It is well recognized that a dynamic biofilm develops upon plant biomass in the herbivore gastrointestinal tract, but this component of the microbiome has not previously been specifically sampled, or directly compared with the biodiversity present in the planktonic fraction of digesta. In this study, the digesta collected from four sheep fed two different diets was separated into three fractions: the planktonic phase, and the microbial populations either weakly or tightly adherent to plant biomass. The community DNA prepared from each fraction was then subjected to both ribosomal intergenic spacer analysis (RISA) and denaturing gradient gel electrophoresis (DGGE). Both types of analysis showed that dietary factors influence community structure, and that the adherent fractions produced more complex profiles. The RIS-clone libraries prepared from the planktonic and adherent populations were then subjected to restriction fragment length polymorphism (RFLP) and DNA sequence analyses, which resulted in a far greater degree of discrimination among the fractions. Although many of the sequenced clones from the adherent populations were assigned to various clusters within the low G+C Gram-positive bacteria, the clone libraries from animals consuming an all-grass diet were largely comprised of novel lineages of Clostridium, while in animals consuming the starch-containing diet, Selenomonas and Ruminococcus spp. were the dominant low G+C Gram-positive bacteria. Additionally, the libraries from hay-fed animals also contained clones most similar to asaccharolytic Clostridia, and other Gram-positive bacteria that specialize in the transformation of plant phenolic compounds and the formation of cinnamic, phenylacetic and phenylpropionic acids. These results reveal, for the first time, the phylogeny of adherent subpopulations that specialize in the transformation of plant lignins and other secondary compounds, which potentiate polysaccharide hydrolysis by other members of the biofilm.

thermophilic, glucose-fermenting, strictly anaerobic, rod-shaped bacterium, strain SEBR 6459T (T = type strain), was isolated from an African oil-producing well. This organism was identified as a member of the genus Thennotoga on the basis of the presence of the typical outer sheath-like structure (toga) and 16s rRNA signature sequences and its ability to grow on carbohydrates (glucose, arabinose, fructose, lactose, maltose, and xylose). Major differences in its 16s rRNA gene sequence, its lower optimum temperature for growth (66"C), its sodium chloride range for growth (0 to 2.8%), its lack of lactate as an end product from glucose fermentation, and its peritrichous flagella indicate that strain SEBR 6459T is not similar to the three previously described Thennotoga species. Furthermore, this organism does not belong to any of the other genera related to the order Thermotogales that have been described. On the basis of these findings, we propose that this strain should be described as a new species, Thermotoga ewi. The type strain of T. e@i is SEBR 6459 (= DSM 9442).The order Thermotogales currently comprises three genera, TheTotoga (17), Thermosipho (18), and Fervidobacterium (19, 35). All of the species belonging to these genera are rod shaped and have a characteristic outer sheath-like structure which can be observed under in situ conditions (34). Members of the recently described genera Geotoga and Petrotoga (9) also possess this morphological feature and, as determined by a 16s rRNA sequence analysis, are distantly related to members of the Thermotogales. Collectively, the five genera mentioned above represent one of the deepest phylogenetic branches in the domain Bacteria (42). These taxa can be differentiated on the basis of their optimum temperatures for growth; Thermotoga species are extreme thermophiles that have optimum temperatures for growth of around 80°C (17, 20, 21, 41), Thermosipho and Fervidobacterium species have optimum temperatures for growth of 65 to 75°C (18, 19, 35) and are regarded as thermophiles, and Geotoga and Petrotoga species are moderate thermophiles having optimum temperatures for growth of less than 60°C (9). Until recently, members of the three genera belonging to the order Therrnotogales (Therrnotoga, Therrnosipho, and Fervidobacterium) have been isolated only from volcanic aquatic environments. Different species have different sodium chloride requirements and optimum temperatures for growth. These differences reflect the restricted ecological habitats (hydrothermal marine environments, hydrothermal terrestrial environments) from which the organisms were isolated. However, Thermotoga species capable of growth at 85°C have recently been isolated from oil wells in the North Sea (39). Davey et al. (9) have also described the isolation of members of two new genera of moderate thermoanaerobes, the genera Petrotoga and Geotoga, from oil fields. These reports have extended our knowl- edge concerning the ecological habitats of these bacteria. In this paper, we describe the results...

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Dairy cows grazing high-digestibility pastures exhibit pronounced diurnal variation in ruminal pH, with pH being below values considered optimal for digestion. Using a dual-flow continuous culture system, the hypothesis that minimizing diurnal variation in pH would improve digestion of pasture when pH was low, but not at a higher pH, was tested. Four treatments were imposed, with pH either allowed to exhibit normal diurnal variation around an average pH of 6.1 or 5.6, or maintained at constant pH. Digesta samples were collected during the last 3 d of each of four, 9-d experimental periods. A constant pH at 5.6 compared with a constant pH of 6.1 reduced the digestibility of organic matter (OM), neutral detergent (NDF), and acid detergent fiber (ADF) by 7, 14, and 21%, respectively. When pH was allowed to vary (averaging 5.6), digestion of OM, NDF, and ADF were reduced by 15,30, and 36%, respectively, compared with pH varying at 6.1. There was little difference in digestion parameters when pH was either constant or varied with an average pH of 6.1. However, when average pH was 5.6, maintaining a constant pH significantly increased digestion of OM, NDF, and ADF by 5, 25, and 24% compared with a pH that exhibited normal diurnal variation. These in vitro results show that gains in digestibility and potential milk production can be made by minimizing diurnal variation in ruminal pH, but only when ruminal pH is low (5.6). However, larger gains in productivity can be achieved by increasing average daily ruminal pH from 5.6 to 6.1.

In two experiments, each with 32 cross-bred ([Merino x Border Leicester] x Poll Dorset) wether lambs (26 to 33 kg weight range), animals were randomly assigned to one of four treatments. A mixture of lucerne chaff:oaten chaff was used as a basal diet, offered in different ratios. Animals were allowed to consume on a free-access basis in Exp. 1 or 90% of ad libitum intake in Exp. 2 in order to provide a low- (6.5 MJ ME/d) and medium- (9.5 MJ ME/d) quality basal diet, respectively. Isoenergetic amounts of lipid supplements, fish meal (80 g DM), canola meal (84 g DM), and soy meal (75 g DM) were tested in Exp. 1. In Exp. 2, fish meal (9% DM), unprotected rapeseed (7% DM), and protected canola seed (6% DM) were fed as supplements. At the end of 53-d (Exp. 1) or 46-d (Exp. 2) experimental periods, lambs were slaughtered at a commercial abattoir and at 24 h postmortem longissimus thoracis (LT) muscle was collected for the analysis of fatty acid (FA) composition of structural phospholipid and storage triglyceride fractions. Fish meal diet increased LT muscle long-chain n-3 FA content by 27% (P < 0.02) in Exp. 1 and 30% (P < 0.001) in Exp. 2 compared with lambs fed the basal diet, but fish meal decreased (P < 0.01) the n-6 FA content only in Exp. 1. Soy meal and protected canola seed diets increased (P < 0.01) LT muscle n-6 FA content but did not affect long-chain n-3 FA content. Longissimus thoracis muscle long-chain n-3 FA were mainly deposited in structural phospholipid, rather than in storage triglyceride. In both Exp. 1 and Exp. 2, the ratio of n-6:n-3 FA in LT muscle was lowest (P < 0.01) in lambs fed fish meal supplement compared with all other treatments. Protected canola seed diet increased the ratio of n-6:n-3 FA (P < 0.01) and PUFA:saturated fatty acid (P < 0.03) content from those animals fed the basal, fish meal, and unprotected rapeseed diets in Exp. 2. This was due to an increase in muscle n-6 FA content, mainly linoleic acid, of both phospholipid (P < 0.001) and triglyceride (P < 0.01) fractions and not to an increase in muscle n-3 FA content. The results indicate that by feeding fish meal supplement, the essential n-3 FA can be increased while lowering the ratio of n-6:n-3 content in lamb meat to an extent that could affect nutritional value, attractiveness, and the economic value of meat.

SUMMARYThe effect of maternal undernutrition between the 30th and 96th day of pregnancy on placental growth in single-bearing ewes was determined separately in two experiments in consecutive years (1986 and 1987) at Mount Derrimut Field Station, Deer Park, Australia. In the first experiment, placental growth measured on the 96th day of pregnancy was reduced (473 v. 596 g) by maternal undernutrition, while in the second, placental growth was increased (600 v. 496 g). Foetal weight and its linear dimensions on the 96th day of pregnancy were not significantly affected by maternal u dernutrition in either experiment. The only significant difference in the animals and experimental conditions between the two studies was the liveweight of the ewes at mating, which was c. 10 kg higher in the second experiment compared with the first (P < 0·01). Body condition score and the change in condition score during the respective nutritional treatments were not significantly different between the two experiments. The results suggest that maternal liveweight, possibly body reserves not quantifiable by condition score, protects and even enhances placental growth during a period of maternal undernutrition.

The extents and sites of digestion of organic matter (OM), and its constituent watersoluble carbohydrates, organic acids, pectin, cellulose, hemicellulose and crude protein have been studied in sheep prepared with re-entrant duodenal cannulas and fed four fresh herbage diets, Ruanui perennial ryegrass, Tama Westerwolds ryegrass, Pitau white clover and Fakir sainfoin, at each of two levels of intake.The water-soluble carbohydrate, organic acids and pectin of all diets were almost completely digested within the rumen. Some 10% of water soluble carbohydrate reached the duodenum on each diet, though this may not have been of dietary origin. Only on legume diets, where pectin concentration was higher, did measurable amounts of pectin reach the intestine, accounting for some 5 % of the pectin.Hemicellulose and cellulose digestibilities differed between diets, being lowest for sainfoin, and next lowest for clover. Between 79 and 94 % of digestible hemicellulose was digested in the stomach, but diet and intake had no significant effect on this partition. Of the digestible cellulose, 87-97 % was digested in the stomach.Digestibility of N was lowest for sainfoin and highest for Tama ryegrass. There were no significant differences between herbage species or intake in the percentage of digested N digested in the stomach or intestines. The tannin contained in sainfoin had no effect on nitrogen digestion. Data from this and other studies reported in the literature were examined as a basis for establishing prediction equations whereby the partition of digestion of the major carbohydrate and nitrogenous constituents in stomach and intestines might be estimated from data obtainable from standard digestibility trials. Regressions were developed for predicting the amounts of OM, cellulose, and hemicellulose digested in the stomach. There are not yet enough suitable data available to predict the amount of nitrogen entering the small intestine.