Chronic inflammation induced by endotoxin from a dysbiotic gut microbiota contributes to the development of obesity-related metabolic disorders. Modification of gut microbiota by a diet to balance its composition becomes a promising strategy to help manage obesity. A dietary scheme based on whole grains, traditional Chinese medicinal foods, and prebiotics (WTP diet) was designed to meet human nutritional needs as well as balance the gut microbiota. Ninety-three of 123 central obese volunteers (BMI ≥ 28 kg m−2) completed a self-controlled clinical trial consisting of 9-week intervention on WTP diet followed by a 14-week maintenance period. The average weight loss reached 5.79 ± 4.64 kg (6.62 ± 4.94%), in addition to improvement in insulin sensitivity, lipid profiles, and blood pressure. Pyrosequencing of fecal samples showed that phylotypes related to endotoxin-producing opportunistic pathogens of Enterobacteriaceae and Desulfovibrionaceae were reduced significantly, while those related to gut barrier-protecting bacteria of Bifidobacteriaceae increased. Gut permeability, measured as lactulose/mannitol ratio, was decreased compared with the baseline. Plasma endotoxin load as lipopolysaccharide-binding protein was also significantly reduced, with concomitant decrease in tumor necrosis factor-α, interleukin-6, and an increase in adiponectin. These results suggest that modulation of the gut microbiota via dietary intervention may enhance the intestinal barrier integrity, reduce circulating antigen load, and ultimately ameliorate the inflammation and metabolic phenotypes.
A group-specific PCR-based denaturing gradient gel electrophoresis (DGGE) method was developed and combined with group-specific clone library analysis to investigate the diversity of the Clostridium leptum subgroup in human feces. PCR products (length, 239 bp) were amplified using C. leptum cluster-specific primers and were well separated by DGGE. The DGGE patterns of fecal amplicons from 11 human individuals revealed host-specific profiles; the patterns for fecal samples collected from a child for 3 years demonstrated the structural succession of the population in the first 2 years and its stability in the third year. A clone library was constructed with 100 clones consisting of 1,143-bp inserts of 16S rRNA gene fragments that were amplified from one adult fecal DNA with one forward universal bacterial primer and one reverse group-specific primer. Eighty-six of the clones produced the 239-bp C. leptum cluster-specific amplicons, and the remaining 14 clones did not produce these amplicons but still phylogenetically belong to the subgroup. Sixty-four percent of the clones were related to Faecalibacterium prausnitzii (similarity, 97 to 99%), 6% were related to Subdoligranulum variabile (similarity, ϳ99%), 2% were related to butyrateproducing bacterium A2-207 (similarity, 99%), and 28% were not identified at the species level. The identities of most bands in the DGGE profiles for the same adult were determined by comigration analysis with the 86 clones that harbored the 239-bp group-specific fragments. Our results suggest that DGGE combined with clone library analysis is an effective technique for monitoring and analyzing the composition of this important population in the human gut flora.The human gastrointestinal tract harbors a highly diverse microbial community, which plays important roles in host nutrition, immunology, health, and disease. In addition to Bacteroides spp. and the Clostridium coccoides cluster, the Clostridium leptum cluster (3) is one of the most predominant populations in the human fecal microflora (2,6,11,12,30). Including species that belong to the genera Clostridium, Eubacterium, and Ruminococcus, the C. leptum cluster contains numerous butyrate-producing and fibrolytic species (6, 22). The metabolic activities of these organisms have a significant effect on the health of the human colon. Members of this cluster are highly oxygen sensitive (22) and difficult to culture (9), so the development of molecular methods to specifically study the diversity of this population and to monitor changes in the cluster after intervention is critical.Probes (17,27,29) and primers (18) specific for the C. leptum group have been designed and used to enumerate this population by fluorescent in situ hybridization (FISH), dot blot hybridization, and real-time PCR. However, these group-specific methods can provide information only on the abundance of the whole population in the human fecal microflora, and they provide few details concerning the composition of the population at the species level.Another way to st...
We first used human flora-associated (HFA) piglets, a significantly improved model for research on human gut microbiota, to study the effects of short-chain fructo-oligosaccharides (scFOS) on the gut bacterial populations. Ten neonatal HFA piglets were assigned to receive basal diets alone or supplemented with scFOS (0.5 g/kg body weight/day) from 1 to 37 days after birth (DAB). The impact of scFOS on the fecal bacterial populations of the piglets before (12 DAB), during (17 DAB), and after (25 and 37 DAB) weaning were monitored by PCR-denaturing gradient gel electrophoresis and real-time quantitative PCR. The Bifidobacterium genus was stimulated consistently, except during weaning, confirming the bifidogenic property of scFOS. At 12 DAB, the Clostridium leptum subgroup was decreased and two unknown Bacteroides-related species were increased; at 25 DAB, the C. leptum subgroup and Subdoligranulum variabile-like species were elevated, whereas one unknown Faecalibacterium-related species was suppressed; and at 37 DAB, the Bacteroides genus was decreased. The results showed that effects of scFOS on non-bifidobacteria varied at different developmental stages of the animals, warranting further investigation into the host-development-related effects of prebiotics on the gut microbiota and the host physiology using the HFA piglets as a model for humans.
ABSTRACT. Seventeen out of 24 human flora-associated (HFA) piglets died after oral administration of whole fecal flora from an apparently healthy human donor. The bacteria isolated from the organs of the infected piglets were identified as Klebsiella pneumoniae by bacteriological and biochemical tests and 16S rRNA gene sequence analysis. The identical K. pneumoniae strain was also isolated from the donor's fecal flora. All three neonatal piglets inoculated with K. pneumoniae from the donor's fecal flora developed severe diarrhea, with 2 eventually dying. This strongly suggests that the opportunistic pathogen K. pneumoniae from the human donor caused the fatal infection in the HFA piglets. The results underscore the importance of safety evaluation of the human donor's fecal flora for HFA piglet development. KEY WORDS: human flora-associated piglets, Klebsiella pneumoniae, sepsis.
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