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Infection and Immunity, July 2004, p. 3914-3924, Vol. 72, No. 7
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.7.3914-3924.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Relative Importance of Heat-Labile Enterotoxin in the Causation of Severe Diarrheal Disease in the Gnotobiotic Piglet Model by a Strain of Enterotoxigenic Escherichia coli That Produces Multiple Enterotoxins{dagger}

Emil M. Berberov,1 You Zhou,1 David H. Francis,2 Michael A. Scott,3,{ddagger} Stephen D. Kachman,4 and Rodney A. Moxley1*

Departments of Veterinary and Biomedical Sciences,1 Biometry, University of Nebraska—Lincoln, Lincoln, Nebraska 68583,4 Department of Veterinary Science, South Dakota State University, Brookings, South Dakota 57007,2 Department of Veterinary Pathobiology, University of Missouri—Columbia, Columbia, Missouri 652113

Received 29 December 2003/ Returned for modification 16 March 2004/ Accepted 1 April 2004

Enterotoxigenic Escherichia coli (ETEC) strains that produce multiple enterotoxins are important causes of severe dehydrating diarrhea in human beings and animals, but the relative importance of these enterotoxins in the pathogenesis is poorly understood. Gnotobiotic piglets were used to study the importance of heat-labile enterotoxin (LT) in infection with an ETEC strain that produces multiple enterotoxins. LT– ({Delta}eltAB) and complemented mutants of an F4+ LT+ STb+ EAST1+ ETEC strain were constructed, and the virulence of these strains was compared in gnotobiotic piglets expressing receptors for F4+ fimbria. Sixty percent of the piglets inoculated with the LT– mutant developed severe dehydrating diarrhea and septicemia compared to 100% of those inoculated with the nalidixic acid-resistant (Nalr) parent and 100% of those inoculated with the complemented mutant strain. Compared to piglets inoculated with the Nalr parent, the mean rate of weight loss (percent per hour) of those inoculated with the LT– mutant was 67% lower (P < 0.05) and that of those inoculated with the complemented strain was 36% higher (P < 0.001). Similarly, piglets inoculated with the LT– mutant had significant reductions in the mean bacterial colony count (CFU per gram) in the ileum; bacterial colonization scores (square millimeters) in the jejunum and ileum; and clinical pathology parameters of dehydration, electrolyte imbalance, and metabolic acidosis (P < 0.05). These results indicate the significance of LT to the development of severe dehydrating diarrhea and postdiarrheal septicemia in ETEC infections of swine and demonstrate that EAST1, LT, and STb may be concurrently expressed by porcine ETEC strains.

* Corresponding author. Mailing address: 111 Veterinary Basic Sciences, University of Nebraska—Lincoln, Lincoln, NE 68583-0905. Phone: (402) 472-8460. Fax: (404) 472-9690. E-mail: rmoxley1{at}unl.edu.

{dagger} This work is a contribution of the University of Nebraska Agricultural Research Division, Lincoln (journal series no. 13995).

Editor: V. J. DiRita

{ddagger} Present address: Department of Veterinary Pathology, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824-1314.

Infection and Immunity, July 2004, p. 3914-3924, Vol. 72, No. 7
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.7.3914-3924.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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