Biohydrogenation of unsaturated fatty acids in continuous culture fermenters during digestion of orchardgrass or red clover with three levels of ground corn supplementation.

Diet digestibility and outputs of biohydrogenation intermediates were assessed in a continuous culture of ruminal microorganisms. Orchardgrass or red clover harvested and frozen during spring or fall served as the primary substrates for fermentation. During 10-d incubations, fermenters were fed thawed forage (50 g of DM/d), forage (42 g/d) plus 8 g/d of corn, or forage (34 g/d) plus 16 g/d of corn. Effluents from the last 3 d of incubation were composited for analyses. Starch input increased from 5 to 27% of DM as corn input increased from 0 to 16 g/d. Corn input reduced (P < 0.01) pH, increased (P < 0.01) microbial DM yield, and increased (P = 0.01) digestibility of DM, NDF, CP, and nonstructural carbohydrates. Overall, apparent hydrogenation (percentage) of cis9-18:1, 18:2n-6, and 18:3n-3 was greater (P < 0.05) with orchardgrass than clover. Hydrogenation of cis9-18:1 and 18:2n-6 increased (P = 0.01), but hydrogenation of 18:3n-3 decreased (P = 0.01) linearly due to corn input, regardless of forage. As a result, output of trans11, cis15-18:2 also decreased (P = 0.01). Average output of cis9,trans11-18:2 was greater (P = 0.01) for clover (1.3 mg/d) compared with orchardgrass (0.6 mg/d), but corn input with either forage increased (P = 0.01) cis9,trans11-18:2 output by 205%. Output of trans11-18:1 was greater (P = 0.01) from orchardgrass compared with clover (174 vs. 90 mg/d), but corn increased (P = 0.01) trans11-18:1 output only from clover fermentations. Output of trans10-18:1 was greater (P = 0.01) in response to orchardgrass compared with clover (10 vs. 4 mg/d), but corn addition doubled the output regardless of forage type. Output of trans10,cis12-18:2, which did not differ due to forage type, increased (P = 0.01) twofold in response to corn. Cis9,cis11-18:2 was a primary conjugated isomer produced from forage fermentations, but its output decreased (P = 0.03) in response to corn input. When inputs of 18:2n-6 plus 18:3n-3 were less than 0.9% of total DM (clover), hydrogenation was low (87%). When 18:2n-6 plus 18:3n-3 inputs were from 1.2 to 1.5% of total DM (orchardgrass), hydrogenation averaged 96%. Despite greater hydrogenation, incremental additions of cis9-18:1 and 18:2n-6 from corn grain increased (P < 0.05) outputs of trans10-18:1, trans11-18:1, trans10,cis12-18:2, cis9,trans11-18:2, and trans,trans-18:2 in effluent. Results suggest that forage species alone or in combination with corn grain can alter hydrogenation and profiles of intermediates to varying degrees.

Influence of yeast culture on ruminal microbial metabolism in continuous culture.

A continuous culture study was conducted to evaluate the effect of two different yeast cultures on ruminal microbial metabolism. The treatments were a) control lactation ration, b) yeast culture 1 (YC1, Diamond-V XP) and c) yeast culture 2 (YC2, A-Max), both fed at an equivalent of 57 g/head per day. The results showed that both yeast culture products increased dry matter (DM) digestion, propionic acid production, and protein digestion compared with the control. Yeast culture 1 demonstrated an increase in molar percentage of propionic acid, a reduction in acetic acid, and a lower mean nadir (daily low) pH compared with YC2. Ruminal cultures treated with YC digested more protein and contributed less bypass N than control. Supplementing YC2 resulted in a tendency for higher microbial N/kg DM digestion than YC1. Yeast culture 1 resulted in production of rumen microbes containing less protein and more ash than YC2. These results support previous research findings that yeast culture does influence microbial metabolism, and specific yeast cultures may have different modes of action.

Degradation of two protein sources at three solids retention times in continuous culture.

Effects of solids retention times (SRT) of 10, 20, and 30 h on protein degradation and microbial metabolism were studied in continuous cultures of ruminal contents. Liquid dilution rate was constant across all retention times at .12 h(-1) (8.3 h mean retention time). Two semipurified diets that contained either soybean meal (SBM) or alfalfa hay (ALFH) as the sole nitrogen source were provided in amounts that decreased as SRT was increased. Digestion coefficients for DM, NDF, and ADF increased with increasing SRT. Digestion coefficients for nonstructural carbohydrates were higher in the SBM diet than in the ALFH diet but were not affected by SRT. Protein degradation in the ALFH diet averaged 51% and was unaffected by retention time. In the SBM diet, digestion of protein was 77, 78, and 96% at 10-, 20-, and 30-h retention times, respectively. Microbial efficiency decreased with increasing SRT and was greater for the SBM than for the ALFH diet. Efficiencies ranged from 30.6 to 35.7 and 20.8 to 29.2 g of N/kg of digested DM for the SBM and ALFH diets, respectively, as SRT decreased from 30 to 10 h. The diaminopimelic acid content of the microbes increased as SRT increased, indicating that changes in microbial species occurred owing to passage rates. From these results, we concluded that the digestibility decreases associated with increased ruminal turnover rates may be less for nonstructural carbohydrates and protein than for the fiber fractions.

Effects of concentrations of peptides on microbial metabolism in continuous culture.

Effects of peptide concentrations on microbial metabolism were investigated using a basal diet in which peptides replaced urea as a N source at levels of 0, 10, 20, and 30% of total N. The basal diet contained 17.8% CP and 46% nonstructural carbohydrates. Each diet was provided to continuous cultures of rumen contents operating at liquid and solids flow rates of 12 and 4.5%/h, respectively. Production of microbial CP and digestion of DM and protein were affected quadratically by peptide addition, with the highest values for each variable on the diet containing 10% peptides. Fiber digestion decreased linearly with peptide addition. The depressed fiber digestion and microbial CP production at peptide concentrations > 10% seemed related to a linear decrease in ammonia concentration in the fermenters as peptide levels increased. Peptide-amino acid N recovered in the supernatant fluid did not increase with increasing peptides, because peptide uptake by the microbes increased as peptide concentrations increased. Even though the efficiency of conversion of peptide N to microbial CP increased with increasing peptides, there was no change in grams of microbial N produced per kilogram of OMD. We suggest that in diets high in nonstructural carbohydrates, excessive peptides can depress protein digestion and ammonia concentrations, resulting in decreased OMD, fiber digestion, and total microbial CP production.

Effect of form of nitrogen on growth of ruminal microbes in continuous culture.

A study was conducted to determine the effect of various forms of N on the growth of ruminal microbes in a continuous culture system with solids and liquid dilution rates comparable to those of a high-producing dairy cow. Nitrogen forms were isolated soy protein, soy peptides, individual amino acids (AA) blended to profile soy protein, and urea, which were fed alone and in combinations so that the total N provided was 1.6% of the diet DM. The 100% soy protein treatment resulted in reduced digestion of N and nonstructural carbohydrate compared with other N forms, and outflow of bacterial N/24 h was less than when peptides were fed. This suggested that proteolysis rather than peptide uptake was the rate-limiting step in N utilization in this study. Non-urea N forms increased ADF digestion, total VFA production and the molar percentages of isobutyrate, isovalerate, and valerate compared to urea, which reflected the contribution of carbon skeletons of AA. When combinations of N forms were used, each form contributed an equal quantity of N, 50% of the total treatment, which was .8% of the diet DM. Combinations of N forms did not enhance, and in most cases reduced, ADF and NDF digestion when compared with individual N forms, and no combinations increased microbial growth over that of the individual forms. These results confirm that N forms other than ammonia are needed not only for maximum microbial growth, and they further demonstrate a need for non-protein N for the fiber digestion. In addition, results of this study suggest a requirement for a minimum level of peptide or AA N, which was met only when individual N forms were fed.

Interactions of fiber and nonstructural carbohydrates on lactation and ruminal function.

Four Holstein cows averaging 147 DIM and fitted with ruminal and duodenal cannulas were used in a 4 x 4 Latin square trial to determine diet effects on DMI, milk production, and ruminal metabolism. Diets contained either rapidly or slowly degraded NDF, referred to as low fill and high fill, respectively, combined with two percentages of nonstructural carbohydrate. Treatments were 39% nonstructural carbohydrate (low or high fill) and 29% nonstructural carbohydrate (low or high fill). Intake of DM was not affected by either fill or nonstructural carbohydrate. Ruminal NDF digestibilities averaged 43.1 and 35.6% for the low fill and high fill diets, respectively. Ruminally digested nonstructural and total carbohydrate increased, but milk production decreased, as nonstructural carbohydrate increased from 29 to 39% in diets. Liquid and solid ruminal passage rates, as measured by Co-EDTA and Yb, respectively, were reduced by either 39% nonstructural carbohydrate or low fill diets. Lower microbial N flow to the duodenum and lower efficiency of microbial growth also were observed for diets with 39% nonstructural carbohydrate. The combination of 39% nonstructural carbohydrate and rapidly degraded fiber gave the highest DM and nonstructural carbohydrate digestion in the rumen but resulted in low microbial N synthesis per day and the least microbial N per kilogram of OM digested.

Balancing carbohydrates and proteins for optimum rumen microbial yield.

Establishing conditions under which rumen fermentation will be optimized requires an understanding of the nutrient requirements of the mixed microbial population. The major nutrients required by rumen microbes are carbohydrates and proteins, but the most suitable sources and quantities needed to support maximum growth have not been determined. Digestion of proteins results in the production of peptides, which can accumulate in the rumen. Peptides are further hydrolyzed to amino acids, some of which are deaminated, producing ammonia. Although peptides, amino acids, and ammonia all may individually serve as sources of N for various microbes, the total population achieves the highest growth rate on mixtures of all three sources. In a somewhat analogous manner, carbohydrates are digested by exoenzymes to oligosaccharides that are available for crossfeeding by the mixed microbial population. Based on data from both in vitro and in vivo studies, there is general agreement that rate of digestion of carbohydrates is the major factor controlling the energy available for microbial growth; in addition, rate of digestion of total carbohydrate is directly related to proportion of starches, pectins, and sugars. Proteins affect both total fermentation and production of microbial DM per unit of carbohydrate fermented. It appears that the quantity of ruminally available protein needed to optimize microbial growth may, under some conditions, be as high as 14 to 15% of diet DM.

Rumen digestive physiology and microbial ecology.

The rumen is a dynamic, continuous fermentation compartment that provides a suitable environment for a variety of species of anaerobic bacteria, protozoa, and fungi. These microorganisms have a complex series of interactions with the feeds supplied to the host, with some using particulate matter as both sources of nutrients and sites of sequestration to avoid being washed from the rumen by the rapid flow of fluids. Because of the ability to use soluble nutrients and to reproduce rapidly, other microbes associate primarily with the liquid phase of the rumen contents. Due to the metabolic activity of all microbial populations, feeds are converted to microbial matter and fermentation end products, which serve as nutrients for the ruminant. Optimum feed utilization by ruminants is dependent on achieving maximum rumen fermentation and flow of microbial protein to the duodenum. At this time, it is clear that the major nutrients required by the microbial populations include both fibrous and nonfibrous sources of carbohydrates and nitrogen in the form of ammonia, amino acids, and peptides. In spite of five decades of research, the exact quantities and sources of these nutrients that will result in optimum rumen fermentation rates and microbial yields are only partially known.

Ruminal digestion and microbial utilization of diets varying in type of carbohydrate and protein.

Three ruminally and duodenally cannulated, lactating Holstein cows were used in a 3 x 3 Latin square experiment to study the effects of differing levels of nonstructural carbohydrate and degradable intake protein on ruminal digestibility and microbial protein production. Three diets were formulated to contain 1) 38 and 13.2%, 2) 31 and 11.8%, and 3) 24 and 9% nonstructural carbohydrate and degradable intake protein as percentages of the DM, respectively. Dry matter intakes were similar for all diets (21.9, 21.1, and 18.3 kg/d for diets 1, 2, and 3, respectively). Likewise, microbial efficiency, as estimated from purine analysis, was unaffected by diet and averaged 24 g of microbial N/kg of OM digested for all treatments. Ruminal digestion of OM averaged 66.6, 65.1, and 55.7% for diets 1, 2, and 3, respectively, resulting in lower microbial N flow per day for diet 3 (317, 333, and 202 g, respectively). Digestion of nonstructural carbohydrate and CP followed similar trends as did OM digestion, whereas NDF digestion remained similar across all diets. These results indicate that nonstructural carbohydrate greater than 24% and ruminally degradable protein greater than 9% of DM will enhance microbial protein flow from the rumen.

Impact of carbohydrate and protein levels on bacterial metabolism in continuous culture.

Diets formulated with three levels of nonstructural carbohydrate (54, 37, and 25% of DM), with various concentrations of degradable intake protein ranging from 19 to 4% of DM, were fermented in continuous cultures to ascertain the effects of ratio of nonstructural carbohydrate to degradable intake protein on bacterial metabolism. Fermenters were maintained at a dilution rate of 12%/h with a solids retention time of 24 h. Regardless of degradable intake protein level, bacterial efficiency (g of bacterial N/kg of DM digested) and VFA production (mM/d) were lower for diets with 25% nonstructural carbohydrate compared with the 37 and 54% nonstructural carbohydrate diets. In response to widening nonstructural carbohydrate:degradable intake protein ratios, bacterial efficiencies at all nonstructural carbohydrate levels declined quadratically from 34.2 to 10.3 with the lowest efficiencies on the 25% nonstructural carbohydrate diets. Bacterial protein production, DM digestion, NDF digestion, and VFA production (mM/d) increased linearly in response to dietary protein. The enhanced NDF and DM digestion, VFA production, and bacterial efficiencies observed with the narrower ratios of nonstructural carbohydrate:degradable intake protein support the theory that level of both degradable intake protein and nonstructural carbohydrate should be considered in order to enhance ruminal digestion and bacterial N production.

Effects of fish meal protein supplementation on milk yield and composition and blood constituents of dairy cows.

Ten holstein and 10 Ayrshire cows were fed diets containing undegradable intake protein from either fish meal or corn gluten meal. Cows were introduced to diets 10 d before projected calving date and individually fed blended rations until 60 d postpartum. Diets were balanced for NE1, CP, and degradable and undegradable protein. Source of undegradable protein did not affect total or FCM yields, DM intake, or milk protein percentage. Cows on fish meal diets tended to lose less BW than those on the corn gluten meal supplement (5.3 vs. 10.3% loss of initial BW). Fish meal supplementation resulted in decreased milk fat and SNF percentage (3.2 vs. 4.2% and 8.37 vs. 8.65%, respectively), but diet did not affect total milk fat, protein, or SNF yield. Plasma nonesterified fatty acids decreased and serum insulin increased with increasing weeks postpartum but were not affected by diet. In this study, no significant advantage was found to using fish meal as a source of undegradable intake protein and feed cost was higher when it was used.

Effects of fish meals on rumen bacterial fermentation in continuous culture.

Effects of various forms of fish meal on microbial metabolism were investigated in continuous cultures of rumen contents. Five diets were formulated to contain 12% ruminally degradable protein and 47 to 48% nonstructural carbohydrate. Soybean meal was the major protein source in the control diet, whereas in the other four diets, various fish meals were substituted for 6% of total diet DM. Fish meals were: fish meal containing 34.4% FFA, fish meal containing 34.4% FFA with CaCl2 added, fish meal containing 65.6% FFA, and fish meal defatted using 1:1 ethanol:ether extraction. The five treatments were fermented with pH either held constant at 6.2 or not controlled. When pH was maintained at 6.2, the inclusion of any fish meal except defatted fish meal reduced the acetate:propionate ratio, decreased protein digestion, and reduced microbial N produced/per kilogram DM digested when compared with the soybean control. When not controlled, pH decreased after feeding to 6.0 or lower. Under these conditions, the soybean control had a lower acetate:propionate ratio and lower NDF digestion than all diets containing fish meal. In this study, oil-containing fish meal affected microbial metabolism more negatively when the fermentation pH was held at 6.2 than when the pH was 6.0 or lower.

Chemical factors involved in ruminal fiber digestion.

In the United States, cattle are commonly fed diets containing cereal grains. The presence of starch and sugars reduces fiber digestion, which may in turn depress intake. In this paper, chemical constraints that may be responsible for the decrease in fiber digestion are explored. A major factor appears to be rumen pH. Moderate depression in pH, to approximately 6.0, results in a small decrease in fiber digestion, but numbers of fibrolytic organisms are usually not affected. Further decreases to 5.5 or 5.0 result in depressed growth rates and decreased fibrolytic microbes, and fiber digestion may be completely inhibited. Proliferation of organisms on readily fermentable carbohydrates may increase the need for total nitrogen as both ammonia and amino acids. The value of amino acids to cellulolytic organisms appears to be primarily as sources of isobutyric, isovaleric, and 2-methylbutyric acids. This reinforces the need to establish dietary requirements for nonprotein nitrogen, degradable protein, and isoacids. Other factors affecting fiber digestion, such as inhibition of cellulytic enzymes and plant concentrations of lignins and phenyl propanoids, are also discussed.

Glucogenic and hormonal responses to abomasal casein and ruminal volatile fatty acid infusions in lactating goats.

To determine responses to abomasal protein infusion and ruminal acetate: propionate ratios, four lactating Toggenburg goats fed hourly a 70% roughage and 30% concentrate diet were used in a Latin-square design with a factorial arrangement of treatments. Either acetate or propionate was infused ruminally and casein or saline infused abomasally. Estimated net energy and volume of the infusates were similar for all treatments. To examine the effects of treatments on glucose metabolism, 2-carbon-14 propionate was infused ruminally and 6-hydrogen-3 glucose was infused intravenously for 9 and 5 h, respectively. Although glucose concentration in plasma was higher and propionate turnover greater with propionate treatment, percentage of glucose derived from propionate, amount of propionate coverted to glucose, and glucose turnover remained unchanged. No differences in glucose metabolism due to the abomasal casein infusion were evident. To determine the effects of treatment on insulin, glucagon, growth hormone, and prolactin in plasma, samples were collected at 10-min intervals for 3 h at 0400 and 1600 h. No diurnal variation or consistent peaks were observed for any of the hormones nor were treatment effects on plasma concentrations of insulin, growth hormone, or prolactin evident. Glucagon concentration was higher with casein treatment; however, no relationship existed between glucagon in plasma and glucogenic parameters measured.

Survey of nutritional management practices and metabolic disorders in West Virginia dairy herds.

Sixteen farmers, representing three major dairy counties in West Virginia, were interviewed to determine current dairy management practices. Herd health records were used to determine the incidence of metabolic and digestive disorders. All feedstuffs used on these farms were sampled for chemical analyses. The total number of cows on the 16 farms surveyed was 1870 with an average herd size of 117 and 6387 kg milk produced per cow per year. Fourteen of the 16 farmers used forage testing services, obtained professional advice on feeding and nutrition, and attempted to feed according to production. Eleven of sixteen farmers increased grain 2 weeks prior to calving and all gradually adjusted the intake of grain to cows recently freshened. Ration imbalance problems included both excesses and deficiencies of energy, protein, phosphorus, and calcium. Excess calcium, phosphorous, and crude protein were correlated (P less than .01, r2 = .74) with the incidence of fat cows. The incidence of metabolic disorders in these herds was low to average and the most prevalent management problem encountered was low butter fat.

Effects of particle size and formaldehyde treatment of soybean meal on milk production and composition for dairy cows.

Twenty-four lactating dairy cattle were used to study the effects of formaldehyde treatment and reduction of particle size of soybean meal on milk production and composition. Cows were fed diets containing one of the following soybean meals: 1) untreated, coarse; 2) untreated, fine; 3) formaldehyde-treated, fine. Formaldehyde reduced the solubility of soybean meal protein from 22.7 to 2.9%. Grinding soybean meal reduced particle size from 842 to 249 micron. Formaldehyde treatment did not affect intake or milk production but resulted in higher efficiency of milk production (1.43 versus 1.46 kg fat-corrected milk/kg dry matter intake). Milk protein (3.08 versus 2.85%) and solids-not-fat (8.51 versus 8.35%) contents were reduced by formaldehyde. Cows fed fine soybean meal had higher dry matter intakes (22.0 versus 20.8 kg dry matter/day) and gained more body weight; however, milk production was not affected by particle size. Grinding soybean meal reduced production efficiency (1.47 versus 1.42 kg milk/kg dry matter intake) and milk fat (3.70 versus 3.33%). Lack of production response and reduced milk protein from formaldehyde treatment suggests possible overprotection of protein. Fine grinding of soybean meal appeared to favor body weight gain rather than milk production.

Effect of low and high fill diets on dry matter intake, milk production, and reproductive performance during early lactation.

Feedstuffs analyzed for rate and extent of ruminal neutral detergent fiber disappearance were used to formulate two diets that differed for predicted time required for ruminal clearance of neutral detergent fiber. Diets with slow and fast estimated disappearance rate of neutral detergent fiber were termed and high and low fill, respectively. For both diets, crude protein, acid detergent fiber, neutral detergent fiber, net energy of lactation, and soluble protein were similar. Twenty-eight lactating cows were limit fed the rations 2 wk prior to calving, then fed for ad libitum consumption until 8 wk after calving. Cows fed low fill produced more milk (30.3 vs. 26.3 kg) and milk protein (.97 vs. .78 kg) and had higher incidence of short estrous cycles and fewer acyclic cows by 8 wk postpartum than cows offered high fill. Dry matter intake, fat-corrected milk yield, daily fat production, and solids-not-fat did not differ between diets. Rumen fermentation measurements for pH and ammonia-nitrogen concentrations were also not different between diets. Cows fed low fill tended to have higher rates of solids and liquid turnover and lower total dry matter in the rumen compared with cows fed high fill. Low ruminal pH on both diets as well as other physiological mechanisms may have been responsible for failure of rates of neutral detergent fiber disappearance to affect intake of dry matter.

Effect of dietary concentration of total nonstructural carbohydrate on energy and nitrogen metabolism and milk production of dairy cows.

Two complete blended diets with a ratio of concentrate: silage dry matter of 60:40 were fed to 12 Holstein cows in the first 12 wk of lactation in an incomplete changeover arrangement of treatments. Diets differed (dry basis) in content of total nonstructural carbohydrate (24.9% versus 32.9%), neutral detergent fiber (37.0% versus 32.1%), and hemicellulose (19.6% versus 15.7%) but were similar in amounts of lignin, crude protein, soluble nitrogen, and acid detergent insoluble nitrogen. The diet with more total nonstructural carbohydrate was associated with greater dry matter intake as a percentage of body weight and greater yields of milk and solids-not-fat. Cellulose digestibility and mean rumen ammonia concentration were lower with this diet. Despite similar protein solubilities, the diet with more total nonstructural carbohydrate contained more rumen degradable nitrogen (80% versus 60%) but similar amounts of rumen degradable dry matter (82% versus 79%). The metabolizable energy of this diet was used more efficiently for the combined functions of maintenance and production, and net energy for lactation was larger (2.2 versus 1.9 Mcal/kg dry matter), as measured calorimetrically.