[en] There exists experimental evidence that necrotoxigenic Escherichia coli (NTEC) strains producing the cytotoxic necrotising factor 1 cause intestinal and extra-intestinal disease in piglets. On the other hand, no experimental model has been developed with NTEC strains producing the cytotoxic necrotising factor 2. In all, 14 colostrum-restricted calves were orally challenged with two strains isolated from the faeces of a diarrheic calf (B20a) or from the heart blood of a septicaemic calf (1404). All calves had diarrhoea which lasted until euthanasia in eight of them. In those calves, diarrhoea was correlated with the faecal excretion of the challenge strains. At necropsy, vascular congestion of the intestinal mucosa, hypertrophy of the mesenteric lymph nodes (MLN) and some congestion of the lungs were observed. Bacteriology confirmed the colonisation of the intestine by the challenge strains which were also recovered from the heart blood, the lungs and/or the liver. Histological sections confirmed enterocolitis, lymphadenitis and limited bronchopneumonia. In the intestinal tissue sections, bacteria testing positive in an in situ DNA hybridisation assay with a CNF2 probe were observed. Those results were confirmed by immunohistochemistry with a polyclonal anti-O78 and a monoclonal anti-F17b antisera. Three of the five control calves receiving either saline or a CNF(-), F17a strain (25KH09) had no clinical signs or lesions. The other two presented a profuse liquid diarrhoea but those calves were positive for the presence of K99(+) E. coli. In this model, both NTEC2 strains were thus, able to colonise the intestine, to cause long-lasting diarrhoea and to invade the blood stream with localisation in various internal organs in colostrum-restricted conventional newborn calves.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Van Bost, Sigrid
Roels, S.
Mainil, Jacques ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Bactériologie et pathologie des maladies bactériennes
Language :
English
Title :
Necrotoxigenic Escherichia Coli Type-2 Invade and Cause Diarrhoea During Experimental Infection in Colostrum-Restricted Newborn Calves
Baehler A., Moxley R. (2000) Escherichia coli O157: H7 induces attaching-effacing lesions in large intestinal mucosal explants from adult cattle. FEMS Microbiol. Lett. 185:239-242.
Blanco J., Cid D., Blanco J.E., Blanco M., Ruiz Santa Quitera J.A., De la Fuente R. (1996) Serogroups, toxins and antibiotic resistance of Escherichia coli strains isolated from diarrhoeic lambs in Spain. Vet. Microbiol. 49:209-217.
Blanco J.E., Blanco M., Blanco J., Mora A., Balaguer L., Mourino M., Juarez A., Jansen W.H. (1996) O serogroups, biotypes and eae genes in Escherichia coli strains isolated from diarrheic and healthy rabbits. J. Clin. Microbiol. 34:3101-3107.
Blanco M., Blanco J.E., Mora A., Blanco J. (1998) Prevalence and characteristics of necrotoxigenic Escherichia coli CNF1+ and CNF2+ in healthy cattle. Res. Microbiol. 149:47-53.
Caprioli A., Falbo V., Roda L.G., Ruggeri F.M., Zona C. (1983) Partial purification and characterisation of an Escherichia coli toxic factor that induces morphological cell alterations. Infect. Immun. 39:1300-1306.
Cid D., Blanco M., Blanco J.E., Ruiz Santa Quiteria J.A., De la Fuente R., Blanco J. (1996) Serogroups, toxins and antibiotic resistance of Escherichia coli strains isolated from diarrheic goat kids in Spain. Vet. Microbiol. 53:349-353.
Comayras C., Tascas C., Pérès S.Y., Ducommun B., Oswald E., De Rycke J. (1997) Escherichia coli cytolethal distending toxin blocks the HeLa cell cycle at G2/M transition by preventing cdc2 protein kinase dephosphorylation and activation. Infect. Immun. 65:5088-5095.
De Rycke J., Guillot J.F., Boivin B. (1987) Cytotoxins in non-enterotoxigenic strains of Escherichia coli isolated from faeces of diarrheic calves. Vet. Microbiol. 15:137-150.
De Rycke J., Plassiart G. (1990) Toxic effects for lambs of cytotoxic necrotising factor from Escherichia coli. Res. Vet. Sc. 49:349-354.
De Rycke J., Gonzales E.A., Blanco J., Oswald E., Blanco M., Boivin R. (1990) Evidence for two types of cytotoxic necrotizing factor in human and animal clinical isolates of Escherichia coli. J. Clin. Microbiol. 28:694-699.
De Rycke J., Mazars P., Nougayrede J.P., Tasca C., Boury M., Herault F., Valette A., Oswald E. (1996) Mitotic block and delayed lethality in HeLa epithelial cells exposed to Escherichia coli BM2-1 producing cytotoxic necrotizing factor type-1. Infect. Immun. 64:1694-1705.
De Rycke J., Milon A., Oswald E. (1999) Necrotoxigenic Escherichia coli (NTEC): Two emerging categories of human and animal pathogens. Vet. Res. 30:221-233.
Falbo V., Famiglietti M., Caprioli A. (1992) Gene block encoding production of cytotoxic necrotizing factor 1 and hemolysin in Escherichia coli isolates from extra-intestinal infections. Infect. Immun. 60:2182-2187.
Falzano L., Fiorentini C., Donelli G., Michel E., Kocks C., Cossart P., Cabanie L., Oswald E., Boquet P. (1993) Induction of phagocytic behaviour in human epithelial cells by Escherichia coli CNF1. Mol. Microbiol. 9:1247-1254.
Fournout S., Dozois C., Odin M., Desautels C., Pérès S., Herault F., Daigle F., Segafredo C., Lafitte J., Oswald E., Fairbrother J.M., Oswald I.P. (2000) Lack of a role of cytotoxic necrotizing factor 1 toxin from Escherichia coli in bacterial pathogenicity and host cytokine response in infected germ free piglets. Infect. Immun. 68:839-847.
Gonzales E.A., Blanco J. (1989) Serotypes and antibiotic resistance of verotoxigenic (VTEC) and necrotizing (NTEC) Escherichia coli strains isolated from calves with diarrhoea. FEMS Microbiol. Lett. 60:31-36.
Lewis F.A., Wells M. (1992) Detection of virus in infected human tissue by in situ hybridisation. In Situ Hybridisation , Wilkinson, D.G. (Ed.). I.R.L. Press, Oxford; 121-136.
Lintermans P.F., Pohl P., Deboeck F., Bertels A., Schlicker C., Vandekerckhove J., Van Damme J., Van Montagu M., De Greve H. (1988) Isolation and nucleotide sequence of the F17A gene encoding the structural protein of the F17 fimbriae in bovine enterotoxigenic Escherichia coli. Infect. Immun. 56:1475-1784.
Lintermans P.F., Pohl P., Bertels A., Charlier G., Vandekerckhove J., Van Damme J., Schoup J., Schlicker C., Korhonen T., De Greve H., Van Montagu M. (1988) Characterization and purification of the F17 adhesin on the surface of bovine enteropathogenic and septicemic Escherichia coli. Am. J. Vet. Res. 49:1794-1799.
Mainil J., Moseley S., Schneider R., Sutch K., Casey T., Moon H. (1986) Hybridization of bovine Escherichia coli isolates with gene probes for four enterotoxins (STaP, STaH, STb, LT) and one adhesion factor (K99). Am. J. Vet. Res. 47:1145-1148.
Mainil J., Herault F., Oswald E. (1997) Presence of pap-, sfa-, and afa-related sequence in necrotoxigenic Escherichia coli isolates from cattle: Evidence for new variants of the AFA family. Can. J. Vet. Res. 61:193-199.
Mainil J., Gerardin J., Jacquemin E. (2000) Identification of the F17 fimbrial subunit- and adhesin-encoding (f17A and f17G) gene variants in necrotoxigenic Escherichia coli from cattle, pigs and humans. Vet. Microbiol. 73:327-335.
Oswald E., De Rycke J., Lintermans P., Van Muylen K., Mainil J., Daube G., Pohl P. (1991) Virulence factors associated with cytotoxic necrotizing factor type-2 in bovine diarrheic and septicemic strains of Escherichia coli. J. Clin. Microbiol. 29:2522-2527.
Oswald E., Sugai M., Labigne A., Wu H.C., Fiorenti C., Boquet P., O'Brien A.D. (1994) Cytotoxic necrotizing factor type-2 produced by virulent Escherichia coli modifies the small GTP-binding proteins Rho involved in assembly of actin stress fibers. Proc. Natl. Acad. Sci. U.S.A. 91:3814-3818.
Oswald E., Pohl P., Jacquemin E., Lintermans P., Van Muylens K., O'Brien A.D., Mainil J. (1994) Specific DNA probes to detect Escherichia coli strains producing cytotoxic necrotising factor type-1 or type-2. J. Med. Microbiol. 40:428-434.
Pérès S.Y., Marches O., Daigle F., Nougareyde J.-P., Herault F., Tasca C., De Rycke J., Oswald E. (1997) A new cytolethal distending toxin (CDT) from Escherichia coli producing CNF2 blocks HeLa cell division in G2/M phase. Mol. Microbiol. 24:1095-1107.
Pohl P., Lintermans P., Van Muylen K., Schotte M. (1982) Colibacilles entérotoxinogènes du veau possédant un antigène d'attachement différent de l'antigène K99. Ann. Med. Vet. 126:569-571.
Pohl P., Lintermans P., Mainil J., Kaeckenbeeck A., Bertels A. (1987) Etude des phénotypes et des facteurs de virulence des Escherichia coli Att25. Ann. Med. Vet. 131:429-439.
Pohl P., Oswald E., Van Muylen K., Jacquemin E., Lintermans P., Mainil J. (1993) Escherichia coli producing CNF1 and CNF2 cytotoxins in animals with different disorders. Vet. Res. 24:311-315.
Pohl P., Imberechts H., Marin M., Schlicker C., Stockmans F. (1997) Prévalence des gènes codant pour les cytotoxines nécrosantes (CNF1 et CNF2) chez des Escherichia coli isolées de bovins malades ou asymptomatiques. Ann. Med. Vet. 141:161-164.
Rogers D., Anderson A., Hunsaker B. (1996) Lung and nosal lesions caused by swine chlamidial isolates in gnotabiotic pigs. I. Vet. Diagn. Invest. 8:45-55.
Runnels P., Moon H., Schneider A. (1980) Development of resistance with host age to adhesion of K99+ Escherichia coli to isolated intestinal epithelial cells. Infect. Immun. 28:298-300.
Smith H.W. (1974) A search for transmissible pathogenic characters in invasive strains of Escherichia coli: The discovery of a plasmid-controlled lethal character closely associated, or identified with colicin V. J. Gen. Microbiol. 83:95-111.
Smith H.W. (1975) Observations on Escherichia coli infection in calves. Proceedings of the First Seminar on Pathology in the CEC Programme of Coordination of Research in Beef Production , Rutter, J.M. (Ed.). Perinatal III-Health in Calves; 47-59.
Wray C., Piercy D.W.T., Carrol P.J., Cooley W.A. (1993) Experimental infection of neonatal pigs with CNF toxin-producing strains of Escherichia coli. Res. Vet. Sci. 54:290-298.