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Methane (CH 4) production of ruminants typically increases with increased dry matter intake (DMI). However, few studies have observed the effects of feeding multiple times a day and its effects on diurnal variation in CH 4 production and energy balance in late-lactation dairy cattle. A study using headbox-style indirect calorimetry and 12 multiparous (225 ± 16.2 d in milk; mean ± SD) lactating Jersey cows was conducted to determine the effects of feeding twice daily on diurnal variation in CH 4 production and total energy balance. A crossover design with 14-d periods (10 d of adaption and 4 d of collection) was used to compare 2 treatments. Treatments consisted of either once a day feeding (1×; 100% of feed given at 1000 h) or twice a day feeding (2×; 50% of feed given at 1000 h and the final 50% at 2000 h) with a common diet fed in both treatments. Dry matter intake was not different between treatments, with a mean of 16.9 ± 0.88 kg/d. Once a day feeding tended to have greater milk yield compared with twice a day feeding (21.2 vs. 20.4 ± 1.59 kg/d, respectively). Milk fat and milk protein percentage were not different, with means of 6.18 ± 0.20% and 3.98 ± 0.08%, respectively. Total CH 4 production did not differ between treatments, with a mean of 402.1 ± 20.8 L/d. Similarly, CH 4 per unit of milk yield and DMI was not different between treatments, with means of 20.5 ± 1.81 and 23.8 ± 1.21 L/kg, respectively. Feeding frequency did not affect diurnal variation of hourly CH 4 production, with a mean of 17.1 ± 0.74 L/h. A trend was observed for a treatment × hour interaction. Methane production per hour increased after the second feeding for cattle fed twice versus once daily. Gross energy, digestible energy, metabolizable energy, and balance (milk plus tissue) per kilogram of DMI did not differ by feeding frequency, with means of 4.41 ± 0.01, 3.05 ± 0.03, 2.63 ± 0.03, and 1.32 ± 0.08 Mcal/ kg of DM, respectively. Metabolizable energy for maintenance was 146 kcal/kg of metabolic body weight, with an efficiency of converting metabolizable energy to net energy balance (milk plus tissue) of 76%. Nitrogen balance did not differ among treatments, with a mean balance of 17.3 ± 13.0 g/d. Therefore, total CH 4 production and energy maintenance were not affected by feeding frequency. However, CH 4 was variable throughout the day, and caution should be exercised when collecting CH 4 samples at a limited number of time points because this may under-or overestimate total production.

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The addition of fat and calcium sulfate to diets fed to ruminants has resulted in a reduction in methane production, but the effects on energy balance have not been studied. A study using indirect calorimetry and 16 multiparous (8 Holstein and 8 Jersey; 78 ± 15 d in milk; mean ± standard deviation) lactating dairy cows was conducted to determine how mitigating methane production by adding corn oil or calcium sulfate to diets containing reduced-fat distillers grains affects energy and nitrogen balance. A replicated 4 × 4 Latin square design with 35-d periods (28 d of adaption and 4 d of collections) was used to compare 4 different dietary treatments. Treatments were composed of a control (CON) diet, which did not contain reduced-fat distillers grain and solubles (DDGS), and treatment diets containing 20% (dry matter basis) DDGS (DG), 20% DDGS with 1.38% (dry matter basis) added corn oil (CO), and 20% DDGS with 0.93% (dry matter basis) added calcium sulfate (CaS). Compared with CON, dry matter intake was not affected by treatment, averaging 29.6 ± 0.67 kg/d. Milk production was increased for diets containing DDGS compared with CON (26.3 vs. 27.8 ± 0.47 kg/d for CON vs. DDGS, respectively), likely supported by increased energy intake. Compared with CON, energy-corrected milk was greater in DG and CO (30.1 vs. 31.4, 31.7, and 31.0 ± 0.67 kg/d for CON, DG, CO, and CaS, respectively). Compared with CON, the addition of calcium sulfate and corn oil to diets containing DDGS reduced methane production per kg of dry matter intake (22.3, 19.9, and 19.6 ± 0.75 L/kg per d for CON, CO, and CaS, respectively). Similarly, methane production per kilogram of energycorrected milk was reduced with the addition of calcium sulfate and corn oil to diets containing DDGS (14.2, 12.5, and 12.4 ± 0.50 L/kg per d for CON, CO, and CaS, respectively). Compared with CON and CaS, the intake of digestible energy was greater for DG and CO treatments (57.7, 62.1, 62.0, and 59.0 ± 1.38 Mcal/d for CON, DG, CO, and CaS, respectively). Intake of metabolizable energy was greater in all treatments containing DDGS compared with CON (50.5 vs. 54.0 ± 1.08 Mcal/d for CON vs. DDGS, respectively). Net balance (milk plus tissue energy) per unit of dry matter was greater in CO (containing DDGS and oil) than CON (1.55 vs. 1.35 ± 0.06 Mcal/kg for CO vs. CON, respectively). Tissue energy was greater in DG and CO compared with CON (6.08, 7.04, and 3.16 ± 0.99 Mcal/d for DG, CO, and CON, respectively.Results of this study suggest that the addition of oil and calcium sulfate to diets containing DDGS may be a viable option to reduce methane production and in the case of oil also improve net energy balance in lactating dairy cows.