Dietary starch source and protein degradability in diets containing sucrose: Effects on ruminal measures and proposed mechanism for degradable protein effects

A feeding study was conducted to evaluate ruminal effects of starch source (STA) and rumen-degradable dietary protein (RDP) in diets with added sucrose. The experimental design was an incomplete Latin square with three 21-d periods, 8 ruminally cannulated lactating cows, and a 2 × 2 factorial arrangement of treatments. Treatments were STA (dry ground corn or high-moisture corn) as more slowly and more rapidly fermenting starch sources, respectively, and relative amount of RDP (+RDP: added protein from soybean meal; −RDP: heat-treated expeller soybean product partially substituted for soybean meal). Diets were formulated to be isonitrogenous and similar in starch and neutral detergent fiber concentrations. Dry matter (DM) intake was 1 kg greater with +RDP compared with −RDP diets. For ruminal digesta measures made 2 h postfeeding, weight of digesta DM was unaffected by treatment; total kilograms of wet digesta and kilograms of liquid tended to be greater with +RDP than with −RDP, and no effect was observed of STA × RDP. Digesta DM percentage was greater with −RDP than with +RDP. At 2 h postfeeding, ruminal pool sizes (mol) of lactate and total AA were larger and those of total organic acids (OA) and ammonia tended to be larger with +RDP than with −RDP; no effects of STA or STA × RDP were detected. Rumen-degradable protein effects on lactate and OA pool sizes may be due to a protein-mediated increase in fermentation rate of carbohydrate. Organic acid concentrations at 2 h postfeeding did not show the same response pattern or significance as the pool size data; high-moisture corn tended to be greater than dry ground corn and no effect was observed for RDP or STA × RDP. Concentration and pool size for OA were more weakly correlated [coefficient of determination (R²) = 0.66] than was the case for other ruminal analytes (R² >0.80). Organic acid pool size and kilograms of digesta liquid were strongly correlated (R² = 0.79), whereas concentration and kilograms of liquid were much less so (R² = 0.21). The correlation of OA moles with kilograms of liquid likely relates to the homeostatic mechanism of water flux across the rumen wall to reduce the osmotic gradient with blood as intraruminal moles of solute change. This action compresses the range of ruminal OA concentrations. With kilograms of ruminal liquid differing across individual measurements, the ruminal OA concentration data are not on the equivalent basis required to be reliably useful for assessing the effect of treatments. Further evaluation of protein effects on carbohydrate fermentation and of methods that allow accurate comparison of treatments for their effect on ruminal OA production are warranted.     

Effects of physically effective fiber on digestive processes and milk fat content in early lactating dairy cows fed total mixed rations

Data from recent research studies were analyzed quantitatively, and the random effect of experiment was assessed to define the physiological responses of dairy cows in early lactation to intake of physically effective neutral detergent fiber (peNDF). All studies were conducted with lactating Holstein cows (84.8 ± 3.54 days in milk) in Latin square designs, and feeds were offered ad libitum as total mixed rations (TMR). The peNDF was estimated by 2 measurement techniques, the NDF content of TMR multiplied by amount of dry matter (DM) retained on a 1.18-mm screen (peNDF_{> 1.18}) and NDF content of TMR multiplied by the proportion of DM retained by 19- and 8-mm Penn State Particle Separator screens (peNDF_{> 8}). Other factors, including concentrations of NDF, forage NDF, non-fiber carbohydrates, the amount of digestible organic matter of forages (FDOM), and the intake of ruminally degradable starch (RDSI) from grain in the diet were also investigated. The studied animal response variables included feed intake, ruminal fermentation, chewing activity, fiber digestibility, and milk production and composition. The ruminal pH (day mean) in this study ranged from 5.30 to 6.59. Using peNDF_{> 1.18} approach, the requirements for physically effective fiber in high-yielding dairy cows fed TMR in an ad libitum intake were estimated to be about 19% of ration DM or 4.1 kg/d or 0.6 kg/100 kg of body weight to maintain a ruminal pH of about 6.0. When peNDF was measured as peNDF_{> 8}, ruminal pH responded in a quadratic fashion but the confidence of estimation was lower (R² = 0.27) compared with the peNDF_{> 1.18} approach (R² = 0.67). Results of these data analyses showed that peNDF_{> 1.18} provided a satisfactory estimation of the mean ruminal pH (R² = 0.67) and NDF digestibility (R² = 0.56). Furthermore, peNDF_{> 1.18} was poorly, although positively, correlated to daily chewing (R² = 0.17), and rumination (R² = 0.24) activity. On the other hand, results from these analyses showed that milk parameters are less sensitive to the effects of dietary peNDF than other variables, such as ruminal pH, chewing activity, and fiber digestibility. Dietary FDOM correlated positively (moderately) to ruminal pH (R² = 0.24), daily chewing (R² = 0.23), and rumination (R² = 0.29) activity, whereas the daily RDSI from grain correlated negatively to ruminal pH (R² = 0.55) and positively to total volatile fatty acids (R² = 0.27). Inclusion of FDOM and RDSI from grain along with peNDF_{> 1.18} in the models that predict rumen pH further improved the accuracy of prediction. This approach appeared to further complement the concept of peNDF that does not account for differences in ruminal fermentability of feeds.     

Effects of spontaneous heating on forage protein fractions and in situ disappearance kinetics of crude protein for alfalfa-orchardgrass hays packaged in large round bales

During 2006 and 2007, forages from 3 individual hay harvests were used to assess the effects of spontaneous heating on concentrations of crude protein (CP), neutral detergent insoluble CP (NDICP), acid detergent insoluble CP (ADICP), and in situ disappearance kinetics of CP and NDICP for large round bales of mixed alfalfa (Medicago sativa L.) and orchardgrass (Dactylis glomerata L.). Over the 3 harvests, 96 large round bales were made at preset bale diameters of 0.9, 1.2, or 1.5 m and at moisture concentrations ranging from 9.3 to 46.6%. Internal bale temperatures were monitored daily during an outdoor storage period. The change in concentrations of NDICP (poststorage − prestorage) increased with heating degree days (HDD) >30°C in a relationship best explained with a nonlinear model {Y = 24.9 - [22.7 × (e^{−0.000010 × x × x})]; R² = 0.892} that became asymptotic at +24.9 percentage units of CP, thereby indicating that NDICP increases rapidly within bales that heat spontaneously. When maximum internal bale temperature (MAX) was used as the independent variable, the best regression model was quadratic and the coefficient of determination was still relatively high (R² = 0.716). The change in concentrations of ADICP (poststorage − prestorage; ΔADICP) also increased with HDD and was best fitted to a nonlinear model {Y = 14.9 - [15.7 × (e^{−0.0000019 × x × x})]} with a very high coefficient of determination (R² = 0.934). A similar quartic response was observed for the regression of ΔADICP on MAX (R² = 0.975). Increases in ΔADICP as a result of heating (HDD or MAX) were paralleled by concurrent increases in hemicellulose at relatively low increments of heating, but the inverse relationship was observed as hemicelluloses likely became reactive and concentrations decreased in more severely heated hays. Changes in ruminal disappearance rate of CP were best fitted to cubic models for regressions on both HDD (R² = 0.939) and MAX (R² = 0.876); these changes represented an approximate 50% rate reduction in severely heated hays relative to prestorage controls. Within ranges of heating most commonly encountered under field conditions, changes in rumen-degradable protein decreased in a primarily linear relationship with HDD or MAX. However, the mean change in rumen-degradable protein for the 4 most severely heated hays was only −2.6 percentage units of CP, which represents a minimal reduction from prestorage controls and is far less than the maximum of −7.9 percentage units of CP observed with less-severe heating. Interpretation of these results was complicated by poor recovery of NDICP from our most severely heated hays following machine rinsing of 0-h Dacron bags; theoretically, and by definition, this unrecovered pool of NDICP is assumed to be entirely degradable in the rumen. It remains unclear whether these responses could be corroborated in vivo or by other analytical techniques, or whether the magnitude of HDD or MAX for our most severely heated hays exceeds the reliable limits for estimating RDP via in situ methodology.     

Dietary modulation of the expression of genes involved in short-chain fatty acid absorption in the rumen epithelium is related to short-chain fatty acid concentration and pH in the rumen of goats

We have tested the hypothesis that increased concentrate intake induces mRNA abundance alterations of genes involved in short-chain fatty acid (SCFA) absorption in the rumen epithelium of goats and these changes of mRNA abundance are probably related to ruminal SCFA concentration and ruminal pH. Goats (n = 12) were randomly allocated to 2 groups and fed either a low-concentrate (LC) diet (10% concentrate; n = 6) or a medium-concentrate (MC) diet (35% concentrate; n = 6) in 2 equal portions daily. Goats were fed separately with their respective diet for 3 wk. The goats were slaughtered 6 h after the morning feeding on d 22. In vivo, goats in the MC treatment exhibited a greater ruminal SCFA concentration (81.9 mM) compared with those in the LC treatment (58.0 mM), and the pH decreased from 7.03 to 6.63. Correspondingly, mRNA expression of candidates for SCFA⁻/HCO₃⁻ exchangers, namely downregulated in adenoma (DRA), putative anion transporter 1 (PAT1), and anion exchanger 2 (AE2) were increased in the MC group. Further, upregulation in monocarboxylate transporter 1 (MCT1) and monocarboxylate transporter 4 (MCT4) mRNA abundances was observed in the MC group. The expression of genes that help the rumen epithelial cells to maintain intracellular pH, including Na⁺/H⁺ exchanger 1 (NHE1), NHE2, NHE3, vacuolar H⁺ ATPase subunit B (vH⁺ATPase), and Na⁺/K⁺ ATPase pump subunit α1 (Na⁺/K⁺ATPase) were also enhanced in the MC group relative to the LC group. During in vitro studies with isolated rumen epithelial cells from goats, exposure to a pH of 6.8 increased MCT1 mRNA expression after 24 h of culture, whereas the mRNA expression of AE2 was downregulated. The presence of SCFA (20 mM) in the medium increased DRA, PAT1, AE2, MCT1, and Na⁺/K⁺ATPase mRNA expression. The expression of vH⁺ATPase was decreased by increased SCFA concentration. The mRNA expression of MCT4 did not vary in vitro with pH (6.8) or SCFA (20 mM). The expression of DRA was increased by synergistic effects of higher SCFA concentration and lower pH, similar to that of MCT1. Thus, diet-dependent rumen epithelial mRNA expression changes in genes involved in SCFA absorption are probably related to ruminal SCFA concentration and pH.     

Relationships between dry matter content,ensiling, ammonia-nitrogen,and ruminal in vitro starch digestibility in high-moisture corn samples

The objectives of the study were (1) to determine relationships between high-moisture corn (HMC) dry matter (DM), ammonia-N [% of crude protein (CP)], and soluble CP concentrations, and pH, with 7-h ruminal in vitro starch digestibility (ivStarchD), and (2) to evaluate the effect of ensiling on pH, ammonia-N, soluble CP, and ivStarchD measurements in HMC. A data set comprising 6,131 HMC samples (55 to 80% DM) obtained from a commercial feed analysis laboratory was used for this study. Month of sample submittal was assumed to be associated with length of the ensiling period. Data for month of sample submittal were analyzed using Proc Mixed in SAS (SAS Institute Inc., Cary, NC) with month as a fixed effect. Regressions to determine linear and quadratic relationships between ivStarchD and ammonia-N, soluble CP, pH, and DM content were performed using Proc Mixed. The ivStarchD increased by 9 percentage units from October to August of the following year. Similar results were observed for ammonia-N and soluble CP with increases from 1.8 to 4.6% of CP and 31.3 to 46.4% of CP, respectively, from October to August of the following year. Ammonia-N was positively related to ivStarchD (R² = 0.61). The DM content of HMC at silo removal was negatively related (R² = 0.47) to ivStarchD with a decrease of 1.6 percentage units in ivStarchD per 1-percentage-unit increase in DM content. The pH of HMC was negatively related to ammonia-N (R² = 0.53), soluble CP (R² = 0.57), and ivStarchD (R² = 0.51). Combined, ammonia-N, DM, soluble CP, and pH provided a good prediction of ivStarchD (adjusted R² = 0.70). Increasing pH, ammonia-N, soluble CP, and ivStarchD values indicate that HMC may need up to 10 mo of ensiling to reach maximum starch digestibility. Ammonia-N, DM content, soluble CP concentration, and pH are good indicators of ruminal in vitro starch digestibility for high-moisture corn.     

Perturbations of plasma metabolites correlated with the rise of rumen endotoxin in dairy cows fed diets rich in easily degradable carbohydrates

Feeding dairy cows diets high in easily degradable carbohydrates increases the incidence of rumen and systemic metabolic disorders; however, the triggering factor is not well understood. In this study, dairy cows were fed 4 different amounts of barley grain-based concentrate at 15, 30, 45, and 60% (dry matter basis) of a total mixed ration to determine whether alterations in the rumen environment would be associated with perturbations of the plasma profile of selected metabolites. In addition, associations among free rumen endotoxin and several plasma metabolites were determined. The study was a replicated 4 × 4 Latin square design with 8 rumen-cannulated lactating dairy cows (60 ± 15 d in milk). Multiple rumen fluid and blood plasma samples were collected and analyzed for pH and rumen fluid endotoxin and for concentrations of glucose, insulin, cholesterol, β-hydroxybutyrate (BHBA), nonesterified fatty acids (NEFA), and lactate in the plasma. Rumen pH decreased below 6.0, from 8 to 12 h after the morning feeding, with the augmentation of the proportion of concentrate in the diet of ≥30%. Feeding diets with >30% concentrate resulted in a rise of free endotoxin in the rumen fluid (8.87 ± 0.39 μg/mL). Inclusion of 60% concentrate in the total mixed ration was associated with enhanced concentrations of glucose (64.5 ± 1.0 mg/dL) and lactate (540.9 ± 36.5 μmol/L) and lowered cholesterol (265.5 ± 13.7 mg/dL), BHBA (449.1 ± 47.4 μmol/L), and NEFA (138.8 ± 19.1 μEq/L) in the blood plasma. The regression analysis revealed that greater concentrations of plasma lactate and lower concentrations of cholesterol, BHBA, and NEFA were related to the rise of rumen endotoxin. Interestingly, 93% of the increase in the plasma lactate was explained by the rise of rumen endotoxin. Moreover, the analysis revealed inverse relationships of rumen endotoxin with plasma cholesterol (R² = 0.47), BHBA (R² = 0.37), and NEFA (R² = 0.50) and a biphasic response of plasma insulin (R² = 0.58). Taken together, feeding dairy cows diets rich in rumen-degradable carbohydrates and low in fiber led to lower rumen pH and a large accumulation of rumen endotoxin; the latter was correlated with perturbations of plasma metabolites allied to carbohydrate and lipid metabolism.     

Using the Small Ruminant Nutrition System to develop and evaluate an alternative approach to estimating the dry matter intake of goats when accounting for ruminal fiber stratification

The first objective of this research was to assess the ability of the Small Ruminant Nutrition System (SRNS) mechanistic model to predict metabolizable energy intake (MEI) and milk yield (MY) when using a heterogeneous fiber pool scenario (GnG1), compared with a traditional, homogeneous scenario (G1). The second objective was to evaluate an alternative approach to estimating the dry matter intake (DMI) of goats to be used in the SRNS model. The GnG1 scenario considers an age-dependent fractional transference rate for fiber particles from the first ruminal fiber pool (raft) to an escapable pool (λ_r), and that this second ruminal fiber pool (i.e., escapable pool) follows an age-independent fractional escape rate for fiber particles (k_e). Scenario G1 adopted only a single fractional passage rate (k_p). All parameters were estimated individually by using equations published in the literature, except for 2 passage rate equations in the G1 scenario: 1 developed with sheep data (G1-S) and another developed with goat data (G1-G). The alternative approach to estimating DMI was based on an optimization process using a series of dietary constraints. The DMI, MEI, and MY estimated for the GnG1 and G1 scenarios were compared with the results of an independent dataset (n = 327) that contained information regarding DMI, MEI, MY, and milk and dietary compositions. The evaluation of the scenarios was performed using the coefficient of determination (R²) between the observed and predicted values, mean bias (MB), bias correction factor (C_b), and concordance correlation coefficient. The MEI estimated by the GnG1 scenario yielded precise and accurate values (R² = 0·82; MB = 0.21 Mcal/d; C_b = 0.98) similar to those of the G1-S (R² = 0.85; MB = 0.10 Mcal/d; C_b = 0.99) and G1-G (R² = 0.84; MB = 0.18 Mcal/d; C_b = 0.98) scenarios. The results were also similar for the MY, but a substantial MB was found as follows: GnG1 (R² = 0.74; MB = 0.70 kg/d; C_b = 0.79), G1-S (R² = 0.71; MB = 0.58 kg/d¹; C_b = 0.85) and G1-G (R² = 0.71; MB = 0.65 kg/d; C_b = 0.82). The alternative approach for DMI prediction provided better results with the G1-G scenario (R² = 0.88; MB = −71.67 g/d; C_b = 0.98). We concluded that the GnG1 scenario is valid within mechanistic models such as the SRNS and that the alternative approach for estimating DMI is reasonable and can be used in diet formulations for goats.     

Rumen pH and fermentation characteristics in dairy cows supplemented with Megasphaera elsdenii NCIMB 41125 in early lactation

This study investigated the effect of using the lactate-utilizing bacterium Megasphaera elsdenii NCIMB 41125 as a probiotic supplement on rumen fermentation and pH in dairy cows in the immediate postcalving period. Fourteen multiparous rumen-fistulated Holstein cows, blocked according to 305-d milk yield in the previous lactation, were used in a randomized complete block design. From d 1 to 28 postcalving, cows were fed ad libitum a total mixed ration with a forage to concentrate ratio of 392:608 and a starch concentration of 299 g/kg of dry matter. Treatments consisting of a minimum of 10¹⁰ cfu of Megasphaera elsdenii NCIMB 41125 or autoclaved M. elsdenii (placebo) were administered via the rumen cannula on d 3 and 12 of lactation (n = 7 per treatment). Mid-rumen pH was measured every 15 min, and eating and ruminating behaviors were recorded for 24 h on d 2, 4, 6, 8, 11, 13, 15, 17, 22, and 28. Rumen fluid for volatile fatty acid and lactic acid analysis was collected at 11 time points on each of d 2, 4, 6, 13, and 15. Yields of milk and milk protein and lactose were similar, but milk fat concentration tended to be higher in cows that received the placebo. Time spent eating and ruminating and dry matter intake were similar across treatments. Ruminal lactic acid concentrations were highly variable between animals, and no cases of clinical acidosis were observed. Both treatment groups had rumen pH <5.6 for more than 3 h/d (a commonly used threshold to define subacute ruminal acidosis), but the length of time with rumen pH <5.6 was markedly reduced in the days immediately after dosing and fluctuated much less from day to day in cows that received M. elsdenii compared with those that received the placebo. Ruminal total volatile fatty acid concentrations were similar across treatments, but the acetate:propionate ratio tended to be smaller in cows that received M. elsdenii. Despite the lack of a measurable treatment effect on ruminal lactic acid concentration, supplementation of early lactation dairy cows with lactate-utilizing M. elsdenii altered the rumen fermentation patterns in favor of propionate, with potential benefits for energy balance and animal productivity.     

Technical note: Evaluation of markers for estimating duodenal digesta flow and ruminal digestibility: Acid detergent fiber,sulfuric acid detergent lignin,and n-alkanes

The amount of digesta flowing to the duodenum is a relevant measurement for the evaluation of nutrient supply to ruminants, which is usually estimated in animals fitted with a duodenal T-type cannula using internal or external markers. This study evaluated acid detergent fiber (ADF) compared with external (C₃₂n-alkane) and internal [sulfuric acid lignin (ADL) and n-alkanes C₃₁ and C₃₃] markers for estimating duodenal flow and(or) ruminal digestibility of dry matter (DM) in cattle and sheep. In the first assay, 4 duodenally cannulated Holstein steers housed in metabolism cages, dosed with C₃₂n-alkane, and fed Avena strigosa plus concentrate and increasing levels of tannin extract to reduce ruminal digestibility, were used in a Latin square design. The mobile-bag technique was used to measure the intestinal disappearance of ADL and ADF from forage (Avena strigosa, Pennisetum purpureum, Cynodon dactylon, and Medicago sativa) and concentrate (corn grain, soybean meal, and sunflower meal) samples that were previously incubated in the rumen of additional fistulated steer for 12, 24, 36, or 48 h. The ADF concentration in residues recovered in the feces was strongly related to the ADF concentration in residues at the duodenum (R² = 0.93, standard deviation = 30.0, n = 901). This relationship showed a lower precision for ADL fraction (R² = 0.88, standard deviation = 12.6, n = 590). In a second assay, duodenal flow and ruminal DM digestibility were calculated from the duodenal and fecal concentration of either marker. We observed a significant effect of marker type on ruminal DM digestibility values, and the effect of tannin treatments was observed only when ADF or ADL was used as the marker. The lowest residual error was obtained for ADF. Ruminal DM digestibility was, on average, higher for C₃₁ and C₃₃n-alkanes, and the use of dosed C₃₂n-alkane resulted in a negative value. In the third assay, a data set of 235 individual observations was compiled from digestibility trials to compare ADF and ADL as markers for estimating duodenal digesta flow in wethers (n = 204) and cattle (n = 31). We observed a strong relationship between markers (R² = 0.84 in sheep and R² = 0.88 in cattle), but variance analysis within trials indicated that ADF was more precise than ADL. In conclusion, in digestibility trials in which fecal output was measured and spot samples of the duodenal digesta were obtained, duodenal flow and ruminal digestibility of the DM may be estimated from the relationship between the ADF concentration in feces and that in the duodenal digesta of ruminants.     

Severity of ruminal acidosis in primiparous holstein cows during the periparturient period

The objectives of this study were: 1) to determine the effect of providing additional prepartum concentrate on the occurrence and severity of ruminal acidosis (RA) and lactational performance during the periparturient period in primiparous cows; and 2) to characterize the occurrence and severity of RA during the periparturient period. We hypothesized that providing additional concentrate prepartum would reduce postpartum RA. Fourteen ruminally cannulated Holstein heifers were paired by expected calving date and body condition score. The heifers were assigned to 1 of 2 prepartum feeding regimens: 1) a control treatment consisting of a far-off diet (forage:concentrate, F:C = 80:20) fed from d −60 to d −25 and a close-up diet (F:C = 54:46) fed from d −24 until parturition; or 2) a high-concentrate (HC) feeding program consisting of 4 prepartum diets, HC-1 (F:C = 68:32) fed from d −60 to d −43, HC-2 (F:C = 60:40) fed from d −42 to d −25, HC-3 (F:C = 52:48) fed from d −24 to d −13, and HC-4 (F:C = 46:54) fed from d −12 until parturition. All cows received the same lactation diet postpartum. Ruminal pH was measured continuously from d −5 to d +5, and for 3 consecutive days starting on d +17 ± 1.2, d +37 ± 1.4, and d +58 ± 1.5 relative to parturition using an indwelling ruminal pH system. Ruminal acidosis was considered to occur when ruminal pH was <5.8 (total RA). Ruminal acidosis was further partitioned into: 1) mild RA (5.8 > ruminal pH > 5.5), 2) moderate RA (5.5 > ruminal pH > 5.2), and 3) acute RA (ruminal pH < 5.2). Feeding additional concentrate prepartum did not reduce postpartum RA. In fact, cows fed the HC treatment had more daily episodes of acute RA than cows fed the control treatment. Day relative to parturition affected the occurrence and severity of RA; RA increased following parturition and was sustained thereafter. The DM intake during the last 5 d of gestation was lower for cows fed the HC treatment compared with cows fed the control treatment, but lactational performance was not affected. We conclude that, under the conditions imposed, feeding additional concentrate prepartum does not reduce postpartum RA. Furthermore, the incidence and severity of RA increases immediately postpartum, emphasizing the need to develop and implement feeding strategies that reduce this risk.     

Ruminal degradability of dry matter,crude protein,and amino acids in soybean meal,canola meal,corn, and wheat dried distillers grains

Different protein sources, such as canola meal (CM) or dried distillers grains (DDG), are currently used in dairy rations to replace soybean meal (SBM). However, little data exists comparing their rumen degradation in a single study. Therefore, the objective of this study was to compare the ruminal degradation of dry matter (DM), crude protein (CP), and AA of SBM, CM, high-protein corn DDG (HPDDG), and wheat DDG plus solubles (WDDGS). In situ studies were conducted with 4 rumen-fistulated lactating Holstein cows fed a diet containing 38% grass hay and 62% corn-based concentrate. Each protein source was incubated in the rumen of each cow in nylon bags for 0, 2, 4, 8, 16, 24, and 48 h to determine DM and CP rumen degradation kinetics, whereas additional bags were also incubated for 16 h to evaluate AA ruminal disappearance. Rumen DM and CP degradability was calculated from rumen-undegraded residues corrected or not for small particle loss. Data were fitted to an exponential model to estimate degradation parameters and effective degradability (ED) was calculated with a passage rate of 0.074 h⁻¹. The WDDGS and SBM had higher uncorrected ED (DM = 75.0 and 72.6%; CP = 84.8 and 66.0%, respectively) than CM and HPDDG (DM = 57.2 and 55.5%; CP = 59.3 and 48.2%, respectively), due to higher soluble fraction in WDDGS and a combination of higher potentially degradable fraction and rate of degradation in SBM. Correction for small particle loss from bags, higher for WDDGS than for the other protein sources, decreased estimated ED but did not alter feed ranking. The ruminal disappearance of AA after 16 h of incubation reflected the overall pattern of CP degradation between protein supplements, but the ruminal disappearance of individual AA differed between protein supplements. Overall, these results indicate that, in the current study, (1) SBM and WDDGS were more degradable in the rumen than CM and HPDDG, and (2) that small particle loss correction is relevant but does not alter this ranking.     

Subacute ruminal acidosis challenge changed in situ degradability of feedstuffs in dairy goats

This study investigated the effects of wheat-induced subacute ruminal acidosis (SARA) on rumen bacterial populations and in situ degradabilities of NDF, starch, and crude protein of feeds. Four multiparous dairy goats (BW = 60 ± 3.3 kg) fitted with ruminal cannulas were assigned to a 2 × 2 crossover design (28-d treatment periods separated by a 7-d washout interval). The treatment diets consisted of 2 levels of cracked wheat: 0 (control, corn based concentrate) and 35% (diet-induced SARA, wheat-based concentrate), with a constant forage- (45% alfalfa hay and 5% corn silage of DM) to-concentrate (50% of DM) ratio. Results indicate that diets with a 35% wheat decreased ruminal pH (6.21 vs. 5.98) and increased the duration (1.13 vs. 4.72 h/d) and area (0.12 vs. 0.78 pH × h/d) of ruminal pH below 5.6 and induced SARA. The SARA increased ruminal total volatile fatty acid concentration, from 105.0 to 123.8 mM, and decreased the acetate molar proportion (62.8 vs. 56.6 mol/100 mol) and the acetate-to-propionate ratio (3.5 vs. 2.8). Compared with the control group, SARA decreases the relative abundance of Fibrobacter succinogenes (−59.3%) and Ruminococcus flavefaciens (−68.4%), whereas it increased Succinimonas amylolytica (198.1%) and Ruminobacter amylophilus (125.2%). The SARA decreased 24- and 48-h dry matter (DM) and neutral detergent fiber (NDF) degradabilities of corn silage. The 48-h degradabilities of DM (51.0 vs. 48.2%) and NDF (40.3 vs. 36.0%) in alfalfa hay were not affected by SARA, but the SARA tended to reduce the 24-h DM (49.6 vs. 46.3%) and NDF (37.8 vs. 33.2%) degradabilities. The effective ruminal degradabilities of DM and NDF in alfalfa hay and corn silage were reduced during SARA. In situ degradability parameters of DM and starch of wheat were not affected by SARA, but starch degradability of corn (9.5 vs. 13.3%/h) increased. The SARA reduced in situ 12-h degradabilities of DM and crude protein of soybean meal and extruded soybean without affecting the degradabilities of the other protein supplements (corn gluten meal, cottonseed meal, corn dried distillers grains with solubles, rapeseed meal, and wheat germ meal). These results indicated that the cracked wheat-induced SARA reduced the degradation of NDF in roughages and that of protein in soybean meal (−19.8%) and extruded soy (−18.9%) and increased the starch degradability in corn, due to the increased amylolytic bacteria and decreased cellulolytic bacteria counts in the rumen.     

Ruminal temperature may aid in the detection of subacute ruminal acidosis

The objective of this study was to investigate the relationship between ruminal pH and ruminal temperature and to develop a predictive equation that can aid in the diagnosis of subacute ruminal acidosis (SARA). Six rumen-fistulated lactating Holstein dairy cows (639 ± 51 kg body weight) were used in the study. Cows were randomly allocated to 1 of 2 dietary treatments: control (% of dry matter, 40% corn silage, 27% mixed haylage, 7% alfalfa hay, 18% protein supplement, 4% ground corn, and 4% wheat bran) or SARA total mixed ration (% of dry matter, 31% corn silage, 20% mixed haylage, 5% alfalfa hay, 15% protein supplement, 19% ground wheat, and 10% ground barley) and were fed daily at 0700 and 1300 h. The experiment consisted of 1 wk of adaptation followed by 1 wk of treatment. Ruminal pH and ruminal temperature were simultaneously and continuously recorded every minute for 4 d per week using the same indwelling electrode. Subacute-acidotic cows spent more time (min/d) below ruminal pH 5.6 and a greater time above 39.2°C than control cows. Ruminal pH nadir had a negative relationship with its corresponding ruminal temperature (R² = 0.77). Therefore, ruminal temperature may have potential to predict ruminal pH and thus aid in the diagnosis of SARA.     

Gold nanoparticle modified conducting polymer of 4-(2,5-di(thiophen-2-yl)-1H-pyrrole-1-l) benzenamine for potential use as a biosensing material

Gold nanoparticle (AuNP) modified conducting polymer of 4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl)benzenamine (SNS-NH₂) was used as the biosensing platform for glucose analysis. Electrochemical measurements were carried out by following the consumed oxygen due to the enzymatic reaction of glucose oxidase (GOx) at −0.7 V vs Ag/AgCl. Optimisation of pH, enzyme loading, stability experiments were carried out. Effect of NP was investigated by monitoring the signal responses at different AuNP sizes and amounts. A linear relation of y = 1.597x + 0.264 (R² = 0.993) was found for glucose concentrations between 0.002 and 5.0 mM. The analytical characteristics of the system were also evaluated for glucose determination in flow injection analysis (FIA) mode. Finally, the system was checked for glucose detection on real samples.     

Prediction of coagulation properties, titratable acidity, and pH of bovine milk using mid-infrared spectroscopy

This study investigated the potential application of mid-infrared spectroscopy (MIR 4,000-900 cm⁻¹) for the determination of milk coagulation properties (MCP), titratable acidity (TA), and pH in Brown Swiss milk samples (n = 1,064). Because MCP directly influence the efficiency of the cheese-making process, there is strong industrial interest in developing a rapid method for their assessment. Currently, the determination of MCP involves time-consuming laboratory-based measurements, and it is not feasible to carry out these measurements on the large numbers of milk samples associated with milk recording programs. Mid-infrared spectroscopy is an objective and nondestructive technique providing rapid real-time analysis of food compositional and quality parameters. Analysis of milk rennet coagulation time (RCT, min), curd firmness (a₃₀, mm), TA (SH°/50 mL; SH° = Soxhlet-Henkel degree), and pH was carried out, and MIR data were recorded over the spectral range of 4,000 to 900 cm⁻¹. Models were developed by partial least squares regression using untreated and pretreated spectra. The MCP, TA, and pH prediction models were improved by using the combined spectral ranges of 1,600 to 900 cm⁻¹, 3,040 to 1,700 cm⁻¹, and 4,000 to 3,470 cm⁻¹. The root mean square errors of cross-validation for the developed models were 2.36 min (RCT, range 24.9 min), 6.86 mm (a₃₀, range 58 mm), 0.25 SH°/50 mL (TA, range 3.58 SH°/50 mL), and 0.07 (pH, range 1.15). The most successfully predicted attributes were TA, RCT, and pH. The model for the prediction of TA provided approximate prediction (R² = 0.66), whereas the predictive models developed for RCT and pH could discriminate between high and low values (R² = 0.59 to 0.62). It was concluded that, although the models require further development to improve their accuracy before their application in industry, MIR spectroscopy has potential application for the assessment of RCT, TA, and pH during routine milk analysis in the dairy industry. The implementation of such models could be a means of improving MCP through phenotypic-based selection programs and to amend milk payment systems to incorporate MCP into their payment criteria.     

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.     

Effects of spontaneous heating on estimates of total digestible nutrients for alfalfa-orchardgrass hays packaged in large round bales

Large round or large square hay packages are more likely to heat spontaneously during storage than hay packaged in conventional (45 kg) bales, and the effects of this phenomenon on the associated energy estimates for these hays can be severe. Our objectives for this project were to assess the relationship between estimates of total digestible nutrients (TDN) and spontaneous heating and to describe any important differences in energy estimates that may result specifically from 2 methods of estimating truly digestible fiber (TD-Fiber). Using the summative approach to estimate TDN, TD-Fiber can be estimated from inputs of protein-corrected neutral detergent fiber (NDFn) and acid detergent lignin (TD-FiberLIG) or from NDFn and 48-h neutral detergent fiber digestibility (TD-FiberNDFD). Throughout 2006 and 2007, mixed alfalfa (Medicago sativa L.)-orchardgrass (Dactylis glomerata L.) hays from 3 individual harvests were obtained from the same 8.2-ha research site near Stratford, Wisconsin. Both options for estimating TD-Fiber (TD-FiberLIG or TD-FiberNDFD) were then used independently via the summative approach to estimate the total TDN concentrations (TDN-LIG or TDN-NDFD, respectively) within these hays. Estimates of both TDN-LIG and TDN-NDFD then were related to heating degree days >30°C accumulated during storage by various regression techniques. Changes (poststorage - prestorage) in TDN-LIG that occurred during storage (ΔTDN-LIG) were best fitted with a nonlinear decay model in which the independent variable was squared [Y = (11.7 × e^{−0.0000033×x×x}) − 11.6; R² = 0.928]. For changes in TDN-NDFD (ΔTDN-NDFD), a quadratic regression model provided the best fit (Y = 0.0000027x² − 0.010x + 0.4; R² = 0.861). Generally, ΔTDN-LIG estimates were 2.0 to 4.0 percentage units lower than ΔTDN-NDFD estimates when heating exceeded 500 HDD. For regressions on maximum internal bale temperature, both ΔTDN-LIG (Y = −0.38x + 16.3; R² = 0.954) and ΔTDN-NDFD (Y = −0.25x + 10.2; R² = 0.848) were best fitted by linear models with heterogeneous (P < 0.001) slopes and intercepts. In both cases, coefficients of determination were high, suggesting that simple measures of spontaneous heating are excellent predictors of energy losses in heated forages. Regardless of method, reductions in TDN were associated primarily with losses of nonfiber carbohydrate, which is known to occur via oxidation of sugars during spontaneous heating. For heated forages, some discrepancy between TDN-LIG and TDN-NDFD existed because the relationship between NDFD and spontaneous heating was shown previously to be very poor, resulting in minimal changes for estimates of TD-FiberNDFD as a consequence of heating. In contrast, TD-FiberLIG declined in close association with heating, largely because TD-FiberLIG was sensitive to changes in concentrations of both NDFn and acid detergent lignin. Discrepancies between TDN-LIG and TDN-NDFD were exacerbated further when neutral detergent fiber rather than NDFn was used to estimate TD-FiberNDFD. Estimates of TDN declined by as much as 13.0 percentage units within severely heated hays, and this is a serious consequence of spontaneous heating.     

Determination of resveratrol in grains,hulls and leaves of common and tartary buckwheat by HPLC with electrochemical detection at carbon paste electrode

A reverse-phase high-performance liquid chromatographic method for the determination of trans-resveratrol with spectrophotometric detection (306 nm) and amperometric detection at carbon paste electrode (E = + 1,2 V) was developed and tested on real samples of grains, hulls and leaves of six varieties of common buckwheat (Fagopyrum esculentum Möench) and two varieties of tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.). Optimal conditions for the determination of trans-resveratrol were as follows: column Kromasil C-18 (7 μm), 125 × 4 mm; mobile phase acetonitrile: diluted BR buffer pH 7 (50:50, 30:70 for grains and hulls and 20:80 for leaves); flow rate 1 ml min⁻¹. Under these conditions, the limit of detection of trans-resveratrol (L_D) was 3.5 × 10⁻⁸ mol l⁻¹ (R² = 0.9986) for electrochemical detection and 3.2 × 10⁻⁸ mol l⁻¹ (R² = 0.9993) for spectrophotometric detection.     

The effects of forage proportion and rapidly degradable dry matter from concentrate on ruminal digestion in dairy cows fed corn silage-based diets with fixed neutral detergent fiber and starch contents

This study investigated the effects of the forage-to-concentrate (F:C) ratio and the rate of ruminal degradation of carbohydrates from the concentrate on digestion in dairy cows fed corn silage-based diets. Six cows with ruminal cannulas were assigned to 6 treatments in a 6 × 6 Latin square. Treatments were arranged in a 3 × 2 factorial design. Three proportions of neutral detergent fiber from forage [FNDF; 7.6, 13.2, and 18.9% of dry matter (DM)] were obtained by modifying F:C (20:80, 35:65, and 50:50). These F:C were combined with concentrates with either high or low content of rapidly degradable carbohydrates. The dietary content of rapidly degradable carbohydrates from the concentrate was estimated from the DM disappearance of concentrate after 4 h of in sacco incubation (CRDM). Thus, 2 proportions of CRDM were tested (20 and 30% of DM). Wheat and corn grain were used as rapidly and slowly degradable starch sources, respectively. Soybean hulls and citrus pulp were used as slowly and rapidly degradable fiber sources, respectively. Concentrate composition was adjusted to maintain dietary starch and neutral detergent fiber contents at 35.9 and 28.9% of DM, respectively. There was no effect of the interaction between F:C and CRDM on DM intake (DMI), ruminal fermentation, chewing activity, and fibrolytic activity. When F:C decreased, DMI increased, the mean ruminal pH linearly decreased, and the pH range linearly increased from 0.95 to 1.27 pH unit. At the same time, the acetate-to-propionate ratio decreased linearly. Decreasing F:C linearly decreased the average time spent chewing per kilogram of DMI from 35.2 to 19.5 min/kg of DMI and decreased ruminal liquid outflow from 11.6 to 9.2 L/kg of DMI, suggesting a decrease in the salivary flow. Increasing CRDM decreased DMI and increased the time during which pH was below 6.0 (3.1 vs. 4.8 h), the pH range (0.90 vs. 1.33), and the initial rate of pH drop. It also increased the volatile fatty acid range (35 vs. 59 mM), thus suggesting an increased rate of fermentation. It also decreased the acetate-to-propionate ratio (2.9 vs. 1.8). Increasing CRDM barely affected the average time spent chewing per kilogram of DMI and the ruminal liquid outflow. These results suggest that rumen pH is controlled by different mechanisms when F:C is decreased or when CRDM is increased. Consequently, FNDF is a good predictor of the chewing time, whereas CRDM is a good predictor of the pH range and volatile fatty acid profiles. Finally, considering both FNDF and CRDM improves the prediction of mean pH.     

Comparison of sterol composition between Tuber fermentation mycelia and natural fruiting bodies

Truffle, belonging to Tuber genera, is a nutritious and sterol-rich edible fungus, and sterol is a potential health beneficial compound. A comparison of Tuber sterol composition indicates that the total sterol contents in the fermentation mycelia (i.e., 10.5 mg g⁻¹) (n = 3) were approximately 3.2-5.6 times higher than that of the fruiting bodies (p < 0.05) with the addition of soybean flour into the basal fermentation media. Moreover, the phytosterol profile of fermentation mycelia could be significantly improved by adding soybean flour into the fermentation media. After the addition of soybean flour, stigmasterol and β-sitosterol appeared in the fermentation mycelia and the contents of total phytosterols (2279 μg g⁻¹) (n = 3), brassicasterol (943 μg g⁻¹) (n = 3), and campesterol (418 μg g⁻¹) (n = 3) were all increased significantly (p < 0.05). Moreover, the total contents of sterols and phytosterols in the fermentation mycelia cultured in the soybean media were much higher than those in the fruiting bodies (i.e., 1883-3240 and 479-1832 μg g⁻¹, respectively) (n = 3, p < 0.05). This work confirms the potentiality of Tuber fermentation mycelia as the alternative resource for its fruiting bodies from the viewpoint of sterols production.