Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1995 Nov;63(11):4350–4357. doi: 10.1128/iai.63.11.4350-4357.1995

Development of an improved animal model of shigellosis in the adult rabbit by colonic infection with Shigella flexneri 2a.

G H Rabbani 1, M J Albert 1, H Rahman 1, M Islam 1, D Mahalanabis 1, I Kabir 1, K Alam 1, M Ansaruzzaman 1
PMCID: PMC173619  PMID: 7591070

Abstract

Rabbits are not usually susceptible to intestinal Shigella infection without extensive pretreatment, including starvation and administration of antimicrobial, antimotility, and toxic agents (carbon tetrachloride). Most animals infected this way die rapidly and do not always develop colonic lesions and signs of dysentery. We describe here a successful experimental infection in the adult rabbit which does not require preparatory treatment and which reproduced characteristic features of human shigellosis. Unstarved, untreated adult rabbits were infected by direct inoculation of virulent Shigella flexneri 2a (10 ml of 10(7) bacteria per ml) into the proximal colon after ligation of the distal cecum (cecal bypass). Within 24 h of infection, most inoculated animals consistently developed clinical dysentery, characterized by liquid stool mixed with mucus and blood, leukocytosis, anorexia, and weight loss. Histologically, there were edema, exudation, superficial ulceration, and polymorphonuclear infiltrations in the lamina propria; crypt abscess formation; focal hemorrhages; and the presence of immunohistochemically stained S. flexneri in the colonic mucosa. Successful bacterial colonization was indicated by the isolation of the challenge strain of S. flexneri 2a from the colonic contents. None of the control rabbits challenged with nonvirulent S. flexneri or without cecal bypass developed dysentery or colitis. We conclude that successful Shigella infection can be induced by direct colonic inoculation with virulent S. flexneri 2a in adult rabbits without starvation and pretreatment. The colitis is dependent on the virulence of the bacteria and on the cecal bypass. This model should be useful in studies of the immunopathogenesis of Shigella infection.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ARM H. G., FLOYD T. M., FABER J. E., HAYES J. R. USE OF LIGATED SEGMENTS OF RABBIT SMALL INTESTINE IN EXPERIMENTAL SHIGELLOSIS. J Bacteriol. 1965 Mar;89:803–809. doi: 10.1128/jb.89.3.803-809.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aghdassi E., Plapler H., Kurian R., Raina N., Royall D., Jeejeebhoy K. N., Cohen Z., Allard J. P. Colonic fermentation and nutritional recovery in rats with massive small bowel resection. Gastroenterology. 1994 Sep;107(3):637–642. doi: 10.1016/0016-5085(94)90109-0. [DOI] [PubMed] [Google Scholar]
  3. Ahmed Z. U., Sarker M. R., Sack D. A. Protection of adult rabbits and monkeys from lethal shigellosis by oral immunization with a thymine-requiring and temperature-sensitive mutant of Shigella flexneri Y. Vaccine. 1990 Apr;8(2):153–158. doi: 10.1016/0264-410x(90)90139-d. [DOI] [PubMed] [Google Scholar]
  4. Anand B. S., Malhotra V., Bhattacharya S. K., Datta P., Datta D., Sen D., Bhattacharya M. K., Mukherjee P. P., Pal S. C. Rectal histology in acute bacillary dysentery. Gastroenterology. 1986 Mar;90(3):654–660. doi: 10.1016/0016-5085(86)91120-0. [DOI] [PubMed] [Google Scholar]
  5. Bennish M. L., Azad A. K., Yousefzadeh D. Intestinal obstruction during shigellosis: incidence, clinical features, risk factors, and outcome. Gastroenterology. 1991 Sep;101(3):626–634. doi: 10.1016/0016-5085(91)90518-p. [DOI] [PubMed] [Google Scholar]
  6. Butler T., Islam M. R., Bardhan P. K. The leukemoid reaction in shigellosis. Am J Dis Child. 1984 Feb;138(2):162–165. doi: 10.1001/archpedi.1984.02140400044010. [DOI] [PubMed] [Google Scholar]
  7. Butler T., Rahman H., Al-Mahmud K. A., Islam M., Bardhan P., Kabir I., Rahman M. M. An animal model of haemolytic--uraemic syndrome in shigellosis: lipopolysaccharides of Shigella dysenteriae I and S. flexneri produce leucocyte-mediated renal cortical necrosis in rabbits. Br J Exp Pathol. 1985 Feb;66(1):7–15. [PMC free article] [PubMed] [Google Scholar]
  8. Carlin N. I., Lindberg A. A. Monoclonal antibodies specific for Shigella flexneri lipopolysaccharides: clones binding to type IV, V, and VI antigens, group 3,4 antigen, and an epitope common to all Shigella flexneri and Shigella dysenteriae type 1 stains. Infect Immun. 1987 Jun;55(6):1412–1420. doi: 10.1128/iai.55.6.1412-1420.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Carlin N. I., Rahman M., Sack D. A., Zaman A., Kay B., Lindberg A. A. Use of monoclonal antibodies to type Shigella flexneri in Bangladesh. J Clin Microbiol. 1989 Jun;27(6):1163–1166. doi: 10.1128/jcm.27.6.1163-1166.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DUTTA N. K., HABBU M. K. Experimental cholera in infant rabbits: a method for chemotherapeutic investigation. Br J Pharmacol Chemother. 1955 Jun;10(2):153–159. doi: 10.1111/j.1476-5381.1955.tb00074.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dickinson R. J., Gilmour H. M., McClelland D. B. Rectal biopsy in patients presenting to an infectious disease unit with diarrhoeal disease. Gut. 1979 Feb;20(2):141–148. doi: 10.1136/gut.20.2.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. DuPont H. L., Hornick R. B., Dawkins A. T., Snyder M. J., Formal S. B. The response of man to virulent Shigella flexneri 2a. J Infect Dis. 1969 Mar;119(3):296–299. doi: 10.1093/infdis/119.3.296. [DOI] [PubMed] [Google Scholar]
  13. FORMAL S. B., DAMMIN G. J., SCHNEIDER H., LABREC E. H. Experimental Shigella infections. II. Characteristics of a fatal enteric infection in guinea pigs following the subcutaneous inoculation of carbon tetrachloride. J Bacteriol. 1959 Dec;78:800–804. doi: 10.1128/jb.78.6.800-804.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. FORMAL S. B., LABREC E. H., SCHNEIDER H. PATHOGENESIS OF BACILLARY DYSENTERY IN LABORATORY ANIMALS. Fed Proc. 1965 Jan-Feb;24:29–34. [PubMed] [Google Scholar]
  15. FRETER R. Experimental enteric Shigella and Vibrio infections in mice and guinea pigs. J Exp Med. 1956 Sep 1;104(3):411–418. doi: 10.1084/jem.104.3.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harig J. M., Soergel K. H., Komorowski R. A., Wood C. M. Treatment of diversion colitis with short-chain-fatty acid irrigation. N Engl J Med. 1989 Jan 5;320(1):23–28. doi: 10.1056/NEJM198901053200105. [DOI] [PubMed] [Google Scholar]
  17. Hentges D. J. Influence of pH on the inhibitory activity of formic and acetic acids for Shigella. J Bacteriol. 1967 Jun;93(6):2029–2030. doi: 10.1128/jb.93.6.2029-2030.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hentges D. J. Inhibition of Shigella flexneri by the normal intestinal flora. I. Mechanisms of inhibition by Klebsiella. J Bacteriol. 1967 Apr;93(4):1369–1373. doi: 10.1128/jb.93.4.1369-1373.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hentges D. J., Maier B. R. Inhibition of Shigella flexneri by the Normal Intestinal Flora III. Interactions with Bacteroides fragilis Strains in Vitro. Infect Immun. 1970 Oct;2(4):364–370. doi: 10.1128/iai.2.4.364-370.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Islam M. M., Azad A. K., Bardhan P. K., Raqib R., Islam D. Pathology of shigellosis and its complications. Histopathology. 1994 Jan;24(1):65–71. doi: 10.1111/j.1365-2559.1994.tb01272.x. [DOI] [PubMed] [Google Scholar]
  21. Katakura S., Reinholt F. P., Kärnell A., Huan P. T., Trach D. D., Lindberg A. A. The pathology of Shigella flexneri infection in rhesus monkeys: an endoscopic and histopathological study of colonic lesions. APMIS. 1990 Apr;98(4):313–319. doi: 10.1111/j.1699-0463.1990.tb01038.x. [DOI] [PubMed] [Google Scholar]
  22. Keusch G. T. Shigella infections. Clin Gastroenterol. 1979 Sep;8(3):645–662. [PubMed] [Google Scholar]
  23. Kinsey M. D., Formal S. B., Dammin G. J., Giannella R. A. Fluid and electrolyte transport in rhesus monkeys challenged intracecally with Shigella flexneri 2a. Infect Immun. 1976 Aug;14(2):368–371. doi: 10.1128/iai.14.2.368-371.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kärnell A., Reinholt F. P., Katakura S., Lindberg A. A. Shigella flexneri infection: a histopathologic study of colonic biopsies in monkeys infected with virulent and attenuated bacterial strains. APMIS. 1991 Sep;99(9):787–796. doi: 10.1111/j.1699-0463.1991.tb01261.x. [DOI] [PubMed] [Google Scholar]
  25. LABREC E. H., FORMAL S. B. Experimental Shigella infections. IV. Fluorescent antibody studies of an infection in guinea pigs. J Immunol. 1961 Nov;87:562–572. [PubMed] [Google Scholar]
  26. Labrec E. H., Schneider H., Magnani T. J., Formal S. B. EPITHELIAL CELL PENETRATION AS AN ESSENTIAL STEP IN THE PATHOGENESIS OF BACILLARY DYSENTERY. J Bacteriol. 1964 Nov;88(5):1503–1518. doi: 10.1128/jb.88.5.1503-1518.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mason D. Y., Cordell J. L., Abdulaziz Z., Naiem M., Bordenave G. Preparation of peroxidase: antiperoxidase (PAP) complexes for immunohistological labeling of monoclonal antibodies. J Histochem Cytochem. 1982 Nov;30(11):1114–1122. doi: 10.1177/30.11.6183312. [DOI] [PubMed] [Google Scholar]
  28. Mathan M. M., Mathan V. I. Ultrastructural pathology of the rectal mucosa in Shigella dysentery. Am J Pathol. 1986 Apr;123(1):25–38. [PMC free article] [PubMed] [Google Scholar]
  29. Oberhelman R. A., Kopecko D. J., Salazar-Lindo E., Gotuzzo E., Buysse J. M., Venkatesan M. M., Yi A., Fernandez-Prada C., Guzman M., León-Barúa R. Prospective study of systemic and mucosal immune responses in dysenteric patients to specific Shigella invasion plasmid antigens and lipopolysaccharides. Infect Immun. 1991 Jul;59(7):2341–2350. doi: 10.1128/iai.59.7.2341-2350.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rabassa A. A., Rogers A. I. The role of short-chain fatty acid metabolism in colonic disorders. Am J Gastroenterol. 1992 Apr;87(4):419–423. [PubMed] [Google Scholar]
  31. Roediger W. E. Role of anaerobic bacteria in the metabolic welfare of the colonic mucosa in man. Gut. 1980 Sep;21(9):793–798. doi: 10.1136/gut.21.9.793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rout W. R., Formal S. B., Giannella R. A., Dammin G. J. Pathophysiology of Shigella diarrhea in the rhesus monkey: intestinal transport, morphological, and bacteriological studies. Gastroenterology. 1975 Feb;68(2):270–278. [PubMed] [Google Scholar]
  33. SERENY B. Acquired natural immunity following recovery from keratoconjunctivitis shigellosa. J Hyg Epidemiol Microbiol Immunol. 1959;3:292–305. [PubMed] [Google Scholar]
  34. Salam M. A., Bennish M. L. Therapy for shigellosis. I. Randomized, double-blind trial of nalidixic acid in childhood shigellosis. J Pediatr. 1988 Nov;113(5):901–907. doi: 10.1016/s0022-3476(88)80029-5. [DOI] [PubMed] [Google Scholar]
  35. Sansonetti P. J., Arondel J., Fontaine A., d'Hauteville H., Bernardini M. L. OmpB (osmo-regulation) and icsA (cell-to-cell spread) mutants of Shigella flexneri: vaccine candidates and probes to study the pathogenesis of shigellosis. Vaccine. 1991 Jun;9(6):416–422. doi: 10.1016/0264-410x(91)90128-s. [DOI] [PubMed] [Google Scholar]
  36. Speelman P., Kabir I., Islam M. Distribution and spread of colonic lesions in shigellosis: a colonoscopic study. J Infect Dis. 1984 Dec;150(6):899–903. doi: 10.1093/infdis/150.6.899. [DOI] [PubMed] [Google Scholar]
  37. Spira W. M., Sack R. B., Froehlich J. L. Simple adult rabbit model for Vibrio cholerae and enterotoxigenic Escherichia coli diarrhea. Infect Immun. 1981 May;32(2):739–747. doi: 10.1128/iai.32.2.739-747.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Takeuchi A., Formal S. B., Sprinz H. Exerimental acute colitis in the Rhesus monkey following peroral infection with Shigella flexneri. An electron microscope study. Am J Pathol. 1968 Mar;52(3):503–529. [PMC free article] [PubMed] [Google Scholar]
  39. Treem W. R., Ahsan N., Shoup M., Hyams J. S. Fecal short-chain fatty acids in children with inflammatory bowel disease. J Pediatr Gastroenterol Nutr. 1994 Feb;18(2):159–164. doi: 10.1097/00005176-199402000-00007. [DOI] [PubMed] [Google Scholar]
  40. Vernia P., Caprilli R., Latella G., Barbetti F., Magliocca F. M., Cittadini M. Fecal lactate and ulcerative colitis. Gastroenterology. 1988 Dec;95(6):1564–1568. doi: 10.1016/s0016-5085(88)80078-7. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES