Amino acid composition of rumen bacteria and protozoa in cattle

Because microbial crude protein (MCP) constitutes more than 50% of the protein digested in cattle, its AA composition is needed to adequately estimate AA supply. Our objective was to update the AA contributions of the rumen microbial AA flowing to the duodenum using only studies from cattle, differentiating between fluid-associated bacteria (FAB), particle-associated bacteria (PAB), and protozoa, based on published literature (53, 16, and 18 treatment means were used for each type of microorganism, respectively). In addition, Cys and Met reported concentrations were retained only when an adequate protection of the sulfur groups was performed before the acid hydrolysis. The total AA (or true protein) fraction represented 82.4% of CP in bacteria. For 10 AA, including 4 essential AA, the AA composition differed between protozoa and bacteria. The most noticeable differences were a 45% lower Lys concentration and 40% higher Ala concentration in bacteria than in protozoa. Differences between FAB and PAB were less pronounced than differences between bacteria and protozoa. Assuming 33% FAB, 50% PAB, and 17% of protozoa in MCP duodenal flow, the updated concentrations of AA would decrease supply estimates of Met, Thr, and Val originating from MCP and increase those of Lys and Phe by 5 to 10% compared with those calculated using the FAB composition reported previously. Therefore, inclusion of the contribution of PAB and protozoa to the duodenal MCP flow is needed to adequately estimate AA supply from microbial origin when a factorial method is used to estimate duodenal AA flow. Furthermore, acknowledging the fact that hydrolysis of 1 kg of true microbial protein yields 1.16 kg of free AA substantially increases the estimates of AA supply from MCP.     

Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture

Eight continuous culture fermentors inoculated with ruminal liquor from heifers fed a 50:50 alfalfa hay:concentrate diet (17.6% crude protein, 28.0% neutral detergent fiber) were used in 3 replicated periods to study the effects of cinnamaldehyde (CIN) and garlic oil (GAR) on rumen microbial fermentation. Treatments were no additive (negative control); 1.25 mg/L (MON) and 12.5 mg/L (MON10) of the ionophore antibiotic monensin (positive control); 31.2 mg/L CIN (CIN) and 312 mg/L (CIN10) of CIN; and 31.2 mg/L GAR (GAR) and 312 mg/L (GAR10) of GAR (Allium sativa). The MON10 caused expected changes in microbial fermentation patterns (a decrease in fiber digestion, ammonia N concentration, and proportions of acetate and butyrate; an increase in the proportion of propionate; and a trend to increase small peptide plus AA N concentration). The CIN decreased the proportion of acetate and branch-chained volatile fatty acids (VFA) and increased the proportion of propionate; CIN10 decreased the proportion of acetate and increased the proportion of butyrate compared with the control. The GAR10 increased the proportion of propionate and butyrate and decreased the proportion of acetate and branch-chained VFA compared with the control. The GAR10 also increased the small peptide plus amino acid N concentration, although no effects were observed on large peptides or ammonia N concentrations. The CIN and GAR10 resulted in similar effects as monensin, with the exception of the effects on the molar proportion of butyrate, which suggests that they might have a different mode of action in affecting in vitro microbial fermentation.     

Essential oil and monensin affect ruminal fermentation and the protozoal population in continuous culture

The interaction of monensin and essential oil was hypothesized to suppress protozoa and methane production while maintaining normal rumen function. The objective of this study was to determine the effects of feeding monensin (MON) and CinnaGar (CIN, a commercial blend of cinnamaldehyde and garlic oil; Provimi North America, Brookville, OH) on ruminal fermentation characteristics. Continuous culture fermentors (n = 4) were maintained in 4 experimental periods in a 4 × 4 Latin square design. Four dietary treatments were arranged in a 2 × 2 factorial: (1) control diet, 37 g/d of dry matter (40 g/d at ～92.5% dry matter) of a 50:50 forage:concentrate diet containing no additive; (2) MON at 11 g/909 kg of dry matter; (3) CIN at 0.0043% of dry matter; and (4) a combination of MON and CIN at the levels in (2) and (3). Treatment had no effects on protozoal populations, concentration of NH₃N, total N flow of effluent, production of total volatile fatty acids, or flows of conjugated linoleic acid and total C18 fatty acids. The MON decreased acetate:propionate ratio and biohydrogenation of both total C18 and 18:1 cis-9 but increased protozoal generation time, concentration of peptide, and flow of 18:1 trans-11. The MON tended to decrease protozoal counts in effluent and flow of 18:0 but tended to increase propionate production. The CIN decreased true organic matter digestibility and protozoal N flow of effluent but increased nonammonia, nonmicrobial N flow. The CIN tended to decrease protozoal counts, microbial N flow, and neutral detergent fiber digestibility but tended to increase biohydrogenation of total C18, 18:2, and 18:3. The CIN tended to increase isovalerate production. The MON and CIN tended to interact for increased methane production and bacterial N flow. A second experiment was conducted to determine the effects of MON and CIN on protozoal nitrogen and cell volume in vitro. Four treatments included (1) control (feed only), (2) feed + 0.0043% dry matter CIN, (3) feed + 2.82 μM MON, and (4) feed + CIN + MON at the same levels as in (2) and (3). With no interactions, MON addition decreased percentage of protozoa that were motile and tended to decrease cell volume at 6 h. The CIN did not affect cell count or other indicators of motility or volume at either 3 or 6 h. Under the conditions of our study, we did not detect an additive response for MON and CIN to decrease protozoal counts or methane production. A 3-dimensional method is suggested to better estimate protozoal cell volume.     

Effects of Cordyceps militaris on the growth of rumen microorganisms and in vitro rumen fermentation with respect to methane emissions

This experiment was designed to investigate the effects of different concentrations (0.00, 0.10, 0.15, 0.20, 0.25, and 0.30 g/L) of dried Cordyceps militaris mushroom on in vitro anaerobic ruminal microbe fermentation and methane production using soluble starch as a substrate. Ruminal fluids were collected from Korean native cattle, mixed with phosphate buffer (1:2), and incubated anaerobically at 38°C for 3, 6, 9, 12, 24, 36, 48, and 72 h. The addition of C. militaris significantly increased total volatile fatty acid and total gas production. The molar proportion of acetate was decreased and that of propionate was increased, with a corresponding decrease in the acetate:propionate ratio. As the concentration of C. militaris increased from 0.10 to 0.30 g/L, methane and hydrogen production decreased. The decrease in methane accumulation relative to the control was 14.1, 22.0, 24.9, 39.7, and 40.9% for the 0.10, 0.15, 0.20, 0.25, and 0.30 g/L treatments, respectively. Ammonia-N concentration and numbers of live protozoa decreased linearly with increasing concentrations of C. militaris. The pH of the medium significantly decreased at the highest level of C. militaris compared with the control. In conclusion, C. militaris stimulated mixed ruminal microorganism fermentation and inhibited methane production in vitro. Therefore, C. militaris could be developed as a novel compound for antimethanogenesis.     

Effects of time at suboptimal pH on rumen fermentation in a dual-flow continuous culture system

Ruminal pH varies considerably during the day, achieving values below 6.0 when cows consume large amounts of concentrates. Low ruminal pH has negative effects on ruminal fermentation. However, previous studies have indicated that rumen bacteria may resist short periods of low ruminal pH, and it is not clear how long this period may be before rumen microbial fermentation is negatively affected. Seven dual-flow continuous culture fermenters (1,320 mL) were used in 3 replicated periods with the same diet (97 g of dry matter/d of a 60:40 forage-to-concentrate diet, 18.3% crude protein, 35.9% neutral detergent fiber), temperature (39°C), and solid (5%/h) and liquid (10%/h) dilution rates to study the effects of increasing time at suboptimal pH on rumen microbial fermentation and nutrient flow. Treatments were a constant pH of 6.4 and 6 different intervals of time during the day (4, 8, 12, 16, 20, 24 h) at suboptimal pH (5.5), with the rest of the day being at pH 6.4. Polynomial equations were derived using the Mixed procedure of SAS, and linear, quadratic and cubic terms were left in the equation if P < 0.10. True organic matter digestion decreased with increasing time at suboptimal pH and was best described by a cubic regression (TOMD = 58.5 − 2.15x + 0.16x² −0.0037x³; R² = 0.74). Digestion of NDF (DNDF = 55.1 − 1.00x; R² = 0.75) and digestion of ADF (DADF = 56.2 − 1.33x; R² = 0.78) decreased linearly with increasing time at suboptimal pH. Total VFA had a cubic response (VFA = 112.7 − 2.09x + 0.17x² − 0.0054x³; R² = 0.82). The proportion of acetate decreased linearly (acetate = 58.7 − 0.61x; R² = 0.79). The propionate proportion increased (propionate = 17.6 + 2.09 × −0.044x²; R² = 0.85) and branched-chain VFA decreased (BCVFA = 4.45 −0.51x + 0.014x²; R² = 0.75) quadratically. The ammonia N concentration (NH₃-N = 5.85 − 0.13x; R² = 0.46) and flow (NH₃-N flow = 0.18 − 0.0039x; R² = 0.43) decreased linearly as the time at suboptimal pH increased. Crude protein degradation (CPd = 41.9 − 1.60x + 0.060x²; R² = 0.71), efficiency of microbial protein synthesis (EMPS = 26.6 − 0.33x + 0.021x²; R² = 0.77), microbial N flow (MN flow = 1.38 − 0.036x + 0.0015x²; R² = 0.77), and dietary N flow (DN flow = 1.49 + 0.041x − 0.0015x²; R² = 0.65) had a quadratic response. The flow of essential, nonessential, and most individual AA increased linearly with increasing time at suboptimal pH. The effects of pH on rumen fermentation appear to start as soon as pH drops to suboptimal pH.     

Effects of BioChlor and Fermenten on microbial protein synthesis in continuous culture fermenters

Meta analysis models were constructed from a data-set of 15 continuous culture fermenter trials and 118 observations on studies with either BioChlor (n = 23 observations) or Fermenten (n = 95) included at 10 and 3%, respectively, of dietary dry matter (DM) to evaluate effects of the ingredients BioChlor and Fermenten (B/F) on rumen function. Digestibility of crude protein was significantly increased by 11% with B/F treatment. This was reflected in significant increases in digestibility of DM and organic matter (OM) by 3.6 and 7.9%, respectively. Increased amounts of sugar in the diet in the presence of B/F tended to reduce digestibility of non-structural carbohydrates (NSC); however, the net effect on NSC digestion was small. There was no effect of treatment on most individual volatile fatty acids (VFA) or total VFA production. Propionate production, however, was significantly reduced in treated fermenters. The main effect of B/F as well as of starch and soluble fiber when combined with the treatment was to increase propionate production; however, the interaction between B/F treatment and sugar decreased propionate production markedly, resulting in a net decrease. The acetate-to-propionate ratio increased by 6% with B/F, largely as a result of the decrease in propionate. Production of nonammonia nitrogen was 1% less in B/F-treated fermenters, and interactions between treatment and starch, sugar, or soluble fiber were significant. Treated fermenters produced 15.7% more microbial nitrogen, in association with a significant 37% increase in rumen protein digestion. Interactions between treatment and starch, soluble fiber, or sugar influenced these results. The interaction of B/F and sugar resulted in a decrease in undegradable protein N and an increase in microbial nitrogen production. Ammonia nitrogen concentrations were increased by 24.6% in treated fermenters. Efficiency of microbial nitrogen production from DM, OM, or carbohydrate was significantly increased by B/F. Sugar content increased efficiency of microbial protein production/kg of OM digested or carbohydrate digested in the presence of treatment by >10 times the increase that was attributable to the interaction of treatment with starch. Treatment with B/F reduced moles of VFA produced/kg of microbial nitrogen produced by 16%. This effect was also substantially influenced by interactions between B/F and sugar. If the fermenter results are representative of those in vivo, milk production responses to treatment with B/F will depend on amounts of starch, soluble fiber, and, particularly, sugar in diets. Milk production responses will also depend on the quality of protein in the diet and the comparative benefit that increased flux of microbial nitrogen provides. Increased digestibility of OM should allow additional ruminant production benefits.     

Investigation of ammonium lactate supplementation on fermentation end products and bacterial assimilation of nitrogen in dual-flow continuous culture

Supplements investigated throughout the present study are produced by fermenting lactose that is present in whey to lactate, yielding products differing in ammonium relative to lactate concentrations and in physical form (liquid or dry). Trials 1 and 2 investigated Lacto-Whey (LW; Fermented Nutrition Corp., Luxemburg, WI) and GlucoBoost (GB; Fermented Nutrition Corp.), respectively, using dual-flow continuous culture systems (n = 4), each with a 4 × 4 Latin square design. A greater proportion of nonprotein nitrogen was present in GB than in LW. In trial 1, the treatment with LW was isonitrogenously dosed against soybean meal (SBM) as a control (no LW) and factorialized with either a wheat- or corn-based concentrate (55% inclusion rate, dry matter basis). We hypothesized that LW would increase propionate production and that the combination of +LW with wheat would increase bacterial assimilation of NH₃-N into cellular N. No differences were observed for total volatile fatty acid (VFA) production per day. However, treatment × time interactions revealed that +LW increased lactate concentration at 0, 0.5, and 1 h and tended to increase molar percentage of propionate at 1 and 1.5 h postfeeding, documenting the immediate availability of lactate converted to propionate in the +LW treatments. The main effect of corn increased the proportion of bacterial N derived from NH₃-N. Trial 2 was designed to investigate GB; isonitrogenous treatments included an SBM control, crystal GB, liquid GB (LGB), and LGB with yeast culture, which were dosed twice daily. We hypothesized that GB would increase propionate production and bacterial assimilation of NH₃-N; the combination of LGB and yeast culture was expected to have a positive additive effect, yielding the greatest VFA production and bacterial NH₃-N assimilation. No differences were observed for total VFA production; however, LGB decreased molar percentage of acetate and increased propionate and butyrate molar percentages. There were no differences in non-NH₃-N flow or microbial N flow. Under the conditions of our studies, lactate in LW and GB was fermented extensively to propionate, and microbial protein synthesis in these treatments was comparable with that in SBM controls.     

Metabolism of small peptides in rumen fluid. Accumulation of intermediates during hydrolysis of alanine oligomers,and comparison of peptidolytic activities of bacteria and protozoa

Oligopeptides of L -alanine up to ala₅, were incubated in vitro in either strained rumen fluid or suspensions of mixed rumen bacteria. The disappearance of substrates and formation of products were measured for 40 min, by which time ala₃, ala₄ and ala₅ were almost totally hydrolysed. Ala₂ was more slowly hydrolysed, and accumulated in incubations with the other peptides. The pattern of formation of ala₂ but not ala₃ from ala₄, and ala₃ and ala₂ but not ala₄ from ala₅, suggested that the peptides were being hydrolysed by a dipeptidyl peptidase mechanism. Three different sheep receiving different diets gave similar results. Neither substrates nor products appeared to be accumulated intracellularly, except for ala₂, and then only if protozoa were present. Protozoa were more active than bacteria in ala₂ hydrolysis, whereas bacteria had greater activities with higher homologues. Similar preferences were observed with glycine peptides, although unlike alanine peptides gly₃ and gly₅ were more slowly degraded than the dimer. These experiments suggest that protozoa are of importance in the accumulation and hydrolysis of dipeptides, whereas bacteria are responsible for the breakdown of larger molecules by a dipeptidyl peptidase mechanism that does not appear to involve accumulation within the cell.     

Effects of supplemental yeast (Saccharomyces cerevisiae) culture on rumen development, growth characteristics, and blood parameters in neonatal dairy calves

Yeast (Saccharomyces cerevisiae) culture was added to a texturized calf starter at 0 (control), 1, or 2% of dry matter to determine effects on intake, growth, blood parameters, and rumen development. Seventy-five Holstein calves (38 male; 37 female) were started on the experiment at 2 +/- 1 d of age and were studied for 42 d. Starter intake was measured, and fecal scoring was conducted daily. Growth and blood parameter measurements were recorded at weekly intervals. A subset of 6 male calves (2 per treatment) was euthanized at 5 wk of age, and rumen tissue was sampled for rumen epithelial growth measurements. An additional 6 male calves were euthanized at 6 wk of age for rumen epithelial growth measurements. Inclusion of yeast culture at 2% of the starter ration significantly increased starter and total dry matter intake, average daily gain, and daily hip width change when compared with the control treatment. Average daily gain was improved by 15.6% for the 2% yeast treatment. Daily change in hip height was also significantly greater for calves receiving 2% supplemental yeast culture than for calves receiving 1%. No significant treatment differences were observed for any other variables. These data suggest that the addition of yeast culture in a dairy calf starter at 2% enhances dry matter intake and growth and slightly improves rumen development in dairy calves.     

Analysis of rumen microbial populations in lactating dairy cattle fed diets varying in carbohydrate profiles and Saccharomyces cerevisiae fermentation product

The rumen microbial ecosystem is a critical factor that links diets to bovine physiology and productivity; however, information about dietary effects on microbial populations has generally been limited to small numbers of samples and qualitative assessment. To assess whether consistent shifts in microbial populations occur in response to common dietary manipulations in dairy cattle, samples of rumen contents were collected from 2 studies for analysis by quantitative real-time PCR (qPCR). In one study, lactating Holstein cows (n = 8) were fed diets in which a nonforage fiber source replaced an increasing proportion of forages and concentrates in a 4 × 4 Latin square design, and samples of ruminal digesta were collected at 9-h intervals over 3 d at the end of each period. In the second study, lactating Holstein cows (n = 15) were fed diets with or without the inclusion of a Saccharomyces cerevisiae fermentation product (SCFP) in a crossover design. In this study, rumen liquid and solid samples were collected during total rumen evacuations before and after feeding in a 42-h period. In total, 146 samples of ruminal digesta were used for microbial DNA isolation and analysis by qPCR. Validated primer sets were used to quantify total bacterial and anaerobic fungal populations as well as 12 well-studied bacterial taxa. The relative abundance of the target populations was similar to those previously reported. No significant treatment effects were observed for any target population. A significant interaction of treatment and dry matter intake was observed, however, for the abundance of Eubacterium ruminantium. Increasing dry matter intake was associated with a quadratic decrease in E. ruminantium populations in control animals but with a quadratic increase in E.ruminantium populations in cows fed SCFP. Analysis of sample time effects revealed that Fibrobacter succinogenes and fungal populations were more abundant postfeeding, whereas Ruminococcus albus tended to be more abundant prefeeding. Seven of the target taxa were more abundant in either the liquid or solid fractions of ruminal digesta. By accounting for the total mass of liquid and solid fractions in the rumen and the relative abundance of total bacteria in each fraction, it was estimated that 92% of total bacteria were found in the solid digesta fraction.     

Comparison of fermentation of diets of variable composition and microbial populations in the rumen of sheep and Rusitec fermenters. II. Protozoa population and diversity of bacterial communities

Four ruminally and duodenally cannulated sheep and 8 Rusitec fermenters were used to determine the effects of dietary characteristics on microbial populations and bacterial diversity. The purpose of the study was to assess how closely fermenters can mimic the differences between diets found in vivo. The 4 experimental diets contained forage to concentrate (F:C) ratios of 70:30 (high forage; HF) or 30:70 (high concentrate; HC) with either alfalfa hay (A) or grass hay (G) as the forage. Total bacterial numbers were greater in the rumen of sheep fed HF diets compared with those fed HC diets, whereas the opposite was found in fermenters. The numbers of cellulolytic bacteria were not affected by F:C ratio in any fermentation system, but cellulolytic numbers were 2.7 and 1.8 times greater in sheep than in fermenters for HF and HC diets, respectively. Neither total bacterial nor cellulolytic numbers were affected by the type of forage in sheep or fermenters. Decreasing F:C ratio increased total protozoa and Entodiniae numbers in sheep by about 29 and 25%, respectively, but it had no effect in fermenters. Isotrichidae and Ophryoscolecinae numbers in sheep were not affected by changing F:C ratio, but both disappeared completely from fermenters fed HC diets. Total protozoa and Entodiniae numbers were greater in sheep fed A diets than in those fed G diets, whereas the opposite was found in fermenters. Results indicate that under the conditions of the present study, protozoa population in Rusitec fermenters was not representative of that in the rumen of sheep fed the same diets. In addition, protozoa numbers in fermenters were 121 and 226 times lower than those in the sheep rumen for HF and HC diets, respectively. The automated ribosomal intergenic spacer analysis of the 16S ribosomal DNA was used to analyze the diversity of liquid- and solid-associated bacteria in both systems. A total of 170 peaks were detected in the automated ribosomal intergenic spacer analysis electropherograms of bacterial pellets across the full set of 64 samples, from which 160 were detected in at least 1 individual from each system (sheep or fermenter). Diversity of liquid-associated bacterial pellets was greater with G diets in fermenters but seemed to be unaffected by diet in sheep. Bacterial diversity in solid-associated bacteria pellets was greater for G diets compared with A diets in sheep and fermenters. Different conditions in the fermenters compared with sheep rumen might have caused a selection of some bacterial strains.     

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.     

Isolation and characterization of a hexokinase mutant of Schwanniomyces castellii: Consequences on cell production in continuous culture

A mutant of Schwanniomyces castellii with reduced glucose phosphorylation and with practically no phosphorylation of fructose was investigated. Carbon catabolite represion of α-glucosidase and amylases was reduced. Repression of β-galactosidase was normal. We have compared in continuous culture this mutant strain with wild type and another previously described mutant. The relationship between the specific rate of glucose consumption (Qs) and residual glucose concentration (s) in an inverse mode, suggests that there may be two types of transport of glucose. Mutation at the phosphorylation level causes apparent modification of the kinetic parameters of glucose uptake rate. The consequence of mutation at the phosphorylation level on biomass production was discussed.     

Effects of 2-hydroxy-4-(methylthio) butanoic acid (HMB) on microbial growth in continuous culture

2-Hydroxy-4-(methylthio) butanoic acid (HMB) positively affects milk composition and yield, potentially through ruminal actions. Four continuous culture fermenters were used to determine the optimal concentration of HMB for digestibility of organic matter (OM), neutral detergent fiber (NDF), acid detergent fiber (ADF), and hemicellulose and synthesis of microbial N. A highly degradable mix of hay and grain was used as a basal diet to simulate a typical lactation diet. Three concentrations of HMB (0, 0.055, and 0.110%) and one concentration of dl-Met (0.097%) were infused into the fermenters according to a 4 x 4 Latin square design. Digesta samples were collected during the last 3 d of each of the four 10-d experimental periods. Digestibility of OM, hemicellulose, and NDF was largely insensitive to treatment. Digestibility of ADF showed a quadratic effect to supplementation of HMB, with 0.055% having lower digestibility than 0 or 0.110%. Total production of VFA was not influenced by HMB supplementation, but differences in concentration and production of individual VFA were seen. Isobutyrate increased linearly with increasing HMB supplementation. Propionate concentration decreased linearly with increased HMB supplementation, but propionate production showed a quadratic trend (P = 0.13). A higher concentration of acetate was detected for dl-Met compared with the highest HMB concentration. There were trends (P < 0.15) for dl-Met to decrease the production of isobutyrate and to lower the concentration of butyrate when compared with HMB. Microbial efficiency was not different among treatments. The proportion of bacterial N produced from NH3-N decreased linearly with increasing HMB, and bacteria receiving dl-Met synthesized more N from NH3-N than those receiving HMB. These data suggest that supplementation of HMB may have a sparing effect on branched chain volatile fatty acids because the fatty acids are not needed to provide carbon for synthesis of valine, isoleucine and leucine with ammonia. Comparisons of bacterial community structure in the fermenter effluent samples using PCR amplicons containing the ribosomal intergenic spacer region and its flanking partial 16S ribosomal RNA gene showed no distinct banding patterns, though treatments tended to group together. Both Met and HMB affect the rumen microbial population, but Met supplied as dl-Met does not act identically to that supplied as HMB.     

Epimural bacterial community structure in the rumen of Holstein cows with different responses to a long-term subacute ruminal acidosis diet challenge

Subacute ruminal acidosis (SARA) is a prevalent metabolic disorder in cattle, characterized by intermittent drops in ruminal pH. This study investigated the effect of a gradual adaptation and continuously induced long-term SARA challenge diet on the epimural bacterial community structure in the rumen of cows. Eight rumen-cannulated nonlactating Holstein cows were transitioned over 1 wk from a forage-based baseline feeding diet (grass silage-hay mix) to a SARA challenge diet, which they were fed for 4 wk. The SARA challenge diet consisted of 60% concentrates (dry matter basis) and 40% grass silage-hay mix. Rumen papillae biopsies were taken at the baseline, on the last day of the 1-wk adaptation, and on the last day of the 4-wk SARA challenge period; ruminal pH was measured using wireless sensors. We isolated DNA from papillae samples for 16S rRNA gene amplicon sequencing using Illumina MiSeq. Sequencing results of most abundant key phylotypes were confirmed by quantitative PCR. Although they were fed similar amounts of concentrate, cows responded differently in terms of ruminal pH during the SARA feeding challenge. Cows were therefore classified as responders (n = 4) and nonresponders (n = 4): only responders met the SARA criterion of a ruminal pH drop below 5.8 for longer than 330 min/d. Data showed that Proteobacteria, Firmicutes, and Bacteroidetes were the most abundant phyla, and at genus level, Campylobacter and Kingella showed highest relative abundance, at 15.5 and 7.8%, respectively. Diversity analyses revealed a significant increase of diversity after the 1-wk adaptation but a decrease of diversity and species richness after the 4-wk SARA feeding challenge, although without distinction between responders and nonresponders. At the level of the operational taxonomic unit, we detected diet-specific shifts in epimural community structure, but in the overall epimural bacterial community structure, we found no differences between responders and nonresponders. Correlation analysis revealed significant associations between grain intake and operational taxonomic unit abundance. The study revealed major shifts in the 3 dominating phyla and, most importantly, a loss of diversity in the epimural bacterial communities during a long-term SARA diet challenge, in which 60% concentrate supply for 4 wk was instrumental rather than the magnitude of the drop of ruminal pH below 5.8.     

Short communication: Intestinal digestibility of amino acids in fluid- and particle-associated rumen bacteria determined using a precision-fed cecectomized rooster bioassay

Microbial protein represents the majority of metabolizable protein absorbed by ruminant animals. Enhanced understanding of the AA digestibility of rumen microbes will improve estimates of metabolizable protein. The objective of this experiment was to determine the digestibility of AA in fluid- (FAB) and particle-associated bacteria (PAB) using the precision-fed cecectomized rooster bioassay. Bacteria were isolated from 4 ruminally cannulated lactating Holstein cows by differential centrifugation, including particle suspension in 0.1% Tween-80 for increased removal of PAB from ruminal digesta. Samples of FAB and PAB were fed to 9 cecectomized roosters to determine standardized digestibility of AA. Total AA digestibility was 76.8 and 75.5% for FAB and PAB, respectively, but did not differ. Differences existed in AA digestibilities within bacterial type when compared with the mean essential AA digestibility value. Compared with previous literature estimates of AA digestibility in microbes (mean = 76%; range = 57–87%) and relative to National Research Council estimates of total AA from rumen bacteria (80%), the precision-fed cecectomized rooster assay is an acceptable in vivo model to determine AA digestibility of rumen bacteria.     

Modulation of rumen pH by sodium bicarbonate and a blend of different sources of magnesium oxide in lactating dairy cows submitted to a concentrate challenge

With the objective of evaluating the potential effects of sodium bicarbonate or a magnesium-based product on rumen pH and milk performance of dairy cattle exposed to a dietary challenge, 30 lactating Holstein cows (648 ± 67 kg of body weight; 44.4 ± 9.9 kg/d of milk yield; 155 ± 75 d in milk) were blocked by parity (9 primiparous and 21 multiparous) and randomly distributed to 3 treatment groups. One group received a total mixed ration (TMR) that acted as a control (CTR), a second group (SB) received the same TMR but with an additional supplementation of 0.8% of sodium bicarbonate, and a third group (MG) received the same TMR as CTR but an additional supplementation of 0.4% of a magnesium-based product (pHix-Up, Timab, Dinard, France). After 1 wk of exposure to this TMR, all 3 rations were supplemented with 1 kg/d of barley, which was then increased 1 kg/wk until reaching 3 kg/d of barley during wk 4 of the study. Every kilogram of barley replaced 1 kg of forage in the diet. Individual feed intake and behavior were monitored using electronic feed bins. Seven cows per treatment were equipped with an intraruminal bolus that recorded pH every 15 min. As the severity of the barley challenge increased, dry matter intake decreased, but this decrease was more pronounced in SB cows than in MG cows, with an intermediate response for CTR cows. The MG cows produced more milk when challenged with 2 or 3 kg/d of additional barley than when challenged with 1 kg/d, whereas CTR cows produced less milk with the 3 kg/d challenge compared with 1 or 2 kg/d, and the SB cows maintained milk production. Milk fat content decreased with barley challenges, with CTR cows experiencing a more severe decrease than SB cows, which maintained stable butterfat values throughout the study, and MG cows showed a decline in milk fat content only with the 3 kg/d of additional barley. Meal size was also reduced as the severity of barley challenge increased, and this reduction was more modest in MG cows than in SB cows. The number of daily meals consumed by SB and MG cows was more constant than that recorded in CTR cows. Cows on the CTR and SB treatments showed a marked decrease in rumen pH with the 3 kg/d of additional barley, whereas MG cows maintained stable rumen pH during the barley challenges and had greater average rumen pH (5.93 ± 0.04) than CTR cows (5.83 ± 0.04) with the 3 kg/d of additional barley; SB cows showed intermediate values (5.85 ± 0.04). Last, MG cows spent less time (32.3 ± 6.1%) with rumen pH ≤5.8 when exposed to the 3 kg/d of barley challenge than CTR and SB cows (50.7 ± 5.02%). In conclusion, supplementation with MG prevents the decline in dry matter intake and milk production induced by a rumen challenge, whereas supplementation with SB prevents the decay in milk production but does not prevent the decrease in feed intake. These changes were probably due to the ability of the MG treatment to prevent a reduction in rumen pH when challenging cows with 3 kg/d of additional barley in the ration.     

Effect of a monensin controlled-release capsule on rumen and blood metabolites in Florida Holstein transition cows

The objective of this study was to determine the effect of a monensin controlled-release capsule administered intraruminally at dry-off on rumen volatile fatty acids, NH3, lactate, pH, and energy blood metabolites in transition dairy cows fed Florida typical diets. In March 2003, 24 cows (10 primiparous and 14 multiparous) dried-off 50 to 70 d before expected parturition were randomly assigned to a treatment (n=12, oral capsule of monensin) or a control group (n=12, no capsule). Both groups received the same diet and were exposed to the same environment and management conditions. At assignment, at 21 d before expected parturition, at calving, and at 7, 14, and 21 d postpartum, blood samples were taken and body condition scores were determined. At 10 d postpartum, rumen and blood samples were obtained in the morning before the first feeding and at 2, 4, and 6 h after feeding. Serum nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), and glucose were measured. Rumen samples were analyzed for concentrations of acetic, propionic, butyric, L- and D-lactic acids, and NH3. Data for rumen and blood metabolites were analyzed by ANOVA, mixed models for repeated measures. Volatile fatty acids were not different between groups. Multiparous treated cows had a significant reduction in rumen NH3 at 6 h after feeding. Treatment with monensin significantly increased body condition score at calving in multiparous cows. During the postpartum period, NEFA and BHBA were noticeably lower in treated than in control primiparous cows. This difference was not observed in multiparous cows.     

亚麻纺织厂除尘设备的维护与安全生产

阐述除尘系统及各除尘部件的工作原理、运行中易出现的故障,并分析故障原因,提出解决方法。同时强调了除尘系统故障状态运行的危险性。