Interactions of alfalfa hay and sodium propionate on dairy calf performance and rumen development

The objective of this experiment was to investigate the effects of different levels of alfalfa hay (AH) and sodium propionate (Pro) added to starter diets of Holstein calves on growth performance, rumen fermentation characteristics, and rumen development. Forty-two male Holstein calves (40 ± 2 kg of birth weight) were used in a complete randomized design with a 3 × 2 factorial arrangement of treatments. Dietary treatments were as follows: (1) control = concentrate only; (2) Pro = concentrate with 5% sodium propionate [dry matter (DM) basis]; (3) 5% AH = concentrate + 5% alfalfa hay (DM basis); (4) 5% AH + Pro = concentrate + 5% alfalfa hay + 5% sodium propionate (DM basis); (5) 10% AH = concentrate + 10% alfalfa hay (DM basis); and (6) 10% AH + Pro = concentrate + 10% alfalfa hay + 5% sodium propionate (DM basis). All calves were housed in individual pens bedded with sawdust until 10 wk of age. They were given ad libitum access to water and starter throughout the experiment and were fed 2 L of milk twice daily. Dry matter intake was recorded daily and body weight weekly. Calves from the control, 10% AH, and 10% AH + Pro treatments were euthanized after wk 10, and rumen wall samples were collected. Feeding of forage was found to increase overall dry matter intake, average daily gain, and final weight; supplementing sodium propionate had no effect on these parameters. Calves consuming forage had lower feed efficiency than those on the Pro diet. Rumen fluid in calves consuming forage had higher pH and greater concentrations of total volatile fatty acids and molar acetate. Morphometric parameters of the rumen wall substantiated the effect of AH supplementation, as plaque formation decreased macroscopically. Overall, the interaction between forage and sodium propionate did not affect calf performance parameters measured at the end of the experiment. Furthermore, inclusion of AH in starter diets positively enhanced the growth performance of male Holstein calves and influenced both the macroscopic and microscopic appearances of the rumen wall. These benefits, however, were small when only sodium propionate was offered.     

Effects of 3-nitrooxypropanol on methane emission,digestion, and energy and nitrogen balance of lactating dairy cows

The objective was to measure effects of 3-nitrooxypropanol (3NP) on methane production of lactating dairy cows and any associated changes in digestion and energy and N metabolism. Six Holstein-Friesian dairy cows in mid-lactation were fed twice daily a total mixed ration with maize silage as the primary forage source. Cows received 1 of 3 treatments using an experimental design based on two 3 × 3 Latin squares with 5-wk periods. Treatments were a control placebo or 500 or 2,500 mg/d of 3NP delivered directly into the rumen, via the rumen fistula, in equal doses before each feeding. Measurements of methane production and energy and N balance were obtained during wk 5 of each period using respiration calorimeters and digestion trials. Measurements of rumen pH (48 h) and postprandial volatile fatty acid and ammonia concentrations were made at the end of wk 4. Daily methane production was reduced by 3NP, but the effects were not dose dependent (reductions of 6.6 and 9.8% for 500 and 2,500 mg/d, respectively). Dosing 3NP had a transitory inhibitory effect on methane production, which may have been due to the product leaving the rumen in liquid outflow or through absorption or metabolism. Changes in rumen concentrations of volatile fatty acids indicated that the pattern of rumen fermentation was affected by both doses of the product, with a decrease in acetate:propionate ratio observed, but that acetate production was inhibited by the higher dose. Dry matter, organic matter, acid detergent fiber, N, and energy digestibility were reduced at the higher dose of the product. The decrease in digestible energy supply was not completely countered by the decrease in methane excretion such that metabolizable energy supply, metabolizable energy concentration of the diet, and net energy balance (milk plus tissue energy) were reduced by the highest dose of 3NP. Similarly, the decrease in N digestibility at the higher dose of the product was associated with a decrease in body N balance that was not observed for the lower dose. Milk yield and milk fat concentration and fatty acid composition were not affected but milk protein concentration was greater for the higher dose of 3NP. Twice-daily rumen dosing of 3NP reduced methane production by lactating dairy cows, but the dose of 2,500 mg/d reduced rumen acetate concentration, diet digestibility, and energy supply. Further research is warranted to determine the optimal dose and delivery method of the product.     

Production of volatile fatty acids in the rumen and cecum-colon of steers as affected by forage:concentrate and forage physical form

Contribution of cecal and ruminal VFA to metabolizable energy was investigated in steers with cannulas in both the rumen and cecum. Animals were fed ad libitum so that data would be applicable to the lactating dairy cow. Diets assigned within a 4 x 4 Latin square were: 20% long alfalfa hay and 80% concentrate; 15% pelleted alfalfa, 5% hay and 80% concentrate; 80% hay and 20% concentrate; 60% pellets, 20% hay and 20% concentrate. Intake of DM was unaffected by diet. Cecal fluid pH, osmolality, and concentrations of valerate and isovalerate were unaffected by diet. Concentrations of total VFA, acetate, propionate, butyrate, and lactate in the cecum increased with proportion of grain in the diet. The high grain diets depressed cecal ammonia concentration and acetate to propionate ratio. Acetate production in the cecum was higher with the high grain diets whereas that in the rumen was lower. Production of propionate and butyrate in both the cecum and rumen was unaffected by diet. Cecal VFA provided 8.6% of metabolizable energy intake, on average. Contribution of ruminal VFA to total metabolizable energy was affected by diet, accounting for 72, 51, 74, and 52% of metabolizable energy from the 20% hay, 20% pelleted alfalfa, 80% hay, and 80% pelleted alfalfa, respectively. Cecal VFA were an important source of energy for ad libitum-fed steers; this contribution would undoubtedly increase with increasing feed intake.     

Effects of antibiotics and oil on microbial profiles and fermentation in mixed cultures of ruminal microorganisms

Ionophores and supplemental fat are fed to lactating cows to improve feed efficiency. Their effect on rumen fermentation is similar, but less is known about their impact on rumen microbes. The objective of this study was to determine the effects of monensin (M), bacitracin (B), and soybean oil (O) on microbial populations. Mixed cultures of rumen microbes were incubated in 5 dual-flow continuous fermentors and fed 13.8 g of alfalfa hay pellets daily (DM basis) for 16 d. All fermentors were allowed to stabilize for 4 d. From d 5 to 10, two fermentors received O (5% of diet DM), one fermentor received M (22 mg/kg), and one received B (22 mg/kg). From d 11 to 16, the 2 fermentors receiving O also received either M (OM) or B (OB) and O was included in the fermentors receiving M (MO) and B (BO). One fermentor served as the control and received 100% alfalfa pellets throughout the experiment. Each run was replicated 3 times. Samples were taken at 2 h after the morning feeding on d 4, 10, and 16 and were analyzed for bacterial populations using terminal restriction fragment length polymorphism. Volatile fatty acid concentration, methane production, and pH in the control cultures were not affected by time and remained similar during the entire experiment. The M and O treatments reduced molar concentration of acetate, increased concentration of propionate, and decreased methane production. Bacitracin did not alter acetate or propionate concentration, but reduced methane production. All 3 treatments (M, B, and O) altered the fragment patterns of microbial profiles. In contrast, treatments MO, OM, BO, and OB had little effect on culture fermentation despite differences in the patterns of microbial fragments. The terminal restriction fragment length polymorphism data suggest that microbial adaptation to the in vitro system in the control fermentor occurred within 4 d.     

Alfalfa pellet-induced subacute ruminal acidosis in dairy cows increases bacterial endotoxin in the rumen without causing inflammation

A study was conducted to determine if subacute ruminal acidosis (SARA) induced by feeding alfalfa pellets results in increases in free bacterial lipopolysaccharide (LPS) in rumen fluid and peripheral blood, and acute phase proteins in plasma, and to determine the effect of alfalfa pellet-induced SARA on feed intake, rumen fermentation characteristics, milk production and composition, and blood metabolites. Eight lactating Holstein cows, 4 of which were ruminally cannulated, were used in a 6-wk experiment and were fed once daily at 0900 h. During wk 1, cows received a diet containing 50% of DM as concentrate and 50% of DM chopped alfalfa hay. Between wk 2 and wk 6, alfalfa hay was gradually replaced with alfalfa pellets at the rate of 8% per week to reduce rumen pH. Rumen pH was monitored continuously in the ruminally cannulated cows using indwelling pH probes. Rumen fluid and peripheral blood were sampled 15 min before feed delivery and at 6 h after feed delivery. Based on adopted threshold of SARA of at least 180 min/d below pH 5.6, SARA was induced from wk 3 onwards. Replacing 40% of alfalfa hay with alfalfa pellets quadratically increased the DMI from 18.1 kg/d in wk 1 to 23.4 kg/d in wk 6. This replacement linearly decreased milk yield (32.7 vs. 35.9 kg/d) and milk fat percentage and yield (2.32 vs. 3.22%, and 0.77 vs. 1.14 kg/d, respectively), but increased milk protein percentage and yield (3.80 vs. 3.04%, and 1.23 vs. 1.07 kg/d, respectively). This gradual replacement also linearly increased the daily averages of total volatile fatty acids (90 to 121.9 mM), acetate (53.9 to 66.8 mM), propionate (21.5 to 39.6 mM), and osmolality (277.7 to 293.8 mmol/kg) in the rumen and decreased the acetate to propionate ratio from 2.62 to 1.73. Replacing alfalfa hay with alfalfa pellets linearly increased blood lactate from 1.00 mM in wk 1 to a peak of 3.46 mM in wk 5. Induction of SARA in this study increased free rumen LPS concentration from 42,122 endotoxin unit (EU)/mL in wk 1 to 145,593 EU/mL in wk 6. However, this increase was not accompanied by an increase in LPS (<0.05 EU/mL) and in acute phase proteins serum amyloid-A, haptoglobin, and LPS-binding protein in peripheral circulation. Results suggest that SARA induced by alfalfa pellets increased LPS in the rumen without causing translocation of LPS and an immune response.     

Short communication: Comparison of pH,volatile fatty acids,and microbiome of rumen samples from preweaned calves obtained via cannula or stomach tube

The objective of this study was to compare rumen samples from young dairy calves obtained via a stomach tube (ST) or a ruminal cannula (RC). Five male Holstein calves (46 ± 4.0 kg of body weight and 11 ± 4.9 d of age) were ruminally cannulated at 15 d of age. Calves received 4 L/d of a commercial milk replacer (25% crude protein and 19.2% fat) at 12.5% dry matter, and were provided concentrate and chopped oats hay ad libitum throughout the study (56 d). In total, 29 paired rumen samples were obtained weekly throughout the study in most of the calves by each extraction method. These samples were used to determine pH and volatile fatty acids (VFA) concentration, and to quantify Prevotella ruminicola and Streptococcus bovis by quantitative PCR. Furthermore, a denaturing gradient gel electrophoresis was performed on rumen samples harvested during wk 8 of the study to determine the degree of similarity between rumen bacteria communities. Rumen pH was 0.30 units greater in ST compared with RC samples. Furthermore, total VFA concentrations were greater in RC than in ST samples. However, when analyzing the proportion of each VFA by ANOVA, no differences were found between the sampling methods. The quantification of S. bovis and P. ruminicola was similar in both extraction methods, and values obtained using different methods were highly correlated (R² = 0.89 and 0.98 for S. bovis and P. ruminicola, respectively). Fingerprinting analysis showed similar bacteria band profiles between samples obtained from the same calves using different extraction methods. In conclusion, when comparing rumen parameters obtained using different sampling techniques, it is recommended that VFA profiles be used rather than total VFA concentrations, as total VFA concentrations are more affected by the method of collection. Furthermore, although comparisons of pH across studies should be avoided when samples are not obtained using the same sampling method, the comparison of fingerprinting of a bacteria community or a specific rumen bacterium is valid.     

Rates of production of acetate, propionate, and butyrate in the rumen of lactating dairy cows given normal and low-roughage diets

Five lactating dairy cows with a permanent cannula in the rumen were given (kg DM/d) a normal diet (7.8 concentrates, 5.1 hay) or a low-roughage (LR) diet (11.5 concentrates, 1.2 hay) in two meals daily in a two-period crossover design. Milk fat (g/kg) was severely reduced on diet LR. To measure rates of production of individual volatile fatty acids (VFA) in the rumen, 0.5 mCi 1-(14)C-acetic acid, 2-(14)C-propionic acid, or 1-(14)C-n-butyric acid were infused into the rumen for 22 h at intervals of 2 to 6 d; rumen samples were taken over the last 12 h. To measure rumen volume, we infused Cr-EDTA into the rumen continuously, and polyethylene glycol was injected 2 h before the morning feed. Results were very variable, so volumes measured by rumen emptying were used instead. Net production of propionic acid more than doubled on LR, but acetate and butyrate production was only numerically lower. Net production rates pooled across both diets were significantly related to concentrations for each VFA. Molar proportions of net production were only slightly higher than molar proportions of concentrations for acetate and propionate but were lower for butyrate. The net energy value (MJ/d) of production of the three VFA increased from 89.5 on normal to 109.1 on LR, equivalent to 55 and 64% of digestible energy, respectively. Fully interchanging, three-pool models of VFA C fluxes are presented. It is concluded that net production rates of VFA can be measured in non-steady states without the need to measure rumen volumes.     

Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems

Two experiments were conducted to determine the effects of several essential oil active compounds on rumen microbial fermentation. In the first experiment, 4 doses (5, 50, 500, and 5,000 mg/L) of 5 essential oil compounds were evaluated using in vitro 24-h batch culture of rumen fluid with a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were control (CON), eugenol (EUG), guaiacol, limonene, thymol (THY), and vanillin. After 24 h, the pH was determined, and samples were collected to analyze ammonia N and volatile fatty acids (VFA). The highest dose of all compounds decreased total VFA concentration and increased the final pH. Eugenol at 5 mg/L tended to reduce the proportion of acetate and the acetate to propionate ratio, at 50 and 500 mg/L tended to reduce ammonia N concentration, and at 500 mg/L reduced the proportion of propionate and branched-chain VFA concentration, without affecting total VFA concentration. All other treatments had minor effects or changes occurred only after total VFA concentration decreased. In the second experiment, 8 dual-flow continuous culture fermenters (1,320 mL) were used in 3 replicated periods (6 d of adaptation and 3 d of sampling) to study the effects of THY and EUG on rumen microbial fermentation. Fermenters were fed 95 g/d of DM of a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were CON, 10 mg/L of monensin (positive control), and 5, 50, or 500 mg/L of THY and EUG, and were randomly assigned to fermenters within periods. During the last 3 d of each period, samples were taken at 0, 2, 4, and 6 h after the morning feeding and analyzed for peptides, amino acids, and ammonia N concentrations, and total and individual VFA concentrations. Monensin changed the VFA profile as expected, but inhibited nutrient digestion. Eugenol and THY decreased total VFA concentration and changed the VFA profile, and only 5 mg/L of THY tended to reduce the proportion of acetate, increased the proportion of butyrate, and increased the large peptides N concentration without decreasing total VFA concentration. Most of these essential oil compounds demonstrated their antimicrobial activity by decreasing total VFA concentration at high doses. However, EUG in batch fermentation and 5 mg/L of THY in continuous culture modified the VFA profile without decreasing total VFA concentration, and EUG in batch fermentation decreased ammonia N concentration.     

In vitro evaluation of cashew nut shell liquid as a methane-inhibiting and propionate-enhancing agent for ruminants

Cashew nut shell liquid (CNSL) containing antibacterial phenolic compounds was evaluated for its potency as a feed additive for ruminants. In experiment 1, ruminal responses to CNSL supplementation were assessed using a batch culture system. Rumen fluid from cattle was diluted with artificial saliva and incubated for 18 h in a batch culture with a mixed diet containing a 30:70 hay:concentrate diet to which raw or heated CNSL was added at a final concentration of 500 μg/mL. In experiment 2, a Rusitec, using rumen fluid from the same cattle, was operated over a period of 7 d during which only raw CNSL was tested at concentrations of 0, 50, 100, or 200 μg/mL, and variations in fermentation and bacterial population were assessed. In experiment 3, a pure culture study was conducted using selected bacteria to determine their susceptibility to CNSL. In experiment 1, methane production was inhibited by raw CNSL (56.9% inhibition) but not by heated CNSL. Total volatile fatty acid concentration was not affected, whereas increased concentrations of propionate and decreased concentrations of acetate and butyrate were observed using either raw or heated CNSL. These changes were more obvious when raw CNSL was tested. In experiment 2, raw CNSL inhibited methanogenesis and increased propionate production in a dose-dependent manner, showing maximum methane inhibition (70.1%) and propionate enhancement (44.4%) at 200 μg/mL supplementation. Raw CNSL increased total volatile fatty acid concentration and dry matter digestibility. Raw CNSL also appeared to induce a dramatic shift in the population of rumen microbiota, based on decreased protozoa numbers and changes in quantitative PCR assay values for representative bacterial species. In experiment 3, using pure cultures, raw CNSL prevented the growth of hydrogen-, formate-, and butyrate-producing rumen bacteria, but not the growth of bacteria involved in propionate production. Based on these data, raw CNSL, rich in the antibacterial phenolic compound anacardic acid, is a potential candidate feed additive with selective activity against rumen microbes, leading to fermentation that results in decreased methane and enhanced propionate production.     

Effects of ruminal ammonia and butyrate concentrations on reticuloruminal epithelial blood flow and volatile fatty acid absorption kinetics under washed reticulorumen conditions in lactating dairy cows

The effect of reticuloruminal epithelial blood flow on the absorption of propionate as a volatile fatty acid (VFA) marker in 8 lactating Holstein cows was studied under washed rumen conditions. The cows were surgically prepared with ruminal cannulas and permanent catheters in an artery and mesenteric, right ruminal, and hepatic portal veins. The experiment was designed with 2 groups of cows: 4 cows adapted to high crude protein (CP) and 4 to low CP. All cows were subjected to 3 buffers: butyric, ammonia, and control in a randomized replicated 3 × 3 incomplete Latin square design. The buffers (30 kg) were maintained in a temporarily emptied and washed rumen for 40 min. The initial concentration of VFA was 84.2 mmol/L. Butyrate was increased from 4 to 36 mmol/L in butyric buffer by replacement of acetate, and ammonia (NH₃) was increased from 2.5 to 22.5 mmol/L in ammonia buffer by replacement of NaCl. Increasing amounts of deuterium oxide (D₂O) were added to the buffers as the order of buffer sequence increased (6, 12, and 18 g of D₂O). Ruminal clearance of D₂O was used to estimate epithelial blood flow. To increase accuracy of the epithelial blood flow estimates, data of ruminal liquid marker (Cr-EDTA), and initial and final buffer volumes were fitted to a dynamic simulation model. The model was used to estimate ruminal liquid passages, residual liquid, and water influx (saliva and epithelia water) for each combination of cow and buffer (n = 24). Epithelial blood flow increased 49 ± 11% for butyric buffer compared with control. The ruminal disappearance of propionate (marker VFA) was affected by buffer and followed the same pattern as for epithelial blood flow. The correlation between ruminal disappearance of propionate and epithelial blood flow (r = 0.56) indicates that the removal of propionate can be limited by epithelial blood flow. The ruminal disappearance of propionate increased 30 ± 12% for the butyric compared with ammonia buffer and 12.5 ± 8% when compared with control. The net portal flux of propionate increased 32 ± 6% in butyric compared with control. In conclusion, rumen epithelial blood flow is positively correlated with ruminal disappearance of propionate and affects the kinetics of ruminal VFA absorption.     

Sodium bicarbonate and alfalfa hay additions to wheat silage diets fed to lactating dairy cows

Two experiments were conducted to examine dietary effects of .8% sodium bicarbonate and 1.4 kg/d of alfalfa hay on performance and rumen metabolism of lactating dairy cows fed 50% wheat silage and 50% concentrate (dry basis). In Experiment 1 with 12 midlactation Holsteins in a 4 X 4 Latin square design, intake, milk production, and milk composition were not affected by treatment. Dietary sodium bicarbonate and alfalfa hay did not alter blood, rumen, or fecal pH. Rumen volatile fatty acid pattern was not affected by sodium bicarbonate, but addition of hay resulted in higher molar percentage propionate and lower acetate: propionate ratios. In Experiment 2 with 32 early lactation cows (20 Holsteins and 12 Jerseys) in a complete randomized block design, supplementation of sodium bicarbonate, alfalfa hay, or both did not affect intake, milk production, or milk composition in the first 8 wk of lactation. Blood, rumen, and fecal pH were not affected by treatment. Dietary sodium bicarbonate did not alter ruminal volatile fatty acid profile, whereas addition of hay increased molar proportion acetate and decreased molar proportion butyrate. A shift in rumen fermentation was observed across treatments from wk 1 through 8 postpartum with molar proportions of acetate and butyrate increasing and molar proportion of propionate decreasing.     

Corn Silage Partially Replacing Short Alfalfa Hay to Optimize Forage Use in Total Mixed Rations for Lactating Cows

We theorized that adding corn silage to a total mixed ration with alfalfa hay as the sole dietary forage would improve nutrient intake and chewing activity and thereby improve rumen fermentation and milk production. The objective of this research was to determine the effects of partial replacement of short alfalfa [physically effective (pe) neutral detergent fiber (NDF) >1.18 mm (peNDF_>1.18) = 33.2%] with corn silage (CS, peNDF_>1.18 = 51.9%) in yellow grease-supplemented total mixed rations on feed intake, chewing behavior, rumen fermentation, and lactation performance by dairy cows. Four multiparous (138 ± 3 d in milk) and 4 primiparous (115 ± 10 d in milk) Holstein cows were used in a 4 × 4 Latin square design experiment with four 21-d periods. Each period had 14 d of adaptation and 7 d of sampling, and parity was the square. Treatments were diets [dry matter (DM) basis] with 1) 40% alfalfa hay (ALF), 2) 24% alfalfa hay + 16% CS (CS40), 3) 20% alfalfa hay + 20% CS (CS50), and 4) 16% alfalfa hay + 24% CS (CS60). Diets had a forage-to-concentrate ratio of 40:60 on a DM basis. Cows had greater intake of DM and thus greater intakes of net energy for lactation, NDF, and peNDF when CS partially replaced alfalfa hay. Replacing alfalfa hay with CS increased daily eating and chewing times in all cows, and increased rumen pH at 4 h postfeeding in multiparous cows. Apparent total-tract digestibility coefficients for crude protein (CP) and NDF were not different among cows fed ALF, CS40, and CS50, but were lower for CS60 than for ALF. Energy-corrected milk yield was greater for CS40 and CS60 than for ALF. Milk protein yield was increased when CS replaced 40, 50, and 60% of alfalfa hay. Milk lactose was greater only for CS60, but milk lactose yield was greater for CS50 and CS60 than for ALF. Milk percentage and yield of fat did not differ among treatments. Therefore, CS partially replacing short alfalfa hay increased DM intake, consequently increased net energy for lactation and physically effective fiber intakes, and thus, improved milk and milk protein and lactose yields.     

Effects of fumarate on ruminal ammonia accumulation and fiber digestion in vitro and nutrient utilization in dairy does

The objective of this study was to evaluate effects of fumarate on ruminal ammonia accumulation and fiber digestion in vitro and on feed intake and nutrient utilization in dairy does. Batch cultures of mixed rumen microorganisms were used to study effects of different concentrations of fumarate on fermentation with various N sources (ammonia as ammonium bicarbonate, casein amino acids, casein peptides, gelatin peptides) and feeds (bermudagrass hay, mixed diet of 60% bermudagrass hay plus 40% concentrate) for 6 and 24 h, respectively. Substrates were grouped into pairs for separate incubations. Monosodium fumarate was added to incubation tubes to achieve final concentrations of 0, 5, and 10 mM fumarate. More ammonia accumulated at the end of incubation with added ammonium bicarbonate. Ammonia concentration was higher for peptide compared with amino acid incubation, and for casein peptide compared with gelatin peptide. Addition of fumarate linearly decreased ammonia for all N sources and for feed substrates. For all substrate types, fumarate treatment increased acetate, propionate, and total volatile fatty acids (VFA), decreased acetate to propionate ratio, and tended to reduce branched-chain VFA. Digestion of feed neutral detergent fiber (NDF) by rumen microorganisms was improved by fumarate along with elevated endoglucanase and xylanase activities. In an animal metabolism experiment, 8 dairy does (4 per treatment) were used in a completely randomized design for 21 d. Does were fed a hay plus concentrate diet without (control) or with fumarate (6 g/head per day) supplementation to determine feed intake, whole-tract nutrient digestibility, and N utilization. Fumarate treatment did not affect weight change or feed intake but increased whole-tract digestion of gross energy, crude protein, and cellulose. Digested N was increased by fumarate supplementation; however, N retention was unaffected. Plasma glucose concentration was elevated with fumarate but urea N concentration remained unchanged. Fumarate addition had significant effects on rumen microbial fermentation by decreasing ammonia and branched-chain VFA, and by increasing acetate and propionate, and NDF digestion. These effects were reflected in the improvement in whole-tract gross energy, crude protein, and cellulose digestion and elevated plasma glucose concentration when dairy does were supplemented with fumarate.     

Effect of Shredding Alfalfa on Cellulolytic Activity,Digestibility, Rate of Passage,and Milk Production

A series of trials was performed with sheep and goats to compare mechanically shredded with conventionally (control) harvested alfalfa hay. Shredded hay tended to contain less CP and more cell wall constituents than the control; digestibility of NDF in vitro also was increased by shredding. Intake of DM and digestibilities of NDF and ADF were increased in sheep fed shredded hay. Digestibilities of DM and CP were similar between treatments. Total tract digestibilities determined using acid detergent lignin and Yb as internal and external markers were similar to those obtained by total collection. There was no effect of shredding on rate of passage and total mean retention time of hay in the digestive tract of sheep. Ruminal pH and VFA were not altered by shredding of hay. However, production of 4% FCM and milk protein content was increased in goats fed shredded hay. Shredded hay suspended in the rumen in situ had higher particle-associated carboxymethylcellulase activity throughout the incubation period than did control hay. The enzyme activity was maximal at 6 h and declined thereafter for both hays. The pattern of particle-associated carboxymethylcellulase activity was similar to that of NDF digestion in vitro.     

Rumen volatile fatty acid molar proportions,rumen epithelial gene expression,and blood metabolite concentration responses to ruminally degradable starch and fiber supplies

The objective of this work was to characterize rumen volatile fatty acid (VFA) concentrations, rumen epithelial gene expression, and blood metabolite responses to diets with different starch and fiber sources. Six ruminally cannulated yearling Holstein heifers (body weight = 330 ± 11.3 kg) were arranged in a partially replicated Latin square experiment with 4 treatments consisting of different starch [barley (BAR) or corn (CRN)] and fiber [timothy hay (TH) or beet pulp (BP)] sources. Treatments were arranged as a 2 × 2 factorial. Beet pulp and TH were used to create relative changes in apparent ruminal fiber disappearance, whereas CRN and BAR were used to create relative changes in apparent ruminal starch disappearance. Each period consisted of 3 d of diet adaptation and 15 d of dietary treatment. In situ disappearance of fiber and starch were estimated from bags incubated in the rumen from d 10 to 14. From d 15 to 17, rumen fluid was collected every hour from 0500 to 2300 h. Rumen fluid samples were pooled by animal/period and analyzed for pH and VFA concentrations. On d 18, 60 to 80 papillae were biopsied from the epithelium and preserved for gene expression analysis. On d 18, one blood sample per heifer was collected from the coccygeal vessel. In situ ruminal starch disappearance rate (7.30 to 8.72%/h for BAR vs. 7.61 to 10.5%/h for CRN) and the extent of fiber disappearance (22.2 to 33.4% of DM for TH vs. 34.4 to 38.7% of DM for BP) were affected by starch and fiber source, respectively. Analysis of VFA molar proportions showed a shift from propionate to acetate, and valerate to isovalerate on TH diets compared with BP. Corn diets favored propionate over butyrate in comparison to BAR diets. Corn diets also had higher molar proportions of valerate. Expression of 1 gene (SLC9A3) were increased in BP diets and 2 genes (BDH1 and SLC16A4) tended to be increased in TH diets. Plasma acetate demonstrated a tendency for a starch by fiber interaction with BAR-BP diets having the highest plasma acetate, but other metabolites measured were not significant. These results suggest that TH has the greatest effect on shifts in VFA molar proportions and epithelial transporters, but does not demonstrate shifts in blood metabolite concentrations.     

Alfalfa containing the glyphosate-tolerant trait has no effect on feed intake, milk composition, or milk production of dairy cattle

The objective of this experiment was to assess if feeding glyphosate-tolerant alfalfa affects feed intake, milk composition, or milk production of dairy cows. One alfalfa (Medicago sativa), variety expressing the CP4 EPSPS protein and grown in southeastern Washington State was harvested at the late vegetative stage as hay. Three commercial conventional varieties of alfalfa hay of similar nutrient composition and harvested in the same geographic region were fed to cows as controls. The commercial hays were selected to be similar in crude protein [18% of dry matter (DM)] and neutral detergent fiber (40% of DM) to the glyphosate-tolerant hay. Sixteen multiparous Holstein cows were fed diets containing alfalfa hay (39.7% of diet DM) from either the glyphosate-tolerant alfalfa, or 1 of the 3 conventional varieties. Diets contained at least 15.7% crude protein and 29% neutral detergent fiber. Experimental design was a replicated 4 × 4 Latin square. Periods were 28 d and feed intake, milk yield, and milk composition were summarized over the last 14 d of each period. Daily milk yield (38.0 kg) and 4% fat-corrected milk (34.7 kg) were not affected by treatment. Milk fat (3.44%) and milk true protein (2.98%) were also not affected by source of hay. Milk lactose (4.72%) and soldis-not-fat (8.5%) did not differ due to treatment. Dry matter intake was similar across treatments (24.4 kg/d). These results are consistent with data from feeding trials with other glyphosate-tolerant crops and previously reported compositional comparisons of glyphosate-tolerant alfalfa with controls. Milk production, milk composition, feed intake, and feed efficiency were not affected by feeding diets that contained nearly 40% glyphosate-tolerant alfalfa hay to lactating dairy cows.     

Effects of acetate,propionate, and pH on volatile fatty acid thermodynamics in continuous cultures of ruminal contents

To investigate the effects of acetate, propionate, and pH on thermodynamics of volatile fatty acids (VFA) in the rumen, a dual-flow continuous culture study was conducted to quantify production of major VFA, interconversions among the VFA, and H₂ and CH₄ emissions in a 4 × 4 Latin square design. The 4 treatments were (1) control: pH buffered to an average of 6.75; (2) control plus 20 mmol/d of infused acetate (InfAc); (3) control plus 7 mmol/d of infused propionate (InfPr); and (4) a 0.5-unit decline in pH elicited by adjustment of the buffer (LowpH). All fermentors were fed 40 g of a pelleted diet containing whole alfalfa pellets and concentrate mix pellets (50:50) once daily. After 7 d of treatment, sequential, continuous infusions of [2-¹³C] sodium acetate (3.5 mmol/d), [U-¹³C] sodium propionate (2.9 mmol/d), and [1-¹³C] sodium butyrate (0.22 mmol/d) were carried out from 12 h before feeding for 36 h. Filtered liquid effluent (4 mL) was sampled at 0, 2, 4, 6, 8, 12, 16, and 22 h after feeding, and assessed for VFA concentrations, with another filtered sample (20 mL) used to quantify aqueous concentrations of CH₄ and H₂. Headspace CH₄ and H₂ gases were monitored continuously. Ruminal microbes were isolated from the mixed effluent samples, and the microbial community structure was analyzed using the 16S rRNA amplicon sequencing technique. The digestibility of neutral detergent fiber, acid detergent fiber, and starch and microbial C sequestrated from VFA were not affected by treatments. The LowpH treatment increased net propionate production and decreased H₂ and CH₄ headspace emissions, primarily due to shifts in metabolic pathways of VFA formation, likely due to the observed changes in bacterial community structure. Significant interconversions occurred between acetate and butyrate, whereas interconversions of other VFA with propionate were relatively small. The InfAc and InfPr treatments increased net acetate and propionate production, respectively; however, interconversions among VFA were not affected by pH, acetate, or propionate treatments, suggesting that thermodynamics might not be a primary influencer of metabolic pathways used for VFA formation.     

The effects of feeding 3-nitrooxypropanol on methane emissions and productivity of Holstein cows in mid lactation

The objective of the current study was to determine the effects of adding 3-nitrooxypropanol to the diet of lactating Holstein cows on methane emissions, rumen fermentation, ruminal microbial profile, and milk production. Twelve ruminally cannulated Holstein cows in midlactation were used in a crossover design study with 28-d periods. Cows were fed a diet containing 38% forage on a dry matter basis with either 2,500 mg/d of 3-nitrooxypropanol (fed as 25 g of 10% 3-nitrooxypropanol on silicon dioxide) or 25 g/d of silicon dioxide (control). After a 21-d diet adaptation period, dry matter intake (DMI) and milk yield were recorded daily. Rumen fluid and digesta were collected on d 22 and 28 for volatile fatty acid analysis and microbial profiling. Enteric methane emissions were measured on d 23 to 27 using the sulfur hexafluoride tracer gas technique. Feeding 3-nitrooxypropanol did not affect DMI; however, methane production was reduced from 17.8 to 7.18 g/kg of DMI. No change in milk or milk component yields was observed, but cows fed 3-nitrooxypropanol gained more body weight than control cows (1.06 vs. 0.39 kg/d). Concentrations of total volatile fatty acids in ruminal fluid were not affected by treatment, but a reduction in acetate proportion and a tendency for an increase in propionate proportion was noted. As such, a reduction in the acetate-to-propionate ratio was observed (2.02 vs. 2.36). Protozoa counts were not affected by treatment; however, a reduction in methanogen copy count number was observed when 3-nitrooxypropanol was fed (0.95 vs. 2.69 × 10⁸/g of rumen digesta). The data showed that feeding 3-nitrooxypropanol to lactating dairy cows at 2,500 mg/d can reduce methane emissions without compromising DMI or milk production.     

Effect of supplemental tallow on performance of dairy cows fed diets with different corn silage:alfalfa silage ratios

A study was conducted to investigate the response to supplemental tallow of lactating cows fed basal diets with different alfalfa silage:corn silage ratios. We postulated that supplemental tallow will have decreasing negative effects on rumen fermentation, dry matter intake (DMI), and milk fat percentage as the dietary ratio of alfalfa silage:corn silage is increased. Eighteen Holstein cows averaging 134 +/- 14 d in milk were used in a replicated 6 x 6 Latin square design with 21-d periods. Treatments were arranged as a 2 x 3 factorial with 0 or 2% tallow (DM basis) and three forage treatments: 1) 50% of diet DM as corn silage, 2) 37.5% corn silage and 12.5% alfalfa silage, and 3) 25% corn silage and 25% alfalfa silage. Cows were allowed ad libitum consumption of a total mixed ration. Diets were formulated to contain 18% crude protein and 32% neutral detergent fiber. No fat x forage treatment interactions were observed. Fat supplemented cows had lower DMI and produced more milk with less milk fat content relative to non-supplemented cows. Concentration of trans-octadecenoic acids was higher in milk fat of tallow-supplemented cows. Tallow supplementation had no effect on ruminal pH and acetate:propionate ratio, but tended to decrease total volatile fatty acid (VFA) concentration in the rumen. Increasing the proportion of alfalfa silage increased DMI, milk fat percentage, and milk fat yield regardless of the fat content of the diet. Total VFA concentration and acetate:propionate ratio in the rumen were increased in response to higher levels of alfalfa in the diets. These results suggest that replacing corn silage with alfalfa silage did not alleviate the negative response of dairy cows to tallow supplementation at 2% of diet DM.