Comparative studies of microbial populations in the rumen, duodenum, ileum and faeces of lactating dairy cows.

AIMS: Understanding factors that influence the composition of microbial populations of the digestive system of dairy cattle will be key in regulating these populations to improve animal performance. Although rumen microbes are well studied, little is known of the dynamics and role of microbial populations in the small intestine of cows. Comparisons of fingerprints of microbial populations were used to investigate the effects of gastrointestinal (GI) segment and animal on community structure. METHODS AND RESULTS: Samples from four lactating dairy cows with ruminal, duodenal and ileal cannulae were collected. Terminal-restriction fragment length polymorphism (T-RFLP) comparisons of small subunit rRNA genes revealed differences in microbial populations between GI segments (P < 0.05). No significant differences in either methanogen populations or microbial community profiles between animals were observed. Quantitative PCR was used to assay relative changes in methanogen numbers compared to procaryote rRNA gene numbers, and direct microscopic counts were used to enumerate total procaryote numbers of the duodenal and ileal samples. CONCLUSIONS: T-RFLP comparisons illustrate significant changes in microbial diversity as digesta passes from one segment to another. Direct counts indicate that microbial numbers are reduced by eight orders of magnitude from the rumen, through the abomasum, and into the duodenum (from c. 10(12) to c. 3.6 x 10(4) cells per ml). Quantitative PCR analyses of rRNA genes indicate that methanogens are present in the duodenum and ileum. SIGNIFICANCE AND IMPACT OF THE STUDY: The contribution of microbial populations of the small intestine to the nutrition and health of cattle is seldom addressed but warrants further investigation.

Development of a mechanistic model to represent the dynamics of particle flow out of the rumen and to predict rate of passage of forage particles in dairy cattle.

A mechanistic and dynamic model was developed to represent physiological aspects of particle dynamics in the reticulo-rumen (RR) and to predict rate of passage out of the RR (Kp) of forage particles quantitatively. The model consists of 2 conceptual pools with 3 spatial compartments of particles; the compartment the particle enters is based on functional specific gravity (FSG). The model assumes 2 major pressure gradient-driven flows of particles out of the RR through the reticulo-omasal orifice between 2 consecutive primary reticular contractions. One is associated with the second phase of primary reticular contraction and involves propulsion of particles in the vicinity of the honeycomb structure of the reticulum from the RR. The second flow involves movement of particles in the reticulum without selection by size. Particle outflow rate was assumed to be proportional to liquid outflow rate. The passage coefficient, defined as the ratio of particle to liquid outflow rate, was estimated for each particle group by an equation derived from the probability of passage based on FSG and particle size. Particles retained on a 1.18-mm screen were defined as large particles. When the model was evaluated with 41 observations in an independent database, it explained 66% of the variation in observed Kp of forage particles with a root mean square prediction error of 0.009. With 16 observations that also included measurements of liquid passage rate, the model explained 81 and 86% of the variation in observed Kp liquid and Kp forage, respectively. An analysis of model predictions using a database with 455 observations indicated that the assumptions underlying the model seemed to be appropriate to describe the dynamics of forage particle flow out of the RR. Sensitivity analysis showed that probability of a particle being in the pool likely to escape is most critical in the passage of large forage particles, whereas the probability of being in the reticulum as well as in the likely to escape pool is important in the passage of small forage and concentrate particles. The FSG of a particle is more important in determining the fate of a particle than its size although they are correlated, especially for forage particles. We conclude that this model can be used to understand the factors that affect the dynamics of particle flow out of the RR and predict Kp of particles out of the RR in dairy cattle.

A mechanistic model for predicting the nutrient requirements and feed biological values for sheep.

The Cornell Net Carbohydrate and Protein System (CNCPS), a mechanistic model that predicts nutrient requirements and biological values of feeds for cattle, was modified for use with sheep. Published equations were added for predicting the energy and protein requirements of sheep, with a special emphasis on dairy sheep, whose specific needs are not considered by most sheep-feeding systems. The CNCPS for cattle equations that are used to predict the supply of nutrients from each feed were modified to include new solid and liquid ruminal passage rates for sheep, and revised equations were inserted to predict metabolic fecal N. Equations were added to predict fluxes in body energy and protein reserves from BW and condition score. When evaluated with data from seven published studies (19 treatments), for which the CNCPS for sheep predicted positive ruminal N balance, the CNCPS for sheep predicted OM digestibility, which is used to predict feed ME values, with no mean bias (1.1 g/100 g of OM; P > 0.10) and a low root mean squared prediction error (RMSPE; 3.6 g/100 g of OM). Crude protein digestibility, which is used to predict N excretion, was evaluated with eight published studies (23 treatments). The model predicted CP digestibility with no mean bias (-1.9 g/100 g of CP; P > 0.10) but with a large RMSPE (7.2 g/100 g of CP). Evaluation with a data set of published studies in which the CNCPS for sheep predicted negative ruminal N balance indicated that the model tended to underpredict OM digestibility (mean bias of -3.3 g/100 g of OM, P > 0.10; RMSPE = 6.5 g/100 g of OM; n = 12) and to overpredict CP digestibility (mean bias of 2.7 g/100 g of CP, P > 0.10; RMSPE = 12.8 g/100 g of CP; n = 7). The ability of the CNCPS for sheep to predict gains and losses in shrunk BW was evaluated using data from six studies with adult sheep (13 treatments with lactating ewes and 16 with dry ewes). It accurately predicted variations in shrunk BW when diets had positive N balance (mean bias of 5.8 g/d; P > 0.10; RMSPE of 30.0 g/d; n = 15), whereas it markedly overpredicted the variations in shrunk BW when ruminal balance was negative (mean bias of 53.4 g/d, P < 0.05; RMSPE = 84.1 g/d; n = 14). These evaluations indicated that the Cornell Net Carbohydrate and Protein System for Sheep can be used to predict energy and protein requirements, feed biological values, and BW gains and losses in adult sheep.

Production and economic potentials of cattle in pasture-based systems of the western Amazon region of Brazil.

Our objectives were to evaluate strategies to improve productivity and economic returns from beef and dual-purpose cattle systems based on data collected on one dual-purpose (Bos taurus x Bos indicus) and two beef (Nellore) cattle farms in the western Amazon region of Brazil. Forage chemical composition and digestion rates of carbohydrate fractions of grazed Brachiaria decumbens and Brachiaria brizantha cv. Marandu grasses and Pueraria phaseoloides (tropical kudzu) legume were measured monthly during a 9-mo period from the end of one dry season to the end of the subsequent rainy season. Measurements of milk and growth responses to grazing these forages were used to predict animal productivity responses to dietary nutrient availability throughout an annual cycle. The ME available for gain in our simulations was always more limiting than metabolizable protein. The predicted ME available for gain was 0.50 kg/d for steers grazing B. brizantha and 0.40 kg/d for finishing steers grazing B. decumbens. Grasses contained more NDF and neutral detergent insoluble protein and less ME (P < 0.05) in the rainiest months than in the less rainy season, which resulted in 20% less predicted weight gain by growing steers (P < 0.05). Supplementation with sorghum grain was required to increase milk production and growth by 25 or 50% per animal, respectively, but this strategy was less profitable than current forage-only diets. Greater productivity of land and labor from higher stocking indicated greater net margins for beef production, but not for milk. This study suggested that more intensive beef production by judicious fertilization of grass-legume pastures and greater stocking density is the preferable strategy for owners of these cattle systems to improve economic returns under current conditions. It also might help decrease the motivation for additional forest clearing.

The effect of a ruminal nitrogen (N) deficiency in dairy cows: evaluation of the cornell net carbohydrate and protein system ruminal N deficiency adjustment.

Twenty-four multiparous and fifteen first lactation Holstein cows averaging 263 days in milk and weighing 614 kg were fed diets adequate or deficient in ruminal nitrogen (N), based on predictions of the Cornell Net Carbohydrate and Protein System (CNCPS). After adjustment to a low crude protein (CP) total mixed rations (TMR; 12.6% CP), the cows were allocated to 13 blocks based on lactation number, milk production, body condition score, and body weight. Within each block, cows were randomly assigned to one of the 3 treatment (TRT) diets (9.4, 11.1 and 14.1% CP for TRT 1, 2, and 3, respectively). All diets contained the same proportion of high moisture corn, chopped grass hay, and minerals, with urea substituted for corn silage as needed to reach the three CP levels. The TRT diets were then fed to the cows for 4 wk. Milk production was significantly affected by TRT: 15.5, 18.8, and 21.7 kg/d for TRT diets 1, 2, and 3, respectively. DMI was increased significantly as the percentage of CP increased from 9.4 to 14.1% CP: 17.6, 20.0, and 21.2 kg/d for TRT diets 1,2, and 3, respectively. CNCPS predictions for production (with and without the N adjustment for ruminal N deficiency) of metabolizable protein (MP) allowable milk were compared with observed milk production. Using the average individual weekly cow data from all 3 TRT, we found that the CNCPS accounted for 72 and 68% of the variation in MP allowable milk without and with the N deficiency adjustment, respectively. The overall mean bias without the N adjustment was 3.3 kg of milk (over prediction model bias of 14.6%, P < 0.001), and the N adjustment reduced the model over-prediction bias to 0.01 kg of milk (P = 0.96).

Raising livestock in resource-poor communities of the North West Province of South Africa--a participatory rural appraisal study.

A participatory research model was used in six village communities in the Central Region of the North West Province of South Africa in order to achieve the following broad objectives: to obtain information on the challenges owners face in raising livestock in these areas and to evaluate the livestock owners' level of knowledge of internal parasites in their animals. Information obtained at participatory workshops clearly indicated a need for improvements in water supply, schools, job creation, and health services. Lack of pasture for grazing livestock was also cited as being important. Other most frequently mentioned livestock problems included 'gall sickness' (a vaguely defined condition not necessarily referring to anaplasmosis), parasites (both external and internal), chicken diseases and ingestion of plastic bags discarded in the environment. When livestock owners were questioned during individual interviews, most were able to identify the presence of parasites in either the live or dead animal. However, it seems likely that this is limited to the identification of tapeworms. It was found that most livestock owners use a combination of treatments, ranging from traditional to folklore to commercial. There were some difficulties in using the participatory methods since it was the first time that the facilitators and the communities had been exposed to them. Many communities had difficulty in dealing with the concept of finding solutions within the community, which is such an integral part of participatory methods.

The amino acid profiles of the whole plant and of four plant residues from temperate and tropical forages.

This study compared the amino acid (AA) profile of five residues (original forage, borate-phosphate buffer residue (BPR), neutral detergent fiber residue with (NDF+) and without (NDF-) sodium sulfite, and acid detergent fiber residue (ADF). Fourteen grasses and legumes from tropical and temperate regions were used in this study. The use of sodium sulfite did not affect the NDF concentration, but the NDF insoluble protein was lower (P < 0.05) in the NDF+ than in the NDF- (3.9 vs 4.5% DM, respectively). For all of the amino acids tested, the amino acid content, expressed as a percentage of CP, was lower in the ADF residue than in the original forage. There were no differences in the amino acid concentrations of the NDF- and NDF+ extracts (P > 0.05). Only in the case of methionine was there a difference in the amount of amino acid when the original forage was compared with the BPR (1.84 vs 1.45 % CP). When the AA profile of each residue was corrected for the AA content of the ADF, no difference was observed between the AA profile of the original forage and of the BPR (P > 0.05). Similar to the result without correction for the amino acids in ADF, the AA profiles of the NDF+ and NDF- fractions were similar (P > 0.05). From this result, we infer that the sodium sulfite had similar effects on all AA in the NDF residue that we tested. There were differences in amino acid concentrations in the original forage and the NDF residues for several amino acids (Met, Cys, Lys, Thr, Arg, Ile, Leu, and Phe) (P < 0.05). When the amino acid values of the original forage and the BPR were used with animal data in the Cornell Net Carbohydrate and Protein System model, few differences in animal predicted performance were evident. These findings suggest that the AA profile of the original forage can be used to predict the AA profile of the undegraded intake protein instead of using the borate-phosphate buffer residue for amino acid analyses. This would simplify obtaining feed amino acid values for use in the Cornell Net Carbohydrate and Protein System.

Rate and extent of digestion of the ethanol-soluble and neutral detergent-insoluble fractions of corn grain.

The objectives of this study were to partition corn grain into three digestible fractions and to measure the rate of disappearance of these fractions in vitro. Seventeen corn grain samples with varied fiber concentrations were extracted with either 80% ethanol or neutral detergent to obtain estimates of the pool size and digestion kinetics of the A, B1, B2, and C fractions. The carbohydrate soluble in 80% ethanol averaged only 2.6+/-.3% of the DM, although 80% ethanol extracted 7.1+/-1.2% of DM of corn grain. The ethanol-soluble fraction of corn grain contained protein, ether-extractable compounds, and a small amount of ash in addition to carbohydrate. Because of this chemical heterogeneity and because of the small size of the ethanol-soluble fraction, it was not possible to determine the digestion rate of this fraction by measuring gas production. The NDF content of the corn grain was 10.6+/-.7% of DM and was highly digestible (94.6+/-1.4%). The digestible NDF contributed 9.5% of the total gas production from corn grain. Because the size, digestibility, and digestion rate of the digestible NDF fraction varied little among corn grain samples, it is not necessary to routinely analyze the digestion kinetics of the digestible NDF fraction of dried corn grain. An average gas production curve of this fraction can be used as a base to subtract from the total gas production curve to generate the gas production curve of the neutral detergent-soluble fraction for dried, ground corn grain samples.

Evaluation of tropical grasses for milk production by dual-purpose cows in tropical Mexico.

Two experiments using the Cornell Net Carbohydrate and Protein System were conducted to characterize the carbohydrate and protein fractions and corresponding rates of digestion of 15 tropical pasture grasses and to evaluate their ability to support milk production by dual-purpose cows. In the first experiment, ranges in carbohydrate and protein fractions of 15 grasses at 35 to 42 d of regrowth were: neutral detergent fiber (NDF) 63.5 to 74.9% of DM; permanganate lignin 4.7 to 7.8% of NDF; CP 5.5 to 11.9% of DM; and soluble protein 15.1 to 44.1% of crude protein (CP). The ranges of rates of digestion expressed as percent per hour were neutral detergent solubles (7.5 to 27.4); NDF (3.8 to 8.4); and neutral detergent insoluble protein (2.9 to 9.5). Predictions of the amount of milk that could be produced based on the amount of metabolizable energy supplied by the diet decreased 35% when NDF increased from 60 to 80%, and increased 88% when the rate of digestion of NDF increased from 3 to 6%/h. The milk production that could be sustained based on metabolizable protein in the diet doubled as CP increased from 4 to 12%. In the second experiment, nitrogen fertilization reduced NDF 7.3% and increased CP 84% without changing protein solubility, resulting in increased rumen nitrogen and metabolizable protein balances. With all forages, the Cornell Net Carbohydrate and Protein System predicted that availability of metabolizable protein would limit milk production. Predicted microbial growth was limited by ruminally available protein rather than by available carbohydrate.

Use of in vitro gas production models in ruminal kinetics.

Physiological systems models for ruminant animals are used to predict the extent of ruminal carbohydrate digestion, based on rates of intake, digestion, and passage to the lower tract. Digestion of feed carbohydrates is described in these models by a first-order rate constant. Recently, an in vitro gas production technique has been developed to determine the digestion kinetics in batch fermentation, and nonlinear mathematical models have been fitted to the cumulative gas production data from these experiments. In this paper, we present an analysis that converts these gas production models to an effective first-order rate constant that can be used directly in rumen systems models. The analysis considers the digestion of an incremental mass of substrate entering the rumen. The occurrence of passage is represented probabilistically, and integration through time gives the total mass of substrate and total rate of digestion in the rumen. To demonstrate the analysis, several gas production models are fitted to a sample data set for corn silage, and the effective first-order rate constants are calculated. The rate constants for digestion depend on ruminal passage rate, an interaction that arises from the nonlinearity of the gas production models.

Phenotypic and phylogenetic characterization of ruminal tannin-tolerant bacteria.

The 16S rRNA sequences and selected phenotypic characteristics were determined for six recently isolated bacteria that can tolerate high levels of hydrolyzable and condensed tannins. Bacteria were isolated from the ruminal contents of animals in different geographic locations, including Sardinian sheep (Ovis aries), Honduran and Colombian goats (Capra hircus), white-tail deer (Odocoileus virginianus) from upstate New York, and Rocky Mountain elk (Cervus elaphus nelsoni) from Oregon. Nearly complete sequences of the small-subunit rRNA genes, which were obtained by PCR amplification, cloning, and sequencing, were used for phylogenetic characterization. Comparisons of the 16S rRNA of the six isolates showed that four of the isolates were members of the genus Streptococcus and were most closely related to ruminal strains of Streptococcus bovis and the recently described organism Streptococcus gallolyticus. One of the other isolates, a gram-positive rod, clustered with the clostridia in the low-G+C-content group of gram-positive bacteria. The sixth isolate, a gram-negative rod, was a member of the family Enterobacteriaceae in the gamma subdivision of the class Proteobacteria. None of the 16S rRNA sequences of the tannin-tolerant bacteria examined was identical to the sequence of any previously described microorganism or to the sequence of any of the other organisms examined in this study. Three phylogenetically distinct groups of ruminal bacteria were isolated from four species of ruminants in Europe, North America, and South America. The presence of tannin-tolerant bacteria is not restricted by climate, geography, or host animal, although attempts to isolate tannin-tolerant bacteria from cows on low-tannin diets failed.

Predicting forage indigestible NDF from lignin concentration.

We used chemical composition and in vitro digestibility data from temperate and tropical forages to develop relationships between indices of lignification and forage indigestible NDF. Neutral detergent fiber indigestibility increased nonlinearly as the lignin concentration of the NDF increased. Differences in estimated indigestible NDF using equations developed for a specific forage class (C3 and C4 grasses and legumes) were small and are probably not biologically significant when compared to those estimated from a common equation. Selected equations were compared with the Cornell Net Carbohydrate and Protein System (CNCPS) for the prediction of ADG. The linear equation (2.4 times NDF lignin content) used by the CNCPS and the Beef NRC had some of the largest errors due to mean bias. A log-log model [4.37 x (lignin/NDF)(.84)] provided the best combination of low total prediction error, low mean bias, and minimal error due to regression bias when permanganate lignin was used. A similar equation based on sulfuric acid lignin [6.17 x (lignin/NDF)(.77)] also met the above criteria. These equations then were evaluated with the CNCPS model against animal growth data from diets ranging in forage quality. Regardless of the equation used for predicting unavailable fiber, the CNCPS underpredicted daily gain, with mean biases ranging from -.10 to -.22 kg/d. Regression bias ranged from .13 to .14 kg/d and the coefficients differed from unity (P = .0001). The new equations gave numerically lower energy allowable ADG by steers compared to the linear equation currently used by the CNCPS model. The estimates were lower due to a higher predicted indigestible NDF, which resulted in a lower estimated forage energy value.

Ensiling effects of the ethanol fractionation of forages using gas production.

We studied the use of gas curve subtraction to distinguish between two fractions soluble in neutral detergent solution. Samples of unfractionated (whole) forage, residue insoluble in 90% ethanol, and isolated NDF were fermented in vitro, and gas production was monitored. The gas volume associated with the ethanol solubles (A fraction) was determined as the difference between the gas from the whole forage and from the ethanol residue. The gas yield associated with the fraction insoluble in 90% ethanol but soluble in neutral detergent solution (B fraction) was determined by subtracting the isolated NDF gas curve from the corresponding ethanol residue curve. This experiment included three forages (alfalfa, bromegrass, and orchardgrass) harvested at two maturities and preserved by freeze-drying or ensiling to form a 3 x 2 x 2 factorial arrangement. Ensiling reduced the rate of gas formation from the A fraction by approximately 30% (P < .01). Ensiling increased (P < .05) the size of the A fraction (2 to 10% of DM) but did not change the volume of gas produced (P > .05). The gas yield from the B1 fraction was reduced 40% (P < .05) by ensiling with no significant change in rate. Curve subtraction of gas production profiles may be used to obtain rate estimates for multiple neutral detergent soluble pools. The separation of the neutral detergent solubles into two pools clarified the effects caused by ensiling. Changes due to ensiling on the rate of gas produced were associated with the A fraction, and the effects on final gas volume were associated with the B1 fraction.

Modeling ruminal pH fluctuations: interactions between meal frequency and digestion rate.

A steady periodic analysis of ruminal carbohydrate digestion was developed to predict the effects of diet and frequency of eating on ruminal pH fluctuation. Tests of the model against previous data showed that pH fluctuations were too large when previously published rates of carbohydrate digestion were used but were improved using rates from an in vitro gas production system, which were lower. With the original digestion rates, the minimum meal frequency to maintain steady-state conditions in the rumen increased from 4 to 12 meals/d as dietary effective neutral detergent fiber (NDF) decreased from 34 to 6% of dry matter (DM); with the revised rates, the minimum frequency was 3 to 6 meals/d, respectively. The minimum effective NDF to maintain a pH value above 6.0 increased from 14 to 23% of DM as meal frequency decreased from steady state to 2 meals/d using the original rates; with the revised rates, the minimum effective NDF was slightly smaller, increasing from 13 to 21% of DM, respectively. Effects of DM intake and body weight on pH fluctuation were minor, and dietary buffers, when used at rates less than 1%, did not reduce fluctuation. Different methods of calculating mean ruminal pH yielded different results for the effect of meal frequency on mean pH.

Manure and microbes: public and animal health problem?

Most environmental concerns about waste management either have focused on the effects of nutrients, especially N and P, on water quality or have emphasized odor problems and air quality. Microbes from manure are often low on the priority list for control and remediation, despite the fact that several outbreaks of gastroenteritis have been traced to livestock operations. The pathogens discussed in this paper include protozoans (Cryptosporidium parvum, Giardia spp.), bacteria (Listeria monocytogenes, Escherichia coli O157:H7, Salmonella spp., and Mycobacterium paratuberculosis), and some enteric viruses. Clinical symptoms, prospects for zoonotic infection, and control methods other than the use of antimicrobials are considered. Recommendations to avoid disease transmission include taking steps to ensure the provision of clean, unstressful environments to reduce disease susceptibility and the careful handling and spreading of manure from animals at high risk for infection, especially young calves. Composting and drying of manure decrease the number of viable pathogens. Environmental controls, such as filter strips, also reduce the risk of water contamination.

Development and evaluation of equations for prediction of feed intake for lactating Holstein dairy cows.

Improved prediction equations for dry matter intake (DMI) of Holstein cows that consume high energy diets were developed using regression techniques applied to a comprehensive database. The equations for predicting DMI, which were dependent on parity, accounted for the effects of milk yield, milk protein, body weight (BW), BW change, days pregnant, ambient temperature, relative humidity, and night cooling. A simplified prediction equation of DMI for farm application was developed and based on milk protein yield and BW at calving. An ambient temperature and a lag adjustment factor for early lactation were developed to improve accuracy of prediction of DMI of dairy cows in early lactation. The developed equations for DMI were evaluated against six independent data files. These equations accounted for 55 to 98% of the variation of the weekly group DMI of the independent validation data. The remainder of the variation in intake was attributed to diet, management, and undescribed animal factors. The equations developed in this study had a mean proportional bias of 5.6% and a mean square prediction error of 5.45 kg2/d. Predicted intake using the new equations was within 3 to 8% of actual intake. The new equations must be applied to situations in which Holstein dairy cows are fed highly digestible diets because dietary fill effects are not considered in these equations. The relationship of milk protein yield and DMI warrants further investigation.

Evaluation of alternative equations for prediction of intake for Holstein dairy cows.

Six prediction equations for dry matter intake (DMI) were evaluated for accuracy with independent data. The equations were selected based on ease of parameter measurement and practical on-farm use. The equations were assessed for accuracy of predicting individual weekly DMI for primiparous (n = 105) and multiparous (n = 136) cows; three-fourths of these cows were supplemented with a sustained-release form of bovine somatotropin (bST). Large variations in accuracy were identified across the six prediction equations for effects of parity and bST. Prediction accuracy of all equations for cows in wk 1 to 24 of lactation was better for primiparous cows than for multiparous cows. Precision of prediction equations was poor for cows in wk 8 through 12 of lactation and for cows in > 40 wk of lactation. The equation for DMI with the best accuracy measured by a low total lactation mean square prediction error was the modified equation of the National Research Council: DMI (kilograms per day) = -0.293 + 0.372 x fat-corrected milk (kilograms per day) + 0.0968 x body weight 0.75 (kilograms). However, the overall mean bias (predicted minus observed) of the prediction of weekly DMI of a single cow was high for all equations, including the modified equation of the National Research Council. For wk 2, 4, 8, 10, and 20 of lactation, the mean bias for the modified equation was +6, +3.4, -1.3, -2.1, and -2.8 kg/d. The accuracy of prediction was lower for cows treated biweekly with bST. High yielding cows and cows treated biweekly with bST had higher milk yields in relation to body weight, and standardized prediction equations for DMI were less accurate.

The effect of preservation method on the neutral detergent soluble fraction of forages.

Fermentation of neutral detergent solubles (NDS) was assessed using a 3 x 3 x 3 factorial arrangement. Three forage species (alfalfa, bromegrass, and orchardgrass) were collected at three maturities and preserved either by freeze drying, oven drying at 50 degrees C, or by ensiling. Each feed sample and its isolated NDF were fermented in vitro and gas production was monitored. Gas yield from NDS was determined as the difference between gas from the unfractionated forage and from its respective NDF. The forages ranged from 23 (immature alfalfa) to 68% NDF (mature orchardgrass). The silages were well fermented with a final pH of < or = 4.5. Increasing maturity decreased the final gas volume but did not change the rate of gas production from the NDS fraction. There was little difference in gas production between freeze-dried and oven-dried forage samples. Ensiling decreased gas yield from the unfractionated forage. The rate of gas production from the NDS fraction of the ensiled forages decreased an average of .05 h-1 compared with the freeze-dried sample. Gas yield from the NDS fraction decreased (from the freeze-dried sample) between 7 and 36% upon ensiling. The curve substraction approach can be used to evaluate the effects of ensiling on the neutral detergent-soluble fraction of forages.

Molecular beacons: trial of a fluorescence-based solution hybridization technique for ecological studies with ruminal bacteria.

Molecular beacons are fluorescent probes developed for solution rather than membrane hybridization. We have investigated the utility of these probes to study rumen microbial ecology. Two cellulolytic species, Ruminococcus albus and Fibrobacter succinogenes, were tested. Membrane and solution hybridizations gave similar results in competition experiments with cocultures of R. albus 8 and F. succinogenes S85.