Prevalence and characterization of verotoxigenic-Escherichia coli isolates from pigs in Malaysia

BMC Veterinary Research

Kwai Lin Thong²  Chew Chieng Yeo³  Peck Toung Ooi¹  Lai Kuan Tan²  Wing Sze Ho² 

[1] Department of Clinical Veterinary Studies, Faculty of Veterinary Medicine, University Putra Malaysia, 43400, Serdang, Selangor Darul Ehsan, Malaysia;Laboratory of Biomedical Science and Molecular Microbiology UMBIO Research Cluster, University of Malaya, 50603, Kuala Lumpur,Malaysia;Faculty of Agriculture and Biotechnology, Universiti Sultan Zainal Abidin, 21300, Kuala Terengganu, Malaysia

关键词: Swine;    Pig farms;    Multidrug resistance;    Postweaning diarrhea (PWD);    Shiga-toxin (Stx)-producing E. coli (STEC);    Verotoxigenic-E. coli (VTEC);   

Others  :  1119529
DOI  :  10.1186/1746-6148-9-109

 received in 2013-03-27, accepted in 2013-05-29,  发布年份 2013

【 摘 要 】

Background

Postweaning diarrhea caused by pathogenic Escherichia coli, in particular verotoxigenic E. coli (VTEC), has caused significant economic losses in the pig farming industry worldwide. However, there is limited information on VTEC in Malaysia. The objective of this study was to characterize pathogenic E. coli isolated from post-weaning piglets and growers with respect to their antibiograms, carriage of extended-spectrum beta-lactamases, pathotypes, production of hemolysins and fimbrial adhesins, serotypes, and genotypes.

Results

PCR detection of virulence factors associated with different E. coli pathotypes (ETEC, EPEC, EHEC, and VTEC) revealed that VTEC was the only pathotype identified from six swine farms located at north-western Peninsular Malaysia. A low prevalence rate of VTEC was found among the swine samples (n = 7/345) and all 7 VTEC isolates were multidrug resistant. Five of these isolates from different hosts raised in the same pen were likely to be of the same clone as they shared identical sero-pathotypes (O139:H1, VT2e/α-hly/F18), resistance profiles and DNA fingerprinting profiles. Two other serotypes, O130: H26 (n = 1) and O168: H21 (n = 1) carrying virulence factors were also identified. O168: H21 is possibly a new serotype as this has not been previously reported.

Conclusions

The occurrence of VTEC with infrequently encountered serotypes that are multidrug resistant and harbouring virulence factors may be of public health concern. The detection of possible clones in this study also showed that the combination of different typing tools including phenotyping and genotyping methods is useful for molecular epidemiologic surveillance and studies.

【 授权许可】

   
2013 Ho et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Flores A: New way to control E. coli in weaned pigs. Agric Res 2004, 52:9.
• [2]Fairbrother JM, Nadeau E, Gyles CL: Escherichia coli in postweaning diarrhea in pigs: an update on bacterial types, pathogenesis, and prevention strategies. Anim Health Res Rev 2005, 6:17-39.
• [3]Botteldoorn N, Heyndrickx M, Rijpens N, Herman L: Detection and characterization of verotoxigenic Escherichia coli by a VTEC/EHEC multiplex PCR in porcine faeces and pig carcass swabs. Res Microbiol 2003, 154:97-104.
• [4]Kaper JB, Nataro JP, Mobley HL: Pathogenic Escherichia coli. Nat Rev Microbiol 2004, 2:123-140.
• [5]Strockbine NA, Wells JG, Bopp CA, Barrett TJ: Overview of detection and subtyping methods. In Escherichia coli O157:H7 and other Shiga toxin-producing E. coli strains. Edited by Kaper JB, O’Brien AD. Washington, D.C: American Society for Microbiology; 1998:331-356.
• [6]DesRosiers A, Fairbrother JM, Johnson RP, Desautels C, Letellier A, Quessy S: Phenotypic and genotypic characterization of Escherichia coli verotoxin-producing isolates from humans and pigs. J Food Prot 2001, 64:1904-1911.
• [7]von Muffling T, Smaijlovic M, Nowak B, Sammet K, Bulte M, Klein G: Preliminary study of certain serotypes, genetic and antimicrobial resistance profiles of verotoxigenic Escherichia coli (VTEC) isolated in Bosnia and Germany from cattle or pigs and their products. Int J Food Microbiol 2007, 117:185-191.
• [8]Nagy B, Arp LH, Moon HW, Casey TA: Colonization of the small intestine of weaned pigs by enterotoxigenic Escherichia coli that lack known colonization factors. Vet Pathol 1992, 29:239-246.
• [9]Vu Khac H, Holoda E, Pilipcinec E, Blanco M, Blanco JE, Mora A, Dahbi G, López C, González EA, Blanco J: Serotypes, virulence genes, and PFGE profiles of Escherichia coli isolated from pigs with postweaning diarrhoea in Slovakia. BMC Vet Res 2006, 2:10. BioMed Central Full Text
• [10]Nagy B, Fekete PZ: Enterotoxigenic Escherichia coli (ETEC) in farm animals. Vet Res 1999, 30:259-284.
• [11]Yu KX, Thong KL: Multiplex PCR for simultaneous detection of virulence genes in Escherichia coli. Mal J Science 2009, 28:1-14.
• [12]Fields PI, Blom KH, Hughes JH, Helsel LO, Feng P, Swaminathan B: Molecular characterisation of the gene encoding H-antigen in Escherichia coli and development of PCR-restriction fragment length polymorphism test for identification of E. coli O157:H7 and O157:NM. J Clin Microbiol 1997, 35:1066-1070.
• [13]Nataro JP, Kaper JB: Diarrheagenic Escherichia coli. Clin Microbiol Rev 1998, 11:142-201.
• [14]Reid SD, Selander R, Whittam TS: Sequence diversity of flagellin (FliC) alleles in pathogenic Escherichia coli. J Bacteriol 1999, 181:153-160.
• [15]Prapasarakul N, Tummaruk P, Niyomtum W, Tripipati T, Serichantalergs O: Virulence genes and antimicrobial susceptibilities of hemolytic and nonhemolytic Escherichia coli isolated from post-weaning piglets in central Thailand. J Vet Med Sci 2010, 72:1603-1608.
• [16]Zhang W, Zhao M, Ruesch L, Omot A, Francis D: Prevalence of virulence genes in Escherichia coli strains recently isolated from young pigs with diarrhea in the US. Vet Microbiol 2007, 123:145-152.
• [17]Oanh TK, Nguyen VK, Do TN, Goddeeris BM, De Greve H: Escherichia coli strains causing edema disease in northern Vietnam share an identical verotoxin 2e. Trop Anim Health Prod 2010, 42:1797-1804.
• [18]Dirkzwager A, Veldman B, Bikker P: A nutritional approach for the prevention of post weaning syndrome in piglets. Anim Res 2005, 54:231-236.
• [19]Han W, Liu B, Cao B, Beutin L, Kruger U, Liu H, Li Y, Liu Y, Feng L, Wang L: DNA microarray-based identification of serogroups and virulence gene patterns of Escherichia coli isolates associated with porcine postweaning diarrhea and edema disease. Appl Environ Microbiol 2007, 73:4082-4088.
• [20]Beutin L, Kruger U, Krause G, Miko A, Martin A, Strauch E: Evaluation of major types of Shiga toxin 2e-producing Escherichia coli present in food, pigs, and the environment as potential pathogens for humans. Appl Environ Microbiol 2008, 74:4806-4816.
• [21]Müller D, Greune L, Heusipp G, Karch H, Fruth A, Tschäpe H, Schmidt MA: Identification of unconventional intestinal pathogenic Escherichia coli isolates expressing intermediate virulence factor profiles by using a novel single-step multiplex PCR. Appl Environ Microbiol 2007, 73:3380-3390.
• [22]Madoroba E, Van Driessche E, De Greve H, Mast J, Ncube I, Read J, Beeckmans S: Prevalence of enterotoxigenic Escherichia coli virulence genes from scouring piglets in Zimbabwe. Trop Anim Health Prod 2009, 41:1539-1547.
• [23]Burgos Y, Beutin L: Common origin of plasmid encoded alpha-haemolysin genes in Escherichia coli. BMC Microbiol 2010, 10:193. BioMed Central Full Text
• [24]CIWF (Compassion in World Farming): Antibiotics in animal farming: Public health and animal welfare. 2011. Available at http://www.ciwf.org.uk/includes/documents/cm_docs/2011/a/antibiotics_in_animal_farming.pdf webcite. Accessed on October 29, 2012
• [25]Docic M, Bilkei G: Differences in antibiotic resistance in Escherichia coli, isolated from East-European swine herds with or without prophylactic use of antibiotics. J Vet Med B Infect Dis Vet Public Health 2003, 50:27-30.
• [26]Lee SI, Rayamahji N, Lee WJ, Cha SB, Shin MK, Roh MY, Sang HY: Genotypes, antibiogram, and pulsed-field gel electrophoresis profiles of Escherichia coli strains from piglets in Korea. J Vet Diagn Invest 2009, 21:510-516.
• [27]Cornick NA, Vu Khac H: Indirect transmission of Escherichia coli O157:H7 occurs readily among swine but not among sheep. Appl Environ Microbiol 2008, 74:2488-2491.
• [28]Olsvik O, Strockbine NA: PCR detection of heat-stable, heat-labile and Shiga-like toxin genes in Escherichia coli. In Diagnostic molecular microbiology. Edited by Persing DH, Smith TF, Tenover FC, White TJ. Washington, DC: American Society for Microbiology; 1993:271-276.
• [29]Imberechts H, Van Pelt N, De Greve H, Lintermans P: Sequences related to the major subunit gene fedA of F107 fimbriae in porcine Escherichia coli strains that express adhesive fimbriae. FEMS Microbiol Lett 1994, 119:309-314.
• [30]Ojeniyi B, Ahrens P, Meyling A: Detection of fimbrial and toxin genes in Escherichia coli and their prevalence in piglets with diarrhea. The application of colony hybridization assay, polymerase chain reaction and phenotypic assays. J Vet Med B 1994, 41:49-59.
• [31]Coimbra RS, Grimont F, Lenormand P, Burguiere P, Beutin L, Grimont PAD: Identification of Escherichia coli O-serogroups by restriction of the amplified O-antigen gene cluster (rfb-RFLP). Res Microbiol 2000, 151:639-654.
• [32]Prager R, Strutz U, Fruth A, Tschäpe H: Subtyping of pathogenic Escherichia coli strains using flagellar (H)-antigens: serotyping versus fliC polymorphisms. Int J Med Microbiol 2003, 29:477-486.
• [33]Thong KL, Lai KS, Puthucheary SD, Koh YT, Ahmad Yassin RM: Subtyping of Salmonella enterica serovar Muenchen by pulsed-field gel electrophoresis, plasmid profiling and antimicrobial susceptibility testing. Mal J Science 2007, 26:1-13.
• [34]Römling U, Tümmler B: Achieving 100% typeability of Pseudomonas aeruginosa by pulsed-field gel electrophoresis. J Clin Microbiol 2000, 38:464-465.
• [35]Lim KT, Yasin R, Yeo CC, Putucheary SD, Thong KL: Characterization of multidrug resistant ESBL- producing Escherichia coli isolates from hospitals in Malaysia. J Biomed Biotechnol 2009, 2009:165637. 10.1155/2009/165637
• [36]Clinical and Laboratory Standards Institute: Performance standards for antimicrobial susceptibility testing; 20th informational supplement (M100-S20). Wayne PA: Clinical and Laboratory Standards Institute; 2010.
• [37]Jarlier V, Nicolas MH, Fournier G, Philippon A: Extended broad-spectrum beta-lactamases conferring transferable resistance to newer beta-lactam agents in Enterobacteriaceae: hospital prevalence and susceptibility patterns. Rev Infect Dis 1988, 10:867-878.
• [38]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, 42:1099-1108.
• [39]Schmidt H, Beutin L, Karch H: Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933. Infect Immun 1995, 6:1055-1061.