Experimental cecitis in gnotobiotic quails monoassociated with Clostridium butyricum strains isolated from patients with neonatal necrotizing enterocolitis and from healthy newborns

Abstract

Using axenic quails fed a diet containing lactose, we have investigated the potentially pathogenic roles of six Clostridium butyricum strains of human origin. Three strains (CB155-3, CB1002, and CB203-1) isolated from neonatal necrotizing enterocolitis patients and two of three strains (CB19-1 and CB25-2) isolated from healthy newborns led to cecal or crop lesions or both similar to those observed in human neonatal necrotizing enterocolitis: thickening of the cecal wall with gas cysts, hemorrhagic ulcerations, and necrotic areas. The lactose-negative strain (CB46-1) did not develop any lesions. The neuraminidase-producing strain (CB155-3) caused lesions in all monoassociated quails, whereas the other strains caused lesions in 28 to 85% of animals. Removal of dietary lactose suppressed all pathological incidence. These results show that lactose fermentation is a prerequisite in these pathological changes and stress the roles played by both the strain and the host in the expression of C. butyricum enteropathogenicity.Clostridium butyricum is frequently isolated from feces of healthy human newborns (6, 10, 29), but the role played by this strain in neonatal necrotizing enterocolitis (NNE) has been questioned by several authors (6,10,11,13,29,30). Nevertheless, no enteropathogenic toxin has ever been attributed to C. butyricum.Popoff et al. (21) were the first to reproduce, in the ceca of chickens monoassociated with C. butyricum strains, histopathological lesions (congestion, hemorrhage, necrosis, and pneumatosis) similar to those observed in human newborns suffering from NNE. However, because of their large size, chickens are not practical animals to use for further investigation in isolators.Ulcerative (18) and necrotic (3) enteritises have been observed in quails. Their small size and fast growth rate encouraged us to test them as an experimental model for the study of C. butyricum enteropathogenicity. We tested three strains isolated from NNE patients; one strain had already been tested in chickens (21). We have also tested three other strains isolated from healthy newborns. MATERIALS AND METHODS Bacterial strains. Of six C. butyricum strains studied, three were isolated from sick premature human newborns: strain CB1002 from a fatal NNE case, strain CB203-1 from a patient with hemorrhagic colitis, and strain CB155-3 from an NNE patient. The three remaining strains, CB19-1, CB25-2, and CB46-1, were isolated from stools of healthy premature newborns (19). Strain CB1002 was used in a previous study (21). CB155-3 was the sole neuraminidase-producing strain (19), and CB46-1 was the sole lactose-nonfermenting strain. Strains were grown in TY broth (Pasteur Production, Paris, France).Maintenance and inoculation of quails. Quails (Coturnix coturnix subsp. japonica) were obtained by the method of Reynier and Sackteder (23) with the following modifications * Corresponding author. for germfree animals. Eggs from a conventional flock kept in a laying battery were collected twice daily and immediately brushed with 0.3% quate...

Degradation and fermentation of α‐gluco‐oligosaccharides by bacterial strains from human colon: in vitro and in vivo studies in gnotobiotic rats

“…butyricum were found in healthy human newborn babies (Gothefors and Blenkharn 1978;Smith et al 1980;Fontaine et al 1986), a number of studies have suggested that this species has a potentially deleterious effect in alactasic premature babies, because fermentation of undigested lactose leads to excessive production of H, and butyrate. These two metabolites have been associated with the occurrence of neonatal necrotizing enterocolitis (Popoff et al 1985;Bousseboua et al 1989). In our model of trixenic rats, clostridial counts remained high, but participation of this organism in GOS degradation appears doubtful because fermentation products associated with glycolytic activity of CI.…”

Clostridium butyricum : from beneficial to a new emerging pathogen

“…Additionally, Bousseboua et al showed that lactose fermentation was a prerequisite for the expression of C. butyricum enteropathogenicity. Numerous gas cysts were always observed in the caecal wall of quails mono-associated with C. butyricum [50]. Neuraminidase production has also been described as a major virulence factor in C. butyricum strains causing NEC in preterm neonates [51].…”

Diet, length of gestation, and fecal short chain fatty acids in healthy premature neonates

“…High production of SCFAs associated with abnormal C 4 ratio (above 40%) has previously been demonstrated as potentially deleterious in infants 5 as in experimental models. 39,40 Therefore, factors responsible for the increase in butyric acid need to be explored. SCFA concentrations and profiles found during this study are in accordance with our previous study in preterm infants when their gestational age varied from 30 to 36 weeks.…”