Molecular Identification of Virulence Factors of E. coli in Three Strains of ETEC, EHEC and EPEC in Calves and Lambs with Diarrhea in Shiraz

Document Type : Full Research Paper

Authors

1 MSC Student, Department Biology, Science Faculty, Sirjan Islamic Azad University, Shiraz Branch Research Science.

2 azi Vaccine and Serum Research Institute, Branch, Agricultural Research, Education and Extension Organisation (AREEO), Tehran, Iran.

Abstract

Pathogenic E.coli (ETEC, EHEC and EPEC) cause diarrhea in new born calves and lambs are important especially due to mortality rate and high economic losses. In this study 200 rectal swab samples were collected from 1-30 day-old calves and lambs with diarrhea during 2013-2014 in Shiraz. The samples were inoculated on Mac Conkey, EMB and Minca agar media. The presence of various virulence genes of ETEC (k99, F41 fimbria and Sta), EHEC (Stx1, Stx2 and eae) and EPEC (bfpb and eae) was investigated using multiplex PCR. The PCR test results showed that out of 133 E. coli isolates, 12.03% and 3% possessed all three virulence genes of ETEC and EHEC, respectively. The sequencing of K99 gene showed 97-100% homology between the isolates. The variety of ETEC, EPEC and EHEC prevalence has been reported in the world. These variations could be due to managing factors, calves age, colostrum intake and dam vaccination.

Keywords


1.Pourtaghi-Shotorban H., Dahpahlevan V., Shaghayegh A. and Badiee A.(2011).Molecular detection of diarrhea genicentrotoxigenic Escherichia coli (ETEC) from calves fecal samples in Alborzprovince. Journal of veterinary clinical research.Vol, 2, NO,1. pp: 33-38. (In persian)
2.Kiani, Y., Tahamtan, Y., Hoseini, M.H and Hayati,M.(2011). An assessment for comparing virulent factors of enterotoxigenicE.coli k99 isolates by PCR and SDS- PAGE methods. Journal of Microbial World. Vol, 3, No,4. pp:238-244.
3.Lotfollahzadeh, S.,Ziaei Daroonkolai, N., ZahraeiSalehi, T., Poorbakhsh, S.A., MokhberDezfouli, M.R and Afshari, GH.R.(2004).A study on the Presence of Escherichia coli ,coccidia and cryptosporidium in stool samples of under one month age diarrheic calves in Ghaemshahr and Babol and antibiotic Sensitivity Of Isolates. J. Fac. Vet. Med. Univ.Tehran.Vol,59. NO,2. pp:131-136.
4.Boyce,T., Swerdlow,D.Land Griffin, P.M.( 1995). Escherichia coli O157: H7 and the Hemolytic- Uremic Syndrome. The New England Journal of Medicin, Vol, 333, No,6. pp: 364-368.
5.Chen, X., Gao, S., Jiao, X and Liu, X.(2004). Prevalence of serogroups and virulence factors of Escherichia coli strains isolated from pigs with postweaning diarrhoea in eastern China. Veterinary Microbiology, Vol, 103, pp: 13-20.
6. Christopher, M.J., Bhaskaran, S., Rathore, K.S and Waghela, S.(2004). Enterotoxigenic K99+ Escherichia coli attachment to host cell receptors inhibited by recombinant pili protein. Veterinary Microbiology, Vol,101, No,2. pp: 153-160.
7.Dastmalchi ,H. and Ayremlou, N.( 2012). Characterization of Shiga toxin-producing Escherichia coli (STEC) in feces of healthy and diarrheic calves in Urmia region, Iran.I. J. M,Vol, 4, No,2. pp: 63-69.
8.Guler, L., Gunduz, K. and Ok, U.(2008). Virulence factors and antimicrobial susceptibility of Escherihia coli isolated from calvesin Turky. Zoonoses and Public Health,Vol, 55, No,5. pp: 249-257.
9.Kim, H., Samadpour, M., Grimm, L., Clausen, C.R., Besser, T.E., Baylor, M., and et al.(1994).Characteristics of antibiotic-resistant Escherichia coli O157:H7in Washington State, 1984-1991.The Journal of Infectious Diseases,Vol, 170, No,6. pp:1606-1609.
10. Kim,Y.J., Kim,J.H., Hur, J andLee,J.H.( 2010). Isolation of Escherichia coli from piglets in South Korea with diarrhea and characteristics of the virulence genes.The Canadian Journal of Veterinary Research,Vol, 74,No,1. pp:59–64 .
11.Naciri, M., Lefay, M., Mancassola, R., Poirier, P and Chermette, R.(1999). Role of Cryptosporidium parvum as a pathogen in neonatal diarrhoea complex in suckling and dairy calves in France. Veterinary Parasitology, Vol, 85, No,5.pp: 245-257.
12.Nagy, B., Moon, H. WandIsaacson, R.E.(1977).Colonization of porcine intestine by enterotoxigenic Escherichia coli selection of piliated forms invivo, adhesion of piliated forms to epithelial cells in vitro, and incidence of a pilus antigen among porcineenteropathogenic E. coli. Infect Immun ,Vol, 16, No,1. pp: 344-352.
13.Nataro, J.P and Kaper, J.B. (1998). Diarrheagenic Escherichia coli. Clinical Microbiology Review,Vol, 11,No,1. pp: 142-201.
14.Nguyen, T.D., Thanh Vo2, T and Khac, H.V.( 2011). Virulence factors in Escherichia coli isolated from calves with diarrhea in Vietnam. J. Vet. Sci, Vol, 12,No,2. pp:159-164.
15.Novotna, R., Alexa, P., Hamrik, J., Madanat, A., Smola, J. and Cizek,A.(2005). Isolation and characterization Shiga toxin-producing Escherichia coli from sheep and goats in Jordan with evidence of multiresistant serotype O157:H7. Vet. Med. – Czech, Vol, 50,No, 3. pp: 111–118.
16. Ok, M., Guler ,L., Turgut, K., Ok, U., Sen, I., Gunduz, K., Birdane, M. F and Guzelbektes, H.(2008). The studies on the Aetiology of Diarrhoea in neonatal calves and determination of virulence gene markers of Escherichia coli Strains by multiplex PCR. Zoonoses and Public Health,Vol, 56,No,2 .pp: 94-101.
17. Phan, Q and Mshar, P.( 2003). Escherichia coliO157:H7 Gastroenteritis- Conecticut. Conecticut Dipartment of Public Health, Vol, 23,No, 2. pp: 5-8.
18. Riley, L.W., Remis, R.S., Helgerson, S.D., McGee, H.B., Wells, J.G and et al .(1983). Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med ,Vol, 308,No,12 . pp:681-685.
19.Schoonderwoerd, M., Clarke, R.C., Dreume, A.A and Rawluk, S.A.(1988). Colitis in calvesnatural and experimental infection with a verotoxin-producing strain of Escherichia coli O111.NM. Can. J .Vet. Res, Vol, 52, No,4. pp: 484-487.
20.Shabana, I.I. (2014). Escherichia coli Pathotypes Associated With Diarrhea in Human and Domestic Animals. American Journal of Animal and Veterinary Sciences, Vol, 9, No, 3. pp: 155-161.
21.Shams, Z., Tahamtan, Y., Pourbakhsh, A., Hosseiny, M.H., Kargar, M. and Hayati,M.(2012). Detection of enterotoxigenic K99 (F5) and F41 from fecal sample of calves by molecular and serological methods. Comp Clin Pathol,Vol, 21, pp:475–478.
22. Sunderland, S., Sarasola, P., Giles, C.G and Smith, D.G.(2003). Efficacy of danofloxacin 18% injectable solution in the treatment of Escherichia coli diarrhoea in young calves in Europe. Research in Veterinary Science,Vol, 74, No,2. pp: 171-178.
23.Verma, Sh., Kumar, M., Kashyap, S.,Singh,M and Venkatesh,V.(2013). Current Scenario Of Escherichia coli And Its Serotype“o157” In Indian Subcontinent. IJIRSET,Vol, 2, Issue 7,ISSN: 2319-8753.
24.Wani, S.A., Bhat, M.A., Samanta, I., Nishikawa,YandBuchh, A.S.( 2003). Isolation and characterization of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic Escherichia coli (EPEC) from calves and lambs with diarrhoea in India. Letters in Applied Microbiology ,Vol, 37,NO, 2. pp: 121–126.
25.Wani, S.A., Hussain, I., Beg, S.A., Rather, M.A., Kabli, Z.A. Mir, M.A and Nishikawa,Y. (2013). Diarrhoeagenic Escherichia coli and salmonellae in calves and lambs in Kashmir.absence, prevalence and antibiogram. Rev. sci. tech. Off. int. Epiz.,Vol,32, No,3.