Experimental Study on the Transfer of Polychlorinated Biphenyls (PCBs) and Polychlorinated Dibenzo-p-dioxins and Dibenzofurans (PCDD/Fs) into Milk of High-Yielding Cows during Negative and Positive Energy Balance.

Dioxin-like polychlorinated biphenyls (dl-PCBs) as well as polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are a major concern for food safety, especially in fat-containing foods of animal origin, such as milk. Due to the lipophilic character of PCDD/Fs and PCBs, it is of special interest to explore whether the metabolic state of high-yielding cows influences the transfer rates into milk. Five German Holstein cows were orally exposed to a mixture of 17 PCDD/Fs, 12 dl-PCBs, and 6 non-dioxin-like PCBs (ndl-PCBs) for two dosing periods of 28 days each. The first period covered the negative energy balance (NEB) after calving, while the second period addressed the positive energy balance (PEB) in late lactation. Each dosing period was followed by a depuration period of around 100 days. During the NEB phase, the transfer rates of 14 PCDD/Fs and 7 dl-PCBs quantified were significantly (p ≤ 0.1) higher compared to the PEB phase, indicating an influence of the metabolic state on the transfer. Furthermore, the congener-specific transfer rates (0.3-39%) were in the range of the results from former studies. This indicates that the milk yield of the exposed cows is not the only determining factor for the transfer of these congeners into milk.

Feed Clusters According to In Situ and In Vitro Ruminal Crude Protein Degradation.

Effective degradation (ED) of crude protein (CP) was estimated in vitro at 0.02, 0.05 and 0.08 h−1 assumed ruminal passage rates for a total of 40 feedstuffs, for which in situ ED was available and used as reference degradation values. For this, the Streptomyces griseus protease test was used. The differences between in vitro CP degradation and the in situ CP degradation values were lowest in legume grains and highest in cereal by-products and barley. The differences between in situ and in vitro ED were expressed using a degradation quotient (degQ), where degQ = (EDin vitro − EDin situ)/EDin situ. Among the tested feedstuffs, eight specific clusters were identified according to degQ for the assumed passage rates. The feedstuffs clustered in an unspecific way, i.e., feedstuffs of different nutrient composition, origin or treatment did not necessarily group together. Formaldehyde−treated rapeseed meal, soybean meal, wheat, a treated lupin, sunflower meal and barley could not be assigned to any of the clusters. Groupwise degradation (range of degQ for assumed passage rates are given in brackets) was detected in grass silages (−0.17, −0.11), cereal by-products together with sugar beet pulp (−0.47, −0.35) and partly in legume grains (−0.14, 0.14). The clustering probably based on different specific nutrient composition and matrix effects that influence the solubility of feed protein and limit the performance of the protease. The matrix can be affected by treatment (chemically, thermally or mechanically), changing the chemical and physical structure of the protein within the plant. The S. griseus protease test had reliable sensitivity to reflect differences between native feedstuffs and treatments (thermally or chemically) that were found in situ. The in situ results, however, are mostly underestimated. The clustering results do not allow a clear conclusion on the groupwise or feed-specific use of carbohydrate-degrading enzymes as pre- or co-inoculants as part of the S. griseus protease test and need to be tested for its potential to make this test more conform with in situ data.

Linking metabolites in eight bioactive forage species to their in vitro methane reduction potential across several cultivars and harvests.

An in vitro Hohenheim gas test was conducted to analyze the fermentation end-products from 17 cultivars of eight polyphenol containing forage species. The polyphenol composition and proanthocyanidin (PA) structural features of all the cultivars were analyzed with UPLC-MS/MS in leaves of vegetative or generative plants. The samples were incubated with and without polyethylene glycol (PEG, a tannin-binding agent) to separate the tannin-effect on methane (CH4, ml/200 mg DM) production from that of forage quality. Sulla and big trefoil, two particularly PA rich species, were found to have the highest CH4 reduction potential of up to 47% when compared to the samples without PEG. However, concomitant reduction in gas production (GP, ml/200 mg DM) of up to 44% was also observed. An increase in both GP and CH4 production under PEG treatments, confirms the role of tannins in CH4 reduction. Moreover, PA structural features and concentration were found to be an important source of variation for CH4 production from PA containing species. Despite having low polyphenol concentrations, chicory and plantain were found to reduce CH4 production without reducing GP. Additionally, interspecies variability was found to be higher than intraspecies variability, and these results were consistent across growth stages, indicating the findings' representativeness.

Diets for Dairy Cows with Different Proportions of Crude Protein Originating from Red Clover Silage versus Soybean Meal: Ruminal Degradation and Intestinal Digestibility of Amino Acids.

The purpose was to assess the effect of exchanging crude protein (CP) of soybean meal (SBM) with red clover silage (RCS) in total mixed rations (TMR) on ruminal degradation and intestinal digestibility (ID) of essential amino acids (EAA). Four TMR and their individual feed components were studied. The TMR were composed of forage and concentrates (75:25), with proportions of RCS in TMR of 0.15, 0.30, 0.45, and 0.60 on a dry matter basis, resulting in diet groups RCS15, RCS30, RCS45, and RCS60, respectively. The ruminal degradation of EAA was determined using the nylon bag technique. For this, samples of TMR and their individual feed components were ruminally incubated for 16 h. The feed residues of TMR obtained after 16 h of incubation were used for the determination of ID of EAA using the mobile-bag technique. Increasing RCS and reducing SBM proportions linearly increased (p < 0.01) the in situ ruminal degradation of individual EAA from 75.5% to 83.5%. The degradation of EAA followed the trend of CP degradation among TMR, except for Cys that was lower (p < 0.05) than that of CP in RCS60 (79.7% vs. 86.3%). The degradation of EAA in individual feed ingredients not always corresponded to the degradation of CP and was feed dependent. Increasing the proportions of RCS in the TMR linearly reduced (p < 0.001) the ID of EAA (except for Ile) from 78.2% to 67.3%. However, the ID of EAA did not always reflect the ID of CP, and in general, the differences between the ID of CP and EAA increased as RCS increased in the TMR. The ID values of most of the EAA were similar (p > 0.05) to ID of CP in RCS15 and RCS30, while they mostly differed (p < 0.05) in RCS45 and RCS60, and being higher for EAA than CP (except for Cys that was lower than CP, p < 0.05). Similar trends were observed for intestinal absorbable AA, resulting in higher values (p < 0.05) of intestinal absorbable for all EAA than of CP in diet RCS60. In conclusion, increasing levels of RCS in TMR reduced the extent of EAA flow into the small intestine, the ID of EAA, and consequently the intestinal absorbable EAA. Therefore, accurate determination of metabolizable AA must be considered for optimal diet formulation when including high proportions of RCS in diets of high-producing dairy cows.

In situ and in vitro determination of the protein value of feeds for ruminants.

The objective of this study was to estimate the amount of post-rumen crude protein (prCP), a precursor to metabolisable protein for ruminants, of feeds and feed mixtures using an in situ and in vitro method, and to contrast the results of both approaches. For this, 34 samples were examined: 9 feeds, 13 feed mixtures made thereof and 12 feed mixtures provided by commercial dairy farms. As a minor aspect, additivity of protein values from feeds was evaluated by comparing measured and calculated values of feed mixtures. Effective prCP was calculated for assumed rumen passage rates (k) of 0.05 h-1 (prCP5) and 0.08 h-1 (prCP8) by in situ measurements of rumen undegraded CP and fermented organic matter (FOM) assuming an efficiency of 181 g microbial CP/kg FOM. Additionally, effective prCP in vitro was estimated using the modified Hohenheim gas test (modHGT) through incubation in rumen-fluid buffer solution for 8 and 24 h followed by ammonia distillation. In vitro estimations were highly correlated with in situ values for both passage rates (k = 0.05 h-1: p < 0.001, R2 = 0.68; k = 0.08 h-1: p < 0.001, R2 = 0.76). The in vitro method yielded higher values for effective prCP than the in situ approach with 29 g/kg OM (k = 0.05 h-1) and 37 g/kg OM (k = 0.08 h-1) on average for all samples. Small positive associative effects - reflecting non-additivity - were found, averaging at 2.2 g prCP8/kg OM (p < 0.05) for the in situ and 10.7 g prCP8/kg OM (p < 0.001) for the in vitro approach. Due to the need of an assumption of a certain value for microbial efficiency in situ, effective prCP might be more accurately estimated in vitro, accounting for nutrient-specific efficiencies as well as interactions between carbohydrate and protein degradation by rumen microbes. Furthermore, the modHGT highlighted associative effects more pronounced and seems suitable as a routine method due to the comparably low effort and high sample throughput. The potential of the modHGT to determine the protein value of feeds could be demonstrated by our study.

Estimation of diet organic matter digestibility in grazing dairy cows.

Precise and continual information on the energy supply from pasture is mandatory for managing grazing ruminants. Therefore, estimating the organic matter (OM) digestibility from faecal crude protein concentration using the regression equation, OM digestibility [%] = 79.76-107.7 · e(-0.01515 · faecal crude protein [g/kg OM]), is known to be a suitable tool. However, essential information regarding faeces sampling times and the required number of samples are not yet available. We therefore analysed the OM digestibility data of an experiment with dairy cows grazing two pasture types and receiving two concentrate types over 6 d in three independent runs. Both pasture type and grazing day affected the OM digestibility estimates, whereas concentrate type and intake did not, indicating that this method reliably detects small changes in OM digestibility of pasture without being interfered by moderate concentrate supplementation, selective grazing behaviour or differences in feed intake. Likewise, as sampling time did not influence OM digestibility, faeces sampling once daily can be recommended to be sufficient for an accurate estimation of OM digestibility. The variance within pasture type and grazing day amounted for 1.1 percentage units of estimated OM digestibility, which enabled to define the minimum sample number required to detect given differences in OM digestibility with adequate statistical certainty. In conclusion, estimating OM digestibility from faecal crude protein concentration is an applicable and sensitive method to reliably detect differences in the quality of ingested pasture using a limited number of animals. Therefore, instructions for faeces sample collection  were provided.

Assessing the Potential of Diverse Forage Mixtures to Reduce Enteric Methane Emissions In Vitro.

Methane emissions from ruminants are a major contributor to agricultural greenhouse gas emissions. Thus, eight different forage species were combined in binary mixtures with Lolium perenne in increasing proportions, in vitro, to determine their methane reduction potential in ruminants. Species were sampled in two consecutive years where possible. The aims were: a) to determine if mixtures with specific forages, particularly those rich in plant specialized metabolites (PSM), can reduce methane emissions compared to ryegrass monocultures, b) to identify whether there is a linear-dose effect relationship in methane emissions from the legume or herb addition, and c) whether these effects are maintained across sampling years. Results showed that all dicot species studied, including the non-tannin-containing species, reduced methane production. The tannin-rich species, Sanguisorba minor and Lotus pedunculatus, showed the greatest methane reduction potential of up to 33%. Due to concomitant reductions in the forage digestibility, Cichorium intybus yielded the lowest methane emissions per digestible forage unit. Contrary to total gas production, methane production was less predictable, with a tendency for the lowest methane production being obtained with a 67.5% share of the legume or herb partner species. Thus, linear increments in the partner species share did not result in linear changes in methane concentration. The methane reduction potential differed across sampling years, but the species ranking in methane concentration was stable.

Evaluation of the net energy for lactation system and estimation of the energy requirements of dairy cows based on a comprehensive analysis of feeding trials.

Respiration experiments with high-yielding dairy cows in Northern Ireland have shown higher energy maintenance requirements than those used in the requirements standards of, e.g. France, UK, USA and Germany. Therefore, the current net energy for lactation (NEL) system of Germany was tested by comparing measured NEL intake with calculated NEL requirements based on a comprehensive dataset from feeding trials conducted at nine research institutions in Germany, Austria and Switzerland. The relationship between NEL requirements and NEL intake is described by the equation: N E L r e q u i r e m e n t s M J / d = 26 . 6 ± 0 . 4 + 0 . 82 ± 0 . 004 ⋅ N E L i n t a k e M J / d w i t h C o e f f i c i e n t   o f   D e t e r m i n a t i o n   R 2 = 0 . 677 , R o o t   M e a n   S q u a r e   E r r o r   R M S E   = 15 . 9   M J   N E L . The equation indicates a systematic over-estimation of NEL requirements in the lower performance range and an under-estimation at higher energy intake levels. A multiple regression analysis was conducted by calculating metabolisable energy (ME) requirements [MJ/d] using metabolic body size (MBS) [kg0.75], milk energy performance (LE) [MJ/d] and body weight change (BWC) [kg/d]: ​ ​ ​ ​ ​ ​ ​ ME intake ( MEI ) [ MJ ] =0 . 651 ( ± 0 . 004 ) ⋅ MBS+1 . 37 ( ± 0 . 006 ) ⋅ LE + 16 . 6 ( ± 0 . 31 ) ⋅ BWC with R 2 = 0. 717 , RMSE=24 . 0 MJ . These results indicate that the energy maintenance requirements are markedly higher than presumed in the feed evaluation systems commonly in use but confirm the results from Northern Ireland (0.600-0.660 MJ ME/kg0.75 MBS). ME efficiency for lactation is also higher (kL = 1/1.37 = 0.73) than that used in the systems and is also similar to the results of Northern Ireland with 0.64-0.69. The energy contribution of BWC derived by this equation is 12.1 MJ/kg (16.6 · 0.73) and distinctly lower than that of 21-25 MJ/kg presumed by the feeding standards, e.g. in Germany. Further, maintenance requirements were linked to milk yield (energy corrected milk (ECM) [kg/d]), as is practiced in the standard Australian energy system: ​ ​ ​ ​ ​ ​ ​ ( MEI ) [ MJ ] =0 . 640  + 0 . 0070 ⋅  ECM) ] ⋅ MBS+1 . 12) ⋅ LE + 16 . 7 ⋅  BWC with R 2 = 0. 719 , RMSE=24 . 0 MJ . These results demonstrate that maintenance energy requirements are partly dependent on milk yield. A differentiated analysis by stage of lactation showed that the regressions coefficients for MBS, LE and BWC change with lactation month; however, these findings apply especially to the first lactation months (i.e. in phases of intensive mobilisation).

The effect of crude protein content of the diet on renal energy losses in horses.

Renal energy losses of horses are high in comparison with other species. In the present study, more data were obtained on this parameter to improve predictive equations for renal energy losses. Four adult ponies (247-344 kg body weight [BW]) were fed with eight different diets based on first cut hay, second cut hay, early first cut fresh grass, late cut herbs-grass mix, early cut clover-grass mix, sugar beet pulp, rice bran and straw. Feed intake was measured, and urine and faeces were quantitatively collected for 3 × 12 hr at daytime and afterwards 3 × 12 hr at nighttime. Feed was analysed for crude nutrients, gross energy, amino acids and neutral-detergent-insoluble crude protein (NDICP); faeces were analysed for crude nutrients and gross energy; and urine was analysed for nitrogen (N) and gross energy. Renal energy losses per gram dry matter (DM) intake (y; kJ/g DM) were strictly correlated to protein content in DM (x; g/kg DM): y = 0.325 + 0.00431x; r2  = .81; n = 38; p < .001. The data suggest that the intercept represents energy losses by detoxification products such as hippuric acid and the regression coefficient by protein metabolites such as urea.

Relationship of Milk Odd- and Branched-Chain Fatty Acids with Urine Parameters and Ruminal Microbial Protein Synthesis in Dairy Cows Fed Different Proportions of Maize Silage and Red Clover Silage.

The aim of this study was to evaluate the relationship of milk odd- and branched-chain fatty acids (OBCFA) with urinary purine derivates and estimated ruminal microbial crude protein (MCP) synthesis. Forty-four lactating Holstein cows were used in a 4 × 4 Latin square design with 21-day periods comprised of a 13-day adaptation phase to diet followed by an 8-day sampling phase. Differences in estimated MCP yield and milk OBCFA composition were found by feeding total mixed rations containing forage (maize silage, MS; red clover silage, RCS) and concentrates (0.75:0.25) with targeted proportions of RCS to MS of 0.15:0.60, 0.30:0.45, 0.45:0.30, and 0.60:0.15 on a dry matter basis. The MCP was estimated from the total urinary purine derivate (PD) excretion (MCPPD) and intakes of metabolizable energy (MCPME) or digestible organic matter (MCPdOM). The Pearson correlations of individual OBCFA with urinary parameters (uric acid, allantoin, PD and nitrogen) were generally weak (r = -0.37 to 0.55). Yields of individual OBCFA correlated positively with MCPME and MCPdOM (r = 0.21 to 0.55). The prediction of urinary PD concentration was moderate (R2 = 0.64) when including the proportion of iso-C17:0. The prediction of total PD excretion was low (R2 = 0.21) with yields of iso-C15:0, anteiso-C17:0, and iso-C16:0. The prediction of MCPPD was high (R2 = 0.99) when including the iso-C16:0 and cis-9 C17:1 concentrations, while those of MCPME and MCPdOM were low (R2 = 0.37 and 0.36, respectively) when including yields of iso-C15:0, cis-9 C17:1, and iso-C18:0. The correlations and regression analyses demonstrate that the estimated MCP synthesis and urinary PD excretion can be only moderately predicted by yields and concentrations of individual or total OBCFA in cow's milk. However, milk OBCFA can still be seen as a promising, non-invasive method for predicting rumen function and microbial protein supply in dairy cows because MCP flow was not directly measured in this study but instead indirectly estimated probably comprising considerable deviations of the assumed values from the true ones.

Effects of abomasally infused amylase and increasing amounts of corn starch on fecal excretion of starch, total and microbial nitrogen, and volatile fatty acids in heifers1.

The aim of the present study was to study the effect of exogenous amylase on postruminal disappearance of increasing amounts of corn starch being infused into the abomasum of heifers, and to detect a possible limitation of starch digestion in the small intestine. Four rumen-fistulated heifers (2 German Black Pied and 2 Jersey × German Black Pied) with an initial BW of 565 ± 6 kg were fed 5.6 kg DM/d of a diet targeted to contain only a negligible amount of starch. Animals were assigned randomly to a crossover trial with 2 experimental periods lasting 35 d each with 10 d of diet adaption followed by 25 d of sample collection. During the sampling period, each animal was abomasally infused with native corn starch at 5 levels (953, 1,213, 1,425, 1,733, and 1,993 g DM/d) each for a 5-d period with and without exogenous amylase, respectively. At days 6 to 10 the heifers received an abomasal infusion of starch in amounts of 724 g/d. Feces were sampled 4 times a day during the collection periods. Titanium dioxide was ruminally administered (10 g/d) to estimate fecal excretion. Purine bases in feces were determined and used as a marker for microbial N excretion. Fecal excretion of microbial N increased linearly with increasing level of starch infusion (P < 0.001), indicating a constant proportion of the infused starch being fermented in the hindgut. In contrast, the apparent digestibility of starch from the total postruminal tract decreased linearly from 90% to 80% (P < 0.001) when the intestinal starch supply increased from 1 to 2 kg/d. There is strong evidence based on the increasing starch excretion with feces and the indication of a constant proportion of infused starch being fermented in the hindgut for a decreasing efficiency of starch digestion in the small intestine with increasing intestinal supply. Amylase administration increased fecal excretion of butyrate (P = 0.04) and tended to increase isovalerate excretion (P = 0.06). However, amylase did not affect fecal excretion of microbial N or starch, suggesting that pancreatic amylase activity may not be the primarily limiting factor of postruminal starch digestion in heifers when corn starch is abomasally infused in amounts up to 2 kg/d.

Accuracy and application of milk fatty acid estimation with diffuse reflectance near-infrared spectroscopy.

Near infrared spectroscopy (NIRS) has the potential to estimate contents of fatty acids (FA) in milk frequently at-farm or during daily milking routine. In this study, a total of 738 raw milk spectra collected from 33 Holstein cows over a period of 30 weeks were recorded. Reference data on FA composition in milk and in milk fat were analysed in laboratory. Calibration models were calculated for single FA and groups of FA in milk and in milk fat. Validation resulted in sufficient Ratio of Prediction to Deviation (RPD) values for some single FA and in higher RPD values for groups of FA when concentrations of FA in milk were predicted. Since the concentrations of most FA in milk are highly correlated with milk fat content, the prediction of FA contents in milk fat is more meaningful when independent predictions are intended. The accuracy of predicting single FA concentrations in milk fat is rather poor for most FA but still comparable to alternative analysing methods such as MIR analysis. The estimation of different groups of FA in milk fat resulted in an improved accuracy based on higher RPD values, which was sufficient to mirror the development in the different lactation phases. The course of cow individual long chain fatty acid (LCFA) concentration in the early lactation stage can be an indicator for body fat mobilisation. The accurate estimation of the extent and duration of body fat mobilisation in cow individuals was rather difficult with NIR predicted LCFA concentrations and would require a higher measuring frequency than applied in this study.

Effect of replacing maize silage with red clover silage in the diet on milk fatty acid composition in cows.

This study aimed to evaluate the effect of replacing maize silage plus soybean meal with red clover silage (RCS) plus wheat on the fatty acid (FA) profile in the milk fat of cows. Forty-four lactating German Holstein cows were used in a 4 × 4 Latin square design with 21-d periods composed of 13 d of adaptation to diets followed by an 8-d sampling phase. Experimental diets offered as total mixed ration consisted of a constant forage-to-concentrate ratio (75:25) with target proportions of RCS to maize silage of 15:60 (RCS15), 30:45 (RCS30), 45:30 (RCS45), and 60:15 (RCS60) on a dry matter basis. Increasing the level of RCS in the diet was accompanied by a reduction of linoleic acid content in the diet and decreased linearly the proportions of linoleic acid in the milk up to 4%. Proportions of α-linolenic acid in milk increased 2-fold with RCS60 compared with RCS15, which resulted from the linear increase in α-linolenic acid intake with incremental levels of RCS. Vaccenic acid in the milk fat was reduced by 24%. Rumenic acid, a conjugated linoleic acid (cis-9,trans-11 conjugated linoleic acid) considered to be a human health promoter, was also decreased by 22%. Reduced rumenic acid in the milk fat was probably due to a reduced amount of vaccenic acid produced in the rumen and, consequently, to the low amount of vaccenic acid to be desaturated to rumenic acid in the mammary gland by Δ9-desaturase. Oleic acid was enriched in the milk fat, although the dietary concentration of oleic acid decreased. Stearic acid proportions remained constant with increasing levels of RCS. The proportions of total polyunsaturated FA were increased by 12%, and the long-chain FA proportions increased linearly with increasing levels of RCS. Myristic acid was reduced linearly, but palmitic acid remained constant. Saturated FA was reduced linearly by 2%. Branched-chain FA, which are presumed to possess anticarcinogenic properties, were reduced to a small extent only (quadratic effect). We conclude that replacing maize silage with RCS appears to alter milk FA composition by reducing linoleic acid intake and ruminal biohydrogenation. Feeding RCS represents a strategy to increase intake of α-linolenic acid in dairy cows. However, because changes in the FA profile show positive as well as negative effects, no distinct conclusions can be drawn with regard to human health benefits.

Postruminal digestion of starch infused into the abomasum of heifers with or without exogenous amylase administration.

The effect of an exogenous amylase on postruminal digestion of starch infused into the abomasum of cattle was studied. Four rumen-cannulated heifers were fed 5.5 kg DM/d of a diet without starch, and assigned randomly to a crossover design. The experiment consisted of 2 periods lasting 23 d each with 10 d for adaptation to the diet followed by 13 d of abomasal infusion and sample collection. During the first 3 d of each infusion phase, isotonic saline solution was infused (1 liter/h) for measurement of baseline values in feces, followed by daily infusions of 880 g DM corn starch (1 kg/10 liters of water) without or with the addition of 2% of amylase. Titanium dioxide (10 g/d) was ruminally administered for estimation of fecal excretion. Digestion of starch in small intestine was calculated as the difference between the amounts of infused starch, disappeared from hindgut and fecal excretion. The apparent disappearance of starch from the hindgut was estimated based on the increment of microbial nitrogen (N) excretion due to starch infusion (1 g microbial N/100 g fermented starch) compared to baseline values. The concentration of purine bases in feces was used to estimate excretion of microbial N. Microbial N excretion increased with starch infusion (P < 0.05) but was not influenced by amylase (P = 0.81). Starch disappearance from the small intestine was not improved by amylase (P = 0.78) and averaged 85%. Amylase affected neither blood concentration of glucose (P = 0.80) nor of insulin (P = 0.26), but glucagon was lower without (P < 0.0001) than with amylase. The infusion of starch increased fecal excretion of total VFA (acetate, propionate, and butyrate) by 53% (P < 0.05), which indicates increased carbohydrate fermentation in the hindgut and incomplete digestion of starch in the small intestine. However, the excretion of total VFA was not affected by amylase (P = 0.66). Lactate excretion was higher at the second day of starch infusion (P < 0.05) without than with amylase, which suggests lower flow of starch from the small intestine to the hindgut due to a possible effect of amylase addition in animals not adapted to starch digestion. However, lactate excretion returned near to baseline values within 2 d, which was probably due to increase of lactate-utilizing bacteria and the adaptation of the microbial population in the hindgut. Further studies with higher starch levels and addition of amylase are recommended.

Replacing maize silage plus soybean meal with red clover silage plus wheat in diets for lactating dairy cows.

The objectives of this study were to evaluate the effects of replacing maize silage plus soybean meal with red clover silage (RCS) plus wheat on feed intake, diet digestibility, N partitioning, urinary excretion of purine derivatives, and milk production in dairy cows. Forty-four lactating German Holstein cows were used in a 4 × 4 Latin square design with 21-d periods composed of a 13-d adaptation phase followed by an 8-d sampling phase. Experimental diets offered as total mixed ration consisted of a constant forage-to-concentrate ratio (75:25) with targeted proportions of RCS-to-maize silage of 15:60 (RCS15), 30:45 (RCS30), 45:30 (RCS45), and 60:15 (RCS60) on a dry matter (DM) basis. Increasing the proportion of RCS plus wheat in the diet decreased linearly the intake of DM from 22.4 to 19.8 kg/d, and of organic matter from 21.1 to 18.1 kg/d. The apparent total tract digestibility (ATTD) of DM and organic matter did not differ across diets and averaged 68.4 and 70.5%, respectively. However, ATTD of N decreased linearly from 68.5 to 63.2%, whereas ATTD of neutral detergent fiber and acid detergent fiber increased linearly from 50.4 to 59.6% and from 48.4 to 57.7%, respectively, when increasing the proportion of RCS plus wheat. Fecal N excretion increased from 31.6 (RCS15) to 37.2% (RCS60) of N intake, whereas urinary N excretion was the lowest (32.8% of N intake) with RCS45. Hence, N efficiency (milk N/N intake) decreased linearly with incremental levels of RCS plus wheat, being the lowest when feeding RCS60 (25.4%), probably due to increased nonprotein N proportion in total dietary N. Urinary excretion of purine derivatives decreased linearly from 378 to 339 mmol/d, which suggests that increasing levels of RCS plus wheat reduced the microbial crude protein flow at the duodenum. Milk yield and milk protein concentration declined linearly from 35.9 to 30.2 kg/d and from 3.20 to 3.01%, respectively, when increasing the proportion of RCS plus wheat. In conclusion, caution should be taken before introducing high levels of RCS plus wheat in diets of high-yielding dairy cows. However, RCS plus wheat can be included up to 30% of the dairy cow diet (DM basis) without a reduction in lactation performance.

Effect of dietary quebracho tannin extract on milk fatty acid composition in cows.

The aim of this study was to examine the capacity of quebracho tannin extract (QTE) to modulate the fatty acid (FA) profile in the milk fat of cows. Fifty Holstein cows yielding 33.2 ± 8.2 kg/d of milk were divided into 2 groups. The cows were fed a basal diet with a forage-concentrate ratio of 66:34 on a dry matter (DM) basis. Diets tested were control (CON, basal diet without QTE) and basal diet plus 15 or 30 g of QTE/kg of DM (QTE15 and QTE30, respectively). Two treatments could be tested simultaneously and were arranged along 6 periods. The milk FA profile was characterized by increments in the proportion of linoleic (LA) and α-linolenic acid (α-LNA) (QTE15 = 10 and 6.1%; QTE30 = 28 and 25%, respectively) compared to CON, which might indicate reduced ruminal biohydrogenation (BH) of both dietary LA and α-LNA. Vaccenic acid (VA) in the milk fat was reduced (QTE15 8.9% and QTE30 12%) compared to CON, which may be linked to inhibited BH of LA and α-LNA. Rumenic acid (RA), a conjugated LA (cis-9,trans-11 conjugated linoleic acid) and an important human health promoter, was unfortunately decreased (QTE15 8.3% and QTE30 16%) in the milk compared with CON, probably because of inhibited ruminal BH of LA. However, reduced RA in the milk was probably due to reduced availability of VA produced in the rumen and the consequently low VA available to be desaturated to RA in the mammary gland by Δ9-desaturase. The proportions of total polyunsaturated FA were increased with QTE15 and QTE30 by 4.7 and 15% compared to CON, respectively, and the long-chain FA proportions were also increased (QTE15 2.0% and QTE30 8.2%). Moreover, myristic and palmitic acid were reduced by QTE30 (9.6 and 3.3%, respectively) compared to CON, which also contributed to increasing the nutritional quality of milk because they are recognized to increase high-density lipoprotein in humans. Branched-chain FA in milk was reduced with QTE treatments, which indicates inhibited ruminal BH and microbial activity. In general, our findings suggest that dietary QTE have the potential to modulate FA profile of milk fat, and this effect is dosage dependent. Because QTE influenced the FA profile of milk fat both positively and negatively, further research is needed before concluding that QTE may improve the nutritional quality of cow milk fat in human diets.

Technical note: An improved tool to insert lines for abomasal infusion in rumen cannulated cattle.

An improved tool was tested that facilitates the insertion of abomasal infusion lines in ruminally cannulated cattle for postruminal infusion studies. The insertion device was made from a 68 × 25 mm section of 20-mm i.d. stainless steel device with a weight of 125 g. The general procedure to place the tool is to pass the insertion device with the infusion line through the rumen cannula, through the sulcus omasi (reticulo-omasal orifice), and finally into the abomasum. Once inside the omasum, the insertion device is placed inside the abomasum by gently shifting the infusion line from the flexible polyvinyl chloride tubing fitted to the insertion device. The insertion device places itself by self-moving down through the abomasal orifice into the abomasum by means of its own weight. After device placement, its position can be verified by palpating through the wall of the rumen. This way, an infusion line can easily be inserted into the abomasum by any person without introducing the hand into the omasum, avoiding possible injuries to the animal and tool being expulsed of its desired position.

Effect of dietary Quebracho tannin extract on feed intake, digestibility, excretion of urinary purine derivatives and milk production in dairy cows.

The aim of this study was to evaluate the effects of dietary Quebracho tannin extract (QTE) on feed intake, apparent total tract digestibility (ATTD), excretion of urinary purine derivatives (PD) and milk composition and yield in dairy cows. Fifty Holstein cows were divided into two groups. To reach a similar performance of both groups, cows were divided according to their milk yield, body weight, days in milk and number of lactations at the start of the experiment averaging 33.2 ± 8.2 kg/d, 637 ± 58 kg, 114 ± 73 d and 2.3 ± 1.6 lactations, respectively. The cows were fed a basal diet as total mixed ration containing on dry matter (DM) basis 34% grass silage, 32% maize silage and 34% concentrate feeds. Three dietary treatments were tested, the control (CON, basal diet without QTE), QTE15 (basal diet with QTE at 15 g/kg DM) and QTE30 (basal diet with QTE at 30 g/kg DM). Two treatments were arranged along six periods each 21 d (13 d adaptation phase and 8 d sampling phase). The ATTD of DM and organic matter were reduced only in Diet QTE30, whereas both QTE treatments reduced ATTD of fibre and nitrogen (N), indicating that QTE impaired rumen fermentation. Nevertheless, feed intake was unaffected by QTE. In Diet CON, urinary N excretion accounted for 29.8% of N intake and decreased in treatments QTE15 and QTE30 to 27.5% and 17.9%, respectively. Daily faecal N excretion increased in treatments CON, QTE15 and QTE30 from 211 to 237 and 273 g/d, respectively, which amounted to 39.0%, 42.4% and 51.7% of the N intake, respectively. Hence, QTE shifted N excretion from urine to faeces, whereas the proportion of ingested N appearing in milk was not affected by QTE (average 30.7% of N intake). Daily PD excretion as indicator for microbial crude protein (CP) flow at the duodenum decreased in treatment QTE30 compared with Diet CON from 413 to 280 mmol/d. The ratios of total PD to creatinine suggest that urinary PD excretion was already lower when feeding Diet QTE15. While there was no effect of Diet QTE15, treatment QTE30 reduced milk yield, milk fat and protein. Both QTE treatments reduced milk urea concentration, which suggest that ruminal degradation of dietary CP was reduced. In summary, adding QTE at dosages of 15 and 30 g/kg DM to diets of lactating dairy cows to improve feed and protein use efficiency is not recommended.

Response of urinary purine derivatives excretion to different levels of ruminal glucose infusion in heifers.

This study investigated the response of urinary purine derivatives (PD) excretion to increasing levels of intraruminal glucose infusion to evaluate how well this indicator reflects induced changes in microbial crude protein flow. Four rumen-cannulated heifers (482 ± 25 kg body weight) were fed at maintenance energy level with a basal diet (on fresh matter basis) of 4 kg/d hay, 1.5 kg/d concentrate and 60 g/d minerals in two equal meals. The trial comprised a control period (Control I) without glucose infusion followed by four consecutive periods in which all animals received 125 g, 250 g, 500 g or 1000 g/d of glucose, respectively. For this, daily dosages of glucose and urea (90 g/d during all periods) were divided into three portions that were dissolved in water and directly administered into the rumen during morning and afternoon feedings and once during noon. After the highest glucose dosage, a second control period was carried out (Control II). Urinary PD excretion increased with glucose infusion of 125 g/d (71.4 mmol/d) and 1000 g/d (74.2 mmol/d) over the level at Control I (53.9 mmol/d (standard error of the mean (SEM) 3.4; p = 0.012). After withdrawing glucose infusion, PD excretion (79.0 mmol/d) did not return to Control I level (p = 0.001). In contrast, faecal nitrogen (N) excretions linearly increased with incremental glucose infusion (p < 0.001) from 33.9 g/d at Control I to 39.7 g/d (SEM 0.5) at 1000 g/d of glucose and were similar in Control I and II (p = 0.086). The contradicting responses in the excretions of faecal N and urinary PD to increasing glucose infusions highlight the limited accuracy of the PD excretion as a non-invasive indicator when incremental dosages of rapidly fermentable carbohydrates are supplied.

Bioavailability of free lysine and protein-bound lysine from casein and fishmeal in juvenile turbot (Psetta maxima).

In the present study, a linear regression analysis between lysine intake and lysine retention was conducted to investigate the efficiency of lysine utilisation (k(Lys)) at marginal lysine intake of either protein-bound or free lysine sources in juvenile turbot (Psetta maxima). For this purpose, nine isonitrogenous and isoenergetic diets were formulated to contain 2·25-4·12 g lysine/100 g crude protein (CP) to ensure that lysine was the first-limiting amino acid in all diets. The basal diet contained 2·25 g lysine/100 g CP. Graded levels of casein (Cas), fishmeal (FM) and L-lysine HCl (Lys) were added to the experimental diets to achieve stepwise lysine increments. A total of 240 fish (initial weight 50·1 g) were hand-fed all the experimental diets once daily until apparent satiation over a period of 56 d. Feed intake was significantly affected by dietary lysine concentration rather than by dietary lysine source. Specific growth rate increased significantly at higher lysine concentrations (P< 0·001). CP, crude lipid and crude ash contents in the whole body were affected by the dietary treatments. The linear regression slope between lysine retention and lysine intake (k(Lys)) was similar between all the dietary lysine sources. The k(Lys) values for the diets supplemented with Cas, Lys or FM were 0·833, 0·857 and 0·684, respectively. The bioavailability of lysine from the respective lysine sources was determined by a slope-ratio approach. The bioavailability of lysine (relative to the reference lysine source Cas) from FM and Lys was 82·1 and 103 %, respectively. Nutrient requirement for maintenance was in the range of 16·7-23·4 mg/kg(0·8) per d, and did not differ between the treatments. There were no significant differences in lysine utilisation efficiency or bioavailability of protein-bound or crystalline lysine from the respective sources observed when lysine was confirmed to be the first-limiting nutrient.