Effect of carbohydrate sources and cotton seed meal in the concentrate: II. Feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in beef cattle

Four, rumen fistulated crossbred (Brahman × native) beef cattle steers were randomly assigned to receive four dietary treatments according to a 2 × 2 factorial arrangement in a 4 × 4 Latin square design. Factor A was carbohydrate source; cassava chip (CC) or CC + rice bran at a ratio 3:1 (CR3:1) and Factor B was cotton seed meal level (CM); and 109 g CP/kg (LCM) and 328 g CP/kg (HCM) at similar overall CP levels (490 g CP/kg). The animals were fed 5 g concentrate/kg BW, and urea-treated rice straw (UTS) (50 g urea/kg DM) was fed ad libitum. Carbohydrate source did not affect feed intake, nutrient digestibility, blood urea nitrogen, rumen fermentation, or microbial protein synthesis; however, animals fed with CC had a higher population of total viable bacteria than the CR3:1 treatment (P < 0.05). Animals that received HCM had a lower total feed intake while ruminal pH was higher than the LCM fed treatment (P < 0.05). The population of total viable and cellulolytic bacteria in animals that received HCM were lower than the LCM fed treatment (P < 0.05). Moreover, use of HCM in beef cattle diets resulted in lower microbial protein synthesis when compared with the LCM fed treatment (P < 0.05) although efficiency of microbial protein synthesis was nonsignificantly different among treatments. Therefore, cassava chip combined with rice bran can be used in a concentrate diet for beef cattle. It is also noted that a high level of cotton seed meal in the concentrate may impact rumen fermentation and animal performance.     

Effect of brewer's grain on rumen fermentation, milk production and milk composition in lactating dairy cows

Six lactating Holstein cows were divided into two groups (n = 3) and used in a double reversal trial with three periods of 14 days each to evaluate the rumen fermentation, milk production and milk composition of cows fed brewer's grain (BG). The control diets contained 14% chopped Sudangrass hay, 24% corn silage, 18% alfalfa hay cube, 34% concentrate mixture‐1 and 10% concentrate mixture‐2 (wheat bran, soybean meal and cottonseed). In the experimental diet, wet BG replaced the concentrate mixture‐2. The protozoal population, concentration of ammonia‐N and volatile fatty acids in the ruminal fluid did not differ between the control and BG diets. The molar percentage of acetic acid was significantly higher (P < 0.05) with the BG diet at 5 h after feeding. The milk yield, the percentage of protein, lactose, solids not‐fat and somatic cell counts of milk did not differ between the two diets. The percentage of milk fat tended to increase with the BG diet. The BG diet significantly increased the proportions of C18:0 and C18:1 in milk fat (P < 0.01, P < 0.05, respectively) and tended to increase that of conjugated linoleic acid.     

Theoretical turnover rate of the rumen liquid fraction in dairy cows and its relationship to feed intake,rumen fermentation,and milk production

Ruminant animals are able to convert plant materials (grain and the human‐indigestible portion of carbohydrates) to milk and meat. In this conversion, most of the plant materials are digested by rumen fermentation and are changed to short‐chain fatty acids, microbial cells, and methane, which is released into the atmosphere. The relationships among feed, rumen fermentation, and milk production are poorly understood. Here we report a novel indicator of characteristics of rumen fermentation, theoretical turnover rate (TTOR) of the rumen liquid fraction. The TTOR was calculated from the presumed rumen volume (PRV) which is estimated by dividing the methane yield by the methane concentration of rumen fluid. The formula for the TTOR is: TTOR = PRV/body weight^0.75. Our present analyses confirm that the TTOR as an indicator is capable of connecting feed, rumen fermentation, and milk production, because dry matter intake/TTOR showed a strong correlation with milk yield/TTOR. In addition, the TTOR may be related to ruminal pH, as we observed that the ruminal pH decreased as the TTOR increased. We propose that the TTOR is a factor characterizing rumen fermentation and a good indicator of the productivity of ruminants and dysbiosis of the rumen microbiome.     

Effect of propylene glycol and undegradable protein source on rumen fermentation, blood metabolism and milk production in lactating dairy cows

Eight lactating Holstein cows were divided into two groups ( n  = 4) and used in a double reversal trial with three periods of 14 days each, to evaluate diets containing propylene glycol (PG) and ruminally undegradable protein (RUP) blend on milk yield and composition, ruminal fermentation and blood metabolism. The control diet contained 20% chopped Sudangrass hay, 20% cubed alfalfa hay, 12% corn silage, and 48% of the respective concentrate mixtures (dry matter basis). The experimental diet (PG + RUP) partially replaced the concentrate mixture from the control diet with 1.4% PG and 2.1% RUP. Both diets contained about 16% crude protein and 71% total digestible nutrients. Dry matter intake was similar between the two diets. Daily production of milk, milk lactose and milk solids-not-fat increased by 9.0%, 11.3% and 9.3%, respectively ( P  < 0.1), for cows fed diets with PG + RUP; milk composition was unchanged. Although the concentration of ruminal total volatile fatty acids was unchanged, the proportion of propionic acid increased, and the proportion of acetic acid decreased with PG + RUP. The concentration of ruminal ammonia nitrogen and number of ciliate protozoa was not significantly affected by PG + RUP. The concentration of glucose in blood plasma increased, the concentration of urea nitrogen was unaffected, and the concentration of some essential free amino acids decreased with PG + RUP. It is suggested that these changes might be caused predominantly by PG, and the addition of PG may exert a favorable effect on milk production through increased metabolism.     

Terpenes and carbohydrate source influence rumen fermentation, digestibility, intake, and preference in sheep

We hypothesized that toxins and nutrients in foods interact to influence foraging behavior by herbivores. Based on this hypothesis we predicted that 1) terpenes in big sagebrush (Artemisia tridentata) influence intake and preference in sheep for diets varying in sources of nonstructural (barley grain) and structural (sugar beet pulp) carbohydrates, and 2) these effects are due to the differential effects of terpenes on fermentation products and apparent digestibility of each class of carbohydrates. Lambs were fed 2 isoenergetic and isonitrogenous diets with varying proportions of the same ingredients (beet pulp- and barley grain-based diet) or offered a choice between the 2 diets; all feeds were fed without and with terpenes, in consecutive periods. We also compared intake and preference of the beet pulp-and barley-based diets before and after the lambs ate a meal of sagebrush. Finally, we assessed the effect of terpenes on ruminal variables and in vivo digestibility. Lambs ate less when fed beet pulp or when they were offered a choice of diets with terpenes (P < 0.001), and intake of the beet pulp-based diet was the most affected (P < 0.05). Lambs preferred the beet pulp-to the grain-based diet with terpenes, but their preference reversed when terpenes were removed from the diets (P < 0.05). When lambs were offered both diets, intake and preference did not differ (P > 0.20) before eating sagebrush, but they preferred the beet pulp-based diet after eating sagebrush (P < 0.05). Intake of sagebrush did not differ among groups consuming the test diets (P = 0.21). Addition of terpenes to both diets increased the digestibility of DM, NDF, and ADF and decreased concentrations of total VFA and acetate (P < 0.05). Terpenes also depressed butyrate concentration in the barley-based diet (P < 0.05). Propionate concentrations were not affected by terpenes in either feed (P = 0.63). In summary, the predominant type of feed ingredient (beet pulp, grain) ingested with terpenes influenced fermentation products, intake, and preference in lambs. The source of energy from supplements, or other plants in the diet, is likely to influence intake and preference for sagebrush in sheep foraging on rangelands. Moreover, ingesting terpenes from sagebrush may also influence intake and preference for other plant species or supplements.     

复合酶制剂对瘤胃发酵及泌乳早期奶牛生产性能的影响

本试验旨在研究添加不同水平复合酶制剂对瘤胃发酵及奶牛生产性能的影响。试验一以奶牛全混合日粮作为底物进行体外瘤胃发酵试验,分为4组,即对照组不添加酶制剂,试验1、2和3组的酶制剂添加量分别为日粮浓度的0.10%、0.15%和0.20%,每组设9个重复。每个重复准确称取0.500 g底物,在体外发酵产气自动记录装置上发酵48 h,测定其发酵参数和营养物质降解率。结果表明:复合酶制剂显著提高发酵液中总挥发性脂肪酸和乙酸浓度(P<0.05);试验组中粗蛋白(P<0.05)和中性洗涤纤维(P<0.01)降解率显著高于对照组。试验二选择体重、胎次、泌乳天数和产奶量相近的泌乳早期荷斯坦奶牛36头,采用随机区组设计分为4组,即对照组和试验1、2和3组,对照组不添加酶制剂,试验1、2和3组分别添加0.10%、0.15%和0.20%的酶制剂,每组9个重复,试验期8周,测定产奶量和乳成分含量,计算3.5%乳脂校正乳。结果表明:复合酶制剂显著提高3.5%乳脂校正乳产量(P<0.05),0.10%、0.15%和0.20%组比对照组分别提高3.88、4.27和2.26 kg·d^-1。0.15%组的乳脂率显著高于对照组(P<0.05),比对照组高12.7%。结论添加复合酶制剂有利于瘤胃发酵和提高生产性能,且添加量为0.15%时效果较好。     

Effect of concentrate crude protein level on grass silage intake, milk yield and nutrient utilisation by dairy cows in early lactation.

Twenty-one multiparous dairy cows were fed concentrates containing three levels (119, 154 and 191 g/kg DM) of crude protein (CP) during the first ten weeks of lactation. Part of the grain and molassed sugar beat pulp was substituted with 0% (RSM0), 15% (RSM15) or 30% (RSM30) repeseed meal. Wilted grass silage was fed ad libitum after calving. The average response between RSM0 and RSM15 was +1.66 kg milk/d per percentage unit change in concentrate CP content. No further response occurred between RSM15 and RSM30. The positive effect of RSM inclusion was seen throughout the experimental period and was associated with increased plasma non esterified fatty acids (NEFA) and decreased plasma insulin concentration one week after calving, and higher efficiency of metabolisable energy utilisation for milk production. Digestibility of the diet remained unaffected. Milk and plasma urea tended to increase with RSM30 indicating excessive supply of rumen degradable protein. Because of the limited potential of cows to compensate for a deficit in feed protein supply by mobilising tissue protein, a substantial milk yield response can be achieved with a moderate level of protein supplementation during early lactation.     

Effect of feeding Bacillus subtilis natto fermentation product on milk production and composition, blood metabolites and rumen fermentation in early lactation dairy cows

This experiment was conducted to determine the effect of Bacillus subtilis natto fermentation product supplementation on blood metabolites, rumen fermentation and milk production and composition in early lactation dairy cows. Thirty-six multiparous Holstein cows (DIM = 29 ± 6 days, parity = 2.8 ± 1.1) were blocked by DIM and parity and then randomly assigned to three treatments (12 per treatment) in a 9-week trial. Cows in control, DFM1 and DFM2 were fed TMR diets supplemented with 0, 6 and 12 g of B. subtilis natto solid-state fermentation product per day per cow respectively. Plasma non-esterified fatty acids were lower (p = 0.03) in DFM1 and DFM2 compared with control cows (633 and 639 vs. 685 μm). Ruminal propionate increased (23.9 vs. 26.3 and 26.9/100 mol, control vs. DFM1 and DFM2 respectively) and acetate decreased (64.2 vs. 62.7 and 62.1/100 mol, control vs. DFM1 and DFM2 respectively) with increasing B. subtilis natto fermentation product supplementation. DMI of the cows in three treatments was not affected by B. subtilis natto fermentation product supplementation, but milk yield was 3.1 and 3.2 kg/day higher for DFM1 and DFM2 than that for control cows on average across the 9-week trial, and significant differences were observed during weeks 5-9 of the trial, which resulted in 9.5% and 11.7% increase in feed efficiency. B. subtilis natto fermentation product supplementation did not affect milk fat percentage and protein yield but increased (p < 0.05) milk fat yield and lactose percentage (p < 0.01) and tended to decrease protein percentage (p = 0.06). The findings show that B. subtilis natto fermentation product was effective in increasing lactation performance of early lactation dairy cows possibly by altering the rumen fermentation pattern without any negative effects on blood metabolites.     

Effects of level of feed intake and Fusarium toxin-contaminated wheat on rumen fermentation as well as on blood and milk parameters in cows

The aims of this study were to examine the effects of and possible interactions between dry matter (DM) intake and feeding Fusarium toxin-contaminated wheat on ruminal fermentation, serum chemical parameters and milk yield of dairy cows. Fourteen dairy cows equipped with ruminal and duodenal cannulas were analysed. All animals were fed the same ration, the daily feed amounts being adjusted to current performance. On DM basis, the ration consisted of 60% concentrate including 55% wheat [Fusarium-contaminated wheat (mycotoxin period) or control wheat (control period)] and was completed with 40% maize and grass silage. Each cow was fed the contaminated wheat [deoxynivalenol (DON), 8.21 mg/kg DM and zearalenone (ZON), 0.09 mg/kg DM] and the control wheat (0.25 mg DON/kg DM and 51 microg ZON/kg DM). As expected, a higher organic matter (OM) intake decreased the amounts of fermented crude nutrients related to the respective intakes. An increased amount of crude protein degraded (p < 0.05) and a lower molar percentage of propionate in the rumen fluid were observed when feeding the Fusarium toxin-contaminated wheat at increased OM intakes in comparison with the control wheat. The activities of serum aspartate aminotransferase (ASAT; p < 0.001), glutamate dehydrogenase (GLDH; p < 0.01) and gamma glutamyl transferase (gamma-GT; p < 0.01) increased with increasing OM intake and were not related to the mycotoxin contamination of the wheat.     

Effect of increased concentrate allotment before evening grazing on herbage intake,nitrogen utilization and rumen fermentation in dairy cows grazed on perennial ryegrass pasture

Two experiments were conducted to elucidate the effect of increased concentrate allotment before evening grazing on herbage intake, nitrogen utilization and rumen fermentation in dairy cows. In experiment 1, nine lactating cows were grazed in the morning and evening sessions (2.5 h each). The cows were allocated to treatments of three concentrate allotment levels before the evening grazing session by altering proportions to daily total offered; 25%, 50% and 75%. Daily herbage dry matter intake quadratically decreased with increasing the concentrate allotment levels (P < 0.05). Nitrogen utilization was similar among treatments. To investigate diurnal changes in rumen fermentation, a second experiment was conducted where six ruminally cannulated non‐lactating dairy cows grazed in the morning and evening sessions (3 h each) were subjected to the same treatments as experiment 1. Total volatile fatty acid concentration in the rumen linearly increased with increasing the concentrate allotment levels throughout the pre‐evening grazing session to the post‐morning grazing session (P < 0.01). The results indicate that dairy cows reduce daily herbage intake but do not alter nitrogen utilization with increasing concentrate allotment before evening grazing. © 2016 Japanese Society of Animal Science     

Effect of supplemental Bacillus cultures on rumen fermentation and milk yield in Chinese Holstein cows

Two experiments were conducted to study the effect of supplemental 100 g/day of live Bacillus cultures (2 × 10¹¹ cell of Bacillus subtilis and Bacillus licheniformis) on rumen fermentation as well as milk yield and composition in Chinese Holstein cows. In experiment 1, investigating 3 × 10 cows, milk yield and milk protein were increased by using B. licheniformis (p < 0.05) in comparison with an unsupplemented group and the B. subtilis group. Body weight was not significantly affected by Bacillus culture supplementation (p > 0.05). Percentage of milk fat and lactose was not significantly different between treatments (p > 0.05). But milk protein increased with B. licheniformis supplementation (p < 0.05). In experiment 2, carried out with three non‐lactating ruminally and duodenally fistulated cows, results showed that B. licheniformis supplementation increased microbial crude protein flow into duodenum (p < 0.05) and decreased the ammonia nitrogen concentration in ruminal fluid at 0.5 h, 1 h, 3 h, 6 h after morning feeding (p < 0.05). Bacillus licheniformis supplementation increased total VFA and acetate concentration in ruminal fluid at 0.5 h, 1 h, 3 h, 6 h after morning feeding (p < 0.05). Bacillus subtilis had no significant effect on rumen fermentation characteristics, duodenal microbial N flow and ruminal apparent nutrient digestibility (p > 0.05). Bacillus licheniformis increased ruminal apparent nutrient digestibility of neutral detergent fibre, acid detergent fibre, and organic matter (p < 0.05).     

甘露寡糖对泌乳早期奶牛瘤胃发酵及生产性能影响的研究

试验旨在研究甘露寡糖(mannan-oligosaccharides MOS)对泌乳早期奶牛瘤胃发酵及生产性能的影响。选择4头泌乳天数为20 d,日产奶量30 kg左右,体重550 kg左右的经产(二胎)中国荷斯坦泌乳奶牛,随机分为两组,对照组饲喂基础日粮,试验组饲喂在基础日粮中添加60 g的试验日粮,采用2×2交叉试验设计,每期21 d。结果表明:添加甘露寡糖可以显著提高奶牛瘤胃中总的挥发酸和氨态氮的含量(P<0.05),其中瘤胃液中乙酸含量有极显著差异(P<0.01),丙酸、丁酸的含量无显著差异(P>0.05)。对奶样进行分析,添加甘露寡糖可以显著提高乳脂率(P<0.05),并且明显降低乳中体细胞数(P<0.05),产奶量、乳蛋白和乳中尿素氮浓度均无显著差异(P>0.05)。试验表明,在日粮中添加甘露寡糖有助于提高奶牛瘤胃乙酸发酵比例,提高奶牛乳脂率。     

Effect of herbage allowance and concentrate supplementation on dry matter intake, milk production and energy balance of early lactating dairy cows

The objective of this study was to investigate the effect of daily herbage allowance and concentrate supplementation level offered at approximately 40 and 80 days in milk (DIM) and the carryover effects at 120 DIM on the production performance of spring calving dairy cows. Sixty-six (30 primiparous and 36 multiparous) Holstein–Friesian dairy cows (mean calving date — 7 Feb ± 9.9 days) were randomly assigned to a 6 treatment (n = 11) grazing study. The experiment was a randomised block design with a 3 × 2 factorial arrangement of treatments (3 daily herbage allowances (DHA's; approximately 13, 16 and 19 kg DM/cow/day; > 4 cm) and 2 concentrate allowances (0 and 4 kg DM/day). Treatments were imposed from 21 February to 8 May. Following this period (subsequent 4-weeks) animals were offered a daily herbage allowance of 20 kg DM/cow/day and no concentrate. Milk production, total dry matter intake (TDMI), energy balance (EB) and blood metabolites were measured on three occasions — at approximately 40, 80 and 120 days in milk, R1, R2 and the carryover period, respectively. Cows offered a low DHA had a lower post-grazing sward height but increased sward utilisation (> 4 cm) during R1 and R2, there was no difference during the carryover period. Concentrate supplementation increased post-grazing sward height by 11% during R2 but had no effect during R1 and the carryover period. Daily herbage allowance had no effect on milk yield or composition during R1 however a low DHA tended to reduce milk yield in R2. Concentrate supplementation increased milk and solids corrected milk (SCM) yield by 4.1 and 2.8 kg/cow/day, respectively during R1 and also increased R2 milk production performance, this effect extended into the carryover period. Offering a low DHA restricted grass dry matter intake (DMI) during R1 and R2 yet concentrate supplementation significantly increased total DMI (2.3 (R1) and 3.0 (R2) kg DM/cow). Animals offered a low DHA had a significantly lower bodyweight (BW) than those offered a medium or high DHA during P1 and P2. Concentrate supplementation increased BW during P1 and P2 (+ 9 and + 14 kg/cow, respectively). There was no effect of treatment on BW during P3. There was no effect of DHA on EB in R1; during R2 animals offered a low DHA had the lowest EB. Concentrate significantly increased EB in R1 and R2 and increased plasma glucose concentration while it decreased plasma NEFA and BHB concentrations. The results of this study indicate that animals should be offered a low DHA up to 80 DIM after which DHA should be increased however animals should also be supplemented with concentrate during the early post-partum period.     

饲喂高油酸玉米对奶牛瘤胃发酵、产乳量和乳成分的影响

选用 8头经产荷斯坦种乳牛分为对照和 HOAC2组 ,以二乘反转法 ,研究了高油酸玉米 ( HOAC)对奶牛瘤胃发酵、产乳量和乳成分的影响。HOAC组用 HOAC替换了对照组饲料中 3 0 %的玉米 ,2组日粮组成和营养成分基本相同。结果表明 ,HOAC的替换对瘤胃液氨氮浓度、挥发性脂肪酸浓度及其组成无显著影响 ( P>0 .1 ) ,但纤毛虫总数显著增加 ( P<0 .0 5 )。 2组牛干物采食量无显著差异 ,HOAC组产乳量增加 0 .7kg/d,但不显著 ( P>0 .1 ) ,4%脂肪校正乳量有提高趋势 1 .4kg/d( 5 .5 % ,P<0 .1 )。乳蛋白质、乳糖和无脂固形物的含量及其生产量 2组间无明显差异。HOAC组乳脂率有增加趋势 ( P<0 .1 ) ,而乳脂生产量显著提高 ( P<0 .0 5 ) ,但脂肪酸组成无明显变化。以上结果显示 :奶牛饲料中HOAC的使用可改善乳成分 ,并对瘤胃发酵产生一定影响。     

Effect of supplementation of soy sauce oil and Ca salts of fatty acids on rumen fermentation,milk production and conjugated linoleic acid in milk of dairy cows

Eight cows were used to evaluate the effects of supplementation of soy sauce oil (SO) or Ca salts of fatty acids (FA) on rumen fermentation and milk production. The control diet (CO) consisted mainly of hay, corn silage and a concentrate. In the experimental diets, 400 g/day per cow of SO or FA (soybean oil and rapeseed oil) was supplemented to the CO diet. Experimental period for the three treatments was 14 days, and milk samples were taken during the last 2 days and rumen sample was taken on the last day. Dry matter intake was not affected by the treatments. The number of rumen protozoa at 0 h increased by SO and FA diets. Total volatile fatty acids at 2 h after feeding of SO diet was decreased compared to CO. The milk composition yield did not differ among treatments, although the percentages of fat and protein were decreased by SO and FA diets. The proportions of C8–C16 fatty acids in milk fat decreased, and those of C18 increased by SO and FA diets. The proportion of cis‐9, trans‐11 conjugated linoleic acid in milk fat by SO and FA diets increased by 120% and 135%, respectively. In spite of the slight suppression of rumen fermentation by SO diet, negative effects on feed intake and milk production were not detected.     

Sorghum silage supplemented with crambe meal improves dry matter intake and milk production in crossbred Holstein cows

We studied the feed intake and milk production of Holstein × Zebu crossbred cows fed sorghum silage diets and concentrates, with and without the addition of crambe meal. Using a change-over design for a total of 120 days, eight cows were fed two diets (concentrates + silages with and without crambe meal) in four 30-day periods, with four replications (animals). The crambe meal diet increased (P < 0.05) the dry matter intake (DMI), crude protein intake (CPI), and non-fibrous carbohydrates intake (NFCI) as well as milk production (4.3%). No difference was noted (P > 0.05) for the milk composition between the treatments. Crambe meal as an additive in sorghum silage (100 g/kg in natural matter) showed a great potential for introduction in dairy farming as it substantially increases CP intake, does not reduce food intake, raises animal productivity, and does not affect milk composition.     

过瘤胃蛋白和丙二醇对乳牛瘤胃发酵、产乳量及乳成分的影响

用 8头荷斯坦黑白花奶牛 ,在饲粮中添加过瘤胃蛋白和丙二醇 ,研究其对乳牛瘤胃发酵、产乳量及乳成分的影响。结果表明 ,过瘤胃蛋白+丙二醇可显著提高丙酸浓度 (P <0 0 5) ,并且产乳量较对照组有增加的趋势。乳脂率、乳蛋白率、乳糖率及无脂固形物率 ,并未因添加过瘤胃蛋白和丙二醇而受到显著影响 (P >0 0 5)。乳蛋白质量、乳脂量及无脂固形物量 ,过瘤胃蛋白 +丙二醇组较对照组有增加的趋势 ,但差异均不显著 (P >0 0 5)。     

Effect of milking frequency in early lactation on energy metabolism, milk production and reproductive performance of dairy cows

The objective of this study was to evaluate the effect of imposing different milking frequencies for the first 4 weeks of lactation on energy balance, blood metabolic profiles and reproductive performance. The milking frequencies imposed were once daily for the first 4 weeks of lactation and twice daily subsequently (X1), twice daily for the entire lactation (X2) and three times daily for the first 4 weeks of lactation and twice daily subsequently (X3). Two weeks prior to the first expected calving date, sixty-three multiparous, Holstein-Friesian cows, were allocated to treatments based on expected calving date, parity (lactation number = 2 and lactation number > 2), BCS and BW. Mean DMI was lower (P < 0.05) on X1 than X2 (15.7 and 17.1 kg/day) over the first 4 weeks of lactation, but DMI on X3 was not different to either. Milk yield was lower (P < 0.05) on X1 than on X2 and X3 (24.1, 31.9 and 31.4 kg/day) from weeks 1 to 4 and at week 10 of lactation (30.9, 33.9 and 33.7 kg/day) but was not different at week 20. Yield of milk constituents followed the same pattern as milk yield. Milking frequency had no significant effect on milk fat or protein concentration but milk lactose concentration was higher (P < 0.05) on X1 than on X2. Cows on X1 experienced less (P < 0.05) negative energy balance than those on X2 and X3 from weeks 1 to 4, with no difference between X2 and X3. Cows on X1 lost less BW (P < 0.05) from weeks 1 to 4 and lost less BCS up to day 73 of lactation than cows on X2 and X3. Plasma NEFA and BHB concentrations tended to be lower and plasma glucose tended to be higher on X1 than on X2 and X3 from weeks 1 to 4, while plasma insulin was higher on X1 than on X2 and X3 for the first 2 weeks of lactation. There was no difference between treatments in any reproductive performance variable measured. These results indicate that reduced milking frequency decreases milk yield and improves energy balance and metabolic status in early lactation.