Todor STOYANCHEV, Koycho KOEV, Georgi ZHELEV, Plamen MARUTSOV, Krasimira GOSPODINOVA, Vladimir PETROV

Çiğ Sütte Shiga-Toksin Üreten Escherichia coli’nin stx1, stx2, eaeA ve H7 Bakımından Multipleks PCR ile Moleküler İncelenmesi ve Karakterizasyonu

Bulgaristan’da süt sığırı işletmelerinden toplanan çiğ sütlerde Shiga-toksin üreten Escherichia coli (STEC) varlığı değerlendirildi. Toplam 20 bölgesel çiftlikten sadece 4’ü yıllık dışkıyla STEC yayma miktarı temel alınarak süt toplama kaynağı olarak seçildi. Tüm örnekler selektif zenginleştirme sonrasında PCR ile analiz edildi. Pozitif örnekler daha sonra moleküler test ile değerlendirildi. İki örnek (%9.08) stx1 için, 1 örnek (%4.54) stx2 için, 10 örnek (%45.54) eae için ve 12 örnek (%54.54) H7 kodlayan genler için pozitif bulundu.

Molecular Screening and Characterization of Shiga Toxin-producing Escherichia coli By Multiplex PCR Assays for stx1, stx2, eaeA, H7 in Raw Milk

The presence of Shiga-toxin producing Escherichia coli (STEC) was evaluated in bulk milk from dairy cattle farms in Bulgaria. From 20 regionalfarms only 4 were selected as a source for milk sampling on the basis of their annual STEC shedding in feces. All the samples were PCRanalyzed after selective enrichment. Positive samples were detected in subsequent molecular testing as: two of samples (9.08%) werepositive for stx1, one sample (4.54%) for stx2, 10 samples (45.54%) for eae and 12 (54.54%) for H7 coding genes.

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1. Caro I, Fernandez-Barata VM, Alonso-Llamazares A, Garcia-Armesto MR: Detection, occurrence, and characterization of Escherichia coli O157:H7 from raw ewe’s milk in Spain. J Food Prot, 69 (4): 920-924, 2006.
2. Pruimboom-Brees, IM, Morgan TW, Ackermann MR, Nystrom ED, Samuel JE, Cornick NA, Moon HW: Cattle lack vascular receptors for Escherichia coli O157:H7 Shiga toxins. Proc Natl Acad Sci U S A, 97, 10325- 10329, 2000. DOI: 10.1073/pnas.190329997
3. Oliver SP, Boor KJ, Murphy SC, Murinda SE: Food safety hazards associated with consumption of raw milk. Foodborne Pathog Dis, 6, 793- 806, 2009. DOI: 10.1089/fpd.2009.0302
4. Li R, Tan X, Xiao J, Wang H, Liu Z, Zhou M, Bi W, Myamoto T: Molecular screening and characterization of Shiga toxin-producing Escherichia coli in retail foods. Food Control, 60, 180-188, 2016. DOI: 10.1016/j.foodcont.2015.07.045
5. Vimont A, Vernozy-Rozand C, Montet MP, Bavai C, Fremaux B, Delignette-Muller ML: Growth of Shiga-Toxin producing Escherichia coli (STEC) and bovine feces background microflora in various enrichment protocols. Vet Microbiol, 123, 274-281, 2007. DOI: 10.1016/j.vetmic. 2007.02.003
6. Fernandez D, Padola NL: Verocytotoxigenic Escherichia coli: Several aspects and also the dairy farms. Rev Argent Microbiol, 44, 312-323, 2012.
7.Food standard agency: E. coli O157 control of cross-contamination. 2011. https://www.eastdunbarton.gov.uk/filedepot_download/11559/547; Accessed: 17.04.2018.
8. Bos J: Multy-County Outbreak of Escherichia coli O157:H7-Subtype MOE014, MBE104, Among Persons who Consumed Unpasteurized Goat’s Milk-Spring 2008, 2008. https://www.marlerblog.com/uploads/ image/Multi-County%20Outbreak%20of%20E.%20coli%20O157H7%20 among%20persons%20who%20consumed%20unpasteurized%20goat‘s %20mealk,%20Missouri,%20Spring%202008.pdf; Accessed: 17.04.2018.
9. Koev K, Zhelev G, Marutsov P, Gospodinova K, Petrov V: Isolation and primary identification of shiga toxin producing Escherichia coli O157 in dairy cattle. BJVM, 21 (4): 445-450, 2018, DOI: 10.15547/bjvm.1077
10. Kim H, Bhunia AK: SEL, a selective enrichment broth for simultaneous growth of Salmonella enterica, Escherichia coli O157:H7 and Listeria monocytogenes. Appl Environ Microbiol, 74, 4853-4866, 2008. DOI: 10.1128/ AEM.02756-07
11. Gannon VPJ, D’Souza S, Graham T, King RK, Rahn K, Read S: Use of the flagellar H7 gene as target in multiplex PCR assays and improved specificity in identification of enterohaemorrhagic Escherichia coli strains. J Clin Microbiol, 35, 656-662, 1997.
12. Hlavsa MC, Roberts VA, Kahler AM, Hilborn ED, Mecher TR, Beach MJ, Wade TJ, Yoder JS: Outbreaks of illness associated with recreational water-United States, 2011-2012. MMWR Morb Mortal Wkly Rep, 64, 668- 672, 2015.
13. McKee R, Madden RH, Gilmour A: Occurrence of verocytotoxinproducing Escherichia coli in dairy and meat processing environments. J Food Prot, 66 (9): 1576-80, 2003.
14. Brenjchi M, Jamshidi A, Farzaneh N, Bassami MR: Identification of shiga toxin producing Escherichia coli O157:H7 in raw cow milk samples from dairy farms in Mashhad using multiplex PCR assay. Iran J Vet Res, 12 (2): 145-149, 2011. DOI: 10.22099/IJVR.2011.56
15. Garbaj AM, Awad EM, Azwai SM, Abolghait SK, Naas HT, Moawad AA, Gammoudi FT, Barbieri I, Eldaghayes IM: Enterohemorrhagic Escherichia coli O157 in milk and dairy products from Libya: Isolation and molecular identification by partial sequencing of 16S rDNA. Vet World, 9, 1184-1189, 2016. DOI: 10.14202/vetworld.2016.1184-1189
16. Rahimi E, Khamesipour F, Yazdi F, Momtaz H: Isolation and characterization of enterohaemorragic Escherichia coli O157:H7 and EHEC O157:NM from raw bovine, camel, water buffalo, caprine and ovine milk in Iran. Kafkas Univ Vet Fak Derg, 18 (4): 559-564, 2012. DOI: 10.9775/ kvfd.2011.5738