C. Delavaud

Association of leptin gene polymorphisms with serum leptin concentration in dairy cows

Leptin is a hormone produced by adipocytes, and its expression is regulated by body fatness and energy balance. This study describes the association of four leptin gene polymorphisms in dairy cows (R4C, A59V, RFLP1, and BM1500) with circulating leptin concentrations during the periparturient period. A59V is located at a between-species conserved region of leptin, and R4C might have effect on the tertiary structure of the leptin protein because of the presence of an extra cystein. RFLP1 is an intronic SNP and BM1500 is a microsatellite located 3.6 kb downstream of the leptin locus. The four polymorphisms were genotyped in 323 HF heifers with known pedigree. Leptin concentrations were determined biweekly from 30 days before until 80 days after parturition. The effect of genotype on leptin concentrations was modeled by fitting a spline in ASREML describing leptin concentrations as a function of days relative to parturition for each genotype/allele. Surprisingly, associations were found during pregnancy, but not during lactation. This indicates that the polymorphism could be more effective during pregnancy. If further studies demonstrate that more leptin-binding protein (Ob-Re) is present in this stage, it is hypothesized that a structural difference in the leptin protein could cause a sub-optimal binding stringency to Ob-Re. Free leptin could be cleared faster than bound leptin, and this could result in lower leptin concentrations during pregnancy for the polymorphism. The effects found might be ascribed to R4C. However, more study on the Ob-Re receptor, like binding stringencies between R4C and wild-type leptin and glycosylation during pregnancy, would provide more insight in the results found.

Effects of body condition score at calving on indicators of fat and protein mobilization of periparturient Holstein-Friesian cows

The objective was to study the effects of body condition score (BCS) at calving on dairy performance, indicators of fat and protein mobilization, and metabolic and hormonal profiles during the periparturient period of Holstein-Friesian cows. Twenty-eight multiparous cows were classed according to their BCS (0 to 5 scale) before calving as low (BCS ≤ 2.5; n=9), medium (2.75 ≤ BCS ≤ 3.5; n=10), and high (BCS ≥ 3.75; n=9), corresponding to a mean of 2.33, 3.13, and 4.17 points of BCS, and preceding calving intervals of 362, 433, and 640 d, respectively. Cows received the same diets based on preserved grass to allow ad libitum feed intake throughout the study, and lactation diet contained 30% of concentrate (dry-matter basis). Measurements and sampling were performed between wk -4 and 7 relative to calving. No significant effects were observed of BCS group on dry matter intake (kg/d), milk yield, BCS loss, plasma glucose, and insulin concentrations. The high-BCS group had the lowest postpartum energy balance and the greatest plasma concentrations of leptin prepartum, nonesterified fatty acids and β-hydroxybutyrate postpartum, insulin-like growth factor 1, and milk fat content. Milk fat yield was greater for the high- than the low-BCS group (1,681 vs. 1,417 g/d). Low-BCS cows had the greatest concentration of medium-chain fatty acids (e.g., sum of 10:0 to 15:0, and 16:0), and the lowest concentration and secretion of preformed fatty acids (e.g., cis-9 18:1) in milk fat. Milk protein secretion was lowest in the low-BCS group, averaging 924, 1,051, and 1,009 g/d for low-, medium-, and high-BCS groups, respectively. Plasma 3-methylhistidine was greater in wk 1 and 2 postpartum compared with other time points, indicating mobilization of muscle protein. Plasma creatinine tended to be lower and the 3-methylhistidine: creatinine ratio was greater in low- compared with medium- and high-BCS cows, suggesting less muscle mass but more intense mobilization of muscle protein in lean cows. High-BCS cows were metabolically challenged during early lactation due to intense mobilization of body fat. Conversely, limited availability of body fat in low-BCS cows was associated with increased plasma indicators of body protein mobilization during the first weeks of lactation, and lower milk protein secretion. These results should be confirmed using an experimental approach where calving BCS variation would be controlled by design.

Effects of fish oil and additional starch on tissue fatty acid profile and lipogenic gene mRNA abundance in lactating goats fed a diet containing sunflower-seed oil

In dairy cattle, diet supplementation with oils affects the lipid metabolism in body tissues via changes in the partitioning and deposition of fatty acids (FAs) and lipogenic gene expression; however, limited data are available in goats. Eight Alpine goats were fed a grassland hay diet supplemented with 90 g/day of sunflower-seed oil or 90 g/day of sunflower-seed oil and fish oil (2 : 1) plus additional starch. The goats were slaughtered on day 21 of the treatments and samples of the mammary secretory tissue, liver, omental and perirenal adipose tissues (ATs) were collected to characterise their FA composition and the mRNA abundance of lipogenic genes and transcription factors involved in their regulation, and to examine the impact of the diet composition on the same parameters. The results are in agreement with the specific physiological adaptation in the lipid metabolism of body tissues that is likely to occur during late lactation because of the coexistence of an active lipogenesis in the mammary secretory tissue and a significant anabolic activity in the ATs. These latter tissues were characterised by high concentrations of saturated FA and very low polyunsaturated FA (PUFA) levels. The content of PUFA was relatively higher in the mammary secretory tissue, in particular in the case of polyunsaturated C18. The highest PUFA contents were found in the liver, in accordance with the greater mRNA abundances of the genes that encode the necessary enzymes for very long-chain n-3 and n-6 PUFA synthesis. However, substantial differences between n-3 and n-6 pathways would most likely exist in the goat liver. Overall, differences in diet composition induced limited changes in the mRNA abundance of genes involved in lipid metabolism, and these were not associated with the few variations observed in tissue FA composition.

Body fat content and feeding level interact strongly in the short- and medium-term regulation of plasma leptin during underfeeding and re-feeding in adult sheep

Circulating leptin is regulated by food intake in the long, medium and short term; however, little is known about putative remnant effects of these successive regulations at any given time. To clarify this, two experiments were conducted in adult sheep, during which body condition parameters and plasma leptin were measured. During experiment 1, twenty ewes with normal body condition were either well fed (101 % of maintenance energy requirements (MER)) or underfed (41 % MER) for 166 d, then rapidly re-fed (at a mean of 208 % MER) for 3 d. Leptinaemia decreased after 14 d of underfeeding, remained depressed until day 166 and did not increase after 3 d re-feeding, whereas it was increased (+153 %; P < 0.05) by re-feeding the previously well-fed ewes. During experiment 2, twenty-four fat or lean ewes were either well fed (114 % MER) or underfed (52 % MER) for 94 d, and gradually re-fed for 2 d and maintained at a high feeding level (235 % MER) for 9 d. Underfeeding decreased leptinaemia in fat (from 4.19 to 2.63 ng/ml) but not lean ewes, and re-feeding increased leptinaemia after 5 d in lean previously well-fed (+123 %; P < 0.05) but not underfed ewes. In fat ewes, the impact of re-feeding was rapid (+144 %; P < 0.001 at 5 d) in previously well-fed ewes, whereas it was more gradual with a maximum at 11 d (+162 %; P < 0.01) in previously underfed ewes. In conclusion, leptinaemia is modulated by short-term energy intake level in interaction with long-term regulations involving nutritional history and body fatness, suggesting that a biological threshold of adiposity (about 20 %) is necessary to allow short- and medium-term leptin regulation.

Potential of milk fatty acid composition to predict diet composition and authenticate feeding systems and altitude origin of European bulk milk

The aims of this work were to elucidate the potential of using milk fatty acid (FA) concentration to predict cow diet composition and altitude of bulk milk collected in 10 different European countries and to authenticate cow-feeding systems and altitude of the production area using a data set of 1,248 bulk cow milk samples and associated farm records. The predictions based on FA for cow diet composition were excellent for the proportions of fresh herbage [coefficient of determination (R2)=0.81], good for hay, total herbage-derived forages, and total preserved forages (R2>0.73), intermediate for corn silage and grass silage (R2>0.62), and poor for concentrates (R2<0.51) in the cow diet. Milk samples were assigned to groups according to feeding system, level of concentrate supplementation, and altitude origin. Milk FA composition successfully authenticated cow-feeding systems dominated by a main forage (>93% of samples correctly classified), but the presence of mixed diets reduced the discrimination. Altitude prediction reliability was intermediate (R2<0.62). Milk FA composition was not able to authenticate concentrate supplementation level in the diet (<58% of samples correctly classified). Similarly, the altitude origin was not successfully authenticated by milk FA composition (<76% of samples correctly classified). The potential of milk FA composition to authenticate cow feeding was confirmed using a data set representative of the diversity of European production conditions.

Effect of extruded linseeds alone or in combination with fish oil on intake, milk production, plasma metabolite concentrations and milk fatty acid composition in lactating goats

Based on the potential benefits for long-term human health, there is interest in developing sustainable nutritional strategies for lowering medium-chain saturated fatty acids (FA) and increasing specific unsaturated FA in ruminant milk. Dietary supplements of extruded linseeds (EL), fish oil (FO) or a mixture of EL and FO increase cis-9,trans-11 CLA and long-chain n-3 polyunsaturated FA in bovine milk. Supplements of FO cause milk fat depression in lactating cows, but information for dairy goats is limited. A total of 14 Alpine goats were used in a replicated 3×3 Latin square with 28-days experimental periods to examine the effects of EL alone or in combination with FO on animal performance, milk fat synthesis and milk FA composition. Treatments comprised diets based on natural grassland hay supplemented with no additional oil (control), 530 of EL or 340 g/day of EL and 39 g/day of FO (ELFO). Compared with the control, ELFO tended (P=0.08) to lower milk fat yield, whereas EL increased (P<0.01) milk fat content and yield (15% and 10%, respectively). Relative to EL, ELFO decreased (P<0.01) milk fat content and yield (19% and 17%, respectively). Relative to the control and ELFO, EL decreased (P<0.05) milk 10:0 to 16:0 and odd- and branched-chain FA content and increased 18:0, cis-18:1, trans-13 18:1 (and their corresponding ∆-9 (desaturase products), trans-12,cis-14 CLA, cis-13,trans-15 CLA, cis-12,trans-14 CLA and trans-11,cis-13 CLA and 18:3n-3 concentrations. ELFO was more effective for enriching (P<0.05) milk cis-9, trans-11 CLA and trans-11 18:1 concentrations (up to 5.4- and 7.1-fold compared with the control) than EL (up to 1.7- and 2.5-fold increases). Furthermore, ELFO resulted in a substantial increase in milk trans-10 18:1 concentration (5.4% total FA), with considerable variation between individual animals. Relative to the control and EL, milk fat responses to ELFO were characterized by increases (P<0.05) in milk trans-16:1 (Δ9 to 11), trans-18:1 (Δ6 to 11), trans-18:2, CLA (cis-9,trans-11, trans-9,cis-11, trans-8,trans-10 and trans-7,trans-9) and 20- and 22-carbon FA concentrations. Overall, EL resulted in a relatively high cis-9 18:1 concentration and an increase in the 18:3n-3/18:2n-6 ratio, whereas combining EL and FO resulted in substantial increases in trans-FA, marginal enrichment in 20:5n-3 and 22:6n-3 and lower 16:0 concentration changes associated with a decrease in milk fat content. In conclusion, data provide further evidence of differential mammary lipogenic responses to diet in the goat compared with the cow and sheep.

Sunflower-seed oil, rapidly-degradable starch, and adiposity up-regulate leptin gene expression in lactating goats

We conducted experiments to evaluate the effects of lipid supplementation and the nature of starchy concentrate on the regulation of leptin synthesis in lactating goats. Multiparous goats in mid- to late lactation received diets based on different forages and containing plant oil or seeds rich in either 18:1c9, 18:2n-6 or 18:3n-3 corresponding to 3%-7% dry matter (DM) as lipid supplements, or diets based on concentrate as either rapidly or slowly degradable starch. The isoenergetic replacement of a part of the concentrate by either oleic sunflower-seed oil, formaldehyde-treated linseeds, or linseed oil did not modify leptinemia and the leptin mRNA concentration in adipose tissues, suggesting a lack of effect of 18:1c9, 18:3n-3, or their biohydrogenation products. Conversely, leptinemia and the leptin mRNA abundance were increased (by 20% and 140%, respectively, P<0.05) in goats fed sunflower-seed oil under a grassland hay-based diet but not a maize silage-based diet, at similar energy intakes and adiposity. Thus, 18:2n-6 per se may up-regulate leptin gene expression, but the effect could be blunted by other fatty acids formed during the ruminal digestion of sunflower-seed oil when combined with maize silage. Consumption of rapidly but not slowly degradable starch increased (by 17%, P<0.05) leptinemia. Moreover, during lactation, plasma leptin was positively correlated (P<0.05) to adiposity parameters and negatively correlated to fiber intake. The results suggest that leptinemia responds poorly to nutritional factors in lactating goats, thus highlighting the physiological need to sustain hypoleptinemia during lactation.

Concentrate Feeding Strategy in Lactating Dairy Cows: Metabolic and Endocrine Changes with Emphasis on Leptin

This study aimed to evaluate metabolic and endocrine adaptations to energy intake inmultiparous Holstein cows (n = 90; mean 9434 kg energy-corrected milk yield/305 d) over the first 20 wk postpartum and to assess the association of leptinwithmetabolic, endocrine, and zootechnical traits. Concentrates were fed automatically for 24 h at 30% (C30) or 50% (C50) of total dry matter intake (DMI) from wk 1 to 10 postpartum and at linearly reduced amounts thereafter. Roughage was fed for ad libitum intake. The DMI was measured over 24 h; milk yield and body weight (BW), twice/d; milk composition, 4 times/wk; and milk acetone, weekly. Blood samples for determination of metabolite, hormone, and electrolyte concentrations and enzyme activities were obtained at wk 2 prepartum, and at wk 1 to 16 and at wk 20 postpartum from 0730 to 0900. Body condition scores (BCS) and backfat thickness were measured postpartum and during wk 1, 4, 8, 12, 16, and 20. Energy balance (EB) was considerably lower, but milk yield only slightly lower, in C30 than C50. Metabolic stress was more marked in C30 thanC50, expressed by lower, glucose, insulin, insulinlike growth factor-1 (IGF-1), triiodothyronine, milk Received July 20, 2002. Accepted October 2, 2002. Corresponding author: J. W. Blum; e-mail: juerg.blum@itz. unibe.ch. 1Presented in part at the 11th International Conference on Production Diseases in Farm Animals, August 12–16, 2001, Department of Animal Science and Animal Health, The Royal Veterinary and Agricultural University, Fredriksberg, Copenhagen, Denmark [M. Reist, D. Erdin, D. von Euw, K. Tschumperlin, C. Delavaud, Y. Chilliard, H. M. Hammon, N. Kunzi, and J. W. Blum (2001) Concentrate Feeding Strategy in lactating dairy cows: Metabolic and Endocrine Changes with emphasis on Leptin]. 2Accepted as part of thePhD-thesis ofM.Reist by theSwiss Federal Institute of Technology, Zurich, Switzerland, September 2001. 1690 protein, and lactose concentrations, higher nonesterified fatty acid, β-hydroxybutyrate, growth hormone, and milk acetone concentrations, and an accelerated decrease in BCS and backfat thickness. Nevertheless, C30 adapted successfully and thus maintained high milk yields despite negative EB. Leptin concentrations were lower in C30 than in C50 over the first 20 wk postpartum and were positively associated with BCS, EB, BW, cholesterol, albumin, insulin, and IGF-1; negatively associated with DMI and triiodothyronine; and were higher in cows calving in spring than in fall. Leptin is one among several factors involved in the regulation of energymetabolism andmay be important for overall homeostatic and homeorhetic control of metabolism and thus for maintenance of performance. (

Physiological adaptations and ovarian cyclicity of Holstein and Montbéliarde cows under two low-input production systems.

The objective was to study milk production, body reserve mobilization, metabolic and hormonal profiles, and ovarian cyclicity of Holstein-Friesian (HOLS) and Montbéliarde (MONT) cows under two low-input dairy production systems with seasonal spring calving: an extensive (EXT; 12 HOLS and 12 MONT) based on permanent diversified grasslands and zero concentrate, and a semi-extensive (SEMI; 12 HOLS and 10 MONT) based on established temporary grasslands and up to 4 kg/day of concentrate. Individual measurements were performed between -4 and 12 weeks of lactation. Cows in EXT secreted less milk (22.1 v. 24.4 kg/day), protein (660 v. 755 g/day) and energy (67.7 v. 74.4 MJ/day), had greater plasma β-hydroxybutyrate (BHBA) (0.97 v. 0.69 mM), lower glucose (59.0 v. 62.0 mg/dl) and IGF-1 (62 v. 71 ng/ml), lower milk fat concentration in fatty acids originating from de novo synthesis (e.g. ∑ 10:0 to 15:0) and greater concentration of those derived in part from mobilization of fat reserves (e.g. 18:0 and ∑>C16), and showed greater frequency of abnormal ovarian cycles compared with SEMI. Across production systems, HOLS produced more milk (24.7 v. 21.8 kg/day), protein (738 v. 674 g/day) and fat (939 v. 819 g/day), secreted more energy (75.1 v. 67.0 MJ/day), lost more body condition score (BCS) (1.41 v. 1.03) and reached a lower BCS nadir (1.12 v. 1.43), had greater plasma BHBA (0.91 v. 0.75 mM), lower insulin (15.9 v. 17.2 µIU/ml) and tended to have lower glucose (59.6 v. 61.4 mg/dl), had lower milk fat concentration in ∑ 10:0 to 15:0, tended to have higher ∑>C16 and tended to show more abnormal estrous cycles compared with MONT. Ultrasound measurements did not differentiate fat mobilization and were confounded by breed differences of skin thickness. The greater nutrient allowance in SEMI improved indicators of physiological status and ovarian function during early lactation compared with EXT, but did not attenuate body reserve mobilization because cows prioritized milk secretion. HOLS secreted more nutrients than MONT but lost more BCS, which negatively affected nutritional balance and tended to affect ovarian cyclicity during early lactation. Breed by system interactions were not observed except for a few variables.

Plasma leptin, glucose and non-esterified fatty acid variations in dromedary camels exposed to prolonged periods of underfeeding or dehydration.

The involvement of plasma leptin in the adaptation of dromedary camels to harsh conditions such as food or water shortages was studied through 2 experiments. In experiment 1, fourteen female camels were either fed at 68% of maintenance energy requirements (MER) during 112d (n=4) or overfed at 134% of MER during the first 56d and then underfed at 17% of MER the next 56d (OV-UN, n=5), or underfed and then overfed for the same durations and energy intake levels (UN-OV, n=5). Weekly plasma samples showed that leptin, glucose and non-esterified fatty acid (NEFA) concentrations were significantly modulated by energy intake level. NEFA increased sharply but transiently in underfed camels of the UN-OV or OV-UN groups, whereas glucose and leptin concentrations decreased with underfeeding and increased with overfeeding with more significant effects in camels that were previously overfed or underfed, respectively. In experiment 2 twelve female camels were either normally watered (n=6) or dehydrated (n=6) during 23d and then rehydrated during 4d. Dehydration specifically increased blood hematocrit, plasma NEFA and glucose whereas leptin decreased slightly. For both experiments, leptinemia was positively related to hump adipocyte volume. Taken together these results provide new data for a better understanding of lipid and energy metabolism in camels.