An in vivo study was carried to determine the effect of HP-inulin, a high-molecular-weight fraction of chicory-derived inulin, on the human gut microflora composition. Ten healthy volunteers were allowed a free-living diet whereby they also ingested 8 g/d of maltodextrin for 14 days and this was followed by 8 g/d HP-inulin for 14 days. Nine of the ten volunteers completed the trial. The trial was conducted in a double blind manner and faeces were collected periodically such that predominant groups of gut bacteria i.e. total bacterial populations, Bacteroides spp., Bifidobacterium spp., Clostridium perfringens/histolyticum sub-group and lactobacilli/enterococci could be enumerated. To overcome difficulties with culture-based techniques, the bacteria were enumerated using fluorescent in situ hybridisation (FISH). A small but statistically significant increase in bifidobacteria was observed when data from the volunteers were pooled. Similarly, a statistically significant increase was observed in clostridial numbers, although the magnitude of change in this bacterial group was about ten times less than that seen with bifidobacteria. HP-inulin intake had little or no effect on numbers of total bacteria, Bacteroides spp., or lactobacilli and enterococci present in the gut microflora of the volunteers. This study has confirmed the prebiotic nature of HP-inulin. However, in this trial the effects were most marked in those volunteers with low starting levels of bifidobacteria-indicating that there may be a relationship between prebiotic effect and initial bifidobacterial numbers. (C) 2001 Academic Press.