Jingyu Wang^1*, Pan Tang¹, Dan Tan¹, Liqin Wang¹, Sandong Zhang¹, Yuanhao Qiu¹, Rui Dong¹, Wanhua Liu¹, Jingjing Huang¹, Ting Chen¹, Juanjuan Ren¹, Cengshan Li¹, Hung-Jen Liu^2,3,4*

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¹ College of Veterinary Medicine, Northwest A&F University, Yangling, China.
² Institute of Molecular Biology, National Chung Hsing University, Taiwan.
³ Agricultural Biotechnology Center, National Chung Hsing University, Taiwan.
⁴ Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taiwan.
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Abstract: Various virulence-associated genes or pathogenicity island are responsible for determining the pathogenicity of Escherichia coli strains. However, the correlation of the number and combination patterns of virulence-associated genes in Escherichia coli strains with their pathogenicity remains largely unknown. In this work, 581 chicken Escherichia coli strains were isolated from 1045 liver samples of dead chickens from 50 chicken farms at four provinces in China during 2007-2012. Based on the pathogenic test of SPF chickens, 320 chickens pathogenic Escherichia coli isolates were identified as highly (n = 193), intermediate (n = 98) and low pathogenic (n = 29) strains, respectively. Furthermore, the number of virulence genes in the 320 chicken pathogenic and 50 non-pathogenic Escherichia coli strains was examined. Our results reveal that thirteen virulence genes in Escherichia coli strains were detected, and all strains carried at least two or more than two virulence-associated genes. This study also suggests that highly pathogenic E. coli strains simultaneously carried at least 8 to13 virulence genes while intermediate pathogenic strains carried at least 5 to 8 virulence genes. The number of virulence-associated genes detected in highly pathogenic strains showed there were more significant differences than that in low pathogenic strains (P < 0.01). The detection rate of genes irp2, fyuA, and colV in high pathogenic strains was significantly higher than that in low and non-pathogenic strains (P < 0.01). Nine virulence-associated genes irp2, fyuA, iucA, iucD, iutA, papC, iss, tsh, and colV were more often detected in highly and intermediate pathogenic E. coli strains. Taken together, our results provide evidences demonstrating that the pathogenicity of Escherichia coli strains is closely associated with the number and combination patterns of virulence-associated genes.

Keywords: Avian Pathogenic Escherichia coli, Pathogenicity, Virulence-Associated Genes

Cite this paper: Wang, J. , Tang, P. , Tan, D. , Wang, L. , Zhang, S. , Qiu, Y. , Dong, R. , Liu, W. , Huang, J. , Chen, T. , Ren, J. , Li, C. , Liu, H. , (2015) The Pathogenicity of Chicken Pathogenic Escherichia coli Is Associated with the Numbers and Combination Patterns of Virulence-Associated Genes. Open Journal of Veterinary Medicine, 5, 243-254. doi: 10.4236/ojvm.2015.512033.

References

[1]   Lutful Kabir, S.M. (2010) Avian Colibacillosis and Salmonellosis: A Closer Look at Epidemiology, Pathogenesis, Diagnosis, Control and Public Health Concerns. International Journal of Environmental Research and Public Health, 7, 89-114.
http://dx.doi.org/10.3390/ijerph7010089

[2]   Vandekerchove, D., Vandemaele, F., Adriaensen, C., Zaleska, M., Hernalsteens, J.P., De Baets, L., Butaye, P., Van Immerseel, F., Wattiau, P., Laevens, H., Mast, J., Goddeeris, B. and Pasmans, F. (2005) Virulence-Associated Traits in Avian Escherichia coli: Comparison between Isolates from Colibacillosis-Affected and Clinically Healthy Layer Flocks. Veterinary Microbiology, 108, 75-87.
http://dx.doi.org/10.1016/j.vetmic.2005.02.009

[3]   Cheng, D.R., Sun, H.C., Xu, J.S. and Gao, S. (2006) Prevalence of LEE and HPI Pathogenicity Islands of Escherichia coli Isolates from Weaned Piglets in China. Acta Microbiologica Sinica, 46, 368-372.

[4]   Yu, J. and Kaper, J.B. (1992) Cloning and Characterization of the eae Gene of Enterohaemorrhagic Escherichia coli O157:H7. Molecular Microbiology, 6, 411-417.
http://dx.doi.org/10.1111/j.1365-2958.1992.tb01484.x

[5]   Horne, S.M., Pfaff-McDonough, S.J., Giddings, C.W. and Nolan, L.K. (2000) Cloning and Sequencing of the iss Gene from a Virulent Avian Escherichia coli. Avian Diseases, 44, 179-184.
http://dx.doi.org/10.2307/1592522

[6]   Forman, S., Nagiec, M.J., Abney, J., Perry, R.D. and Fetherston, J.D. (2007) Analysis of the Aerobactin and Ferric Hydroxamate Uptake Systems of Yersinia pestis. Microbiology, 153, 2332-2341.
http://dx.doi.org/10.1099/mic.0.2006/004275-0

[7]   Xia, M.Y., Tang, W. and Yue, H. (2010) Progress on Virulence-Associated Factors ColBM and RfaH in Escherichia coli. Progress in Veterinary Medicine, 31, 86-89.

[8]   Carniel, E., Guilvout, I. and Prentice, M. (1996) Characterization of a Large Chromosomal “High-Pathogenicity Island” in Biotype 1B Yersinia enterocolitica. Journal of Bacteriology, 178, 6743-6751.

[9]   Provence, D.L. and Curtiss, R.R. (1994) Isolation and Characterization of a Gene Involved in Hemagglutination by an Avian Pathogenic Escherichia coli strain. Infection and Immunity, 62, 1369-1380.

[10]   Cloud, S.S., K.Rosenberger, J., Fries, P.A., Wilson, R.A. and Odor, E.M. (1985) In Vitro and in Vivo Characterization of Avian Escherichia coli. I. Serotypes, Metabolic Activity, and Antibiotic Sensitivity. Avian Disease, 29, 1084-1093.
http://dx.doi.org/10.2307/1590463

[11]   Rosenberger, J.K., Fries, P.A., Cloud, S.S. and Wilson, R.A. (1985) In Vitro and in Vivo Characterization of Avian Escherichia coli. II. Factors Associated with Pathogenicity. Avian Diseases, 29, 1094-1107.
http://dx.doi.org/10.2307/1590464

[12]   Rosenberger, J.K., Fries, P.A. and Cloud, S.S. (1985) In Vitro and in Vivo Characterization of Avian Escherichia coli. III. Immunization. Avian Diseases, 29, 1108-1117.
http://dx.doi.org/10.2307/1590465

[13]   Janben, T., Schwarz, C., Preikschat, P., Voss, M., Philipp, H.C. and Wieler, L.H. (2001) Virulence-Associated Genes in Avian Pathogenic Escherichia coli (APEC) Isolated from Internal Organs of Poultry Having Died from Colibacillosis. International Journal of Medical Microbiology, 291, 371-378. http://dx.doi.org/10.1078/1438-4221-00143

[14]   Johnson, J.R. and Stell, A.L. (2000) Extended Virulence Genotypes of Escherichia coli Strains from Patients with Urosepsis in Relation to Phylogeny and Host Compromise. Journal of Infectious Diseases, 181, 261-272.
http://dx.doi.org/10.1086/315217

[15]   Sperandio, V., Kaper, J.B., Bortolini, M.R., Neves, B.C., Keller, R. and Trabulsi, L.R. (1998) Characterization of the Locus of Enterocyte Effacement (LEE) in Different Enteropathogenic Escherichia coli (EPEC) and Shiga-Toxin Producing Escherichia coli (STEC) Serotypes. FEMS Microbiology Letters, 164, 133-139.
http://dx.doi.org/10.1111/j.1574-6968.1998.tb13078.x

[16]   Bach, S., de Almeida, A. and Carniel, E. (2000) The Yersinia High-Pathogenicity Island Is Present in Different Members of the Family Enterobacteriaceae. FEMS Microbiology Letters, 183, 289-294.
http://dx.doi.org/10.1111/j.1574-6968.2000.tb08973.x

[17]   Carniel, E. (2001) The Yersinia High-Pathogenicity Island: An Iron-Uptake Island. Microbes Infection, 3, 561-569.
http://dx.doi.org/10.1016/S1286-4579(01)01412-5

[18]   Clermont, O., Bonacorsi, S. and Bingen, E. (2001) The Yersinia High-Pathogenicity Island Is Highly Predominant in Virulence-Associated Phylogenetic Groups of Escherichia coli. FEMS Microbiology Letters, 196, 153-157.
http://dx.doi.org/10.1111/j.1574-6968.2001.tb10557.x

[19]   Jores, J., Rumer, L., Kiessling, S., Kaper, J.B. and Wieler, L.H. (2001) A Novel Locus of Enterocyte Effacement (LEE) Pathogenicity Island Inserted at pheV in Bovine Shiga Toxin-Producing Escherichia coli Strain O103:H2. FEMS Microbiology Letters, 204, 75-79.
http://dx.doi.org/10.1111/j.1574-6968.2001.tb10866.x

[20]   Penteado, A.S., Ugrinovich, L.A., Blanco, J., Blanco, M., Blanco, J.E., Mora, A., Andrade, J.R., Correa, S.S. and Pestana de Castro, A.F. (2002) Serobiotypes and Virulence Genes of Escherichia coli Strains Isolated from Diarrheic and Healthy Rabbits in Brazil. Veterinary Microbiology, 89, 41-51.
http://dx.doi.org/10.1016/S0378-1135(02)00148-7

[21]   Rodriguez-Siek, K.E., Giddings, C.W., Doetkott, C., Johnson, T.J. and Nolan L.K. (2005) Characterizing the APEC Pathotype. Veterinary Research, 36, 241-256.
http://dx.doi.org/10.1051/vetres:2004057

[22]   Hu, Y.X., Ou-Yang, D.M., Yu, X.F., Li, M.L., Liu, Q.T. and He, Z.X. (2008) Analysis of Partial Sequence of Enteropathogenic Escherichia coli Strains with Yersinia HPI irp2. Journal of Chinese Infection Control, 7, 167-169.

[23]   Smith, M.G., Jordan, D., Chapman, T.A., Chin, J.J., Barton, M.D., Do, T.N., Fahy, V.A., Fairbrother, J.M. and Trott, D.J. (2010) Antimicrobial Resistance and Virulence Gene Profiles in Multi-Drug Resistant Enterotoxigenic Escherichia coli Isolated from Pigs with Post-Weaning Diarrhoea. Veterinary Microbiology, 145, 299-307.
http://dx.doi.org/10.1016/j.vetmic.2010.04.004

[24]   La Ragione, R.M., Mclaren, I.M., Foster, G., Cooley, W.A. and Woodward, M.J. (2002) Phenotypic and Genotypic Characterisation of Avian Escherichia coli O86:K61 Isolates Possessing a Gamma-Like Intimin. Applied and Environmental Microbiology, 68, 4932-4942.
http://dx.doi.org/10.1128/AEM.68.10.4932-4942.2002

[25]   Galli, L., Miliwebsky, E., Irino, K., Leotta, G. and Rivas, M. (2010) Virulence Profile Comparison between LEE- Negative Shiga Toxin-Producing Escherichia coli (STEC) Strains Isolated from Cattle and Humans. Veterinary Microbiology, 143, 307-313.
http://dx.doi.org/10.1016/j.vetmic.2009.11.028

[26]   Wang, Y.J., Fan, C. and Li, Y.J. (2004) The Relation between fimC Gene and Pathogenicity of Avian Escherichia coli. Acta Veterinaria et Zootechnica Sinica, 35, 680-684.

[27]   Jin, W.J., Zheng, Z.M., Zhang, Y.Z., Qin, A.J., Shao, H.X., Liu, Y.L., Wang, J. and Wang, Q.D. (2008) Distribution of Virulence-Associated Genes of Avian Pathogenic Escherichia coli Isolates in China. Agricultural Science in China, 7, 1511-1515.
http://dx.doi.org/10.1016/S1671-2927(08)60410-1

[28]   Johnson, T.J., Siek, K.E., Johnson, S.J. and Nolan, L.K. (2006) DNA Sequence of a ColV Plasmid and Prevalence of Selected Plasmid-Encoded Virulence Genes among Avian Escherichia coli Strains. Journal of Bacteriology, 188, 745-758.
http://dx.doi.org/10.1128/JB.188.2.745-758.2006

[29]   Johnson, T.J., Johnson, S.J. and Nolan, L.K. (2006) Complete DNA Sequence of a ColBM Plasmid from Avian Pathogenic Escherichia coli Suggests That It Evolved from Closely Related ColV Virulence Plasmids. Journal of Bacteriology, 188, 5975-5983.
http://dx.doi.org/10.1128/JB.00204-06