Experimental Induction of Escherichia coli Diarrhoea in Weaned Piglets

Luciana Rossi, Simona Vagni, Carlo Polidori, Giovanni Loris Alborali, Antonella Baldi, Vittorio Dell’Orto

Department of Veterinary Sciences and Technology for Food Safety, Università di Milano, Milan, Italy.
Istituto Zooprofilattico Sperimentale della Lombardia ed Emilia Romagna (IZSLER), Brescia, Italy.
Università Telematica San Raffaele Roma, Rome, Italy.

DOI: 10.4236/ojvm.2012.21001                

Abstract

Escherichia coli diarrhoea is a multifactorial condition which usually occurs during the post-weaning and is responsible for economic losses in pig production. One approach, to evaluate if substitute of antibiotic in vivo is effective in controlling postweaning diarrhea in the gastrointestinal tract ecosystem, is to use an appropriate disease model. However, there are still many criticisms related to the incidence and the severity of the diarrhoea in the experimental conditions. The aim of the study was to set up an Escherichia coli challenge model in order to induce a significant percentage of diarrhoea in weaned piglets for the evaluation of innovative compounds in vivo. A total of 35 piglets, weaned at 33±2 days were randomized into 3 groups: control (CG), infected 1 (IG1) and infected 2 (IG2). One day after arrival piglets of IG1 and IG2 were orally inoculated with 3.7 × 10⁸ CFU of Escherichia coli O149. All piglets were fed a high protein ration for 3 days. Daily health status and faeces were recorded by a point scale individually. Challenge strains in faecal samples were evaluated by polymerase chain reaction, serotyping and biochemical identification. Diarrhoea was observed in 96.67% (58.6% severe; 41.4% mild) of all infected piglets and occurred on average 1.3 days after the challenge. The CG group presented one piglet with a transient mild diarrhoea. The E. coli challenge significantly affected the consistency and color of faeces (P < 0.001). The E. coli O149, mainly hemolytic (88%), was isolated in 56% of faecal samples and the 70% of piglets with severe diarrhoea shed E. coli O149 in the faeces. Zootechnical parameters did not show significant differences. The experimental conditions described in this study allowed to effectively induce diarrhoea in weaned piglets. In conclusion a multifactorial approach (infectious, nutritional and management) is necessary to reproduce in vivo diarrhea in piglets.

Keywords

Piglet, Diarrhoea, Escherichia coli; Challenge; Weaning Period

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	T. T. T. Hong, N. Q. Linh, B. Ogle and J. E. Lindberg, “Survey on the Prevalence of Diarrhoea in Pre-Weaning Piglets and on Feeding Systems as Contributing Risk Factors in Smallholdings in Central Vietnam,” Tropical Animal Health and Production, Vol. 38, No. 5, 2006, pp. 397-405. doi:10.1007/s11250-006-4399-z
[2]	A. Thomsson, D. Rantzer, J. Botermans and J. Svedsen, “The Effect of Feeding System at Weaning on Performance, Health and Feeding Behaviour of Pigs of Different Sizes,” Acta Agriculturae Scandinavica, Section A—Animal Science, Vol. 58, No. 2, 2008, pp. 78-83.
[3]	H. Vondruskova, R. Slamova, M. Trckova, Z. Zraly and I. Pavlik, “Alternatives to Antibiotic Growth Promoters in Prevention of Diarrhoea in Weaned Piglets: A Review,” Veterinari Medicina, Vol. 55, 2010, pp. 199-224.
[4]	J. Callesen, D. Halas, F. Thorup, K. E. Bach Knudsen, J. C. Kim, B. P. Mullan, R. H. Wilson and J. R. Pluske, “The Effects of Weaning Age, Diet Composition and Categorization of Creep Feed Intake by Piglets on Diarrhoea and Performance after Weaning,” Livestock Science, Vol. 108, No. 1-3, 2007, pp. 120-123.
[5]	J. M. Fairbrother, E. Nadeau and C. L. Gyles, “Escherichia coli in Postweaning Diarrhea in Pigs: An Update on Bacterial Types, Pathogenesis, and Prevention Strategies,” Animal Health Research Reviews, Vol. 6, No. 1, 2005, pp. 17-39. doi:10.1079/AHR2005105
[6]	M. T. Sorensen, E. M. Vestergaard, S. K. Jensen, C. Lauridsen and S. Hojsgaard, “Performance and Diarrhoea in Piglets Following Weaning at Seven Weeks of Age: Challenge with E. coli O 149 and Effect of Dietary Factors,” Livestock Science, Vol. 123, No. 2-3, 2009, pp. 314- 321.
[7]	F. Madec, N. Bridoux, S. Bounaix, R. Cariolet, Y. Duval-Iflah, D. J. Hampson and A. Jestin, “Experimental Models of Porcine Post-Weaning Colibacillosis and their Relationship to Post-Weaning Diarrhoea and Digestive Disorders as Encountered in the Field,” Veterinary Microbiology, Vol. 72, No. 3-4, 2000, pp. 295-310. doi:10.1016/S0378-1135(99)00202-3
[8]	T. M. Laine, T. Lyytikainen, M. Yliaho, and M. Anttila, “Risk Factors for Post-Weaning Diarrhoea on Piglet Producing Farms in Finland,” Acta Veterinaria Scandinavica, Vol. 50, 2008, p. 21. doi:10.1186/1751-0147-50-21
[9]	M. D. Barton, “Antibiotic Use in Animal Feed and its Impact on Human Health,” Nutrition Research Reviews, Vol. 13, No. 2, 2000, pp. 279-299. doi:10.1079/095442200108729106
[10]	A. K. Sarmah, M. T. Meyer and A. B. A. Boxall, “A Global Perspective on the Use, Sales, Exposure Pathways, Occurrence, Fate and Effects of Veterinary Antibiotics (VAs) in the Environment,” Chemosphere, Vol. 65, No. 5, 2006, pp. 725-759. doi:10.1016/j.chemosphere.2006.03.026
[11]	M. G. Smith, D. Jordan, T. A. Chapman, J. J. C. Chin, M. D Barton, T. N. Do, V. V. Fahy, J. M. Fairbrother and D. J. Trott, “Antimicrobial resistance and Virulence Gene Profiles in Multi-Drug Resistant Enterotoxigenic Escherichia coli Isolated from Pigs with Post-Weaning Diarrhea,” Veterinary Microbiology, Vol. 145, No. 3-4, 2010, pp. 299-307. doi:10.1016/j.vetmic.2010.04.004
[12]	I. Camerlink, L. Ellinger, E. J. Bakker and E. A. Lantinga, “Homeopathy as Replacement to Antibiotics in the Case of Escherichia coli Diarrhoea in Neonatal Piglets,” Homeopathy, Vol. 99, No. 1, 2009, pp. 57-62. doi:10.1016/j.homp.2009.10.003
[13]	C. Domeneghini, A. Di Giancamillo, S. Arrighi and G. Bosi, “Gut-Trophic Feed Additives and their Effects upon the Gut Structure and Intestinal Metabolism. State of the Art in the Pig, and Perspectives towards Humans,” Histology and Histopathology, Vol. 21, No. 1-3, 2006, pp. 273-283.
[14]	C. Corino, M. Musella, G. Pastorelli, R. Rossi, K. Paolone, L. Costanza, A. Manchisi and G. Maiorano, “Influences of Dietary Conjugated Linoleic Acid (CLA) and Total Lysine Content on Growth, Carcass Characteristics and Meat Quality of Heavy Pigs,” Meat Science, Vol. 79, No. 2, 2008, pp. 307-316. doi:10.1016/j.meatsci.2007.10.001
[15]	K. R. Hodgson and M. Barton, “Treatment and Control of Enterotoxigenic Escherichia coli Infections in Pigs,” CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources, Vol. 4, No. 44, 2009, pp. 1-16. doi:10.1079/PAVSNNR20094044
[16]	L. Zhang, Y.-Q. Xu, H.-Y. Liu, T. Lai, J.-L. Ma, J.-F. Wang and Y.-H. Zhu, “Evaluation of Lactobacillus rhamnosus GG Using an Escherichia coli K88 Model of Piglet Diarrhoea: Effects on Diarrhoea Incidence, Faecal Microflora and Immune Responses,” Veterinary Microbiology, Vol. 141, No. 1-2, 2010, pp. 142-148. doi:10.1016/j.vetmic.2009.09.003
[17]	AOAC, “Official Methods of Analysis,” 18th Edition, Association of Official Analytic Chemists, Gaithersburg, 2005.
[18]	DM, “Approvazione dei Metodi di Analisi per il Controllo Ufficiale degli Alimenti per Animali e Soppressione di Altri Metodi Inerenti al Controllo del Medesimo Settore Merceologico,” Gazzetta Ufficiale, No. 31, Suppl. 13, 21 Dicembre 1998.
[19]	AOCS, “Official Methods of the American Oil Chemists Society,” 5th Edition, American Oil Chemists Society, Champaign, 1998.
[20]	G. M. Jensen, K. Frydendahlb, O. Svendsen, C. B. Jorgensen, S. Cirerac, M. Fredholm, J. P. Nielsen and K. Moller, ”Experimental Infection with Escherichia coli 0149: F4ac in Weaned Piglets,” Veterinary Microbiology, Vol. 115, No. 1-3, 2006, pp. 243-249. doi:10.1016/j.vetmic.2006.01.002
[21]	J. Blanco and M. Blanco, “Escherichia coli Enterotoxigenicos, Necrotoxigenicos y Verotoxigenicos de Origin Humano y Bovino,” Servicio de Publicaciones Diputacion Provincial, Lugo, 1993.
[22]	J. E. Blanco, M. Blanco, J. Blanco, A. Mora, L. Balaguer, J. Mouriňo and W. H. Jansen,” O Serogroups, Biotypes and Eae Genes in Escherichia coli Strains Isolated from Diarrheic and Healthy Rabbits,” Journal of Clinical Microbiology, Vol. 34, No. 12, 1996, pp. 3101-3107.
[23]	C. Farina, A. Goglio, G. Conedera, F. Minelli and A. Caprioli, “Antimicrobial Susceptibility of Escherichia coli O157 and Other Enterohaemorrhagic Escherichia coli Isolated in Italy,” European Journal of Clinical Microbiology & Infectectious Disease, Vol. 15, No. 4, 1996, pp. 351-353. doi:10.1007/BF01695674
[24]	Z. Lin, H. Kurazono, S. Yamasaki and Y. Takeda, “Detection of Various Variant Verotoxin Genes in Escherichia coli by Polymerase Chain Reaction,” Microbiology and Immunology, Vol. 37, No. 7, 1993, pp. 543-548.
[25]	H. Russmann, E. Kothe, S. Schmidt, D. Franke, A. Harmsen, A. Caprioli and H. Karch, “Genotyping of Shiga-Like Toxin Genes in Non-O157 Escherichia coli Strains Associated with Haemolytic Uraemic Syndrome,” Journal of Medical Microbiology, Vol. 42, No. 6, 1995, pp. 404-410. doi:10.1099/00222615-42-6-404
[26]	S. Franke, F. Gunzer, L. H. Wieler, G. Baljer and H. Karch, “Construction of Recombinant Shiga-Like Toxin-IIv (SLT-IIv) and Its Use in Monitoring the SLT-IIv Antibody Status in Pigs,” Veterinary Microbiology, Vol. 43, No. 1, 1995, pp. 41-52. doi:10.1016/0378-1135(94)00071-4
[27]	H. Karch, H. Bohm, F. Schimdt, F. Gunzer, S. Aleksic and J. Heesemann, “Clonal Structure and Pathogenicity of Shiga-Like Toxin-Producing, Sorbitol-Fermenting Escherichia coli O157:H” Journal of Clinical Microbiology, Vol. 31, No. 5, 1993, pp. 1200-1205.
[28]	E. Cox, E. Schrauwen, V. Cools and A. Houvenaghel, “Experimental Induction of Diarrhoea in Newly-Weaned Piglets,” Journal of Veterinary Medicine Series A, Vol. 38, No. 1-10, 1991, pp. 418-426. doi:10.1111/j.1439-0442.1991.tb01030.x
[29]	L. Prohászka and F. Baron, “The Predisposing Role of High Dietary Protein Supplies in Enteropathogenic E. coli Infections of Weaned Pigs,” Zentralblatt für Veterinarmedizin Reihe B, Vol. 27, No. 3, 1980, pp. 222-232. doi:10.1111/j.1439-0450.1980.tb01908.x
[30]	S. Macfarlane and G. T. Macfarlane, “Proteolysis and Amino Acid Fermentation,” In: G. R. Gibson and G. T. Macfarlane, Eds., Human Colonic Bacteria: Role in Nutrition, Physiology and Pathology, CRC Press, Boca Raton, 1995, p. 75.
[31]	J. M. Heo, J. C. Kim, C. F., Hansen, B. P. Mullan, D. J. Hampson and J. R. Pluske, “Feeding a Diet with Decreased Protein Content Reduces Indices of Protein Fermentation and the Incidence of Postweaning Diarrhea in Weaned Pigs Challenged with an Enterotoxigenic Strain of Escherichia coli,” Journal of Animal Science, Vol. 87, No. 9 , 2009, pp. 2833-2843. doi:10.2527/jas.2008-1274
[32]	F. O. Opapeju, D. O. Krause, R. L. Payne, M. Rademacher and C. M. Nyachoti, “Effect of Dietary Protein Level on Growth Performance, Indicators of Enteric Health, and Gastrointestinal Microbial Ecology of Weaned Pigs Induced with Postweaning colibacillosis,” Journal of Animal Science, Vol. 87, No. 8, 2009, pp. 2635-2643. doi:10.2527/jas.2008-1310
[33]	J. I. Sarmiento, T. A. Casey and H. W. Moon, “Postweaning Diarrhea in Swine: Experimental Model of Enterotoxigenic Escherichia coli Infection,” American Journal of Veterinary Research, Vol. 49, No. 7, 1988, pp. 1154-1159.
[34]	A. J. Van Dijk, P. M. M. Enthoven, S. G. C. Van den Hoven, M. M. M. H. Van Laarhoven, T. A. Niewold, M. J. A. Nabuurs and A. C. Beynen, “The Effect of Dietary Spray-Dried Porcine Plasma on Clinical Response in Weaned Piglets Challenged with a Pathogenic Escherichia coli,” Veterinary Microbiology, Vol. 84, No. 3, 2002, pp. 207-218. doi:10.1016/S0378-1135(01)00463-1
[35]	M. J. A. Nabuurs, F. G. van Zijderveld and P. W. de Leeuw, “Clinical and Microbiological Field Studies of Diarrhoea in Pigs at Weaning in the Netherlands,” Research in Veterinary Science, Vol. 55, No. 1, 1993, pp. 70-77. doi:10.1016/0034-5288(93)90037-G