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Antagonism against Vibrio cholerae by diffusible substances produced by bacterial components of the human faecal microbiota

S.H. SILVA, E.C. VIEIRA^*, R.S. DIAS and J.R. NICOLI

Departmento de Microbiologia and *Departamento de Bioquímica-Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais and Fundação Ezequiel Dias, Belo Horizonte MG, Brazil

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

Received 22 Feb. 2000; revised version received 2 June 2000; accepted 29 June 2000.

Abstract

Cholera vibrios sometimes survive, probably in low-level silent populations, in the small intestine of chronic carriers or pass through the gastrointestinal tract of a few individuals without causing diarrhoea or colonisation. To understand these situations, the present study used plate cultures (ex-vivo test) to investigate the frequency of appearance of an inhibitory halo against Vibrio cholerae produced by faecal specimens from 92 healthy volunteers (40 females, 52 males) aged 4–61 years. The frequency of inhibitory halo was 20.6% in the whole group. An apparently higher percentage (27.3%) was observed in the age range 20–40 years when compared with the range 4–19 years (10.7%), but not the range 41–61 years (20.0%). Frequency was significantly higher in males (30.8%) than females (7.5%). The dominant microbiota of a volunteer whose faeces produced an inhibitory halo was isolated by plate culture of decimal dilutions in an anaerobic chamber. Potential isolates of 26 apparently different morphologies were associated with germ-free NIH mice. One week later, the inhibitory test showed an antagonistic halo around the faeces from the associated animals, but not from the axenic mice. Of the 26 bacteria isolated, two (Lactobacillus sp. and Peptostreptococcus sp.) produced a compound antagonistic against V. cholerae in an in-vitro assay. When bi-associated with germ-free mice those strains eliminated the vibrio from the intestinal ecosystem in c. 5 days.