A comparison of enzymatic and molecular approaches to characterize the cellulolytic microbial ecosystems of the rumen and the cecum.
We used RNA probes and enzyme activities to compare the cellulolytic microbial ecosystems of the rumen and the cecum. Four rumen- and cecum-cannulated wethers were fed a diet of barley plus hay (60:40). Digesta samples were collected 1 h before feeding and 3, 6, and 9 h after feeding for measurements on microbial populations, and 1 h before feeding and 3 and 6 h after feeding for digestion measurements, pH, and VFA. Polysaccharidase and glycosidase specific activities of solid-adherent microorganisms were measured respectively by the amount of reducing sugars released from xylan or avicel or p-nitrophenol from the p-nitrophenol derivatives of xylose and glucose. The distribution and amounts of the three main cellulolytic bacterial species (Fibrobacter succinogenes, Ruminococcus albus, and Ruminococcus flavefaciens) were determined by dot-blot hybridization using specific 16SrRNA-targeting probes. Enzyme activities were higher in the rumen than in the cecum and before feeding than at 3 h after feeding. The sum of the three cellulolytic bacterial species represented, on average, 4.5% of the total bacterial RNA in the two compartments and did not vary with sampling time. The cellulolytic bacterial community structure was different in the two compartments, with F. succinogenes as the main species in the rumen and R. flavefaciens in the cecum. The lower cellulolytic activity in the cecum than in the rumen could not be ascribed to any difference in the structure of the cellulolytic bacterial community between these two compartments, and other hypotheses related to digestion are proposed.
Michalet-Doreau
B
B
URH-DIM, INRA-Theix, Laboratoire de Microbiologie, INRA-Theix, Genès Champanelle, France. bdoreau@clermont.inra.fr
Fernandez
I
I
Fonty
G
G
D000821:Animal Feed; D000818:Animals; D002404:Catheterization; D002432:Cecum / Q000382:microbiology*; D005232:Fatty Acids, Volatile / Q000032:analysis; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006095:Gram-Positive Cocci / Q000201:enzymology* / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting / Q000662:veterinary; D008297:Male; D015336:Molecular Probe Techniques / Q000662:veterinary; D015347:RNA Probes; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics*; D012417:Rumen / Q000382:microbiology*; D012756:Sheep; D013997:Time Factors
D005232:Fatty Acids, Volatile; D015347:RNA Probes; D012336:RNA, Ribosomal, 16S; D006026:Glycoside Hydrolases