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A method of decontaminating Strongyloides venezuelensis larvae for the study of strongyloidiasis in germ-free and conventional mice

W.A. MARTINS^*,, A.L. MELO, J.R. NICOLI^*, D.C. CARA, M.A. R. CARVALHO^*, M.A. LANA^*, E.C. VIEIRA and L.M. FARIAS^*

*Departamento de Microbiologia, Departamento de Parasitologia, Departamento de Patologia and Departamento de Bioquímica-Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, 30161-970 Belo Horizonte MG, Brazil

Corresponding author: Dr J.R. Nicoli (e.mail: jnicoli{at}mono icb.ufmg.br).

Received 5 July 1999; revised version accepted 15 Sept. 1999.

Abstract

To study the possible influence of intestinal micro-organisms on the course of strongyloidiasis in mice, a method was developed to obtain axenic infective larvae of Strongyloides venezuelensis. Cultured larvae from conventional mice were treated with sodium hypochlorite 0.25% for 10 min, washed in distilled water and then exposed to various combinations of antibiotics for 30 or 60 min. Success was achieved with a combination of penicillin 1805mumg/L and ceftazidime 15mumg/ml. Decontamination of the larvae was determined by aerobic and anaerobic culture and by inoculation into gnotobiotic mice. Viability was established by subcutaneous inoculation of larvae into germ-free and conventional mice. Preliminary results showed that gnotobiotic mice were more susceptible than conventional mice to infection with axenic S. venezuelensis larvae as judged by faecal egg excretion, recovery of worms in the small intestine and histopathological examination of the duodenal mucosa. These results suggest that the normal intestinal flora protects the host against experimental infection with S. venezuelensis.

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