We divided the adhesion process of the predominant cellulolytic rumen bacteria Fibrobacter succinogenes, Ruminococcus flavefaciens, and Ruminococcus albus into four phases: 1) transport of the nonmotile bacteria to the substrate; 2) initial nonspecific adhesion of bacteria to unprotected sites of the substrate that is dominated by constitutive elements of bacterial glycocalyx; 3) specific adhesion via adhesins or ligands formation with the substrate, which can be dominated by several bacterial organelles including cellulosome complexes, fimbriae connections, glycosylated epitopes of cellulose-binding protein (CBP) or glycocalyx, and cellulose-binding domain (CBD) of enzymes; 4) proliferation of the attached bacteria on potentially digestible tissues of the substrate. Each of the phases and its significance in the adhesion process are described. Factors affecting bacterial adhesion are described including: 1) factors related to bacterial age, glycocalyx condition, and microbial competition; 2) factors related to the nature of substrate including, cuticle protection, surface area, hydration, and ionic charge; and 3) environmental factors including pH, temperature, and presence of cations and soluble carbohydrate. Based on the information available from the literature, it appears that each of the predominant rumen bacteria--F. succinogenes, R. flavefaciens, and R. albus--has a specific mechanism of adhesion to cellulose. In F. succinogenes, both the glycosidic residues of the outer membrane CBP and especially of the 180-kDa CBP, and the distinct CBD of EG2 EGF and Cl-stimulated cellobiosidase, may play a role in the adhesion to cellulose. No direct evidence, except scanning electron microscopy observations, yet supports the existence of either cellulosome complex or fimbriae structures involved in the adhesion mechanism of F. succinogenes. At least two mechanisms, cellulosome-like complexes and carbohydrate epitopes of the glycocalyx layer are involved in the specific adhesion of R. flavefaciens to cellulose. Ruminococcus albus possesses at least two mechanisms for specific adhesion to cellulose: a cellulosomal-like mechanism, and a CbpC (Pil)-protein mechanism that probably involves the production of fimbrial-like structures. Indirect and direct studies suggested that carbohydrate epitopes of CBPs and CBD epitope of cellulases may also be involved mostly in the nonspecific phase of adhesion of R. albus.

The potent hepatotoxin and carcinogen aflatoxin B1 (AFB1) is a common mycotoxin contaminant of grains used in animal feeds. Aflatoxin M1 (AFM1) is the major metabolite of AFB1 in mammals, being partially excreted into milk, and is a possible human carcinogen. The maximum permitted concentration of AFM1 in cows’ milk is 0.05 μg/kg in Israel and the European Union. Since milk yield and the carry-over of AFB1 in the feed to AFM1 in the milk are highly correlated, it was considered important to determine the AFM1 carry-over in Israeli-Holstein dairy cows, distinguished by world record high milk production. Twelve such cows were used to determine AFM1 carry-over following daily oral administration of feed containing ~86 μg AFB1 for 7 days. The mean carry-over rate at steady-state (Days 3–7) was 5.8% and 2.5% in mid-lactation and late-lactation groups, respectively. The carry-over appears to increase exponentially with milk yield and could be described by the equation: carry-over% = 0.5154 e0.0521 × milk yield, with r2 = 0.6224. If these data truly reflect the carry-over in the national Israeli dairy herd, the maximum level of AFB1 in feed should not exceed 1.4 μg/kg, a value 3.6 times lower than the maximum residue level currently applied in Israel.

This study measured the effects of including soyhulls as partial roughage replacement in total mixed rations (TMR) fed to 25 pairs of cows during early lactation, on the dry matter (DM) intake, particle kinetics, rumination, in vivo DM and NDF digestibility, milk and FCM yields, and BW changes. The 2 diets used in this study differed in the content of roughage and roughage NDF [23.5 vs. 35.0%, and 12.8 vs. 18.7% in the experimental (EXP) and control (CON) TMR, respectively]. The EXP TMR contained 20.5% less physically effective NDF than the CON TMR (11.7 vs. 14.1% of DM, respectively). These differences were expressed in a greater intake per meal (by 13.3%), a higher rate of meal intake (by 23.2%), a similar number of meals per day, a shorter daily eating duration (by 13%), and a higher total daily DMI (by 7.2%) in the EXP cows as compared with the CON cows. The in vivo DM and NDF digestibility was higher by 4.9 and 22.7%, respectively, in the EXP cows than in the CON cows. The rumination time for the TMR in the EXP cows was 12.7% (54.3 min/d) shorter than in the CON cows, and this was probably related to the difference of 12.4% in physically effective NDF intake between the 2 groups. Patterns of daily rumination and feed consumption throughout an average day showed a delay of approximately 1 to 2 h between the eating and rumination peaks. Particle flow from the rumen of the EXP cows was characterized by a longer rumen mean retention time (by 17.8%) and longer rumination time per kilogram of roughage ingested (by 23.5%) as compared with the CON cows. Thus, favorable conditions for NDF digestion were created in the rumen of the EXP cows, as reflected in their rumen pH values (6.67). The advantage of the EXP cows in intake and digestibility was reflected in a concomitant increase of 7.4% in milk production and of 9.2% in FCM yield as compared with the CON cows. No difference was found between the 2 groups with respect to efficiency of feed utilization for milk production and BW changes.

The aim of the study was to determine the effect of 10 sources of lactic acid bacteria (LAB) on dry matter digestibility (DM-D) and neutral detergent fiber digestibility (NDF-D), in various combinations with starch, in vitro. The soluble starch represented a concentrate feed, whereas silage represented feeding only roughage. The DM-D and NDF-D were determined after 24 and 48 h of incubation to represent effective (24 h) and potential (48 h) digestibility. Addition of LAB was both by direct application of the inoculants to rumen fluid (directly fed microbials) and by the use of preinoculated silages. For each feed combination, tubes without added LAB served as controls. The results indicate that, overall, some LAB inoculants applied at ensiling or added directly to the rumen fluid had the potential to increase the DM-D and NDF-D. The major significant inoculant effect on NDF-D was obtained after 24 h of incubation, whereas the effect after 48 h was mainly nonsignificant. The effective inoculants seemed to minimize the inhibitory effect of the starch on NDF-D within 24 h, perhaps by competition with lactate-producing rumen microorganisms.

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The cellulolytic bacterium Ruminococcus albus 8 adheres tightly to cellulose, but the molecular biology underpinning this process is not well characterized. Subtractive enrichment procedures were used to isolate mutants of R. albus 8 that are defective in adhesion to cellulose. Adhesion of the mutant strains was reduced 50% compared to that observed with the wild-type strain, and cellulose solubilization was also shown to be slower in these mutant strains, suggesting that bacterial adhesion and cellulose solubilization are inextricably linked. Two-dimensional polyacrylamide gel electrophoresis showed that all three mutants studied were impaired in the production of two high-molecular-mass, cell-bound polypeptides when they were cultured with either cellobiose or cellulose. The identities of these proteins were determined by a combination of mass spectrometry methods and genome sequence data for R. albus 8. One of the polypeptides is a family 9 glycoside hydrolase (Cel9B), and the other is a family 48 glycoside hydrolase (Cel48A). Both Cel9B and Cel48A possess a modular architecture, Cel9B possesses features characteristic of the B 2 (or theme D) group of family 9 glycoside hydrolases, and Cel48A is structurally similar to the processive endocellulases CelF and CelS from Clostridium cellulolyticum and Clostridium thermocellum, respectively. Both Cel9B and Cel48A could be recovered by cellulose affinity procedures, but neither Cel9B nor Cel48A contains a dockerin, suggesting that these polypeptides are retained on the bacterial cell surface, and recovery by cellulose affinity procedures did not involve a clostridium-like cellulosome complex. Instead, both proteins possess a single copy of a novel X module with an unknown function at the C terminus. Such X modules are also present in several other R. albus glycoside hydrolases and are phylogentically distinct from the fibronectin III-like and X modules identified so far in other cellulolytic bacteria.

Nonforage byproduct feeds, including distillers dried grains (DDG), corn gluten feed (CGF), wheat bran (WB), beet pulp (BP), soybean hulls (SH), and dried citrus pulp (DCP), were examined for monosaccharide composition and in vitro digestibility by ruminal liquor. The dicotyledonous feeds (BP, SH, and DCP) contained more galactose, pectin, and NDF glucans and less NDF-xylan and NDF-arabinose than the monocotyledonous ones (DDG, CGF, and WB). The lowest values of lignin were found in CGF, SH, and DCP. Digestibility of total carbohydrate was around 90% in CGF, DCP, BP, and SH and around 80% in DDG and WB. Digestibility of total NDF polysaccharides was 86% in CGF and SH, 78--84% in DDG, DCP, and BP, and 56% in WB. In all byproducts, digestibility of NDF glucose and arabinose was higher than that of NDF xylose and uronic acids.