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Using Culture and PCR Technique for Detection of Escherichia coli O157:H7 in Duhok Cheese

Yıl 2015, Cilt: 18 Sayı: 1, 31 - 34, 02.11.2015

Öz

ABSTRACT: This study was aimed to investigate the presence of Escherichia coli O157:H7 in Duhok cheese by culture and Polymerase Chain Reaction (PCR) technique. The cheese samples (n:120) were purchased from different supermarkets and shops from Zakho, Akre, Amedy and Duhok cities during September 2011 to February 2012.

X-gal positive, MUG negative, indole positive colonies were found as 10% (12/120) of samples. These colonies were confirmed by biochemical tests (API E 20 test, latex agglutination tests) and results showed that 7 (5.83%) of E. coli isolates from cheese were positive for O157:H7. These isolates were also confirmed by PCR assay using specific primers for rfbO157 and flicH7. The numbers of positive isolates for rfbO157 and flicH7 were 3.33 and 0.83%, respectively. The PCR results confirmed as 0.83% of samples were E. coli O157:H7.

Key words: Duhok cheese, Escherichia coli O157:H7, Cultural technique, PCR technique

 

Kültür ve PZR Yöntemi ile Duhok Peynirinde Escherichia coli O157:H7'nin Araştırılması

 

ÖZET: Bu çalışmada Duhok peynirinde of E. coli O157:H7'nin varlığı kültür ve Polimeraz Zincir Reaksiyonu tekniği (PZR) ile araştırılmıştır. Peynir örnekleri (n:120) Zakho, Akre, Amedy ve Duhok şehirlerinden Eylül 2011-Şubat 2012 tarihleri arasında yerel market ve bakkallardan temin edilmiştir.

X-gal pozitif, MUG negatif,  indol pozitif olan %10 (12/120) oranında şüpheli koloni tespit edilmiştir. Şüpheli kolonilere biyokimyasal testler uygulanmış (API E 20 ve latex aglütinasyon testi) ve 7 (%5,83) örnekte E. coli O157:H7 tespit edilmiştir. Bu izolatlararfbO157 ve flicH7 primerleri kullanılarak PCR yöntemi ile doğrulama işlemi uygulanmıştır. İzolatlar %3,33 oranında O157 ve 0,83 oranında H7 antijeni pozitif olarak tespit edilmiştir. PCR analizlerinin sonunda örneklerin %0,83'ünün E. coli O157:H7 olduğu gözlenmiştir.

Anahtar Kelimeler: Duhok Peyniri, Escherichia coli O157:H7, Kültür tekniği, PZR tekniği

Kaynakça

  • Abdul-Raouf, U.M., Ammar, M.S., Beuchat, L.R. 1996. Isolation of Escherichia coli O157:H7 from some Egyptian foods. International Journal of Food Microbiology, 29: 423-426.
  • Altekruse, S.F., Timbo, B.B., Mowbray, J.C., Bean, N.H., outbreaks of human illness in the United States, 1972 to 1992: Sanitary manufacturing practices protect consumers. Journal of Food Protection, 61:1405-1407. 1998. Cheese-associated
  • Andreoli, S.P., Trachtman, H., Acheson, D.W.K., Siegler, R.L., Obrig, T.G. 2002. Hemolytic uremic syndrome: epidemiology, pathophysiology, and therapy. Pediatric Nephrology, 17: 293-297.
  • Beutin, L., Krause, G., Zimmermann, S., Kaulfuss, S., Gleier, K. 2004. Characterization of Shiga toxin- producing Escherichia coli strains isolated from human patients in Germany over a 3-year period. Journal ofClinical Microbiology, 42: 1099-1108.
  • Blanco, J., Blanco, M., Blanco, J.E., Mora, A., González, E..A., Bernárdez, M.I., Alonso, M.P., Coira, A., Rodríguez, A., Rey, J., Alonso, J.M., Usera, M.A. 2003. Verotoxin-Producing Escherichia coli in Spain: Prevalence, Serotypes, and Virulence Genes of O157:H7 and non-O157 VTEC in Ruminants, Raw Beef Products, and Humans. Experimental Biology and Medicine, 228: 345-351.
  • Centers for Disease Control and Prevention (CDC). 2007 Escherichia coli O157:H7 infection associated with drinking raw milk- Washington and Oregon, November December 2005. Morbidity Mortality Weekly Report, 56: 165-167. Weagant, Feng, P., S.D., Jinneman, K. 2011. Diarrheagenic Escherichia coli. FDA-BAM.
  • Ferens, W.A., Hovde, C.J. 2011. Escherichia coli O157:H7: Animal Reservoir and Sources of Human Infection. Foodborne Pathogens and Disease, 8: 465-487.
  • Gannon, V.P.J., D’souza, S., Graham, T., King, R.K., Rahn, K., Read, S. 1997. Use of flagellar H7 gene as a target in multiplex PCR assay and improved specificity and identification of enterohaemorrhagic Escherichia coli Microbiology, 35: 656-662. Journal of Clinical
  • Govaris, A., Koidis, P., Papatheodorou, K. 2002. Behavior of Escherichia coli O157:H7 in sour milk, cow’s milk yogurt and ewe’s milk yogurt. Journal of Dairy Research, 69: 655-660.
  • Honish, L., Predy, G., Hislop, N., Chui, L., Kowalewska-Grochowska, K., Trottier, L., Kreplin, C., Zazulak, I. 2005. An outbreak of Escherichia coli O157:H7 hemorrhagic colitis associated with unpasteurized Gouda cheese. Canadian Journal of Public Health, 96: 182-184.
  • Karmali, M.A. 1989. Infection by verotoxin-producing Escherichia coli. Clinical Microbiology Reviews, 2: 15-38.
  • Morgan, D., Newman, C.P., Hutchinson, D.N., Walker, A.M., Rowe, B., Majid, F. 1993.Verotoxin producing associated with the consumption of yoghurt. Epidemiology and Infection, 111: 181-187.
  • Öksüz, Ö., Arici, M., Kurultay, S., Gümüş, T. 2004. Incidence of Escherichia coli O157 in raw milk and white pickled cheese manufactured from raw milk in Turkey. Food Control, 15, 453-456.
  • Paton, A.W., Paton, J.C. 1998. Detection and Characterization of Shiga Toxigenic Escherichia coli by Using Multiplex PCR Assays for stx1, stx2, eaeA, EnterohemorrhagicE. colihlyA, rfbO111 and rfbO157. Journal of Clinical Microbiology, 36(2): 598-602.
  • Rahimi, E., Chaleshtori, S.S., Parsaei, P. 2011. Prevalence Escherichia coli O157 isolated from traditional cheese, ice creamand yoghurt in Iran. African Journal of Microbiology Research, 5(22): 3706- 3710. resistance of
  • Schlesser, J.E., Gerdes, R., Ravishankar, S., Madsen, K., Mowbray, J., Teo, A.Y.L. 2006. Survival of a five-strain cocktail of Escherichia coli O157:H7 during a 60 day aging period of cheddar cheese made from unpasteurized milk. Journal of Food Protection, 69: 90-99.
  • Schrade, J.P., Yager, J. 2001. Implication of milk and milk products in food disease in France and in different industrialized countries. International Journal of Food Microbiology, 67: 1-17.
  • Stampi, S., Caprioli, A., De Luca, G., Quaglio, P., Sacchetti, R., Zanetti, F. Escherichia coli O157 in bovine meat products in Northern Microbiology, 90: 257-262. 2004. Detection of
  • Italy. International Journal of Food
  • Tobias, J., Vutukuru, S.R. 2012. Simple and rapid multiplex PCR for identification of the main human diarrheagenicEscherichia Research. 167 (9):564-570. coli. Microbiological

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Yıl 2015, Cilt: 18 Sayı: 1, 31 - 34, 02.11.2015

Öz

Bu çalışmada Duhok peynirinde of E. coli O157:H7'nin varlığı kültür ve Polimeraz Zincir Reaksiyonutekniği (PZR) ile araştırılmıştır. Peynir örnekleri (n:120) Zakho, Akre, Amedy ve Duhok şehirlerinden Eylül 2011Şubat 2012 tarihleri arasında yerel market ve bakkallardan temin edilmiştir.X-gal pozitif, MUG negatif, indol pozitif olan %10 (12/120) oranında şüpheli koloni tespit edilmiştir. Şüphelikolonilere biyokimyasal testler uygulanmış (API E 20 ve latex aglütinasyon testi) ve 7 (%5,83) örnekte E. coliO157:H7 tespit edilmiştir. Bu izolatlararfbO157 ve flicH7 primerleri kullanılarak PCR yöntemi ile doğrulama işlemiuygulanmıştır. İzolatlar %3,33 oranında O157 ve 0,83 oranında H7 antijeni pozitif olarak tespit edilmiştir. PCRanalizlerinin sonunda örneklerin %0,83'ünün E. coli O157:H7 olduğu gözlenmiştir

Kaynakça

  • Abdul-Raouf, U.M., Ammar, M.S., Beuchat, L.R. 1996. Isolation of Escherichia coli O157:H7 from some Egyptian foods. International Journal of Food Microbiology, 29: 423-426.
  • Altekruse, S.F., Timbo, B.B., Mowbray, J.C., Bean, N.H., outbreaks of human illness in the United States, 1972 to 1992: Sanitary manufacturing practices protect consumers. Journal of Food Protection, 61:1405-1407. 1998. Cheese-associated
  • Andreoli, S.P., Trachtman, H., Acheson, D.W.K., Siegler, R.L., Obrig, T.G. 2002. Hemolytic uremic syndrome: epidemiology, pathophysiology, and therapy. Pediatric Nephrology, 17: 293-297.
  • Beutin, L., Krause, G., Zimmermann, S., Kaulfuss, S., Gleier, K. 2004. Characterization of Shiga toxin- producing Escherichia coli strains isolated from human patients in Germany over a 3-year period. Journal ofClinical Microbiology, 42: 1099-1108.
  • Blanco, J., Blanco, M., Blanco, J.E., Mora, A., González, E..A., Bernárdez, M.I., Alonso, M.P., Coira, A., Rodríguez, A., Rey, J., Alonso, J.M., Usera, M.A. 2003. Verotoxin-Producing Escherichia coli in Spain: Prevalence, Serotypes, and Virulence Genes of O157:H7 and non-O157 VTEC in Ruminants, Raw Beef Products, and Humans. Experimental Biology and Medicine, 228: 345-351.
  • Centers for Disease Control and Prevention (CDC). 2007 Escherichia coli O157:H7 infection associated with drinking raw milk- Washington and Oregon, November December 2005. Morbidity Mortality Weekly Report, 56: 165-167. Weagant, Feng, P., S.D., Jinneman, K. 2011. Diarrheagenic Escherichia coli. FDA-BAM.
  • Ferens, W.A., Hovde, C.J. 2011. Escherichia coli O157:H7: Animal Reservoir and Sources of Human Infection. Foodborne Pathogens and Disease, 8: 465-487.
  • Gannon, V.P.J., D’souza, S., Graham, T., King, R.K., Rahn, K., Read, S. 1997. Use of flagellar H7 gene as a target in multiplex PCR assay and improved specificity and identification of enterohaemorrhagic Escherichia coli Microbiology, 35: 656-662. Journal of Clinical
  • Govaris, A., Koidis, P., Papatheodorou, K. 2002. Behavior of Escherichia coli O157:H7 in sour milk, cow’s milk yogurt and ewe’s milk yogurt. Journal of Dairy Research, 69: 655-660.
  • Honish, L., Predy, G., Hislop, N., Chui, L., Kowalewska-Grochowska, K., Trottier, L., Kreplin, C., Zazulak, I. 2005. An outbreak of Escherichia coli O157:H7 hemorrhagic colitis associated with unpasteurized Gouda cheese. Canadian Journal of Public Health, 96: 182-184.
  • Karmali, M.A. 1989. Infection by verotoxin-producing Escherichia coli. Clinical Microbiology Reviews, 2: 15-38.
  • Morgan, D., Newman, C.P., Hutchinson, D.N., Walker, A.M., Rowe, B., Majid, F. 1993.Verotoxin producing associated with the consumption of yoghurt. Epidemiology and Infection, 111: 181-187.
  • Öksüz, Ö., Arici, M., Kurultay, S., Gümüş, T. 2004. Incidence of Escherichia coli O157 in raw milk and white pickled cheese manufactured from raw milk in Turkey. Food Control, 15, 453-456.
  • Paton, A.W., Paton, J.C. 1998. Detection and Characterization of Shiga Toxigenic Escherichia coli by Using Multiplex PCR Assays for stx1, stx2, eaeA, EnterohemorrhagicE. colihlyA, rfbO111 and rfbO157. Journal of Clinical Microbiology, 36(2): 598-602.
  • Rahimi, E., Chaleshtori, S.S., Parsaei, P. 2011. Prevalence Escherichia coli O157 isolated from traditional cheese, ice creamand yoghurt in Iran. African Journal of Microbiology Research, 5(22): 3706- 3710. resistance of
  • Schlesser, J.E., Gerdes, R., Ravishankar, S., Madsen, K., Mowbray, J., Teo, A.Y.L. 2006. Survival of a five-strain cocktail of Escherichia coli O157:H7 during a 60 day aging period of cheddar cheese made from unpasteurized milk. Journal of Food Protection, 69: 90-99.
  • Schrade, J.P., Yager, J. 2001. Implication of milk and milk products in food disease in France and in different industrialized countries. International Journal of Food Microbiology, 67: 1-17.
  • Stampi, S., Caprioli, A., De Luca, G., Quaglio, P., Sacchetti, R., Zanetti, F. Escherichia coli O157 in bovine meat products in Northern Microbiology, 90: 257-262. 2004. Detection of
  • Italy. International Journal of Food
  • Tobias, J., Vutukuru, S.R. 2012. Simple and rapid multiplex PCR for identification of the main human diarrheagenicEscherichia Research. 167 (9):564-570. coli. Microbiological
Toplam 20 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Bölüm MİKROBİYOLOJİ (Microbiology)
Yazarlar

Özlem Turgay

Sowayla Mıkaeel Bu kişi benim

Yayımlanma Tarihi 2 Kasım 2015
Yayımlandığı Sayı Yıl 2015 Cilt: 18 Sayı: 1

Kaynak Göster

APA Turgay, Ö., & Mıkaeel, S. (2015). Using Culture and PCR Technique for Detection of Escherichia coli O157:H7 in Duhok Cheese. KSÜ Doğa Bilimleri Dergisi, 18(1), 31-34. https://doi.org/10.18016/ksujns.94848
AMA Turgay Ö, Mıkaeel S. Using Culture and PCR Technique for Detection of Escherichia coli O157:H7 in Duhok Cheese. KSÜ Doğa Bilimleri Dergisi. Kasım 2015;18(1):31-34. doi:10.18016/ksujns.94848
Chicago Turgay, Özlem, ve Sowayla Mıkaeel. “Using Culture and PCR Technique for Detection of Escherichia Coli O157:H7 in Duhok Cheese”. KSÜ Doğa Bilimleri Dergisi 18, sy. 1 (Kasım 2015): 31-34. https://doi.org/10.18016/ksujns.94848.
EndNote Turgay Ö, Mıkaeel S (01 Kasım 2015) Using Culture and PCR Technique for Detection of Escherichia coli O157:H7 in Duhok Cheese. KSÜ Doğa Bilimleri Dergisi 18 1 31–34.
IEEE Ö. Turgay ve S. Mıkaeel, “Using Culture and PCR Technique for Detection of Escherichia coli O157:H7 in Duhok Cheese”, KSÜ Doğa Bilimleri Dergisi, c. 18, sy. 1, ss. 31–34, 2015, doi: 10.18016/ksujns.94848.
ISNAD Turgay, Özlem - Mıkaeel, Sowayla. “Using Culture and PCR Technique for Detection of Escherichia Coli O157:H7 in Duhok Cheese”. KSÜ Doğa Bilimleri Dergisi 18/1 (Kasım 2015), 31-34. https://doi.org/10.18016/ksujns.94848.
JAMA Turgay Ö, Mıkaeel S. Using Culture and PCR Technique for Detection of Escherichia coli O157:H7 in Duhok Cheese. KSÜ Doğa Bilimleri Dergisi. 2015;18:31–34.
MLA Turgay, Özlem ve Sowayla Mıkaeel. “Using Culture and PCR Technique for Detection of Escherichia Coli O157:H7 in Duhok Cheese”. KSÜ Doğa Bilimleri Dergisi, c. 18, sy. 1, 2015, ss. 31-34, doi:10.18016/ksujns.94848.
Vancouver Turgay Ö, Mıkaeel S. Using Culture and PCR Technique for Detection of Escherichia coli O157:H7 in Duhok Cheese. KSÜ Doğa Bilimleri Dergisi. 2015;18(1):31-4.