstract

This study was aimed at the investigation of the prevalence of Clostridium perfringens in ground beef and sheep meats and to detect the cpa, cpb, etx, iA, cpe and cpb2 toxin genes by multiplex PCR. Obtained from retail markets and butchers, 100 ground meat samples, 50 of which were beef and 50 of which were sheep meat samples, were examined. Out of the 50 ground beef samples 48 (96 %) and out of the 50 ground sheep meat samples 44 (88 %) were contaminated with C. perfringens. According to microscopic examination and the results of biochemical tests, 262 positive isolates obtained from these samples were identified as C. perfringens. Multiplex PCR results demonstrated that, out of the 262 isolates, 203 (77.4 %) possessed only the cpa gene (type A), while 20 (7.6 %) carried the cpa and cpb2 toxin genes (type A-cpb2), and 6 (2.2 %) the cpa and cpe toxin genes (type A-cpe). Furthermore, it was determined that, out of the 262 C. perfringens isolates, 4 (1.5 %), 10 (3.8 %) and 19 (7.2 %) were of the types B, C and D, respectively.

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References

  • Albini S, Brodard I, Jaussi A, Wollschlaeger N, Frey J, Miserez R, Abril C (2008) Real-time multiplex PCR assays for reliable detection of Clostridium perfringens toxin genes in animal isolates. Vet Microbiol 127:179–185

    Article  CAS  PubMed  Google Scholar 

  • Ali MS, Fung DYC (1991) Occurrence of Clostridium perfringens in ground beef and ground turkey evaluated by three methods. J Food Saf 11:197–203

    Article  Google Scholar 

  • Al-Khaldi SF, Myers KM, Rasooly A, Chizhikov V (2004) Genotyping of Clostridium perfringens toxins using multiple oligonucleotide microarray hybridization. Mol Cell Probes 18:359–367

    Article  CAS  PubMed  Google Scholar 

  • Atwa EI, Abou El-Roos AN (2011) Incidence of Clostridium perfringens in meat products at some Egyptian governorates. Int J Microbiol Res 2:196–203

    Google Scholar 

  • Baums CG, Schotte U, Amtsberg G, Goethe R (2004) Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates. Vet Microbiol 100:11–16

    Article  CAS  PubMed  Google Scholar 

  • Billington SJ, Wieckowski EU, Sarker MR, Bueschel D, Songer JG, McClane BA (1998) Clostridium perfringens type E animal enteritis isolates with highly conserved, silent enterotoxin gene sequences. Infect Immun 66:4531–4536

    CAS  PubMed Central  PubMed  Google Scholar 

  • Brynestad S, Granum PE (2002) Clostridium perfringens and foodborne infections. Int J Food Microbiol 74:195–202

    Article  PubMed  Google Scholar 

  • Çakmak Ö (2001) Ankara’da tüketime sunulan sığır kıymalarında C. perfringens’in varlığı ve kontaminasyon düzeyinin belirlenmesi. Yüksek Lisans tezi, Ankara Üniversitesi, Ankara

  • Çakmak Ö, Bilir Ormancı S, Tayfur M, Erol I (2006) Presence and contamination level of Clostridium perfringens in raw frozen ground poultry and poultry burgers. Turk J Vet Anim Sci 30:101–105

    Google Scholar 

  • Erol I, Goncuoglu M, Ayaz ND, Bilir Ormanci FS, Hildebrandt G (2008) Molecular typing of Clostridium perfringens isolated from turkey meat by multiplex PCR. Lett Appl Microbiol 47:31–34

    Article  CAS  PubMed  Google Scholar 

  • Fach P, Popoff MR (1997) Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test. Appl Environ Microbiol 63:4232–4236

    CAS  PubMed Central  PubMed  Google Scholar 

  • Fisher DJ (2006) Clostridium perfringens beta2 toxin: a potential accessory toxin in gastrointestinal diseases of humans and domestic animals. University of Pittsburgh School of Medicine, Pittsburgh

    Google Scholar 

  • Fisher DJ, Miyamoto K, Harrison B, Akimoto S, Sarker MR, McClane BA (2005) Association of beta2 toxin production with Clostridium perfringens type A human gastrointestinal disease isolates carrying a plasmid enterotoxin gene. Mol Microbiol 56:747–762

    Article  CAS  PubMed  Google Scholar 

  • Garmory HS, Chanter N, French NP, Bueschel D, Songer JG, Titball RW (2000) Occurrence of Clostridium perfringens β2-toxin amongst animals, determined using genotyping and subtyping PCR assays. Epidemiol Infect 124:61–67

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gibert M, Jolivet-Renaud C, Popoff MR (1997) Beta2 toxin, a novel toxin produced by Clostridium perfringens. Gene 203:65–73

    Article  CAS  PubMed  Google Scholar 

  • Gökçe Hİ, Genç O, Sözmen M, Gökçe G (2007) Determination of Clostridium perfringens toxins types in sheeps with suspected enterotoxemia in Kars province. Turk J Vet Anim Sci 31:355–360

    Google Scholar 

  • Guang-Hua W, Xiao-Ling Q (1994) The incidence of C perfringens, S. aureus salmonella and L. monocytogenes in retail meat and meat products in Beijing. Fleischwirtsch 74:288–290

    Google Scholar 

  • Guran HS, Oksuztepe G (2013) Detection and typing of Clostridium perfringens from retail chicken meat parts. Lett Appl Microbiol 57:77–82

    Article  CAS  PubMed  Google Scholar 

  • Heikinheimo A, Korkeala H (2005) Multiplex PCR assay for toxinotyping Clostridium perfringens isolates obtained from Finnish broiler chickens. Lett Appl Microbiol 40:407–411

    Article  CAS  PubMed  Google Scholar 

  • Johansson A, Aspan A, Bagge E, Båverud V, Engström BE, Johansson KE (2006) Genetic diversity of Clostridium perfringens type A isolates from animals, food poisoning outbreaks and sludge. BMC Microbiol 6:47

    Article  PubMed Central  PubMed  Google Scholar 

  • Kalender H, Kılıç A, Atıl E (2007) Enterotoxemia in a cow due to Clostridium perfringens type A. Turk J Vet Anim Sci 31:83–84

    Google Scholar 

  • Kamber U, Gokce HI, Elmali M (2007) Clostridium perfringens and its toxins in minced meat from Kars, Turkey. Food Addit Contam 24:673–678

    Article  CAS  PubMed  Google Scholar 

  • Koç R, Gökçe Hİ (2007) Determination of the toxins and biotypes of C. perfringens in diarrhoeic calves in the Kars district of Turkey. Turk J Vet Anim Sci 31:207–211

    Google Scholar 

  • Kokai-Kun JF, McClane BA (1997) Determination of functional regions of Clostridium perfringens enterotoxin through deletion analysis. Clin Infect Dis 25:S165–S167

    Google Scholar 

  • Krause P, Schmoldt R, Tolgay Z, Yurtyerg A (1972) Mikrobiologische und serologische Untersuchungen an Lebensmitteln in der Türkei. Fleischerei 1:83–86

    Google Scholar 

  • Lindström M, Heikinheimo A, Lahti P, Korkeala H (2011) Novel insights into the epidemiology of Clostridium perfringens type A food poisoning. Food Microbiol 28:192–198

    Article  PubMed  Google Scholar 

  • Lynch M, Painter J, Woodruff R, Braden C (2006) Surveillance for foodborne-disease outbreaks—United States, 1998–2002. Morbid Mortal Wkly Rep 10:1–34

    Google Scholar 

  • Meer RR, Songer JG (1997) Multiplex polymerase chain reaction assay for genotyping Clostridium perfringens. Am J Vet Res 58:702–705

    CAS  PubMed  Google Scholar 

  • Miki Y, Miyamoto K, Kaneko-Hirano I, Fujiuchi K, Akimoto S (2008) Prevalence and characterization of enterotoxin gene-carrying Clostridium perfringens isolates from retail meat products in Japan. Appl Environ Microbiol 74:5366–5372

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Miwa N, Nishina T, Kubo S, Honda H (1997) Most probable numbers of enterotoxigenic Clostridium perfringens in intestinal contents of domestic livestock detected by nested PCR. J Vet Med Sci 59:557–560

    Article  CAS  PubMed  Google Scholar 

  • Nordic committee on food analysis (2009) Clostridium perfringens. Determination in foods, feed and environmental samples. Method no. 95, 5th edn, Oslo

  • Özcan C, Gürçay M (2000) Elazığ ve çevresinde 1994–1998 yılları arasında küçük ruminantlarda enterotoksemi insidensi. Turk J Vet Anim Sci 24:283–286

    Google Scholar 

  • Petit L, Gibert M, Popoff MR (1999) Clostridium perfringens: toxinotype and genotype. Trends Microbiol 7:104–110

    Article  CAS  PubMed  Google Scholar 

  • Phillips D, Jordan D, Morris S, Jenson I, Sumner J (2006) A national survey of the microbiological quality of beef carcasses and frozen boneless beef in Australia. J Food Prot 69:1113–1117

    PubMed  Google Scholar 

  • Sancak YC, Boynukara B, Ağaoğlu S (1993) Van’da tüketime sunulan kıymaların mikrobiyolojik kalitesi. Yüzüncü Yıl Üniversitesi Veteriner Fakültesi Dergisi 44:73–86

    Google Scholar 

  • Sarıgüzel D (2005) Ankara‘da tüketime sunulan hindi kıymalarında Clostridium perfringens‘in varlığı ve elde edilen izolatlarda cpe geninin saptanması. Ankara Üniversitesi Sağlık Bilimleri Enstitüsü, Yüksek Lisans Tezi

  • Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson MA, Roy SL, Jones JL, Griffin PM (2011) Foodborne illness acquired in the United States-major pathogens. Emerg Infect Dis 17:7–15

    Article  PubMed Central  PubMed  Google Scholar 

  • Schalch B, Eisgruber H, Geppert P, Stolle A (1996) Vergleich von vier Routineverfahren zur Bestaetigung von Clostridium perfringens aus Lebensmitteln. Arch Lebensmittelhyg 47:27–30

    Google Scholar 

  • Seyitoğlu Ş, Cengiz Ş, Altun KS, Küçükkalem ÖF, Söztutmaz İ (2012) Investigation of Clostridium perfringens toxins in sheeps in the region of Erzurum by toxin neutralization, ELISA and lateks agglütination tests. J Etlik Vet Microbiol 23:39–43

    Google Scholar 

  • Smedley JG, Fisher DJ, Sayeed S, Chakrabarti G, McClane BA (2004) The enteric toxins of Clostridium perfringens. Rev Physiol Biochem Pharmacol 152:183–204

    Article  CAS  PubMed  Google Scholar 

  • Songer JG (1996) Clostridial enteric diseases of domestic animals. Clin Microbiol Rev 9:216–234

    CAS  PubMed Central  PubMed  Google Scholar 

  • SPSS (2003) SPSS statistical package, version 14.01. Chicago

  • Stagnitta PV, Micalizzi B, De Guzmán AMS (2002) Prevalence of enterotoxigenic Clostridium perfringens in meats in San Luis, Argentina. Anaerobe 8:253–258

    Article  Google Scholar 

  • Tooloei M, Masodei MH (2008) The distribution and prevalence rate of enterotoxaemia in sheep in East Azerbaijan Iran in spring. J Anim Vet Adv 7:1434–1439

    Google Scholar 

  • Turkey Central Research Institute for Field Crop (2013). http://www.tarlabitkileri.gov.tr/gis_web/bultenler/2013_1.pdf. Accessed 29 Dec 2013

  • United Kingdom Food Standards Agency (2010) A UK-wide survey of microbiological contamination of fresh red meats on retail sale. http://www.food.gov.uk/multimedia/pdfs/fsis0110redmeat.pdf. Accessed 25 June 2013

  • Uzal FA (2004) Diagnosis of Clostridium perfringens intestinal infections in sheep and goats. Anaerobe 10:135–143

    Article  CAS  PubMed  Google Scholar 

  • Wahl E, Rømma S, Granum PE (2013) A Clostridium perfringens outbreak traced to temperature-abused beef stew, Norway, 2012. Eurosurveillance 18:28

    Google Scholar 

  • Wen Q, McClane BA (2004) Detection of enterotoxigenic Clostridium perfringens type A isolates in American retail foods. Appl Environ Microbiol 70:2685–2691

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

The present study was supported by the Dicle University Research Fund (DUBAP/10-VF-83). We thank Şenay Erçelik and Zülküf Emre Papatya for their excellent laboratory assistance.

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Correspondence to Husnu Sahan Guran.

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Guran, H.S., Vural, A. & Erkan, M.E. The prevalence and molecular typing of Clostridium perfringens in ground beef and sheep meats. J. Verbr. Lebensm. 9, 121–128 (2014). https://doi.org/10.1007/s00003-014-0866-z

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