Plasma ghrelin and oxytomodulin concentrations in lactating dairy cows receiving abomasal soybean oil, corn starch, and casein infusions

The effects of increased postruminal supply of casein, corn starch, and soybean oil on plasma concentrations of the gastrointestinal hormones ghrelin and oxyntomodulin (OXM) were investigated. Four mid-lactation Holstein cows were used in a 4×4 Latin square. Treatments were continuous abomasal infusions (23h/d) for 7 d of water, soybean oil (500g/d), corn starch (1100g/d), or casein (800g/d). Jugular vein plasma was obtained every 30min for 7h on days 1 and 7. Soybean oil and casein infusion decreased preprandial plasma ghrelin concentration by approximately 20% on both d (time-by-treatment P<0.10); however, dry matter intake (DMI) was depressed only after 7 d of oil infusion. Infusion of soybean oil, corn starch, or casein did not change the plasma OXM concentration (P>0.20). The present data indicate that plasma ghrelin concentration is depressed immediately before feeding by the postruminal infusion of soybean oil and casein, but it is not affected during the postprandial period. Plasma ghrelin concentration was not altered (P>0.20), pre- or postfeeding, by increased postruminal supply of corn starch. In addition, plasma OXM concentration did not respond (P>0.20) to postruminal nutrient infusion. In conclusion, a decrease in DMI when fat is infused could be partially explained by the decrease in prefeeding plasma ghrelin concentration, but a decrease in prefeeding plasma ghrelin concentration is not always associated with a decrease in DMI, as observed for the infusion of casein. Plasma OXM concentration was not affected by postruminal infusion of macronutrients.

Effect of abomasal glucose infusion on plasma concentrations of gut peptides in periparturient dairy cows

Six Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in the portal vein, hepatic vein, mesenteric vein, and an artery were used to study the effects of abomasal glucose infusion on splanchnic plasma concentrations of gut peptides. The experimental design was a randomized block design with repeated measurements. Cows were assigned to one of 2 treatments: control or infusion of 1,500 g of glucose/d into the abomasum from the day of parturition to 29 d in milk. Cows were sampled 12 ± 6 d prepartum and at 4, 15, and 29 d in milk. Concentrations of glucose-dependent insulinotropic polypeptide, glucagon-like peptide 1(7–36) amide, and oxyntomodulin were measured in pooled samples within cow and sampling day, whereas active ghrelin was measured in samples obtained 30 min before and after feeding at 0800 h. Postpartum, dry matter intake increased at a lower rate with infusion compared with the control. Arterial, portal venous, and hepatic venous plasma concentrations of the measured gut peptides were unaffected by abomasal glucose infusion. The arterial, portal venous, and hepatic venous plasma concentrations of glucose-dependent insulinotropic polypeptide and glucagon-like peptide 1(7–36) amide increased linearly from 12 d prepartum to 29 d postpartum. Plasma concentrations of oxyntomodulin were unaffected by day relative to parturition. Arterial and portal venous plasma concentrations of ghrelin were lower postfeeding compared with prefeeding concentrations. Arterial plasma concentrations of ghrelin were greatest prepartum and lowest at 4 d postpartum, giving a quadratic pattern of change over the transition period. Positive portal venous-arterial and hepatic venous–arterial concentration differences were observed for glucagon-like peptide 1(7–36) amide. A negative portal venous–arterial concentration difference was observed for ghrelin pre-feeding. The remaining portal venous–arterial and hepatic venous–arterial concentration differences of gut peptides did not differ from zero. In conclusion, increased postruminal glucose supply to postpartum transition dairy cows reduced feed intake relative to control cows, but did not affect arterial, portal venous, or hepatic venous plasma concentrations of gut peptide hormones. Instead, gut peptide plasma concentrations increased as lactation progressed. Thus, the lower feed intake of postpartum dairy cows receiving abomasal glucose infusion was not attributable to changes in gut peptide concentrations.

Effects of ammonia treatment and undegradable protein supplementation on nutrient digestion of sheep fed on wheat straw based diets.

A study was conducted to investigate the effects of wheat straw ammonisation and supplementation with a rumen undegradable protein (UDP) source on nutrient digestion and nitrogen balance by lambs while diets were supplemented with kibbled carob pods as energy source. Ammonisation increased the crude protein content of wheat straw by nearly 100% and decreased the contents of neutral detergent fibre and acid detergent fibre by 7% and 1.7% respectively. Treating the straw with ammonia resulted in significant (P<0.01) increase in nitrogen (N) intake and intakes of organic matter (OM) and dry matter (DM) tended toward significance (P<0.1). The UDP source had no effect (P>0.05) on DM and OM intakes but resulted in an increase (P<0.05) of N intakes. Both, ammonization and UDP supplementation increased (P<0.01) the DM, OM and N digestibility. In conclusion, the results of this study suggest that ammonisation and UDP supplementation is a practical dietary manipulation option to improve the nutritional status of ruminants fed on roughage-based diets.

The competitive ability of pea-barley intercrops against weeds and the interactions with crop productivity and soil N availability

Grain legumes, such as peas (Pisum sativum L.), are known to be weak competitors against weeds when grown as the sole crop. In this study, the weed-suppression effect of pea–barley (Hordeum vulgare L.)intercropping compared to the respective sole crops was examined in organic field experiments across Western Europe (i.e., Denmark, the United Kingdom, France, Germany and Italy). Spring pea (P) and barley(B) were sown either as the sole crop, at the recommended plant density (P100 and B100, respectively), or in replacement (P50B50) or additive (P100B50)intercropping designs for three seasons (2003–2005). The weed biomass was three times higher under the pea sole crops than under both the intercrops and barley sole crops at maturity. The inclusion of joint experiments in several countries and various growing conditions showed that intercrops maintain a highly asymmetric competition over weeds, regardless of the particular weed infestation (species and productivity), the crop biomass or the soil nitrogen availability. The intercropping weed suppression was highly resilient, whereas the weed suppression in pea sole crops was lower and more variable. The pea–barley intercrops exhibited high levels of weed suppression, even with a low percentage of barley in the total biomass. Despite a reduced leaf area in the case of a low soil N availability, the barley sole crops and intercrops displayed high weed suppression, probably because of their strong competitive capability to absorb soil N. Higher soil N availabilities entailed increased leaf areas and competitive ability for light, which contributed to the overall competitive ability against weeds for all of the treatments. The contribution of the weeds in the total dry matter and soil N acquisition was higher in the pea sole crop than in the other treatments, in spite of the higher leaf areas in the pea crops.

Effect of wheat dwarfing genes on nitrogen-use efficiency

Near isogenic lines (NILs) varying for alleles for reduced height (Rht) and photoperiod insensitivity (Ppd-D1a) in a cvar Mercia background (rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht8c+Ppd-D1a, Rht-D1c, Rht12) were compared at a field site in Berkshire, UK, but within different systems (‘organic’, O, in 2005/06, 2006/07 and 2007/08 growing seasons v. ‘conventional’, C, in 2005/06, 2006/07, 2007/08 and 2008/09). In 2007 and 2008, further NILs (rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht-B1b+Rht-D1b, Rht-D1b+Rht-B1c) in both Maris Huntsman and Maris Widgeon backgrounds were added. The contrasting systems allowed NILs to be tested in diverse rotational and agronomic, but commercially relevant, contexts, particularly with regard to the assumed temporal distribution of nitrogen availability, and competition from weeds. For grain, nitrogen-use efficiency (NUE; grain dry matter (DM) yield/available N; where available N=fertilizer N+soil mineral N), recovery of N in the grain (grain N yield/available N), N utilization efficiency to produce grain (NUtEg; grain DM yield/above-ground crop N yield), N harvest index (grain N yield/above-ground crop N yield) and dry matter harvest index (DMHI; grain DM yield/above-ground crop DM yield) all peaked at final crop heights of 800–950 mm. Maximum NUE occurred at greater crop heights in the organic system than in the conventional system, such that even adding just a semi-dwarfing allele (Rht-D1b) to the shortest background, Mercia, reduced NUE in the organic system. The mechanism of dwarfing (gibberellin sensitive or insensitive) made little difference to the relationship between NUE and its components with crop height. For above-ground biomass: dwarfing alleles had a greater effect on DM accumulation compared with N accumulation such that all dwarfing alleles could reduce nitrogen utilization efficiency (NUtE; crop DM yield/crop N yield). This was particularly evident at anthesis in the conventional system when there was no significant penalty for severe dwarfism for N accumulation, despite a 3-tonne (t)/ha reduction in biomass compared to the tallest lines. Differences between genotypes for recovery of N in the grain were thus mostly a function of net N uptake after anthesis rather than of remobilized N. This effect was compounded as dwarfing, except when coupled with Ppd-D1a, was associated with delayed anthesis. In the organic experiments there was greater reliance on N accumulated before anthesis, and genotype effects on NUE were confounded with effects on N accumulated by weeds, which was negatively associated with crop height. Optimum height for maximizing wheat NUE and its components, as manipulated by Rht alleles, thus depend on growing system, and crop utilization (i.e. biomass or grain production).

Simulating the effects of grassland management and grass ensiling on methane emission from lactating cows

A dynamic, mechanistic model of enteric fermentation was used to investigate the effect of type and quality of grass forage, dry matter intake (DMI) and proportion of concentrates in dietary dry matter (DM) on variation in methane (CH(4)) emission from enteric fermentation in dairy cows. The model represents substrate degradation and microbial fermentation processes in rumen and hindgut and, in particular, the effects of type of substrate fermented and of pH oil the production of individual volatile fatty acids and CH, as end-products of fermentation. Effects of type and quality of fresh and ensiled grass were evaluated by distinguishing two N fertilization rates of grassland and two stages of grass maturity. Simulation results indicated a strong impact of the amount and type of grass consumed oil CH(4) emission, with a maximum difference (across all forage types and all levels of DM 1) of 49 and 77% in g CH(4)/kg fat and protein corrected milk (FCM) for diets with a proportion of concentrates in dietary DM of 0.1 and 0.4, respectively (values ranging from 10.2 to 19.5 g CH(4)/kg FCM). The lowest emission was established for early Cut, high fertilized grass silage (GS) and high fertilized grass herbage (GH). The highest emission was found for late cut, low-fertilized GS. The N fertilization rate had the largest impact, followed by stage of grass maturity at harvesting and by the distinction between GH and GS. Emission expressed in g CH(4)/kg FCM declined oil average 14% with an increase of DMI from 14 to 18 kg/day for grass forage diets with a proportion of concentrates of 0.1, and on average 29% with an increase of DMI from 14 to 23 kg/day for diets with a proportion of concentrates of 0.4. Simulation results indicated that a high proportion of concentrates in dietary DM may lead to a further reduction of CH, emission per kg FCM mainly as a result of a higher DM I and milk yield, in comparison to low concentrate diets. Simulation results were evaluated against independent data obtained at three different laboratories in indirect calorimetry trials with COWS consuming GH mainly. The model predicted the average of observed values reasonably, but systematic deviations remained between individual laboratories and root mean squared prediction error was a proportion of 0.12 of the observed mean. Both observed and predicted emission expressed in g CH(4)/kg DM intake decreased upon an increase in dietary N:organic matter (OM) ratio. The model reproduced reasonably well the variation in measured CH, emission in cattle sheds oil Dutch dairy farms and indicated that oil average a fraction of 0.28 of the total emissions must have originated from manure under these circumstances.

Characterization of a highly biodiverse floodplain meadow using hyperspectral remote sensing within a plant functional trait framework

We assessed the potential for using optical functional types as effective markers to monitor changes in vegetation in floodplain meadows associated with changes in their local environment. Floodplain meadows are challenging ecosystems for monitoring and conservation because of their highly biodiverse nature. Our aim was to understand and explain spectral differences among key members of floodplain meadows and also characterize differences with respect to functional traits. The study was conducted on a typical floodplain meadow in UK (MG4-type, mesotrophic grassland type 4, according to British National Vegetation Classification). We compared two approaches to characterize floodplain communities using field spectroscopy. The first approach was sub-community based, in which we collected spectral signatures for species groupings indicating two distinct eco-hydrological conditions (dry and wet soil indicator species). The other approach was “species-specific”, in which we focused on the spectral reflectance of three key species found on the meadow. One herb species is a typical member of the MG4 floodplain meadow community, while the other two species, sedge and rush, represent wetland vegetation. We also monitored vegetation biophysical and functional properties as well as soil nutrients and ground water levels. We found that the vegetation classes representing meadow sub-communities could not be spectrally distinguished from each other, whereas the individual herb species was found to have a distinctly different spectral signature from the sedge and rush species. The spectral differences between these three species could be explained by their observed differences in plant biophysical parameters, as corroborated through radiative transfer model simulations. These parameters, such as leaf area index, leaf dry matter content, leaf water content, and specific leaf area, along with other functional parameters, such as maximum carboxylation capacity and leaf nitrogen content, also helped explain the species’ differences in functional dynamics. Groundwater level and soil nitrogen availability, which are important factors governing plant nutrient status, were also found to be significantly different for the herb/wetland species’ locations. The study concludes that spectrally distinguishable species, typical for a highly biodiverse site such as a floodplain meadow, could potentially be used as target species to monitor vegetation dynamics under changing environmental conditions.

Contrasting effects of dwarfing alleles and nitrogen availability on mineral concentrations in wheat grain

Background and aim Concentrations of essential minerals in plant foods may have declined in modern high-yielding cultivars grown with large applications of nitrogen fertilizer (N). We investigated the effect of dwarfing alleles and N rate on mineral concentrations in wheat. Methods Gibberellin (GA)-insensitive reduced height (Rht) alleles were compared in near isogenic wheat lines. Two field experiments comprised factorial combinations of wheat variety backgrounds, alleles at the Rht-B1 locus (rht-B1a, Rht-B1b, Rht-B1c), and different N rates. A glasshouse experiment also included Rht-D1b and Rht-B1b+D1b in one background. Results In the field, depending on season, Rht-B1b increased crop biomass, dry matter (DM) harvest index, grain yield, and the economically-optimal N rate (Nopt). Rht-B1b did not increase uptake of Cu, Fe, Mg or Zn so these minerals were diluted in grain. Nitrogen increased DM yield and mineral uptake so grain concentrations were increased (Fe in both seasons; Cu, Mg and Zn in one season). Rht-B1b reduced mineral concentrations at Nopt in the most N responsive season. In the glasshouse experiment, grain yield was reduced, and mineral concentrations increased, with Rht allele addition. Conclusion Effects of Rht alleles on Fe, Zn, Cu and Mg concentrations in wheat grain are mostly due to their effects on DM, rather than of GA-insensitivity on Nopt or mineral uptake. Increased N requirement in semi-dwarf varieties partly offsets this dilution effect.

The possibility of lowering oil content of potato chips by combining atmospheric frying with postfrying vacuum application

Four protocols involving the application of low pressures, either toward the end of frying or after frying, were investigated with the aim of lowering the oil content of potato chips. Protocol 1 involving frying at atmospheric pressure followed by a 3 min draining time constituted the control. Protocol 2 involved lowering of pressure to 13.33 kPa, 40 s before the end of frying, followed by draining for 3 min at the same pressure. Protocol 3 was the same as protocol 2, except that the pressure was lowered 3 s before the end of frying. Protocol 4 involved lowering the pressure to 13.33 kPa after the product was lifted from the oil and holding it at this value over the draining time of 3 min. Protocol 4 gave a product having the lowest oil content (37.12 g oil/100 g defatted dry matter), while protocol 2 gave the product with highest oil content (71.10 g oil/100 g defatted dry matter), followed by those obtained using protocols 1 and 3(68.48 g oil/100 g defatted dry matter and 52.50 g oil/100 g defatted dry matter, respectively). Protocol 4 was further evaluated to study the effects of draining times and vacuum applied, and compared with the control. It was noted that over the modest range of pressures investigated, there was no significant effect of the vacuum applied on the oil content of the product. This study demonstrates that the oil content of potato chips can be lowered significantly by combining atmospheric frying with draining under vacuum.

Synergistic effects of mixing cocksfoot and sainfoin on in vitro rumen fermentation: role of condensed tannins

Some bioactive secondary metabolites in forage legumes can cause digestive interactions, so that the rumen fermentation pattern of a mixture of forages can differ from the average values of its components. The objective of this study was to investigate the potential role of condensed tannins (CT) on the synergistic effects between one grass species, cocksfoot, and one CT-containing legume species, sainfoin, on in vitro rumen fermentation characteristics. Cocksfoot and sainfoin in different proportions (in g/kg, 1000:0, 750:250, 500:500, 250:750 and 0:1000) were incubated under anaerobic conditions in culture bottles containing buffered rumen fluid from sheep. Incubations were carried out using artificial saliva with and without polyethylene glycol (PEG), which binds and thus inactivates CT. Rumen fermentation parameters describing the degradation and the fate of the energetic and nitrogenous substrates were measured at 3.5 and 24 h. At the early fermentation stage, when the sainfoin level increased from 0 to 1000 g/kg, the ammonia concentration in the medium quadratically decreased from 3.20 to 0.53 mmol/l in absence of PEG (P<0.01) but not in its presence. This result demonstrates that sainfoin CT decreased the rumen degradation of the proteins in the whole mixture, including the proteins in cocksfoot, rather than just the proteins in sainfoin. Interestingly, the total gas and methane productions were lower in mixtures incubated in absence of PEG than in presence of PEG (P<0.001) while no significant PEG effect was observed on digestibility. At the late fermentation stage, a positive quadratic effect on dry matter digestibility was detected without PEG (P<0.05), indicating a synergistic action of cocksfoot plus sainfoin on plant substrate degradation due to CT. The presence of PEG increased gas production (P<0.001) and NH3-N concentration in the medium (P<0.001). Our results suggest that CT could allow a better utilization of plant substrates in mixtures by the rumen ecosystem by improving the partitioning of degraded substrates toward lower gas losses, and decreasing the protein degradation.

Incremental effect of a calcium salt of cis-monounsaturated fatty acids supplement on milk fatty acid composition in cows fed maize silage-based diets

In most Western countries, saturated fatty acid (SFA) intake exceeds recommended levels, which is considered a risk factor for cardiovascular disease (CVD). As milk and dairy products are major contributors to SFA intake in many countries, recent research has focused on sustainable methods of producing milk with a lower saturated fat concentration by altering dairy cow diets. Human intervention studies have shown that CVD risk can be reduced by consuming dairy products with reduced SFA and increased cis-monounsaturated fatty acid (MUFA) concentrations. This milk fatty acid profile can be achieved by supplementing dairy cow diets with cis-MUFA-rich unsaturated oils. However, rumen exposure of unsaturated oils also leads to enhanced milk trans fatty acid (TFA) concentrations. Because of concerns about the effects of TFA consumption on CVD, feeding strategies that increase MUFA concentrations in milk without concomitant increases in TFA concentration are preferred by milk processors. In an attempt to limit TFA production and increase the replacement of SFA by cis-MUFA, a preparation of rumen-protected unsaturated oils was developed using saponification with calcium salts. Four multiparous Holstein-Friesian cows in mid-late lactation were used in a 4 × 4 Latin square design with 21-d periods to investigate the effect of incremental dietary inclusion of a calcium salt of cis-MUFA product (Ca-MUFA; 20, 40, and 60 g/kg of dry matter of a maize silage-based diet), on milk production, composition, and fatty acid concentration. Increasing Ca-MUFA inclusion reduced dry matter intake linearly, but no change was observed in estimated ME intake. No change in milk yield was noted, but milk fat and protein concentrations were linearly reduced. Supplementation with Ca-MUFA resulted in a linear reduction in total SFA (from 71 to 52 g/100 g of fatty acids for control and 60 g/kg of dry matter diets, respectively). In addition, concentrations of both cis- and trans-MUFA were increased with Ca-MUFA inclusion, and increases in other biohydrogenation intermediates in milk fat were also observed. The Ca-MUFA supplement was very effective at reducing milk SFA concentration and increasing cis-MUFA concentrations without incurring any negative effects on milk and milk component yields. However, reduced milk fat and protein concentrations, together with increases in milk TFA concentrations, suggest partial dissociation of the calcium salts in the rumen

Evidence of a universal scaling relationship for leaf CO2 drawdown along an aridity gradient

The leaf carbon isotope ratio (δ13C) of C3 plants is inversely related to the drawdown of CO2 concentration during photosynthesis, which increases towards drier environments. We aimed to discriminate between the hypothesis of universal scaling, which predicts between-species responses of δ13C to aridity similar to within-species responses, and biotic homoeostasis, which predicts offsets in the δ13C of species occupying adjacent ranges. The Northeast China Transect spans 130–900 mm annual precipitation within a narrow latitude and temperature range. Leaves of 171 species were sampled at 33 sites along the transect (18 at ≥ 5 sites) for dry matter, carbon (C) and nitrogen (N) content, specific leaf area (SLA) and δ13C. The δ13C of species generally followed a common relationship with the climatic moisture index (MI). Offsets between adjacent species were not observed. Trees and forbs diverged slightly at high MI. In C3 plants, δ13C predicted N per unit leaf area (Narea) better than MI. The δ13C of C4 plants was invariant with MI. SLA declined and Narea increased towards low MI in both C3 and C4 plants. The data are consistent with optimal stomatal regulation with respect to atmospheric dryness. They provide evidence for universal scaling of CO2 drawdown with aridity in C3 plants.

Bio-fortification of potato tubers using foliar zinc-fertiliser

Worldwide, many people are zinc (Zn)-deficient. Dietary Zn intake can be increased by producing crops with higher concentrations of Zn in their edible portions. This can be achieved by applying Zn-fertilisers to varieties with an increased ability to acquire Zn and to accumulate Zn in their edible portions. Potato (Solanum tuberosum L.) is an important food crop and is, therefore, a target for bio-fortification with Zn. Field trials incorporating a core collection of 23 potato genotypes, performed over 4 years (2006 – 2009), indicated significant genotypic effects on tuber Zn concentration and suggested that tuber Zn concentration was influenced by environmental effects, but also found that genotype environment (G E) interactions were not significant. Tuber Zn concentrations averaged 10.8 mg kg–1 dry matter (DM), and the ratio between the lowest and the highest varietal tuber Zn-concentration averaged 1.76. Tuber Zn concentrations could be increased by foliar Zn-fertilisation. Tuber yields of ‘Maris Piper’ were unaffected by foliar applications of < 1.08 g Zn plant–1. The relationship between tuber Zn concentration and foliar Zn application followed a saturation curve, reaching a maximum at approx. 30 mg Zn kg–1 DM at a foliar Zn application rate of 1.08 g plant–1. Despite a 40-fold increase in shoot Zn concentration compared to the unfertilised controls following foliar Zn fertilisation with 2.16 g Zn plant–1, only a doubling in tuber Zn concentration was observed. This suggests that the biofortification of tubers with Zn was restricted by the limited mobility of Zn in the phloem. A significant positive linear relationship between tuber Zn concentration and tuber N concentration supported the hypothesis of co-transport of Zn and N-compounds in the phloem.

Using rain-use efficiency to explore livestock production trends in rangelands in the Transkei, South Africa

We qualitatively describe the condition of communally managed rangelands in the Transkei, South Africa, using GIS and high resolution near-infrared imagery. Using livestock census data from 28 magisterial districts in the Transkei, we explored the trends in livestock biomass from 1923–1998. The area had been subjected to intensive herbivory by domestic livestock during that period, and the high livestock biomass had been blamed for the perceived degradation or ‘overgrazing’ of the region. Our assessment used the concept rain-use efficiency (RUE) (kg dry matter ha–1 mm–1) to determine whether there is evidence of change in the efficiency of the system to produce domestic livestock. We calculated RUE from annual livestock numbers and the mean annual rainfall for each district. We found no evidence of a decline in rain-use efficiency between the two assessment periods (1923–1944, 1945–1998). There was evidence of a shift in the ratio of sheep to goats between 1923 and 1998, with goat numbers increasing (greater than twofold) relative to sheep in eight districts. This trend may be associated with changes in the structure of vegetation. We conclude that this region is not showing evidence of system run down that affects domestic livestock production.

Short communication: plasma concentration of glucose-dependent insulinotropic polypeptide may regulate milk energy production in lactating dairy cows

In dairy cows, an increase in plasma concentration of glucose-dependent insulinotropic polypeptide (GIP) is associated with an increase in metabolizable energy intake, but the role of GIP in energy partitioning of dairy cattle is not certain. The objective of this study was to examine the relationship between plasma GIP concentrations and energy partitioning toward milk production. Four mid-lactation, primiparous, rumenfistulated Holstein-Friesian cows were fed a control diet of 55% forage and 45% concentrate [dry matter (DM) basis] in a 4 × 4 Latin square design with 4-wk periods. The 4 treatments were (1) control diet fed at 1000 and 1600 h, and (2) once-daily (1000 h) feeding, (3) twice daily (1000 and 1600 h) feeding, and (4) 4 times/d (1000, 1600, 2200 and 0400 h) feeding of the control diet plus 1 dose (1.75 kg on a DM basis at 0955 h) into the rumen of supplemental vegetable proteins (Amino Green; SCA NuTec Ltd., Thirsk, UK). Measurements of respiratory exchange and energy balance were obtained over 4 d during the last week of each period while cows were housed in open-circuit respiration chambers. Blood was collected from the jugular vein every 30 min for 12 h, using indwelling catheters, starting at 0800 h on d 20 of each period. Plasma GIP concentration was measured in samples pooled over each 5 consecutive blood samplings. The relationships between plasma GIP, DM intake, heat production, respiratory quotient, milk yield, and milk energy output were analyzed using linear correlation procedures, with metabolizable intake as a partial variant. Plasma GIP concentration was not correlated with heat production, or milk yield, but was positively correlated with milk energy yield (correlation coefficient = 0.67) and negatively correlated with RQ (correlation coefficient = −0.72). The correlations between GIP and RQ and milk energy output do not imply causality, but suggest that a role for GIP may exist in the regulation of energy metabolism in dairy cows.