Characterization of Shiga toxin-producing Escherichia coli (STEC) in feces of healthy and diarrheic calves in Urmia region, Iran
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Abstract
Background and Objectives: Shiga toxin-producing Escherichia coli (STEC) have emerged as human pathogens and contamination of foods of animal origin has been a major public health concern. The aim of the present study was to determine the dissemination of STEC in healthy and diarrheic calves in Urmia region which is located in West Azerbaijan province, Iran.
Materials and Methods: In the current study, a total of 124 Escherichia coli isolates from clinically healthy (n = 73) and diarrheic calves (51) belonging to 6 different farms located in West Azerbaijan province, Iran, were screened by the polymerase chain reaction (PCR) assay for the presence of virulence genes characteristic for STEC, that is, Shiga-toxin producing gene(s) (stx1, stx2), intimin (eaeA) and enterohemolysin (hlyA).
Results: STEC isolates were recovered from 21.92% (16/73) in healthy calves, and 19.6% (10/51) in diarrheic calves. Overall, PCR results showed that 6 (23.1%) isolates carried stx1 gene, 7 (26.92%) possessed stx2 gene while 13 isolates (50%) gave positive amplicon both for stx1 and stx2 genes. All stx positive isolates were assayed further to detect eaeA and hlyA sequences. Seven out of the 26 (26.92%) Shiga toxin gene positive isolates were positive for the eaeA gene, and 15 (57.69%) were positive for the hlyA gene. Both virulence genes (eaeA and hlyA) in the same isolate were observed in 5 (19.23%) of the stx+ isolates. In total, diverse virulence gene profiles were detected, from which isolates with the genetic profile stx1 stx2 hlyA was the most prevalent. In addition, eaeA gene was more evident in isolates from diarrheic calves than in healthy calves.
Conclusion: There was no significant difference in detecting STEC isolates between healthy and diarrheic calves. It seems that calves to be the reservoir of STEC within the herds and calf management may represent specific control points for reducing STEC spread within dairy units.
Frydendahl K. Prevalence of serogroups and virulence genes in Escherichia coli associated with postweaning diarrhoea and edema disease in pigs and a comparison of diagnostic approaches. Vet Microbiol 2002; 85: 169-182.
Boerlin P, Travis R, Gyles CL, Reid-Smith R, Janecko N, Lim H, et al. Antimicrobial resistance and virulence genes of Escherichia coli isolates from swine in Ontario. Appl Environ Microbiol 2005; 71: 6753-6761.
Lockary VM, Hudson RF, Ball CL. Shiga toxin- producing Escherichia coli, Idaho. Emerg Infect Dis 2007; 13: 1262-1264.
Werber D, Behnke SC, Fruth A, Merle R, Menzler S, Glaser S, et al. Shiga toxin-producing Escherichia coli infection in Germany: different risk factors for different age groups. Am J Epidemiol 2007; 165: 425-434.
Paton AW, Paton JC. Detection and characterization of Shiga toxigenic Escherichia coli by using multiplex PCR assays for stx1, stx2, eaeA, enterohemorrhagic E. coli hlyA, rfb , and rfb . J Clin Microbiol 1998; 36: 598-602.
Blanco M, Blanco JE, Mora A, Dahbi G, Alonso MP,Gonzalez EA, et al. Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from cattle in Spain and identification of a new intimin variant gene (eae-ξ). J Clin Microbiol 2004; 42: 645-651.
Mora A, Blanco JE, Blanco M, Alonso MP, Dhabi G, Echeita A, et al. Antimicrobial resistance of Shiga toxin (verotoxin)-producing Escherichia coli O157:H7 and non-O157 strains isolated from humans, cattle, sheep and food in Spain. Res Microbiol 2005; 156: 793-806.
Schmidt H, Beutin L, Karch H. Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933. Infect Immun 1995; 63: 1055-1061.
Caprioli A, Morabito S, Brugere H, Oswald E.Enterohaemorrhagic Escherichia coli: emerging issues on virulence and modes of transmission. Vet Res 2005;36: 289-311.
Renter DG, Sargeant JM. Enterohemorrhagic Escherichia coli O157: epidemiology and ecology in bovine production environments. Anim Health Res Rev 2002; 3: 83-94.
Blanco M, Blanco JE, Blanco J, Mora A, Prado C, Alonso MP, et al. Distribution and characterization of faecal verotoxin-producing Escherichia coli (VTEC) isolated from healthy cattle. Vet Microbiol 1997; 54: 309-319.
Roopnarine RR, Ammons D, Rampersad J, Adesiyun AA. Occurrence and characterization of verocytotoxigenic Escherichia coli (VTEC) strains from dairy farms in Trinidad. Zoonoses Public Health 2007; 54: 78-85.
Cray WC, Jr., Moon HW. Experimental infection of calves and adult cattle with Escherichia coli O157:H7. Appl Environ Microbiol 1995; 61: 1586-1590.
Widiasih DA, Ido N, Omoe K, Sugii S, Shinagawa K.Duration and magnitude of fecal shedding of Shiga toxin-producing Escherichia coli from naturally infected cattle. Epidemiol Infect 2004; 132: 67-75.
Cobbold R, Desmarchelier P. Horizontal transmission of Shiga toxin-producing Escherichia coli within groups of dairy calves. Appl Environ Microbiol 2002; 68:4148-4152.
Schierack P, Romer A, Jores J, Kaspar H, Guenther S, Filter M, et al. Isolation and characterization of intestinal Escherichia coli clones from wild boars in Germany. Appl Environ Microbiol 2009; 75: 695-702.
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 2011; 12: 145-149.
Jafari F, Shokrzadeh L, Hamidian M, Salmanzadeh- Ahrabi S, Zali MR. Acute diarrhea due to enteropathogenic bacteria in patients at hospitals in Tehran. Jpn J Infect Dis 2008; 61: 269-273.
Aslani MM, Ahrabi SS, Alikhani YM, Jafari F, Zali RM, Mani M. Molecular detection and antimicrobial resistance of diarrheagenic Escherichia coli strains isolated from diarrheal cases. Saudi Med J 2008; 29:388-392.
Guler L, Gunduz K, Ok U. Virulence factors and antimicrobial susceptibility of Escherichia coli isolated from calves in Turkey. Zoonoses Public Health 2008;55: 249-257.
Riffon R, Sayasith K, Khalil H, Dubreuil P, Drolet M, Lagace J. Development of a rapid and sensitive test for identification of major pathogens in bovine mastitis by PCR. J Clin Microbiol 2001; 39: 2584-2589.
Cebula TA, Payne WL, Feng P. Simultaneous identification of strains of Escherichia coli serotype O157:H7 and their Shiga-like toxin type by mismatch amplification mutation assay-multiplex PCR. J Clin Microbiol 1995; 33: 248-250.
Osek J. Development of a multiplex PCR approach for the identification of Shiga toxin-producing Escherichia coli strains and their major virulence factor genes. J Appl Microbiol 2003; 95: 1217-1225.
Batchelor M, Knutton S, Caprioli A, Huter V, Zanial M, Dougan G, et al. Development of a universal intimin antiserum and PCR primers. J Clin Microbiol 1999; 37:3822-3827.
Osek J, Gallien P, Protz D. Characterization of shiga toxin-producing Escherichia coli strains isolated from calves in Poland. Comp Immunol Microbiol Infect Dis 2000; 23: 267-276.
Moreira CN, Pereira MA, Brod CS, Rodrigues DP, Carvalhal JB, Aleixo JA. Shiga toxin-producing Escherichia coli (STEC) isolated from healthy dairy cattle in southern Brazil. Vet Microbiol 2003; 93: 179-183.
Osek J, Winiarczyk S. Prevalence of eae and shiga toxin genes among Escherichia coli strains isolated from healthy calves. J Vet Med B Infect Dis Vet Public Health 2001; 48: 67-72.
Shaw DJ, Jenkins C, Pearce MC, Cheasty T, Gunn GJ, Dougan G, et al. Shedding patterns of verocytotoxin- producing Escherichia coli strains in a cohort of calves and their dams on a Scottish beef farm. Appl Environ Microbiol 2004; 70: 7456-765.
Wani SA, Hussain I, Nabi A, Fayaz I, Nishikawa Y.Variants of eae and stx genes of atypical enteropathogenic Escherichia coli and non-O157 Shiga toxin-producing Escherichia coli from calves. Lett Appl Microbiol 2007;45: 610-615.
Arya G, Roy A, Choudhary V, Yadav MM, Joshi CG. Serogroups, atypical biochemical characters, colicinogeny and antibiotic resistance pattern of Shiga toxin-producing Escherichia coli isolated from diarrhoeic calves in Gujarat, India. Zoonoses Public Health 2008; 55: 89-98.
Nguyen TD, Vo TT, Vu-Khac H. Virulence factors in Escherichia coli isolated from calves with diarrhea in Vietnam. J Vet Sci 2011; 12: 159-164.
Wani SA, Bhat MA, Samanta I, Nishikawa Y, Buchh AS.Isolation and characterization of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic Escherichia coli (EPEC) from calves and lambs with diarrhea in India. Lett Appl Microbiol 2003; 37: 121-126.
Mercado EC, Gioffre A, Rodriguez SM, Cataldi A, Irino K, Elizondo AM, et al. Non-O157 Shiga toxin- producing Escherichia coli isolated from diarrheic calves in Argentina. J Vet Med B Infect Dis Vet Public Health 2004; 51: 82-88.
Sandhu KS, Gyles CL. Pathogenic Shiga toxin- producing Escherichia coli in the intestine of calves. Can J Vet Res 2002; 66: 65-72.
Hall GA, Reynolds DJ, Chanter N, Morgan JH, Parsons KR, Debney TG, et al. Dysentery caused by Escherichia coli (S102-9) in calves: natural and experimental disease. Vet Pathol 1985; 22: 156-163.
Fernandez D, Rodriguez EM, Arroyo GH, Padola NL, Parma AE. Seasonal variation of Shiga toxin-encoding genes (stx) and detection of E.a coli O157 in dairy cattle from Argentina. J Appl Microbiol 2009; 106: 1260-1267.
Blanco JE, Blanco M, Alonso MP, Mora A, Dahbi G, Coira MA, et al. Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from human patients: prevalence in Lugo, Spain, from 1992 through 1999. J Clin Microbiol 2004;42: 311-319.
Beutin L, Krause G, Zimmermann S, Kaulfuss S, Gleier K. Characterization of Shiga toxin-producing Escherichia coli strains isolated from human patients in Germany over a 3-year period. J Clin Microbiol 2004;42: 1099-108.
Aidar-Ugrinovich L, Blanco J, Blanco M, Blanco JE, Leomil L, Dahbi G, et al. Serotypes, virulence genes, and intimin types of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) isolated from calves in Sao Paulo, Brazil. Int J Food Microbiol 2007; 115: 297-306.
Blanco M, Padola NL, Kruger A, Sanz ME, Blanco JE, Gonzalez EA, et al. Virulence genes and intimin types of Shiga-toxin-producing Escherichia coli isolated from cattle and beef products in Argentina. Int Microbiol 2004; 7: 269-276.
Maidhof H, Guerra B, Abbas S, Elsheikha HM, Whittam TS, Beutin L. A multiresistant clone of Shiga toxin-producing Escherichia coli O118:[H16] is spread in cattle and humans over different European countries. Appl Environ Microbiol 2002; 68: 5834-5842.
Schmidt H, Geitz C, Tarr PI, Frosch M, Karch H. Non-O157:H7 pathogenic Shiga toxin-producing Escherichia coli: phenotypic and genetic profiling of virulence traits and evidence for clonality. J Infect Dis 1999; 179:115-123.
Irino K, Kato MA, Vaz TM, Ramos, II, Souza MA, Cruz AS, et al. Serotypes and virulence markers of Shiga toxin-producing Escherichia coli (STEC) isolated from dairy cattle in Sao Paulo State, Brazil. Vet Microbiol 2005; 105: 29-36.
Leomil L, Aidar-Ugrinovich L, Guth BE, Irino K, Vettorato MP, Onuma DL, et al. Frequency of Shiga toxin-producing Escherichia coli (STEC) isolates among diarrheic and non-diarrheic calves in Brazil. Vet Microbiol 2003; 97: 103-109.
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| Issue | Vol 4 No 2 (2012) | |
| Section | Articles | |
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| E. coli Iran Shiga toxin calves | ||
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