A statistical approach to identify prevalent virulence factors responsible for post-weaning diarrhoeic piglets

https://doi.org/10.17221/84/2021-VETMEDCitation:

Lin CS, Huang CH, Adi VSK, Huang CW, Cheng YI, Chen JH, Liu YC (2022): A statistical approach to identify prevalent virulence factors responsible for post-weaning diarrhoeic piglets. Vet Med-Czech 67, 430–439.

download PDF

A statistical approach was carried out to identify the prevalent virulence factors responsible for post-weaning diarrhoea (PWD). Healthy piglets’ faecal samples and diarrhoeic piglets’ rectal swab specimens were secured. Twenty-six (26) and 100 independent enterotoxigenic Escherichia coli (ETEC) strains were subsequently isolated. These strains were assessed utilising polymerase chain reaction to identify the encoding genes of six virulence factors: heat-labile enterotoxin (LT; encoded by eltAB), heat-stable enterotoxin A (STa; encoded by estA), heat-stable enterotoxin B (STb; encoded by estB), enteroaggregative E. coli heat-stable enterotoxin 1 (EAST1; encoded by astA), Shiga toxin 2e (Stx2e; encoded by stx2e), and F18 fimbriae (encoded by fedA). The LT and ST secretions were investigated using enzyme-linked immunosorbent assays. From direct observation, no stx2e was evident in the 126 strains. Among the 26 strains retrieved from the healthy piglets, none harboured fedA or secreted LT; 23% (6/26) secreted ST, and 50% (13/26) carried astA. A statistical regression was applied on the 100 E. coli strains retrieved from the diarrhoeic piglets, where fedA was set as the dependent variable and the enterotoxin secretions were set as the independent variables. The results exhibit that the LT secretion was the only significant factor (P < 0.000 1) correlated to fedA in the diarrhoeic piglets; thus, it is concluded that the prevalent virulence factors for PWD were the ECET strain with F18 fimbriae adhesion and LT secretion, but not astA or stx2e.

References:
Baron EJ, Antonson S. Identification of unusual pathogenic gram-negative aerobic and facultatively anaerobic bacteria. Clin Infect Dis. 1997 Mar;24(3):1-9. https://doi.org/10.1093/clinids/24.3.537
 
Beutin L, Kruger U, Krause G, Miko A, Martin A, Strauch E. Evaluation of major types of Shiga toxin 2E-producing Escherichia coli bacteria present in food, pigs, and the environment as potential pathogens for humans. Appl Environ Microbiol. 2008 Aug;74(15):4806-16. https://doi.org/10.1128/AEM.00623-08
 
Cheng D, Sun H, Xu J, Gao S. PCR detection of virulence factor genes in Escherichia coli isolates from weaned piglets with edema disease and/or diarrhea in China. Vet Microbiol. 2006 Jul 20;115(4):320-8. https://doi.org/10.1016/j.vetmic.2006.02.013
 
Choi YS, Lee JK, Jung JT, Jung YC, Jung JH, Jung MO, Choi YI, Jin SK, Choi JS. Comparison of meat quality and fatty acid composition of longissimus muscles from purebred pigs and three-way crossbred LYD pigs. Korean J Food Sci Anim Resour. 2016 Oct 31;36(5):689-96. https://doi.org/10.5851/kosfa.2016.36.5.689
 
Crowley E, Bird P, Fisher K, Goetz K, Boyle M, Benzinger MJ Jr, Juenger M, Agin J, Goins D, Johnson R. Evaluation of the VITEK 2 Gram-negative (GN) microbial identification test card: Collaborative study. J AOAC Int. 2012 May-Jun;95(3):778-85. https://doi.org/10.5740/jaoacint.CS2012_02
 
Cutler SA, Lonergan SM, Cornick N, Johnson AK, Stahl CH. Dietary inclusion of colicin e1 is effective in preventing postweaning diarrhea caused by F18-positive Escherichia coli in pigs. Antimicrob Agents Chemother. 2007 Nov;51(11):3830-5. https://doi.org/10.1128/AAC.00360-07
 
Dubreuil JD. Escherichia coli STb toxin and colibacillosis: Knowing is half the battle. FEMS Microbiol Lett. 2008 Jan;278(2):137-45. https://doi.org/10.1111/j.1574-6968.2007.00967.x
 
Erume J, Berberov EM, Kachman SD, Scott MA, Zhou Y, Francis DH, Moxley RA. Comparison of the contributions of heat-labile enterotoxin and heat-stable enterotoxin b to the virulence of enterotoxigenic Escherichia coli in F4ac receptor-positive young pigs. Infect Immun. 2008 Jul;76(7):3141-9. https://doi.org/10.1128/IAI.01743-07
 
Evans DJ Jr, Evans DG, Gorbach SL. Production of vascular permeability factor by enterotoxigenic Escherichia coli isolated from man. Infect Immun. 1973 Nov 1;8(5):725-30. https://doi.org/10.1128/iai.8.5.725-730.1973
 
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 Jun;6(1):17-39. https://doi.org/10.1079/AHR2005105
 
Frydendahl K, Kare Jensen T, Strodl Andersen J, Fredholm M, Evans G. Association between the porcine Escherichia coli F18 receptor genotype and phenotype and susceptibility to colonisation and postweaning diarrhoea caused by E. coli O138:F18. Vet Microbiol. 2003 May 2;93(1):39-51. https://doi.org/10.1016/S0378-1135(02)00348-6
 
Gyles CL. Shiga toxin-producing Escherichia coli: An overview. J Anim Sci. 2007 Mar;85(13_Suppl):E45-62. https://doi.org/10.2527/jas.2006-508
 
Ho WS, Tan LK, Ooi PT, Yeo CC, Thong KL. Prevalence and characterization of verotoxigenic-Escherichia coli isolates from pigs in Malaysia. BMC Vet Res. 2013 Jun 4;9:109. https://doi.org/10.1186/1746-6148-9-109
 
Kim YJ, Kim JH, Hur J, Lee JH. Isolation of Escherichia coli from piglets in South Korea with diarrhea and characteristics of the virulence genes. Can J Vet Res. 2010 Jan;74(1):59-64.
 
Lee CH, Hu ST, Swiatek PJ, Moseley SL, Allen SD, So M. Isolation of a novel transposon which carries the Escherichia coli enterotoxin STII gene. J Bacteriol. 1985 May;162(2):615-20. https://doi.org/10.1128/jb.162.2.615-620.1985
 
Lee SI, Kang SG, Kang ML, Yoo HS. Development of multiplex polymerase chain reaction assays for detecting enterotoxigenic Escherichia coli and their application to field isolates from piglets with diarrhea. J Vet Diagn Invest. 2008 Jul;20(4):492-6. https://doi.org/10.1177/104063870802000413
 
Marques LRM, Peiris JSM, Cryz SJ, O’Brien AD. Escherichia coli strains isolated from pigs with edema disease produce a variant of shiga-like toxin II. FEMS Microbiol Lett. 1987;44(1):33-8. https://doi.org/10.1111/j.1574-6968.1987.tb02237.x
 
McConnell MM, Smith HR, Willshaw GA, Scotland SM, Rowe B. Plasmids coding for heat-labile enterotoxin production isolated from Escherichia coli O78: Comparison of properties. J Bacteriol. 1980 Jul;143(1):158-67. https://doi.org/10.1128/jb.143.1.158-167.1980
 
Meijerink E, Neuenschwander S, Fries R, Dinter A, Bertschinger HU, Stranzinger G, Vogeli P. A DNA polymorphism influencing alpha(1,2)fucosyltransferase activity of the pig FUT1 enzyme determines susceptibility of small intestinal epithelium to Escherichia coli F18 adhesion. Immunogenetics. 2000 Nov;52(1-2):129-36. https://doi.org/10.1007/s002510000263
 
Nagy B, Fekete PZ. Enterotoxigenic Escherichia coli in veterinary medicine. Int J Med Microbiol. 2005 Oct;295(6-7):443-54. https://doi.org/10.1016/j.ijmm.2005.07.003
 
Ngeleka M, Pritchard J, Appleyard G, Middleton DM, Fairbrother JM. Isolation and association of Escherichia coli AIDA-I/STb, rather than EAST1 pathotype, with diarrhea in piglets and antibiotic sensitivity of isolates. J Vet Diagn Invest. 2003 May;15(3):242-52. https://doi.org/10.1177/104063870301500305
 
Paiva de Sousa C, Dubreuil JD. Distribution and expression of the astA gene (EAST1 toxin) in Escherichia coli and Salmonella. Int J Med Microbiol. 2001 Apr;291(1):15-20. https://doi.org/10.1078/1438-4221-00097
 
Pass MA, Odedra R, Batt RM. Multiplex PCRs for identification of Escherichia coli virulence genes. J Clin Microbiol. 2000 May;38(5):2001-4. https://doi.org/10.1128/JCM.38.5.2001-2004.2000
 
Skerman FJ, Formal SB, Falkow S. Plasmid-associated enterotoxin production in a strain of Escherichia coli isolated from humans. Infect Immun. 1972 Apr;5(4):622-4. https://doi.org/10.1128/iai.5.4.622-624.1972
 
Teng CH, Wu PC, Tang SL, Chen YC, Cheng MF, Huang PC, Ko WC, Wang JL. A large spatial survey of colistin-resistant gene mcr-1-carrying E. coli in rivers across Taiwan. Microorganisms. 2021 Mar 31;9(4):722. https://doi.org/10.3390/microorganisms9040722
 
Toledo A, Gomez D, Cruz C, Carreon R, Lopez J, Giono S, Castro AM. Prevalence of virulence genes in Escherichia coli strains isolated from piglets in the suckling and weaning period in Mexico. J Med Microbiol. 2012 Jan;61(Pt 1):148-56. https://doi.org/10.1099/jmm.0.031302-0
 
Vu-Khac H, Holoda E, Pilipcinec E, Blanco M, Blanco JE, Dahbi G, Mora A, Lopez C, Gonzalez EA, Blanco J. Serotypes, virulence genes, intimin types and PFGE profiles of Escherichia coli isolated from piglets with diarrhoea in Slovakia. Vet J. 2007 Jul;174(1):176-87. https://doi.org/10.1016/j.tvjl.2006.05.019
 
Wachsmuth K, Wells J, Shipley P, Ryder R. Heat-labile enterotoxin production in isolates from a shipboard outbreak of human diarrheal illness. Infect Immun. 1979 Jun;24(3):793-7. https://doi.org/10.1128/iai.24.3.793-797.1979
 
Wasteson Y, Olsvik O. Specific DNA fragments coding for ST1 and LT1 toxins, and K88 (F4) adhesin in enterotoxigenic Escherichia coli. Zentralbl Veterinarmed B. 1991 Aug;38(6):445-52. https://doi.org/10.1111/j.1439-0450.1991.tb00894.x
 
Yamamoto T, Yokota T. Plasmids of enterotoxigenic Escherichia coli H10407: Evidence for two heat-stable enterotoxin genes and a conjugal transfer system. J Bacteriol. 1983 Mar;153(3):1352-60. https://doi.org/10.1128/jb.153.3.1352-1360.1983
 
Yang GY, Guo L, Su JH, Zhu YH, Jiao LG, Wang JF. Frequency of diarrheagenic virulence genes and characteristics in Escherichia coli isolates from pigs with diarrhea in China. Microorganisms. 2019 Sep 2;7(9):308. https://doi.org/10.3390/microorganisms7090308
 
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 Jul 20;123(1-3):145-52. https://doi.org/10.1016/j.vetmic.2007.02.018
 
Zhang W. Progress and challenges in vaccine development against enterotoxigenic Escherichia coli (ETEC)-associated porcine post-weaning diarrhoea (PWD). J Vet Med Res. 2014 Aug;1(2): [13].
 
Zang CZ, Kan SC, Yeh CW, Lin CC, Shieh CJ, Liu YC. Ultrasound-assisted (R)-phenylephrine whole-cell bioconversion by S. marcescens N10612. Ultrason Sonochem. 2015 Sep;26:415-21. https://doi.org/10.1016/j.ultsonch.2015.01.010
 
download PDF

© 2022 Czech Academy of Agricultural Sciences | Prohlášení o přístupnosti