Ege Bölgesi’ndeki sığırların süt ve dışkı örneklerinden Escherichia coli O157:H7 izolasyonu ve verotoksinlerinin belirlenmesi

Öz

Escherichia coli O157:H7 serotipi; insan patojeni olarak ilk kez 1982 yılında ABD’de meydana gelen iki ayrı hemorajik kolitis salgını sonucu tanımlanmaya başlamıştır ve en sık izole edilen enterik patojenlerden biridir. E. coli O157:H7 ile infekte olan kişiler asemptomatik olabilir veya kanlı yada kansız diare, hemorajik kolitis (HC), hemolitik üremik sendrom (HUS) ve trombotik trombositopenik purpura (TTP) semptomları gösterebilir. Shiga-benzeri toxin üreten E. coli (STEC), Vero toxin-üreten E.coli (VTEC) olarak da adlandırılır. İnfeksiyonu gıda kaynaklıdır ve sığırlar STEC infeksiyonunun birincil rezervuarı olarak kabul edilmektedir. STEC infeksiyonlarının patogenezinde rol alan en önemli virülens faktörleri, şigatoksinler, hemolizin, intimin yapışma faktörü, pO157 plazmidi ve tip III sekresyon sistemi olarak belirtilmektedir. Bu çalışmada, incelenen 150 süt ve 150 dışkı örneğinden 2’si süt ve diğer 2’si ise dışkı örneklerinden olmak üzere 4 (%1.3) E. coli O157:H7 izole edildi. Süt örneklerinden izole edilen suşlardan 1’inin VT1 ürettiği belirlendi.

Anahtar Kelimeler

• Erdem ÇİÇEK
• Serap SAVAŞAN
• E. coli O157:H7
• izolasyon
• sığır
• verotoksin
• 2010
• Etlik Veteriner Mikrobiyoloji Dergisi

Isolation of Escherichia coli O157:H7 in milk and feaces of cattle in Aegean Region and determination of their verotoxin

Öz

Escherichia coli O157:H7 was first identified as a human pathogen following two geographically separate outbreaks of hemorrhagic colitis in the United States in 1982. E. coli O157:H7 was one of the most frequently isolated enteric pathogen. Persons infected with E. coli O157:H7 may be asymptomatic or symptoms such as diarrhea, bloody diarrhea, hemorrhagic colitis (HC), hemolytic-uremic syndrome, and thrombotic thrombocytopenic purpura may be observed. Infections caused by Shiga-like toxin-producing E. coli (STEC), also known as Vero toxin-producing E. coli (VTEC), in humans is food-borne and infected cattle act as a primer reservoir for STEC infection. The most important virulence factors which act role in the pathogenesis of STEC infections are reported as shiga toxins, hemolysin, intimin adherence factor and type III secretion system. In this study, 4 (1.3%) E. coli O157:H7, two of them were from milk samples and the other two from faeces samples were isolated from 150 milk samples and 150 faeces samples. We determined that one strain isolated from milk samples produced VT1.

Anahtar Kelimeler

• Cattle
• E. coli O157:H7
• isolation
• verotoxin.

Kaynakça

1. Baets LD, Taelen IVD, Filette MD, Pierard D, Allison L, Greve HD, Hernalsteens JP, Imberechts H, (2004). Genetic Typing of Shiga Toxin 2 Variants of Escherichia coli by PCR-Restriction Fragment Lenght Polymorphism Analysis. Appl Environ Microbiol. 70 (10), 6309-6314.
2. Bekar M, (1997). Enterobacteriaceae Familyası Mikroorganizmaların Genel Karakterleri ve Tanı Yöntemleri. Veteriner Kontrol ve Araştırma Enstitüsü Müdürlüğü, Yayın No. 97–1 Ankara.
3. Blanco J, Blanco M, Blanco JE, Mora A, Alonso MP, Gonzales EA, Bernardez MI, (2001). Epidemiology of Verocytotoxigenic Escherichia coli (VTEC) in Ruminants. In: Verocytotoxigenic Escherichia coli. G. Duffy, P. Garvey, McDowell D.A. eds. Trumbull, Connecticut, USA: Food & Nutrition Press. p.113-148.
4. Boerlin P, Mceven SA, Petzold FB, Wilson JB, Jhonson RP, Gyles CL, (1999). Associations Between Virulence Factors of Shiga Toxin-Producing Escherichia coli and Disease in Humans. J Clin Microbiol. 37 (3), 497-503.
5. Brooks GF, Butel JS, Morse SA, (2004). Enteric Gram Negative Rods (Enterobacteriaceae). Chapter 16 in:Jawetz Melnick and Adelberg’s Medical Microbiology. Twenthy Third Edition, Mcgraw Hill Education, Boston. p. 252-255.
6. Caprioli A, Morabito S, Brugere H, Oswalt E, (2005). Enterohemorrhagic Escherichia coli: Emerging Issues on Virulence and Modes of Transmission. Vet Res. 36, 289-311.
7. Çabalar M, Boynukara B, Gülhan T, Ekin İH, (2001). Prevalence of Rotavirus, Escherichia coli K99 and O157:H7 in Healthy Dairy Cattle Herds in Van, Turkey. Turk J Vet Anim Sci. 25, 191-196.
8. Dean-Nystrom EA, Bosworth BT, Moon HW, O’brıen AD, (1998). Escherichia coli O157:H7 Requires Intimin for Enteropathogenicity in Calves. Infect Immun. 66 (9), 4560–4563.

9. Dunn JR, (2003). The Epidemiology of Shiga – Toxigenic Escherichia coli O157:H7 in Louisiana Dairy Cattle, Beef Catle and White – Tailed Deer. J Wildlife Dis. 40 (2), 361-365
10. Durso LM, Smith D, Hutkins RW, (2004). Measurements of Fitness and Competition in Commensal Escherichia coli and E. coli O 157:H7 Strains. Appl Environ Microbiol. 70 (11), 6466-6472.
11. Franzolin MR, Alves RCB, Keller R, Gomes TAT, Beutin K, Barreto ML, Milroy C, Strina A, Riberio H, Trabusi LR, (2005). Prevalence of Diarrheagenic Escherichia coli in Children with Diarrhea in Salvador, Bahia, Brazil. Mem. Inst. Oswaldo Cruz. 100 (4), 359-363.
12. Friedrich AW, Bielaszewska M, Zhang W, Pulz M, Kuczius T, Ammon A, Karch H, (2002). Escherichia coli Harboring Shigo Toxin 2 Gene Variants: Frequency and Association with Clinical Symptomes. J Infect Dis. 185, 74-84.
13. Gorbach , Barleth, Blacklow, (2004). Infectious diseases. Chapter 70, Escherichia coli and Other Shıga-Toxin Producing E. coli. Lippincott, Philedelphia. p.643-647.
14. Gupro A, Hunter SB, Bidol SA, Dietrich S, Kincaid J, Salehi E, Nicholson L, Genese CA, Weinstein ST, Marengo L, Kimura AC, Brooks JT, (2004). Escherichia coli O157 Cluster Evalation. Emerg Infect Dis. 10 (10), 1856-1858.
15. Halkman AK, Noveir MR, Doğan HB, (2001). Escherichia coli O157:H7 serotipi. Ankara Üniversitesi Ziraat Fakültesi Gıda Mühendisliği Bölümü, Sim Matbaacılık Ltd, Şti, Ankara s:46.
16. Hancock DD, Besser TE, Kinsel ML, Tarr PI, Rice DH, Paros MG, (1994). The prevalence of Escherichia coli O157:H7 in dairy and beef cattle in Washington State. Epidemiol Infect. 113 (2), 199-207.
17. Heuvelink AE, Bleumink B, Van Den Biggelaar FL, Te Gıffel MC, Beumer RR, De Boer E, (1998). Occurence and survival of verocytotoxin-producing Escherichia coli O157 in raw cow’s milk in Netherlands. J Food Protect. 61, 1597-1601.
18. Jin HY, Tao KH, Li YX, Li FO, Li SQ, (2005). Microarry Analysis of Escherichia coli O 157:H7. W J Gastroenterolo. 11 (37), 5811-5815.
19. Kim JY, Kiro SH, Kwon NH, Bae WK, Lim JY, Koo HC, Kim JM, Noh KM, Jong WK, Park KT, Park YH, (2005). Isolation and Identification of Escherichia coli O 157:H7 Using Differend Detection Methots Molecular Determination by Multiplex PCR and RAPD. J Clin Microbiol. 6 (1), 7-19.
20. Kusumato M, Okitsu T, Nishiya Y, Suzuki R, Yamai S, Kawamura Y, (2001). Spontaneous Reactivation of Shiga Toxins in Escherichia coli O157:H7 Cells Caused by Transposon Excision. J Biosci and Bioengineer. 92 (2), 114-120.
21. Maldonado Y, Fiser JC, Nakatsu CH, Bhunia AK, (2005). Cytotoxicity Potential and Genotypic Characterization of Escherichia coli Isolated from Environmental and Food Sources. Appl Environ Microbiol. 71(4), 1890-1898.
22. Orlandi PP, Magalhaes GF, Matos NB, Silva T, Penatti M, Nogueira PA, Peraira da Silva LH, (2006). Etiyology of Diarrheal Infections in Children of Porto Velho ( Rondonia, Western Amazon Region, Brazil). Brazilian J Med Biologic Res. 39 (4), 507-517.
23. Öksüz Ö, Arıcı M, Kurultay S, Gümüs T, (2004). Incidence of Escherichia coli O157 in raw milk and pickled cheese manifactured from raw milk in Turkey. Food Cont. 15, 453-456.
24. Padhye NY, Doyle MP, (1991). Rapid procedure for detecting enterohemorrhagic Escherichia coli O157 in food. Appl Environ Microbiol. 57, 2693-2698.
25. Picozzi C, Foschino R, Heuvelink A, Beumer R, (2005). Phenotypic and genotypic characterization of sorbitol-negative or slow-fermenting (suspected O157) Escherichia coli isolated milk samples in Lombardy region. Lett Appl Microbiol. 40, 491-496.
26. Rangel JM, Sparling PH, Crowe C, Griffin PM, Swerdlow DL, (2005). Epidemiology of Escherichia coli O 157:H7 Outbreaks, United States 1982-2002. Emerg Infect Dis. 11 (4), 603-609.
27. Roldan ML, Chinen I, Otero Jl, Mılıwebsky ES, Alfaro N, (2007). Isolation, characterization and typing of Escherichia coli O157:H7 strains from beef products and milk. Revista Argentina de Microbiología. 39, 113-119.
28. Topçu Aİ, Söyletir G, Doğanay M, (2002). Bakteriyel İnfeksiyonlar. İnfeksiyon Hastalıkları ve Mikrobiyoloji, cilt 1, Nobel Tıp Kitapevi, İstanbul, s.1564-1575.
29. Tsai WL, Miller CE, Richter ER, (2000). Determination of the Sensitivity of a Rapid Escherichia coli O157:H7 Assay for Testing 375-Gram Composite Samples. Appl Environ Microbiol. 66 (9), 4149–4151.
30. Ünlütürk A, Turantaş F, eds., (1998). Gıda Mikrobiyolojisi, 1. Basım, Mengi Tan Basımevi, İzmir. s:605.
31. Wang G, Clark CG, Rodgers FG, (2002). Detection in Escherichia coli of the Genes Encoding the Major Virulance Factors, the Genes Defining the O 157:H7 Serotype and Components of the Type 2 Shiga Toxin Family by Multiplex PCR. J Clin Microbiol. 40 (1), 3619-3619.
32. Wells JG, Shipman LD, Greene KD, Sowers EG, Green JH, Cameron DN, Downes FP, Martin ML, Griffin PM, Ostroff SM, Potter ME, Tauxe RV, Wachsmuth IK, (1991). Isolation of Escherichia coli serotype O157:H7 and other shiga-like-toxin-producing E.coli from dairy cattle. J Clin Microbiol. 29, 985-989.
33. Yang ZM, Kovar J, Kim J, Nietfelt J, Smith DR, Moxley RA, Olson ME, Fey PD, Benson AK, (2004). Identification of Common Subpopulations of Non-Sorbitol-Ferment ing, ß-Glucuronidase- Negative Escherichia coli O 157:H7 from Bovina Production Environments and Human Clinical Samples. Appl Environ Microbiol. 70 (11), 6846-6854.
34. Yılmaz A, Gun H, Yılmaz H, (2002). Frequency of Escherichia coli O157:H7 in Turkish cattle. J Food Protect. 10, 1637-1640.
35. Zhao T, Doyle MP, Shere J, Garber L, (1995). Prevalence of Enterohemorrhagic Escherichia coli O157:H7 in a Survey of Dairy Herds. Appl Environ Microbiol. 61(4), 1290-129.