Combination of milk variables and on-farm data as an improved diagnostic tool for metabolic status evaluation in dairy cattle during the transition period.

Milk composition, particularly milk fatty acids, has been extensively studied as an indicator of the metabolic status of dairy cows during early lactation. In addition to milk biomarkers, on-farm sensor data also hold potential in providing insights into the metabolic health status of cows. While numerous studies have explored the collection of a wide range of sensor data from cows, the combination of milk biomarkers and on-farm sensor data remains relatively underexplored. Therefore, this study aims to identify associations between metabolic blood variables, milk variables, and various on-farm sensor data. Second, it seeks to examine the supplementary or substitutive potential of these data sources. Therefore, data from 85 lactations on metabolic status and on-farm data were collected during 3 wk before calving up to 5 wk after calving. Blood samples were taken on d 3, 6, 9, and 21 after calving for determination of ß-hydroxybutyrate (BHB), nonesterified fatty acids (NEFA), glucose, insulin-like growth factor-1 (IGF-1), insulin, and fructosamine. Milk samples were taken during the first 3 wk in lactation and analyzed by mid-infrared for fat, protein, lactose, urea, milk fatty acids, and BHB. Walking activity, feed intake, and body condition score (BCS) were monitored throughout the study. Linear mixed effect models were used to study the association between blood variables and (1) milk variables (i.e., milk models); (2) on-farm data (i.e., on-farm models) consisting of activity and dry matter intake analyzed during the dry period ([D]) and lactation ([L]) and BCS only analyzed during the dry period ([D]); and (3) the combination of both. In addition, to assess whether milk variables can clarify unexplained variation from the on-farm model and vice versa, Pearson marginal residuals from the milk and on-farm models were extracted and related to the on-farm and milk variables, respectively. The milk models had higher coefficient of determination (R2) than the on-farm models, except for IGF-1 and fructosamine. The highest marginal R2 values were found for BHB, glucose, and NEFA (0.508, 0.427, and 0.303 vs. 0.468, 0.358, and 0.225 for the milk models and on-farm models, respectively). Combining milk and on-farm data particularly increased R2 values of models assessing blood BHB, glucose, and NEFA concentrations with the fixed effects of the milk and on-farm variables mutually having marginal R2 values of 0.608, 0.566, and 0.327, respectively. Milk C18:1 was confirmed as an important milk variable in all models, but particularly for blood NEFA prediction. On-farm data were considerably more capable of describing the IGF-1 concentration than milk data (marginal R2 of 0.192 vs. 0.086), mainly due to dry matter intake before calving. The BCS [D] was the most important on-farm variable in relation to blood BHB and NEFA and could explain additional variation in blood BHB concentration compared with models solely based on milk variables. This study has shown that on-farm data combined with milk data can provide additional information concerning the metabolic health status of dairy cows. On-farm data are of interest to be further studied in predictive modeling, particularly because early warning predictions using milk data are highly challenging or even missing.

Prediction of first test day milk yield using historical records in dairy cows.

The transition between two lactations remains one of the most critical periods during the productive life of dairy cows. In this study, we aimed to develop a model that predicts the milk yield of dairy cows from test day milk yield data collected in the previous lactation. In the past, data routinely collected in the context of herd improvement programmes on dairy farms have been used to provide insights in the health status of animals or for genetic evaluations. Typically, only data from the current lactation is used, comparing expected (i.e., unperturbed) with realised milk yields. This approach cannot be used to monitor the transition period due to the lack of unperturbed milk yields at the start of a lactation. For multiparous cows, an opportunity lies in the use of data from the previous lactation to predict the expected production of the next one. We developed a methodology to predict the first test day milk yield after calving using information from the previous lactation. To this end, three random forest models (nextMILKFULL, nextMILKPH, and nextMILKP) were trained with three different feature sets to forecast the milk yield on the first test day of the next lactation. To evaluate the added value of using a machine-learning approach against simple models based on contemporary animals or production in the previous lactation, we compared the nextMILK models with four benchmark models. The nextMILK models had an RMSE ranging from 6.08 to 6.24 kg of milk. In conclusion, the nextMILK models had a better prediction performance compared to the benchmark models. Application-wise, the proposed methodology could be part of a monitoring tool tailored towards the transition period. Future research should focus on validation of the developed methodology within such tool.

Effects of a dietary supplement enriched in palmitoleic acid on fatty acid composition of follicular fluid, granulosa cell metabolism, and oocyte developmental capacity in early lactation dairy cows.

In high-yielding dairy cows, some fertility traits can be influenced by the fatty acid (FA) composition of the follicular fluid during early lactation. The first objective of the current study was to evaluate the potential of dietary supplements enriched in specific FA to influence the FA composition of follicular fluid lipid classes in early lactation dairy cows. The second objective was to determine the influence of the resulting follicular fluid FA composition on the folliculogenesis, lipid and energy metabolism of granulosa cells, as well as oocyte quality and embryo development. Twenty Holstein multiparous cows in late gestation were randomly assigned to 200 g/d of FA supplements enriched in (1) palmitic acid (control treatment; 82% 16:0; PA) in the rumen or (2) palmitoleic acid (sea buckthorn oil; 27% cis-9 16:1, 28% 16:0, 22% cis-9 18:1, and 11% cis-9,cis-12 18:2; SBT) in the abomasum. The treatment period ranged from 20 ± 5 d precalving to 67 ± 2 d postcalving. Cumulus-oocyte complexes, granulosa cells, and follicular fluid were recovered from 2 sequential sessions of ovum pick-up (OPU-1 and OPU-2) at 46 and 67 ± 2 d postcalving (mean ± standard deviation). On the same days, blood samples were collected. Milk performance was recorded, and feed and milk samples were collected from d 8 to 10 ± 3 (onset of lactation), d 35 to 37 ± 2 (before OPU-1), and d 63 to 65 ± 2 (before OPU-2). Treatments did not affect milk yield or fat concentration throughout the experimental trial. Compared with PA, SBT increased the cis-9 16:1 concentration in milk fat, in plasma esterified lipid classes (phospholipids, cholesterol esters, and triacylglycerols), and in follicular fluid phospholipids and cholesterol esters at OPU-1. Abundance of mRNA for stearoyl-CoA desaturase 1 and 5, and perilipin 2 in granulosa cells was not different between treatments, but an increase in the level of stearoyl-CoA desaturase 5 was observed between the 2 OPU periods. Treatments did not affect oocyte quality and developmental capacity or embryo lipid metabolism when cultivated in vitro. These results suggest that limited modifications in the FA composition of the oocyte microenvironment via dietary lipid supplements enriched in specific FA had no major effects on granulosa cell metabolism and oocyte developmental capacity in early lactation cows.

Invited review: Role of rumen biohydrogenation intermediates and rumen microbes in diet-induced milk fat depression: An update.

To meet the energy requirements of high-yielding dairy cows, grains and fats have increasingly been incorporated in ruminant diets. Moreover, lipid supplements have been included in ruminant diets under experimental or practical conditions to increase the concentrations of bioactive n-3 fatty acids and conjugated linoleic acids in milk and meat. Nevertheless, those feeding practices have dramatically increased the incidence of milk fat depression in dairy cattle. Although induction of milk fat depression may be a management tool, most often, diet-induced milk fat depression is unintended and associated with a direct economic loss. In this review, we give an update on the role of fatty acids, particularly originating from rumen biohydrogenation, as well as of rumen microbes in diet-induced milk fat depression. Although this syndrome seems to be multi-etiological, the best-known causal factor remains the shift in rumen biohydrogenation pathway from the formation of mainly trans-11 intermediates toward greater accumulation of trans-10 intermediates, referred to as the trans-11 to trans-10 shift. The microbial etiology of this trans-11 to trans-10 shift is not well understood yet and it seems that unraveling the microbial mechanisms of diet-induced milk fat depression is challenging. Potential strategies to avoid diet-induced milk fat depression are supplementation with rumen stabilizers, selection toward more tolerant animals, tailored management of cows at risk, selection toward more efficient fiber-digesting cows, or feeding less concentrates and grains.

Distinct blood and milk 18-carbon fatty acid proportions and buccal bacterial populations in dairy cows differing in reticulorumen pH response to dietary supplementation of rapidly fermentable carbohydrates.

Nine Holstein dairy cows were fed diets with increasing proportions of rapidly fermentable carbohydrates (RFCH) to investigate the effect on reticular pH, milk fat content (MFC), 18-carbon fatty acid proportions in blood plasma and milk, and bacterial community in buccal swab samples. Inter-animal variation was expected in terms of reticular pH response upon higher RFCH proportions, which would be reflected in the occurrence or not of milk fat depression (MFD). Moreover, this variation in occurrence of MFD was hypothesized to be related to differences in blood and milk fatty acid proportions and in the bacterial community in buccal samples. Cows were fed a total mixed ration throughout the experiment, which consisted of 4 periods: adaptation (d 0-4) and low (d 5-18), increasing (d 19-24), and high RFCH (d 25-28). During the increasing RFCH period, the standard concentrate (211 g of starch/kg of dry matter) was gradually and partly replaced by a concentrate high in RFCH (486 g of starch/kg of dry matter). The reticular pH was measured using a bolus and the time below pH 6.00 was calculated on a daily basis. On d 13, 14, 25, 27, and 28, plasma and milk samples were collected and analyzed for 18-carbon fatty acid proportions, and buccal swabs were collected for bacterial community analysis based on 16S rRNA gene amplicon sequencing. Inter-animal variation was observed in terms of reticular pH, which allowed us to divide the cows into 2 groups: tolerant (time below pH 6.00 ≤ 0.1 h/d) and susceptible cows (time below pH 6.00 ≥ 1.26 h/d). The lower reticular pH of susceptible cows was accompanied by lower MFC. Both groups already differed in reticular pH and MFC during the low-RFCH period. Furthermore, higher RFCH amounts did not decrease the reticular pH in either of the 2 groups. Nevertheless, MFD was observed in both groups during the high-RFCH period compared with the low-RFCH period. Lower MFC in animals with lower reticular pH or during the high-RFCH period was associated with a shift in 18-carbon fatty acids toward trans-10 at the expense of trans-11 intermediates, which was observed in plasma as well as in milk samples. Moreover, lower MFC was accompanied by shifts in the relative abundance of specific bacteria in buccal samples. Genera Dialister, Sharpea, Carnobacterium, Acidaminococcus, and uncultured genera belonging to the Betaproteobacteria were more abundant in situations with greater trans-10 proportions.

Reducing enteric methane emissions from dairy cattle: Two ways to supplement 3-nitrooxypropanol.

The aim of this work was to determine the effect of 3-nitrooxypropanol (3-NOP) on the enteric methane (CH4) emissions and performance of lactating dairy cows when mixed in with roughage or incorporated into a concentrate pellet. After 2 pretreatment weeks without 3-NOP supplementation, 30 Holstein Friesian cows were divided into 3 homogeneous treatment groups: no additive, 3-NOP mixed in with the basal diet (roughage; NOPbas), and 3-NOP incorporated into a concentrate pellet (NOPconc). The pretreatment period was followed by a 10-wk treatment period in which the NOPbas and NOPconc cows were fed 1.6 g of 3-NOP/cow per day. After the treatment period, a 2-wk washout period followed without 3-NOP supplementation. The CH4 emissions were measured using a GreenFeed unit (C-Lock Inc., Rapid City, SD) installed in a freestall with cubicles during the entire experimental period. On average for the total treatment period and compared with the no-additive group, CH4 production (g/d) was 28 and 23% lower for NOPbas and NOPconc, respectively. Methane yield (g/kg of dry matter intake) and methane intensity (g/kg of milk) were 23 and 24% lower for NOPbas, respectively, and 21 and 22% lower for NOPconc, respectively. No differences were found between NOPbas and NOPconc. Moreover, supplying 3-NOP did not affect total dry matter intake, milk production, or milk composition. The results of this experiment show that 3-NOP can reduce enteric CH4 emissions of dairy cattle when incorporated into a concentrate pellet and that this reduction is not different from the effect of mixing in 3-NOP with the basal diet (roughage). This broadens the possibilities for using 3-NOP in the dairy sector worldwide, as it is not always feasible to provide an additive mixed in with the basal diet.

Supplementation of DHA-Gold pre and/or postnatally to goat kids modifies in vitro methane production and rumen morphology until 6 mo old.

This study aimed to investigate the effect of pre and/or postnatal supplementation of a dry whole cell algae (DHA-Gold) to goat kids, on in vitro methane (CH4) production, animal growth, and rumen morphology at the age of 6 mo. Furthermore, the in vitro retreatment effect of DHA-Gold was evaluated. Twenty pregnant Saanen goats giving birth to 2 male kids were used. Half of these does were supplemented (D+) with 18.2 g/d of DHA-Gold in the last 3 wk of pregnancy, whereas the other half was not (D-). After kidding, one goat kid per doe in both groups was supplemented daily with 0.28 g/kg of body weight of DHA-Gold (k+) until 12 wk, whereas the other goat kids were untreated (k-). This resulted in 4 experimental groups D+k+, D+k-, D-k+, and D-k-. In vitro incubations were performed at the ages of 4 wk, 11 wk, and 6 mo. At the age of 6 mo, goat kids were euthanized and additional incubations were performed supplementing 4 doses of DHA-Gold (0, 0.4, 0.8, and 1.6 mg/mL). Additionally, rumen tissue of the atrium ruminis, ventral rumen, and dorsal blind sac were collected to assess rumen morphology. Rumen inocula of 4-wk-old goat kids supplemented D+ showed lower (P < 0.05) in vitro CH4 production, however, this was mainly due to a reduction in the overall fermentation, while CH4 expressed relatively to total volatile fatty acids (VFA) was higher when goat kids were treated D+ or k+. The detrimental D+ effect on VFA production diminished at 11 wk old but remained a tendency (0.05 < P < 0.1). As for 4 wk D+ as well as k+ supplementation of DHA-Gold stimulated rather than inhibited in vitro CH4 production expressed relative to total VFA. Supplementation of DHA-Gold either D+ or k+ decreased density, width, and surface area of the ruminal papillae. However, no effect on animal growth was observed. Moreover, detrimental effects of D+ or k+ treatment on VFA production or stimulation of relative CH4 production were no longer observed at 6 mo old. Nevertheless, direct exposure of DHA-Gold to 6-mo-old inoculum linearly (P < 0.05) decreased CH4 and VFA production, which tended (P = 0.06) to be greater when using D-rumen inoculum. Accordingly, neither D+ nor k+ DHA-Gold supplementation showed potential for reduction of rumen methanogenesis. Furthermore, this early life intervention could represent some risk for impaired rumen papillae development, which, however, did not impair animal performance.

Promising perspectives for ruminal protection of polyunsaturated fatty acids through polyphenol-oxidase-mediated crosslinking of interfacial protein in emulsions.

Previously, polyunsaturated fatty acids (PUFA) from linseed oil were effectively protected (>80%) against biohydrogenation through polyphenol-oxidase-mediated protein crosslinking of an emulsion, prepared with polyphenol oxidase (PPO) extract from potato tuber peelings. However, until now, emulsions of only 2 wt% oil have been successfully protected, which implies serious limitations both from a research perspective (e.g. in vivo trials) as well as for further upscaling toward practical applications. Therefore, the aim of this study was to increase the oil/PPO ratio. In the original protocol, the PPO extract served both an emulsifying function as well as a crosslinking function. Here, it was first evaluated whether alternative protein sources could replace the emulsifying function of the PPO extract, with addition of PPO extract and 4-methylcatechol (4MC) to induce crosslinking after emulsion preparation. This approach was then further used to evaluate protection of emulsions with higher oil content. Five candidate emulsifiers (soy glycinin, gelatin, whey protein isolate (WPI), bovine serum albumin and sodium caseinate) were used to prepare 10 wt% oil emulsions, which were diluted five times (w/w) with PPO extract (experiment 1). As a positive control, 2 wt% oil emulsions were prepared directly with PPO extract according to the original protocol. Further, emulsions of 2, 4, 6, 8 and 10 wt% oil were prepared, with 80 wt% PPO extract (experiment 2), or with 90, 80, 70, 60 and 50 wt% PPO extract, respectively (experiment 3) starting from WPI-stabilized emulsions. Enzymatic crosslinking was induced by 24-h incubation with 4MC. Ruminal protection efficiency was evaluated by 24-h in vitro batch simulation of the rumen metabolism. In experiment 1, protection efficiencies were equal or higher than the control (85.5% to 92.5% v. 81.3%). In both experiments 2 and 3, high protection efficiencies (>80%) were achieved, except for emulsions containing 10 wt% oil emulsions (<50% protection), which showed oiling-off after enzymatic crosslinking. This study demonstrated that alternative emulsifier proteins can be used in combination with PPO extract to protect emulsified PUFA-rich oils against ruminal biohydrogenation. By applying the new protocol, 6.5 times less PPO extract was required.

Polyunsaturated fatty acids are less effective to reduce methanogenesis in rumen inoculum from calves exposed to a similar treatment early in life.

The aim of this study was to evaluate the dose response on in vitro methane (CH) production of PUFA to which the inoculum donor animals had been exposed early in life. Sixteen Holstein calves (160 ± 3 and 365 ± 2 kg BW) at 6 and 12 mo of age were used as inoculum donors. Half of the calves were given increasing amounts of extruded linseed from birth (22 g/d) until 4 mo of age (578 g/d) first mixed with milk and then included in their concentrate. Linseed oil (LSO) was supplemented in vitro at 5 different doses (0, 0.6, 1.2, 2.4, and 4.8 mg/mL). Supplementation of LSO in the rumen inocula at both ages linearly decreased ( < 0.05) the in vitro CH production. Total in vitro VFA production was not affected by LSO supplementation. Inhibition of CH was smaller when using the rumen inoculum from calves that had received a similar treatment early in life ( < 0.05). Differences in response to in vitro supplementation of a type of fatty acids similar to those applied during early life suggest some "changes" in the functioning of the rumen microbial community.

Postruminal synthesis modifies the odd- and branched-chain fatty acid profile from the duodenum to milk.

Milk odd- and branched-chain fatty acids (OBCFA) have been suggested as potential biomarkers for rumen function. The potential of milk OBCFA as a biomarker depends on whether their profile reflects the profile observed in the duodenum. The objective of this study was to evaluate whether the OBCFA profile in duodenum samples is reflected in plasma and milk. For this, 2 dairy cattle experiments were used. In experiment 1, 4 Holstein cows fitted with rumen and proximal duodenum cannulas were used in a 4×4 Latin square design. The treatments consisted of 2 nitrogen levels (143 vs. 110g of crude protein/kg of dry matter for high and low N, respectively) combined with either 1 of the 2 energy sources (i.e., starch from barley, corn, and wheat or fiber from soybean hulls and dehydrated beet pulp). In experiment 2, 4 Holstein cows fitted with rumen and proximal duodenum cannulas were used in a 3×3 Latin square design, with the treatments consisting of 3 diets: (1) RNB-, a diet with a crude protein content of 122g/kg of dry matter, predicted to provide protein digested in the small intestine according to the requirement of the animals, but with a shortage of rumen degradable protein; (2) RNB- to which 6g/d of niacin was added through inclusion in the mineral and vitamin premix, and (3) RNB- to which urea was added to balance rumen degradable N supply resulting in a CP content of 156g/kg of dry matter. In both experiments, samples of duodenal digesta, plasma, and milk were collected and analyzed for fatty acids. Additionally, lipids in plasma samples were separated in lipid classes and analyzed for fatty acids. The OBCFA profile in milk was enriched in 15:0, iso-17:0, anteiso-17:0, and cis-9-17:1 as compared with duodenal samples, and milk secretions even exceeded duodenal flows, which suggests occurrence of postruminal synthesis, such as de novo synthesis, desaturation, and elongation. The postruminal modification of the OBCFA profile might hamper the application of OBCFA as diagnostic tools of rumen function.

Performance, bioenergetic status, and indicators of oxidative stress of environmentally heat-loaded Holstein cows in response to diets inducing milk fat depression.

Effects of grain type and dietary oil supplement on production performance, energy balance, metabolic heat production, and markers of liver function of heat-loaded lactating dairy cows were evaluated using 8 multiparous Holstein cows (77.0d in milk) in a duplicated 4×4 Latin square design with a 2×2 factorial arrangement of treatments. Experimental diets contained either ground barley or ground corn supplemented with either fish oil or soybean oil at 2% of dietary dry matter. Mean daily maximum temperature, minimum relative humidity, and maximum temperature-humidity index were 35.3°C, 11.3%, and 77.0, respectively. Dietary treatment did not affect rectal temperature (38.9°C), but respiration rate tended to decrease in cows fed fish oil versus soybean oil. Dry matter intake decreased for the fish oil-supplemented diets (21.1 vs. 24.3kg/d), which was negatively correlated with plasma concentrations of alkaline phosphatase (r=-0.45; n=32) and malondialdehyde (r=-0.26; n=32). Actual milk yield (41.9kg/d) and energy-corrected milk yield (36.6kg/d) were not affected by grain type, whereas feeding fish oil decreased milk yield as compared with soybean oil (40.4 vs. 43.4kg/d). Milk fat depression occurred in all dietary treatments, especially when cows were fed fish oil because of the presence of polyunsaturated FA in the diets. trans-10 C18:1 was negatively correlated with milk fat yield (r=-0.38; n=32). Daily milk cis-9,trans-11 C18:2 secretion was 29.6% less in cows fed barley- versus corn-based diets but 31.8% greater in cows fed fish oil as compared with cows fed soybean oil. Because of a lower dry matter intake, metabolic heat production was decreased in cows fed fish oil relative to cows fed soybean oil. Although feeding fish oil versus soybean oil decreased net energy for both maintenance and lactation, net energy balance remained unchanged across treatments. In vivo plasma lipoperoxidation was greater in cows fed fish oil versus soybean oil, which substantiated increased susceptibility of plasma lipoperoxidation when cows were fed fish oil. Plasma concentration of malondialdehyde was positively correlated with plasma aspartate aminotransferase (r=0.38; n=32), which is an indicator of liver function in heat-loaded cows. Results suggest that in heat-loaded cows fed diets supplemented with soybean oil versus fish oil, biosynthesis in the mammary gland was prioritized over anabolism and oxidation in peripheral adipose and muscle tissues regardless of type of grain used.

Protection of polyunsaturated oils against ruminal biohydrogenation and oxidation during storage using a polyphenol oxidase containing extract from red clover.

Polyunsaturated fatty acid (PUFA) are to a large extent subject to biohydrogenation in a ruminal environment, which results to the healthy value of these PUFA being lost upon dietary addition to ruminants. PUFA are also prone to lipid oxidation upon storage. Therefore, it was tested whether emulsions could be protected against in vitro ruminal biohydrogenation and oxidation during storage by using protein extracts rich in polyphenol oxidase, an enzyme responsible for browning of plant tissues. PUFA rich emulsions were made with a protein extract from red clover (Trifolium pratense L.) before adding a synthetic diphenol (4-methylcatechol) to induce protection. Results after in vitro incubation confirmed the hypothesis and indicated the potential to prevent PUFA in linseed or fish oil from ruminal biohydrogenation and oxidation during storage through addition of 4-methylcatechol to the emulsions. Protection depended on the amount of oil present and protein concentrations in the emulsions. Protection efficiency increased with increasing the amounts of diphenol present in the emulsion per unit interfacial surface area. It is suggested that protection is caused by an effective encapsulation by cross-linking of the protein layer at the emulsion interface. For the first time, a method is described to protect PUFA using an enzyme abundantly available in nature, polyphenol oxidase, in combination with 4-methylcatechol.

A simulated mucus layer protects Lactobacillus reuteri from the inhibitory effects of linoleic acid.

Lactobacillus reuteri is a commensal, beneficial gut microbe that colonises the intestinal mucus layer, where it makes close contact with the human host and may significantly affect human health. Here, we investigated the capacity of linoleic acid (LA), the most common polyunsaturated fatty acid (PUFA) in a Western-style diet, to affect L. reuteri ATCC PTA 6475 prevalence and survival in a simulated mucus layer. Short-term (1 h) survival and mucin-agar adhesion assays of a log-phase L. reuteri suspension in intestinal water demonstrated that the simulated mucus layer protected L. reuteri against the inhibitory effects of LA by lowering its contact with the bacterial cell membrane. The protective effect of the simulated mucus layer was further evaluated using a more complex and dynamic model of the colon microbiota (SHIME®), in which L. reuteri survival was monitored during 6 days of daily exposure to LA in the absence (L-SHIME) and presence (M-SHIME) of a simulated mucus layer. After 6 days, luminal L- and M-SHIME L. reuteri plate counts had decreased by 3.1±0.5 and 2.6±0.9 log cfu/ml, respectively. Upon supplementation of 1.0 g/l LA, the decline in the luminal L. reuteri population started earlier than was observed for the control. In contrast, mucin-agar levels of L. reuteri (in the M-SHIME) remained unaffected throughout the experiment even in the presence of high concentrations of LA. Overall, the results of this study indicate the importance of the mucus layer as a protective environment for beneficial gut microbes to escape from stress by high loads of the antimicrobial PUFA LA to the colon, i.e. due to a Western-style diet.

Effect of supplementing coconut or krabok oil, rich in medium-chain fatty acids on ruminal fermentation, protozoa and archaeal population of bulls.

Medium-chain fatty acids (MCFA), for example, capric acid (C10:0), myristic (C14:0) and lauric (C12:0) acid, have been suggested to decrease rumen archaeal abundance and protozoal numbers. This study aimed to compare the effect of MCFA, either supplied through krabok (KO) or coconut (CO) oil, on rumen fermentation, protozoal counts and archaeal abundance, as well as their diversity and functional organization. KO contains similar amounts of C12:0 as CO (420 and 458 g/kg FA, respectively), but has a higher proportion of C14:0 (464 v. 205 g/kg FA, respectively). Treatments contained 35 g supplemental fat per kg DM: a control diet with tallow (T); a diet with supplemental CO; and a diet with supplemental KO. A 4th treatment consisted of a diet with similar amounts of MCFA (i.e. C10:0+C12:0+C14:0) from CO and KO. To ensure isolipidic diets, extra tallow was supplied in the latter treatment (KO+T). Eight fistulated bulls (two bulls per treatment), fed a total mixed ration predominantly based on cassava chips, rice straw, tomato pomace, rice bran and soybean meal (1.5% of BW), were used. Both KO and CO increased the rumen volatile fatty acids, in particular propionate and decreased acetate proportions. Protozoal numbers were reduced through the supplementation of an MCFA source (CO, KO and KO+T), with the strongest reduction by KO. Quantitative real-time polymerase chain reaction assays based on archaeal primers showed a decrease in abundance of Archaea when supplementing with KO and KO+T compared with T and CO. The denaturing gradient gel electrophoresis profiles of the rumen archaeal population did not result in a grouping of treatments. Richness indices were calculated from the number of DGGE bands, whereas community organization was assessed from the Pareto-Lorenz evenness curves on the basis of DGGE band intensities. KO supplementation (KO and KO+T treatments) increased richness and evenness within the archaeal community. Further research including methane measurements and productive animals should elucidate whether KO could be used as a dietary methane mitigation strategy.

The logistic curve as a tool to describe the daily ruminal pH pattern and its link with milk fatty acids.

Daily ruminal pH variation can be summarized by a cumulative logistic curve based on the amount of time below multiple pH points and characterized by 2 parameters (ß(0) and ß(1)). Moreover, rumen pH variation affects the rumen microbiome as well as the biohydrogenation pathways resulting in a modified secretion of milk fatty acids (FA). The aims of this study were to assess the shifts in milk FA due to rumen pH changes and to estimate the relationship between milk FA and the 2 parameters of the logistic curve. The data consisted of milk samples of 2 experiments. In experiment 1, 3 cows were subjected to 5 treatments in which the type and amount of concentrate were changed during 33 d: (1) control diet 1, (2) stepwise replacement of a standard concentrate (CONC) by a CONC rich in rapidly fermentable carbohydrates, (3) increase in the total amount of CONC, (4) treatment with a buffer solution, and (5) control diet 2. A 3×3 Latin square design with 3 cows was used in the second experiment. During the first 14 d of each period, the cows received a control diet with a standard CONC, whereas in the last 7 d the standard CONC was replaced step-by-step by a CONC rich in rapidly fermentable carbohydrates and the amount of CONC was increased. During each period, a different buffer treatment was added to the diet. Milk FA and pH reacted similarly in both experiments: decreasing proportions of iso FA and increasing proportions of odd-chain FA were observed. However, an abrupt change to a 76% CONC diet as for one cow of experiment 1 led to almost a 10-fold increase in C18:1 trans-10 (0.79 vs. 6.75 g/100g of FA). In experiment 2, the stepwise approach of adding CONC and the continuous supplementation of buffer led to minimal increases in C18:1 trans-10 and decreases in rumen pH compared with the diet with standard CONC only. Fatty acid proportions were influenced by the level of rumen pH (ß(1)) or the rumen pH variation (ß(0)), or both. High proportions of C18:1 trans-10 (above 4 g/100g of FA) occurred with low and largely fluctuating pH (low ß(1), low ß(0)), whereas situations with low, stable pH (low ß(1), great ß(0)) did not induce a shift toward the secondary biohydrogenation pathway. C18:1 trans-11 and C18:2 cis-9, trans-11 were only influenced by the pH variation and not by the average pH, whereas iso C14:0 and iso C16:0 FA were only dependent on the average pH and not influenced by diurnal pH variation. Overall, milk FA changes were related to pH changes; however, this relationship is not straightforward and needs further research.

Influence of a marine algae supplementation on the oxidative status of plasma in dairy cows during the periparturient period.

This study was part of a larger study that addressed the effects of marine algae (ALG) supplementation in the ration of high yielding periparturient dairy cows. The objectives were to induce milk fat depression (MFD) in early lactation by feeding docosahexaenoic acid (DHA) from ALG and to determine the effects on milk production, milk components and metabolic status early post partum. This study focuses on the oxidative status in the plasma during the ALG supplementation. Plasma samples were collected from 16 Holstein Friesian cows at the day of parturition and at -1, 2, 4 and 6 weeks relative to calving with half of the cows receiving the ALG supplement (44gDHA/d) from 3 weeks pre partum on. The following parameters were measured in plasma: ferric reducing ability of plasma (FRAP), α-tocopherol level, glutathione peroxidase activity (GSH-Px) and thiobarbituric acid reactive substances (TBARS) concentration. There was a significant effect of time for FRAP and α-tocopherol indicating changes in the plasma oxidative status around parturition. The ALG supplementation was successful in creating a milk fat depression (MFD) but could not improve the energy balance. Feeding of ALG significantly increased lipid peroxidation as measured by TBARS, probably through their high content of n-3 polyunsaturated fatty acids.

The effect of marine algae in the ration of high-yielding dairy cows during transition on metabolic parameters in serum and follicular fluid around parturition.

Sixteen Holstein cows were assigned to 2 groups to evaluate the caloric and metabolic effect of feeding marine algae (ALG) from 3 wk prepartum until 12 wk postpartum. Milk production characteristics and the profiles of hormones and metabolites in the serum were monitored from -7 to 46 d in milk (DIM) and in follicular fluid (FF) from 14 to 46 DIM. All cows received a corn- and grass silage-based partially mixed ration supplemented with concentrate and protein supplement. In the diet of the ALG group, 2 kg of the concentrate was replaced by a concentrate containing ALG (44 g/d of docosahexaenoic acid). Diets were isocaloric (net energy basis) and equal in intestinal digestible protein. The ALG diet increased milk yield (41.2 vs. 38.2 kg/d) and decreased milk fat yield (1.181 vs. 1.493 kg/d) and milk fat content (31.6 vs. 40.7 g/kg). Protein yield (1.336 vs. 1.301 kg/d) was not affected but a tendency toward decreased milk protein content (32.8 vs. 34.7 g/kg) was observed. Marine algae supplementation increased the ß-hydroxybutyric acid (BHBA) concentration in FF of the ALG cows compared with that in the controls (0.992 vs. 0.718 mmol/L). The total protein concentration in FF was decreased in ALG (62.9 vs. 67.6 g/L). Plasma and serum metabolites did not significantly differ between treatments except for a tendency toward a lower concentration of urea in the serum of the control compared with ALG (4.69 vs. 5.13 mmol/L). Based on metabolizable energy calculations, a daily energy-sparing effect of 3.48 Mcal was obtained due to milk fat depression (MFD). The concomitant increase in milk yield suggests that at least part of this spared energy is used to stimulate milk production. Theoretically, 3.48 Mcal of ME could lead to an increase in milk yield of 7.43 kg/d, which is higher than the observed 3 kg/d. However, when evaluating nutrient requirements during MFD in early lactation, we calculated that increased milk production is caused by a propionate-saving effect of 2.71 mol in the udder when milk fat is depressed. Concurrent increased BHBA concentrations in FF in the ALG group cannot be attributed to a worsened energy status of the animals because all other indicators contradict any change in energy balance, indicating that BHBA might not be an appropriate metabolic parameter to estimate the energy balance in early lactating dairy cows during MFD.

Monitoring of vitamin E status of dry, early and mid-late lactating organic dairy cows fed conserved roughages during the indoor period and factors influencing forage vitamin E levels.

Analysis of blood plasma of 60 cows selected on six Flemish organic dairy farms revealed that on average 12% of all samples and on one farm up to 28% of the organic cows showed plasma vitamin E concentrations less than 3.0 µg/ml, which is considered the minimum level to avoid health risks due to vitamin E shortages. Furthermore, this study showed early lactating and dry cows to be more at risk in relation to animals in mid-late lactation. In European organic farming, vitamin supplements are only allowed if granted by the local authority to satisfy daily requirements. Therefore, the vitamin E content of the feedstuffs used on the farms was determined. Grass clover silage (GCS) and mixed silage had significantly more vitamin E than hay, maize or grain (p < 0.05) [mean (SD): 52 (35), 29 (20), 4.5 (1.7), 4.9 (4.4) and 7.1 (3.8) mg/kg DM, respectively]. Apparently, variation in the vitamin E content in the silage samples was huge. Hence, the vitamin E content of ryegrass, white and red clover was determined in a second lab scale experiment and the effects of wilting, DM content and supplementation of ensiling additives were investigated. Fresh ryegrass had a higher vitamin E content than white and red clover (p < 0.05) [156 (11.3), 49.3 (0.67) and 74.3 (5.73) µg/g DM, respectively]. These differences remained after the wilting or ensiling. Supplementation of formic acid or lactic acid bacteria at ensiling had no significant effect on the vitamin E content. Overall, it can be concluded that GCS is the most important source of vitamin E in organic dairy farming. A legal possibility for case-related supplementation should be retained in organic dairy farming as approximately 18% of all dry and early lactating cows were at risk of vitamin E shortage.

Effect of dietary starch or micro algae supplementation on rumen fermentation and milk fatty acid composition of dairy cows.

Two experiments with rumen-fistulated dairy cows were conducted to evaluate the effects of feeding docosahexaenoic acid (DHA; C22:6 n-3)-enriched diets or diets provoking a decreased rumen pH on milk fatty acid composition. In the first experiment, dietary treatments were tested during 21-d experimental periods in a 4 x 4 Latin square design. Diets included a control diet, a starch-rich diet, a bicarbonate-buffered starch-rich diet, and a diet supplemented with DHA-enriched micro algae [Schizochytrium sp., 43.0 g/kg of dry matter intake (DMI)]. Algae were supplemented directly through the rumen fistula. The total mixed ration consisted of grass silage, corn silage, soybean meal, and a standard or glucogenic concentrate. The glucogenic and buffered glucogenic diet had no effect on rumen fermentation and milk fatty acid composition because, unexpectedly, no reduced rumen pH was detected. The algae diet had no effect on rumen pH but provoked decreased butyrate and increased isovalerate molar proportions in the rumen. In addition, algae supplementation affected rumen biohydrogenation of linoleic and linolenic acid as reflected in the modified milk fatty acid composition toward increased conjugated linoleic acid (CLA) cis-9 trans-11, CLA trans-9 cis-11, C18:1 trans-10, C18:1 trans-11, and C22:6 n-3 concentrations. Concomitantly, on average, a 45% decrease in DMI and milk yield was observed. Based on these drastic and impractical results, a second animal experiment was performed for 20 d in which 9.35 g/kg of total DMI of algae were incorporated in the concentrate and supplemented to 3 rumen-fistulated cows. Algae concentrate feeding increased rumen pH, which was associated with decreased rumen short-chain fatty acid concentrations. Moreover, a different shift in rumen short-chain fatty acid proportions was observed compared with the first experiment because molar proportions of butyrate, isobutyrate, and isovalerate increased, whereas acetate molar proportion decreased. The milk fatty acid profile changed as in experiment 1. However, the decrease in DMI and milk yield was less pronounced (on average 10%) at this algae supplementation level, whereas milk fat percentage decreased from 47.9 to 22.0 g/kg of milk after algae treatment. In conclusion, an algae supplementation level of about 10 g/kg of DMI proved effective to reduce the milk fat content and to modify the milk fatty acid composition toward increased CLA cis-9 trans-11, C18:1 trans, and DHA concentrations.