Phylogenetic Grouping of Verotoxigenic Escherichia coli (VTEC) Obtained from Sheep and Broiler Chicken in Northwestern Iran

Verotoxigenic Escherichia coli (VTEC) are major foodborne pathogens with an increasing public health concern. The purpose of this study was to investigate the occurrence and the phylogenetic groups of VTEC isolates from the feces of healthy sheep and broiler chickens at a slaughterhouse in Urmia region, Northwestern Iran. A total of 446 E. coli isolates (97 from sheep and 349 from broiler chickens) were assessed for the occurrence of the Vtx-encoding genes (vtx1 and vtx2) using polymerase chain reaction. Then, all the recovered VTEC isolates were phylogenetically grouped based on the Clermont phylotyping method using three genetic sequences, the so-called chuA, yjaA, and TSPE4.C2. The vtx gene-carrying E. coli was identified in 46.4% (45/97) of sheep-originated isolates and in 8.3% (29/349) of broiler chicken-originated isolates. In general, phylotyping revealed that 74 VTEC isolates segregated in the phylogenetic groups A (32.4%; designated as VTEC-A), B1 (44.6%; VTEC-B1), B2 (9.5%; VTEC-B2), and D (13.5%; VTEC-D). The results also showed that the dissemination of VTEC isolates of sheep and broiler chicken origin varied noticeably in their assignment to B1 and D phylogenetic groups (p

Kuzeybatı İran'da Koyun ve Broyler Tavuğundan Elde Edilen Verotoksijenik Escherichia coli (VTEC) Filogenetik Grubu

Verotoksijenik Escherichia coli (VTEC), artan bir halk sağlığı sorunu olan başlıca gıda kaynaklı patojenlerdir. Çalışmanın amacı, Kuzeybatı İran’ın Urmia bölgesinde kesilen sağlıklı koyun ve broyler tavuklarının dışkılarından VTEC izolatlarının oluşum ve filogenetik gruplarını araştırmaktır. Bu çalışmada, 446 E. coli izolatı (koyundan 97 ve broyler tavuktarından 349), polimeraz zincir reaksiyonu (PCR) kullanılarak Vtx-kodlayan genlerin (vtx1 ve vtx2) oluşumu için test edilmiştir. Daha sonra, tüm geri kazanılmış VTEC izolatları, chuA, yjaA ve TSPE4.C2 olarak adlandırılan üç genetik sekans kullanılarak Clermont filotiplemeye dayanan filogenetik olarak gruplandırılmıştır. Sonuç olarak, koyun kaynaklı izolatların %46,4’ünde (45/97) vtx geni taşıyan E. coli saptanmış ve broylerler için saptanan yüzdeler ise %8,3 (29/349) olmuştur. Genel olarak, filotipleme, 74 VTEC izolatının, filogenetik olarak grup A (%32,4, VTEC-A olarak belirlenmiş), B1 (%44,6; VTEC-B1), B2 (%9,5; VTEC-B2) ve D (%13,5; VTEC-D) olarak ayrıldığını ortaya koymuştur. Buna ek olarak, sonuçlar ayrıca, koyun ve broyler tavuklarının VTEC izolatlarının yayılmasının B1 ve D filogenetik grupları arasında belirgin olarak farklılık gösterdiğini göstermiştir (p

Kaynakça

Alonso, M.Z., Lucchesi, P.M.A., Rodríguez, E.M., Parma, A.E., Padola, N.L., 2012. Enteropathogenic (EPEC) and Shigatoxigenic Escherichia coli (STEC) in broiler chickens and derived products at different retail stores. Food Control 23(2), 351-355. [Crossref]

Bagheri, M., Ghanbarpour, R., Alizade, H., 2014. Shiga toxin and beta-lactamases genes in Escherichia coli phylotypes isolated from carcasses of broiler chickens slaughtered in Iran. International Journal of Food Microbiology 177, 16-20. [Crossref] Bergthorsson, U., Ochman, H., 1998. Distribution of chromosome length variation in natural isolates of Escherichia coli. Molecular Biology and Evolution 15(1), 6-16. [Crossref]

Blanco, M., Blanco, J.E., Mora, A., Rey, J., Alonso, J.M., Hermoso, M., Hermoso, J., Alonso, M.P., Dahbi, G., Gonzalez, E.A., Bernardez, M.I., Blanco, J., 2003. Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from healthy sheep in Spain. Journal of Clinical Microbiology 41(4), 1351-1356. [Crossref]

Carlos, C., Pires, M.M., Stoppe, N.C., Hachich, E.M., Sato, M.I., Gomes, T.A., Amaral, L.A., Ottoboni, L.M., 2010. Escherichia coli phylogenetic group determination and its application in the identification of the major animal source of fecal contamination. BMC Microbiology 10, 161. [Crossref]

Cho, S., Fossler, C.P., Diez-Gonzalez, F., Wells, S.J., Hedberg, C.W., Kaneene, J.B., Ruegg, P.L., Warnick, L.D., Bender, J.B., 2013. Herd-level risk factors associated with fecal shedding of Shiga toxin-encoding bacteria on dairy farms in Minnesota, USA. Canadian Veterinary Journal 54(7), 693-697.

Clermont, O., Bonacorsi, S., Bingen, E., 2000. Rapid and simple determination of the Escherichia coli phylogenetic group. Applied and Environmental Microbiology 66(10), 4555-4558. [Crossref]

Clermont, O., Olier, M., Hoede, C., Diancourt, L., Brisse, S., Keroudean, M., Glodt, J., Picard, B., Oswald, E., Denamur, E., 2011. Animal and human pathogenic Escherichia coli strains share common genetic backgrounds. Infection, Genetics and Evolution 11(3), 654-662. [Crossref]

Dixit, S.M., Gordon, D.M., Wu, X.Y., Chapman, T., Kailasapathy, K., Chin, J.J., 2004. Diversity analysis of commensal porcine Escherichia coli - associations between genotypes and habitat in the porcine gastrointestinal tract. Microbiology 150(Pt 6), 1735-1740. [Crossref]

Escobar-Paramo, P., Grenet, K., Le Menac’h, A., Rode, L., Salgado, E., Amorin, C., Gouriou, S., Picard, B., Rahimy, M.C., Andremont, A., Denamur, E., Ruimy, R., 2004. Large-scale population structure of human commensal Escherichia coli isolates. Applied and environmental microbiology 70(9), 5698-5700. [Crossref]

Farooq, S., Hussain, I., Mir, M.A., Bhat, M.A., Wani, S.A., 2009. Isolation of atypical enteropathogenic Escherichia coli and Shiga toxin 1 and 2f-producing Escherichia coli from avian species in India. Letters in Applied Microbiology 48(6), 692-697.]

Garcia-Aljaro, C., Moreno, E., Andreu, A., Prats, G., Blanch, A.R., 2009. Phylogroups, virulence determinants and antimicrobial resistance in stx(2) gene-carrying Escherichia coli isolated from aquatic environments. Research in Microbiology 160(8), 585-591. [Crossref]

Ghanbarpour, R., Kiani, M., 2013. Characterization of non-O157 shiga toxin-producing Escherichia coli isolates from healthy fat-tailed sheep in southeastern of Iran. Tropical Animal Health and Production 45(2), 641-648. [Crossref]

Ghanbarpour, R., Sami, M., Salehi, M., Ouromiei, M., 2011. Phylogenetic background and virulence genes of Escherichia coli isolates from colisepticemic and healthy broiler chickens in Iran. Tropical Animal Health and Production 43(1), 153-157. [Crossref]

Girardeau, J.P., Dalmasso, A., Bertin, Y., Ducrot, C., Bord, S., Livrelli, V., Vernozy-Rozand, C., Martin, C., 2005. Association of virulence genotype with phylogenetic background in comparison to different seropathotypes of Shiga toxin-producing Escherichia coli isolates. Journal of Clinical Microbiology 43(12), 6098-6107. [Crossref]

Gordon, D.M., Clermont, O., Tolley, H., Denamur, E., 2008. Assigning Escherichia coli strains to phylogenetic groups: multi-locus sequence typing versus the PCR triplex method. Environmental Microbiology 10(10), 2484-2496. [Crossref]

Gyles, C.L., 2007. Shiga toxin-producing Escherichia coli: an overview. Journal of Animal Science 85(13 Suppl), E45-62. [Crossref]

Higgins, J., Hohn, C., Hornor, S., Frana, M., Denver, M., Joerger, R., 2007. Genotyping of Escherichia coli from environmental and animal samples. Journal of Microbiological Methods 70(2), 227-235. [Crossref]

Horcajo, P., Dominguez-Bernal, G., De La Fuente, R., Ruiz-Santa-Quiteria, J.A., Orden, J.A., 2010. Association of vt1c with verotoxin-producing Escherichia coli from goats and sheep. Journal of Veterinary Diagnostic Investigation 22(2), 332-334. [Crossref]

Ishii, S., Meyer, K.P., Sadowsky, M.J., 2007. Relationship between phylogenetic groups, genotypic clusters, and virulence gene profiles of Escherichia coli strains from diverse human and animal sources. Applied and Environmental Microbiology 73(18), 5703-5710. [Crossref]

Kaper, J.B., Nataro, J.P., Mobley, H.L., 2004. Pathogenic Escherichia coli. Nature Reviews Microbiology 2, 123-140. [Crossref]

Maluta, R.P., Fairbrother, J.M., Stella, A.E., Rigobelo, E.C., Martinez, R., de Avila, F.A., 2014. Potentially pathogenic Escherichia coli in healthy, pasture-raised sheep on farms and at the abattoir in Brazil. Veterinary Microbiology 169(1-2), 89-95. [Crossref]

Martins, R.P., da Silva, M.C., Dutra, V., Nakazato, L., Leite Dda, S., 2013. Preliminary virulence genotyping and phylogeny of Escherichia coli from the gut of pigs at slaughtering stage in Brazil. Meat Science 93(3), 437-440. [Crossref]

Mora, A., Herrera, A., Mamani, R., Lopez, C., Alonso, M.P., Blanco, J.E., Blanco, M., Dahbi, G., Garcia-Garrote, F., Pita, J.M., Coira, A., Bernardez, M.I., Blanco, J., 2010. Recent emergence of clonal group O25b:K1:H4-B2-ST131 ibeA strains among Escherichia coli poultry isolates, including CTX-M-9-producing strains, and comparison with clinical human isolates. Applied and Environmental Microbiology 76(21), 6991-6997. [Crossref]

Mora, A., Viso, S., Lopez, C., Alonso, M.P., Garcia-Garrote, F., Dabhi, G., Mamani, R., Herrera, A., Marzoa, J., Blanco, M., Blanco, J.E., Moulin-Schouleur, M., Schouler, C., Blanco, J., 2013. Poultry as reservoir for extraintestinal pathogenic Escherichia coli O45:K1:H7-B2- ST95 in humans. Veterinary Microbiology 167(3-4), 506-512. [Crossref]

Navidinia, M., Peerayeh, S.N., Fallah, F., Bakhshi, B., 2013. Phylogenetic Groups and Pathogenicity Island Markers in Escherichia coli Isolated From Children. Jundishapur Journal of Microbiology 6(10), e8362. [Crossref]

Novais, A., Vuotto, C., Pires, J., Montenegro, C., Donelli, G., Coque, T.M., Peixe, L., 2013. Diversity and biofilm-production ability among isolates of Escherichia coli phylogroup D belonging to ST69, ST393 and ST405 clonal groups. BMC Microbiology 13, 144. [Crossref]

Nowrouzian, F.L., Wold, A.E., Adlerberth, I., 2005. Escherichia coli strains belonging to phylogenetic group B2 have superior capacity to persist in the intestinal microflora of infants. The Journal of Infectious Disease 191(7), 1078-1083. [Crossref]

Obeng, A.S., Rickard, H., Ndi, O., Sexton, M., Barton, M., 2012. Antibiotic resistance, phylogenetic grouping and virulence potential of Escherichia coli isolated from the faeces of intensively farmed and free range poultry. Veterinary Microbiology 154(3-4), 305-315. [Crossref]

Oporto, B., Esteban, J.I., Aduriz, G., Juste, R.A., Hurtado, A., 2008. Escherichia coli O157:H7 and non-O157 Shiga toxin-producing E. coli in healthy cattle, sheep and swine herds in Northern Spain. Zoonoses Public Health 55(2), 73-81. [Crossref]

Osek, J., 2003. Development of a multiplex PCR approach for the identification of Shiga toxin-producing Escherichia coli strains and their major virulence factor genes. Journal of Applied Microbiology 95(6), 1217-1225. [Crossref]

Paton, A.W., Paton, J.C., 2002. Direct detection and characterization of Shiga toxigenic Escherichia coli by multiplex PCR for stx1, stx2, eae, ehxA, and saa. Journal of Clinical Microbiology 40(1), 271-274. [Crossref]

Ramazanzadeh, R., Zamani, S., Zamani, S., 2013. Genetic diversity in clinical isolates of Escherichia coli by enterobacterial repetitive intergenic consensus (ERIC)-PCR technique in Sanandaj hospitals. Iranian Journal of Microbiology 5(2), 126-131.

Rey, J., Blanco, J.E., Blanco, M., Mora, A., Dahbi, G., Alonso, J.M., Hermoso, M., Hermoso, J., Alonso, M.P., Usera, M.A., Gonzalez, E.A., Bernardez, M.I., Blanco, J., 2003. Serotypes, phage types and virulence genes of shiga-producing Escherichia coli isolated from sheep in Spain. Veterinary Microbiology 94(1), 47-56. [Crossref]

Riffon, R., Sayasith, K., Khalil, H., Dubreuil, P., Drolet, M., Lagace, J., 2001. Development of a rapid and sensitive test for identification of major pathogens in bovine mastitis by PCR. Journal of Clinical Microbiology 39(7), 2584-2589. [Crossref]

Saei, H.D., Ayremlou, N., 2012. Characterization of Shiga toxin-producing Escherichia coli (STEC) in feces of healthy and diarrheic calves in Urmia region, Iran. Iranian Journal of Microbiology 4(2), 63-69.

Salehi, M., 2014. Determination of intimin and Shiga toxin genes in Escherichia coli isolates from gastrointestinal contents of healthy broiler chickens in Kerman City, Iran. Comparative Clinical Pathology 23(1), 175-179. [Crossref]

Tahamtan, Y., Hayati, M., Mehdi Namavari, M., 2010. Contamination of sheep carcasses with verocytotoxin producing Escherchia coli during slaughtering. Transboundary and Emerging Diseases 57(1-2), 25-27. [Crossref]

Unno, T., Han, D., Jang, J., Lee, S.N., Ko, G., Choi, H.Y., Kim, J.H., Sadowsky, M.J., Hur, H.G., 2009. Absence of Escherichia coli phylogenetic group B2 strains in humans and domesticated animals from Jeonnam Province, Republic of Korea. Applied and Environmental Microbiology 75(17), 5659-5666. [Crossref]

Vettorato, M.P., de Castro, A.F., Cergole-Novella, M.C., Camargo, F.L., Irino, K., Guth, B.E., 2009. Shiga toxin-producing Escherichia coli and atypical enteropathogenic Escherichia coli strains isolated from healthy sheep of different populations in Sao Paulo, Brazil. Letters in Applied Microbiology 49(1), 53-59. [Crossref]

Wasteson, Y., 2001. Zoonotic Escherichia coli. Acta Veterinaria Scandinavica 95, 79-84.

Zweifel, C., Zychowska, M.A., Stephan, R., 2004. Prevalence and characteristics of Shiga toxin-producing Escherichia coli, Salmonella spp. and Campylobacter spp. isolated from slaughtered sheep in Switzerland. International Journal of Food Microbiology 92(1), 45-53.[Crossref]

Kaynak Göster