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This experiment studied the effect of 3 forms of presentation of linseed fatty acids (FA) on methane output using the sulfur hexafluoride tracer technique, total tract digestibility, and performance of dairy cows. Eight multiparous lactating Holstein cows (initial milk yield 23.4 [+ or -] 2.2 kg/d) were assigned to 4 dietary treatments in a replicated 4 x 4 Latin square design: a control diet (C) consisting of corn silage (59%), grass hay (6%), and concentrate (35%) and the same diet with crude linseed (CLS), extruded linseed (ELS), or linseed oil (LSO) at the same FA level (5.7% of dietary DM). Each experimental period lasted 4 wk. All the forms of linseed FA significantly decreased daily C[H.sub.4] emissions (P< 0.001) but to different extents (-12% with CLS, -38% with ELS, -64% with LSO) compared with C. The same ranking among diets was observed for C[H.sub.4] output expressed as a percentage of energy intake (P < 0.001) or in grams per kilogram of OM intake (P < 0.001). Methane production per unit of digested NDF was similar for C, CLS, and ELS but was less for LSO (138 vs. 68 g/kg of digested NDF, respectively; P < 0.001). Measured as grams per kilogram of milk or fat-corrected milk yield, methane emission was similar for C and CLS and was less for ELS and LSO (P < 0.001), LSO being less than ELS (P < 0.01). Total tract NDF digestibility was significantly less (P < 0.001) for the 3 supplemented diets than for C (-6.8% on average; P < 0.001). Starch digestibility was similar for all diets (mean 93.5%). Compared with C, DMI was not modifled with CLS (P > 0.05) but was decreased with ELS and LSO (-3.1 and -5.1 kg/d, respectively; P < 0.001). Milk yield and milk fat content were similar for LSO and ELS but less than for C and CLS (19.9 vs. 22.3 kg/d and 33.8 vs. 43.2 g/kg, on average, respectively; P < 0.01 and P < 0.001). Linseed FA offer a promising dietary means to depress ruminal methanogenesis. The form of presentation of linseed FA greatly influences methane output from dairy cows. The negative effects of linseed on milk production will need to be overcome if it is to be considered as a methane mitigation agent. Optimal conditions for the utilization of linseed FA in ruminant diets need to be determined before recommending its use for the dairy industry. Key words: dairy cow, digestion, fatty acid, linseed, linseed oil, methane

This work investigated the metabolic basis for the variability of carcass and i.m. adiposity in cattle. Our hypothesis was that the comparison of extreme breeds for adiposity might allow for the identification of some metabolic pathways determinant for carcass and i.m. adiposity. Thus, 23- to 28-mo-old steers of 3 breeds, 2 with high [Angus or Japanese Black x Angus (J. Black cross)] and 1 with low (Limousin) i.m. and carcass adiposity, were used to measure activities or mRNA levels, or both, of enzymes involved in de novo lipogenesis [acetyl-coA carboxylase, fatty acid synthase (FAS), glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme], circulating triacylglycerol (TAG) uptake (lipoprotein lipase), and fatty acid esterification (glycerol-3-phosphate dehydrogenase), as well as the mRNA level of leptin, an adiposity-related factor. In a first study, enzyme activities were assayed in the s.c. adipose tissue (AT), the oxidative rectus abdominis, and the glycolytic semitendinosus muscles from steers finished for 6 mo. Compared with Angus or J. Black cross, Limousin steers had a 27% less (P = 0.003) rib fat thickhess, and 23 and 29% less (P [less than or equal to] 0.02) FAS and G6PDH activities in s.c. AT. In rectus abdominis and semitendinosus, the 75% less (P < 0.001) TAG content was concomitant with 50% less (P < 0.001) G6PDH activity. In a second study, enzyme activities plus mRNA levels were assayed in an oxido-glycolytic muscle, the longissimus thoracis (LT), in the i.m. AT dissected from LT, and in s.c. AT from the same Limousin steers and from Angus steers finished for 10 mo. Compared with Angus, the 50% less (P < 0.001) rib fat thickness in Limousin contrasted with the 1.1- to 5.8-fold greater (P [less than or equal to] 0.02) mRNA levels or activities, or both, of acetyl-coA carboxylase, G6PDH, lipoprotein lipase, and glycerol-3-phosphate dehydrogenase in s.c. AT. Conversely, the 90% less (P < 0.001) TAG content in Limousin LT was concomitant to the 79 and 83% less (P [less than or equal to] 0.002) G6PDH activity and leptin mRNA level. Such differences could arise from a greater number of adipocytes in LT from Angus steers because no difference was found between Limousin and Angus for G6PDH activity and leptin mRNA in i.m. AT. We conclude that FAS and G6PDH in s.c. AT could be involved in differences in carcass adiposity, but this relationship disappeared when the fatness increased strongly. Leptin and G6PDH are related to the expression of marbling whatever the body condition and thus could be relevant indicators of marbling in beef cattle. Key words: adipose tissue, bovine, leptin, lipogenic activity, muscle

An ovine-specific RIA, shown to be reliable for bovine leptin determination, was used to study the effects of breed, body fatness, feeding level, and meal intake on plasma leptin level in adult cattle. Eighteen fat Charolais, fat Holstein, and lean Holstein adult cows were either well-fed (130% of maintenance energy requirements [MER]) or underfed (60% of MER) for 3 wk. The breed tended to have a small effect on plasma leptin level, which was decreased by 70% (P < 0.05) in lean compared to fat Holstein cows. A strong curvilinear relationship was found between mean adipocyte volume and plasma leptin concentrations in well-fed (r = +0.95) and underfed (r = +0.91) cows. Underfeeding caused a significant decrease in plasma leptin levels from 8.0 [+ or -] 3.1 to 6.1 [+ or -] 2.3 ng/mL (P < 0.01). Nine adult Holstein cows initially fed at 130% of MER (control) were underfed to 21% of MER for 7 d, and five of them were refed to 237% of MER for 21 d. Plasma leptin measured 1 h before meal distribution was decreased from 5.9 [+ or -] 0.4 to 3.8 [+ or -] 0.2 ng/mL (P < 0.01) by underfeeding and increased to reach 8.8 [+ or -] 1.0 ng/mL (P < 0.01) after refeeding. It was positively related to plasma glucose (r = +0.52, P < 0.01) and negatively related to plasma NEFA (r = -0.67, P < 0.001). Plasma leptin measured 4 h after meal distribution was positively related to feeding level and to plasma 3-OH-butyrate (r = +0.61, P < 0.005) and negatively related to plasma NEFA (r = -0.56, P < 0.01). Differences between pre- and post-prandial leptin cancentrations showed a decrease after meal intake in control and well-fed cows (-7 and -19%, P < 0.01, respectively) and an increase in underfed cows (+12%, P < 0.01). Leptin response to meal intake was positively related to glucose response (r = +0.66, P < 0.001) and negatively related to 3-OH-butyrate response (r = -0.78, P < 0.001). By using the 'multispecies' commercial RIA, leptin concentrations were lower and we observed similar physiological responses, although less related to other hormones or metabolites. These data provide evidence, first, that a specific RIA for ruminant leptin determination is necessary to better understand leptin regulation, and second, that plasma leptin is strongly related to adipose cell size and positively related to feeding level in adult cattle, and that an effect of meal intake could be mediated by glucose and(or) ketone bodies. Key Words: Body Fat, Breed Differences, Dairy Cows, Feeding Frequency, Leptin, Meals

We examined the ability of the ovine mammary gland to synthesize leptin throughout pregnancy and lactation. Leptin gene expression was assayed by real-time reverse transcription and polymerase chain reaction in mammary gland from ewes at 15, 80, 106, 112, or 141 d of pregnancy and at 0 (30 min after parturition), 3, 48, or 70 d of lactation. Leptin mRNA level was high at the beginning (the first 80 d) and at the end of pregnancy and was lower at mid-pregnancy and throughout lactation. Furthermore, during these periods of mammary leptin expression, the location of leptin protein, as determined by immunohistochemical analysis, changed within mammary tissue. It was located in adipose cells during early stages of pregnancy, in epithelial cells after full cell differentiation just before parturition, and in myoepithelial cells after parturition. These data, compared with published data on leptin receptor gene expression, provide evidence that leptin could be produced by different cell types of the mammary gland and could act as a paracrine factor on mammary cell growth and differentiation via adipose-epithelial cells and myoepithelial-epithelial cell interactions. Key Words: Lactation, Leptin, Mammary Tissue, Pregnancy, Sheep

An in vitro system was used to study the regulation of glucose and acetate utilization by bovine and ovine adipose tissue (AT). Adult nonlactating, nonpregnant cows (n = 5) and ewes (n = 5) were given limited amounts of feed for 8 to 10 d, then overfed for 10 (ewes) or 21 (cows) d. Perirenal AT explants were incubated for 7 d. The effect of 2 mU/ mL of insulin on glucose utilization was significant in bovine but not in ovine AT during the 1st d in vitro, whereas the relationship was reversed from the 4th d of incubation. The effect of insulin on acetate utilization was significant in bovine but not in ovine AT. The addition of dexamethasone (DEX, 100 nM) to the insulin-supplemented medium increased (P < .05) glucose and acetate utilization by ovine AT. However, DEX did not affect substrate utilization by bovine AT in the presence of insulin. The DEX alone decreased glucose and acetate utilization by bovine and ovine AT during the first 4 d of incubation. So, the effects of insulin and(or) DEX on substrate utilization differed with species and incubation time. Key Words: Ruminants, Adipose Tissue, Glucose, Acetate, Insulin, Dexamethasone

An in vitro system was used to study the regulation of lipoprotein lipase (LPL) activity in bovine adipose tissue. The utilization of two energetic and lipogenic substrates, acetate and glucose, and the activity of glucose-6-phosphate dehydrogenase (G6PDH), an enzyme involved in de novo lipogenesis, were also studied. Nine nonlactating, nonpregnant Holstein cows were given limited amounts of feed for 10 d, then they were overfed for 3 to 5 wk. Samples of perirenal adipose tissue were incubated for 24 or 48 h. Insulin (2 mU/mL) increased (P [is less than] .001) daily glucose and acetate utilization and attenuated (P [is less than] .001) the loss of G6PDH activity detected after 1 or 2 d of incubation. Dexamethasone (DEX, 10 nM) added to the insulin-supplemented medium decreased (P [is less than] .02) glucose utilization, but it did not change acetate utilization or G6PDH activity. A higher concentration of DEX (100 nM) potentiated (P [is less than] .004) the ability of insulin to attenuate the decrease in G6PDH activity without changing substrate utilization. Under basal conditions, LPL activity was decreased by approximately 66% after 2 d of incubation. The decline in LPL activity was attenuated by insulin addition (P [is less than] .02) and was further attenuated (P [is less than] .004) by 100 nM of DEX. The addition of 10% fetal bovine serum alone to the medium had no effect on LPL activity, and fetal bovine serum decreased this activity when it was added to the insulin-supplemented medium.