Probe But1243 only hybridized with a minor percentage (<?1%) of the total CMC bacteria. Thus, in total, probes Fibr225, Ral1436, Rfl155, Clos549, and Inc852 hybridized with 75.2�C78.5% of the total CMC bacteria in these samples. The identities of the remaining CMC bacteria (21�C25%) are still unknown (Table?2). No clear pattern could be seen when the percentage values of the CMC bacteria hybridizing with these newly designed probes (Clos549, Inc852, <a href="http://www.selleckchem.com/products/Dasatinib.html
">Dasatinib and But1243) in the Liq were compared with the corresponding populations in the Sol of the rumen digesta of same cows fed same forage (Table?2). In some cases, it was higher in the Liq than in the Sol and in some cases it was higher in the Sol, while in other cases the levels were comparable. When the mean percentages of CMC bacteria hybridizing with http://www.selleckchem.com/
these probes in the rumen digesta of the cows fed different forages were compared (Table?2), the percentage of the CMC bacteria hybridizing with Clo549 was higher with alfalfa (17.9%) than with triticale (14.4%) feeds, and the mean percentage of the CMC bacteria hybridizing with Inc852 in cows fed alfalfa (10.2%) was comparable to the numbers in cows fed triticale (9.9%). Both the bacterial pure cultures of cellulolytic F.?succinogenes, R.?albus, and R.?flavefaciens and bacterial cells in the rumen digesta of cows fed alfalfa or triticale which hybridized with FISH probes designed to target them reacted positively with the CMC staining developed in this study. This indicates that the CMC staining can be used reliably to detect cells with cell-surface-associated CMCase and endoglucanase activities and involved in cellulose digestion in situ. Our results showed that 8.2�C10.1% of the total bacterial numbers in the Liq and Sol of rumen digesta Dinaciclib
of cows fed either alfalfa or triticale were CMC bacteria. No similar study has been carried out previously where the total numbers of bacteria with cell-surface-associated CMCase and endoglucanase activities in the rumen have been determined in situ, making it difficult to compare our results with other data. However, the relative abundances of total numbers of CMC bacteria determined here are close to the theoretical percentages (12%) estimated by Russell et?al. (2009) for total cellulolytic bacterial numbers present in rumen, estimations that were based on kinetic constants of cellulolytic bacterial pure cultures and the rumen. As mentioned earlier, both CMCases and endoglucanases are enzymes that have activity against ��-1,4 glucans. Although possession of CMCase and endoglucanase activities does not imply necessarily that these bacteria can hydrolyze crystalline cellulose, because some may lack the attachment protein necessary for crystalline cellulose digestion (Fields et?al., 1998). However, the overall involvement of bacteria with CMCase and endoglucanase activities in cellulose digestion is not in doubt (Weimer, 1992). Here, direct evidence is provided showing that known cellulolytic populations F.