Authors : M. S. Yahaya , M. Goto , W. Yimiti , Y. Gamo , W. Kim , S. Karita , B. Smerjai , Y. Kawamoto and S. Ogawa
Abstract: This study evaluated the changes of the microbiota on Tinaroo legume (Neonotonia wightii) during fermentation using denaturing gradient gel electrophoresis (DGGE) and Scanning electron microscopy (SEM). As well as determined the effects of the changes on silage fermentation and ruminal degradability. The in situ rumen degradability of the silages, Control (without additive), without the additives of fermented juice of epiphytic lactic acid bacteria (FJLB) and original fresh matter were evaluated at 8, 16, 24, 48 and 72 hours with Nylon bags. To estimate the rate and extent of DM degradability and their degradation characteristics data were fitted to the exponential equation of P = a+b (1-e-ct). The results in this study revealed that addition of sugar prior to incubation of FJLB solution lower its pH values and increased its number LAB (cfu) colony forming units compared to non sugar added FJLB and epiphytic LAB samples. The DGGE bands indicated a substantial differences between samples with the Control silage showing a mixture of dominant and faint bands (group A) on replicates samples no. 1, 2, and 3. The pH (5.69) of the Control silage did not inhibit the development of clostridial type of bacteria resulting in higher population of community of microbiota which resulted in the propionic iso-butyric and butyric types of fermentation in the silo. Similarly, the group C bands present in epiphytic microbiota were absent in FJLB additive and FJLB silage, suggesting a community of microorganism that are only tolerance to higher pH (6.40). But as the pH values were lowered in FJLB additive and FJLB silage during fermentation the group A and C DGGE bands were absent, but these samples exhibited similar prominent DGGE bands in group D, suggesting a similar community of microbiota resistant to low pH. SEM revealed higher different shapes (rod or cocci and oval) of epiphytic microbial dead bodies (debris) adherent on the surfaces and bottoms of leaves and stem of the original fresh matter followed by the Control than observed in the FJLB silages. FJLB silage showed consistent similar shapes (oval) of microbial debris adherent through out the entire surfaces of the leaf and stem. The results from SEM revealed a typical present of bacteria in the silo similar to those found in the rumen and the identity of banding patterns in the DGGE analysis consisted with the changes in the results of the SEM, pH values, Colony forming units, fermentation profiles of silages and ruminal degradability of fresh matter and silages. The characterization of microbial community is considered to be an initial step in measuring the diversity of microorganisms and could lead to a better understanding of their role in natural environment as well as during fermentation process of forages.
M. S. Yahaya , M. Goto , W. Yimiti , Y. Gamo , W. Kim , S. Karita , B. Smerjai , Y. Kawamoto and S. Ogawa , 2004. Epiphytic Microbiota on Tropical Tinaroo Legume (Neonotonia wightii) as Revealed by Denaturing Gradient Gel Electrophoresis (DGGE) and Scanning Electron Microscopy (SEM) and their Effects on Silage Fermentation and Ruminal Degradability . Journal of Animal and Veterinary Advances, 3: 340-347.