François Bocquier

Adipose tissue metabolism and its role in adaptations to undernutrition in ruminants.

Changes in the amount and metabolism of adipose tissue (AT) occur in underfed ruminants, and are amplified during lactation, or in fat animals. The fat depot of the tail of some ovine breeds seems to play a particular role in adaptation to undernutrition; this role could be linked to its smaller adipocytes and high sensitivity to the lipolytic effect of catecholamines. Glucocorticoids and growth hormone probably interact to induce teleophoretic changes in the AT responses to adenosine and catecholamines during lactation. Fat mobilization in dry ewes is related both to body fatness and to energy balance. The in vivo beta-adrenergic lipolytic potential is primarily related to energy balance, whereas basal postprandial plasma non-esterified fatty acids (NEFA) are related to body fatness, and preprandial plasma NEFA is the best predictor of the actual body lipid loss. Several mechanisms seem to be aimed at avoiding excessive fat mobilization and/or insuring a return to the body fatness homeostatic set point. As well as providing the underfed animal with fatty acids as oxidative fuels, AT acts as an endocrine gland. The yield of leptin by ruminant AT is positively related to body fatness, decreased by underfeeding, beta-adrenergic stimulation and short day length, and increased by insulin and glucocorticoids. This finding suggests that the leptin chronic (or acute) decrease in lean (or underfed respectively) ruminants is, as in rodents, a signal for endocrine, metabolic and behavioural adaptations aimed at restoring homeostasis.

Effects of photoperiod and feeding level on adipose tissue and muscle lipoprotein lipase activity and mRNA level in dry non-pregnant sheep

The aim of the present study was to investigate the effects of photoperiod and feeding level on lipid metabolism in ovine perirenal and subcutaneous adipose tissues (AT) and in skeletal and cardiac muscles. Twenty dry non-pregnant ovariectomised ewes were divided into two groups and subjected to either 8 h or 16 h light/d, and underfed at 22 % energy requirements for 7 d. Half of the ewes in each group were slaughtered and the remaining ewes were refed at 190 % energy requirements for 14 d, until slaughtering. Refeeding increased (2.6-4.3-fold) malic enzyme (ME), fatty acid synthase (FAS), glucose-6-phosphate dehydrogenase (G6PDH) and glycerol-3-phosphate dehydrogenase (G3PDH) activities in subcutaneous AT as well as lipoprotein lipase (LPL) activity in perirenal (3.5-fold) and subcutaneous (10-fold) AT and to a lesser extent (1.4-fold) in the skeletal longissimus thoracis and cardiac muscles. Moreover, variations of LPL mRNA level followed variations of LPL activity: refeeding increased perirenal AT- and cardiac muscle-mRNA levels (7.4- and 2-fold respectively). The main finding of this study is that, for a given level of food intake, long days (compared with short days) increased the LPL activity in the longissimus thoracis muscle and, in refed ewes, the activities of LPL and ME in subcutaneous AT. Furthermore, long days increased LPL mRNA level in cardiac muscle and perirenal AT. Thus, our results show that there are direct effects of photoperiod on sheep AT lipogenic potential, as well as on muscle LPL activity, which are not caused by changes in nutrient availability.

Nutritional status induces divergent variations of GLUT4 protein content, but not lipoprotein lipase activity, between adipose tissues and muscles in adult cattle.

Metabolic adaptations to variations in food supply are incompletely understood in ruminant animal adipose tissue (AT) and muscle. To explore this, we studied lipid metabolism and glucose transport potential in one internal and one external AT, as well as in one oxidative and one glycolytic muscle from control, 7 d underfed and 21 d refed adult cows. Refeeding increased (+79 to +307 %) the activities of enzymes involved in de novo lipogenesis (fatty acid synthase, malic enzyme, glucose-6-phosphate dehydrogenase) in perirenal and subcutaneous AT; underfeeding did not modify these variables. Underfeeding decreased the activities of lipoprotein lipase (LPL) in perirenal AT (-70 %) and cardiac muscle (-67 %), but did not modify the activities in subcutaneous AT and longissimus thoracis. Refeeding increased LPL activities in all tissues (+40 to +553 %) to levels comparable with (cardiac muscle) or greater than (AT, longissimus thoracis) those observed in control cows. Such variations in perirenal and cardiac muscle LPL activities did not result from variations in LPL mRNA levels, but suggest a post-transcriptional regulation of LPL in these nutritional conditions. Underfeeding did not modify GLUT4 contents in perirenal AT and muscles, while refeeding increased it only in perirenal AT (+250 %). Our present results contrast with previous results in rats, where LPL is regulated in opposite directions in AT and muscles, and GLUT4 is generally increased by fasting and decreased by refeeding in skeletal muscles. The present results highlight the bovine specificity of the response, which probably arises in part from peculiarities of ruminant animals for nutrient digestion and absorption.

Plasma non-esterified fatty acid response to a β-adrenergic challenge before or after feeding in energy underfed or overfed, dry or lactating cows

The effects of adrenergic challenge (in order to evaluate the adipose tissue lipolytic potential) and time of challenge relative to feeding on the response curve of plasma non-esterified fatty acids (NEFA) in four underfed or overfed non-lactating non-pregnant cows were studied. Basal NEFA and NEFA response to isoproterenol (ISO; 4 nmol/kg body weight) were higher when challenged before than after feeding and higher in underfed than overfed cows. Interaction between feeding level and time of challenge was significant (P < 0·001) for the rising part of NEFA response. Pooled NEFA response curves from several trials (no. = 63 challenges) were analysed in order to obtain a simplified procedure for the prediction of the NEFA response. High correlations were found between the response area or maximal NEFA response and NEFA response at 15 min (r = 0·95 and 0·98, respectively). This simplified procedure was applied on pooled data from 84 challenges in order to evaluate the effects of energy balance, physiological status, body condition score (BCS) and time of challenge relative to feeding. Energy balance had a significant effect on basal plasma NEFA and NEFA response at 15 min after ISO challenge (–7·6 and –14·2 μmol/l when the daily energy balance increased by 1 MJ, respectively). Time of ISO injection relative to feeding had a greater effect on stimulated NEFA than on basal NEFA (308 v. 239 μmol/l). There was a significant effect of BCS (41 μmol/l per unit of BCS) on the basal plasma NEFA level. The NEFA response at 15 min after ISO challenge was lower (387 μmol/l) in lactating cows than in dry cows. The NEFA response to ISO at 15 min could provide an efficient method of studying the adipose tissue lipolytic potential of cattle in vivo.

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.

Effects of underfeeding and refeeding on offals weight in the Barbary ewes

The influence of long-term (161 days) underfeeding and refeeding (154 days) on the weight of offal components, and digestive content, was studied in 26 adult Barbary ewes. Effects on the protein and mineral content of offals were also measured. Animals were split in three groups: one was fed hay at maintenance level (M), another also received vetch-oat hay to cover 0.2 of maintenance energy requirements (L), and the third one was fed at 0.2 of maintenance requirements during 161 days, then refed at 1.3 times of energy requirements with a diet of hay and barley grain (LH) during 154 days. Underfed animals body weight (BW) at slaughter was 31kg. Animals of groups M and LH were slaughtered at similar BW (50kg). Underfeeding reduced empty BW by 38% and the weight of most metabolically active organs: skin (40%), blood (31%), rumen (34%) and liver (19%), but not others (small intestine). Total offals weight was reduced by 27%. Overfeeding of previously underfed animals (LH) induced a significant increase in liver weight in comparison to M group (18%), owing to a higher nutrient supply. The amount of protein in offals and their fat-free weight were decreased by underfeeding, whereas the amount of minerals was significantly modified only in components rich in bone. These results showed that splanchnic organ mass decreases with underfeeding which leads to energy requirements decrease and contributes towards resistance of Barbary ewes to harsh food conditions.

Circannual body reserve dynamics and metabolic profile changes in Romane ewes grazing on rangelands

Throughout an entire year, 41 Romane ewes reared in an extensive rangeland were used to investigate temporal changes in body reserves (BRs) and profiles of related metabolites and metabolic hormones. Ewes were allocated to homogeneous groups according to BW and BCS and were distributed by parity (primiparous [PRIM], n = 21; multiparous [MULT], n = 20) and litter size (LSi; lambing singletons [SING], n = 21 or TWINS, n = 20). The feeding system was based on rotational grazing of rangeland paddocks and progressive supplementation with hay, silage, and barley at late pregnancy during the winter. Individual BW, BCS, plasma NEFA, β-hydroxybutyrate (β-OHB), glucose, insulin, leptin, and triiodothyronine (T3) were monitored at -56, -12, 8, 49, 76, 107, 156, 195, 216, 246, and 301 d relative to lambing. The BR mobilization was observed from late pregnancy to the end of suckling and varied as a function of the ewe energy balance but also because of transitions from fertilized to native rangeland paddocks and by supplementation. Contrarily, BR accretion occurred from weaning, during the dry-off, and until the start of the next pregnancy. Lipolysis was well reflected by NEFA, β-OHB, and T3 kinetics. Mean BW (but not mean BCS) was affected by parity (MULT > PRIM), whereas both BW and BCS were influenced by LSi (SING > TWINS) but only for MULT. The most drastic BW loss was observed during the mid-suckling period (49 d in milk [DIM]) in all ewes. The lack of effects of LSi in PRIM but not in MULT was also evident in the majority of blood plasma kinetics, which were affected (P < 0.0001) by physiological stage in all ewes. A tendency to ketosis (β-OHB) was found in ewes nursing TWINS around lambing, irrespective of parity. Glucose concentrations were greater during suckling and dry-off, and a peak (0.96 ± 0.05 g/L) was attained at 156 DIM in MULT nursing TWINS. The highest plasma leptin concentration was observed during the start and the middle of the next pregnancy in MULT (107-216 DIM; 9.6 ± 0.44 ng/mL). In all ewes the physiological stage affected T3, which was affected by LSi just in MULT (from late pregnancy, MULT by SING > MULT by TWINS; 99.91 vs 85.52 ng/dL) and during suckling (111.7 ± 4.18 ng/dL). Lamb BW was affected at birth and weaning by parity (MULT > PRIM) and LSi (SING > TWINS). Overall, temporal changes in BR were directly affected by the transition of physiological states and feeding levels, whereas individual responses were predetermined by parity. In MULT, the reactivity and magnitude of response was influenced by LSi. The whole set of parameters allowed us to detect sensitive and critical periods throughout the entire annual cycle. We thus identified opportunities for improved nutritional management, for example, during physiological states usually underestimated such as early and mid-pregnancy. This work indicates the applicability of long-term studies about BR dynamics in ruminants as a potential component contributing to farm economic resilience.

In vivo estimation of body composition from the dilution space of deuterium oxide in fat-tailed Barbary ewes

Abstract Sixteen dry ewes of the fat-tailed Barbary breed were used to formulate prediction equations of body water, fat, protein and energy content from the live body weight (BW) and the dilution space of deuterium oxide (SD 2 O). Ewes were injected with 0.5 g D 2 O/kg BW and four blood samples collected after infusion. The D 2 O content of blood water was determined by infra-red spectrometry. Ewes were weighed, body condition scored and then slaughtered. The body water, fat, protein, ash and energy were chemically determined. At slaughter, ewes weighed 43.4±7.0 kg, and contained 26.0±2.6 kg of water and 9.3±4.2 kg of fat. A relative lack of variation in the fat-free empty body (FFEB) was confirmed in this particular breed (water percentage, 74.5±1.6). Adiposity (fat as percent empty BW; EBW) varied between 15.1 and 36.9%. There was a close negative relationship ( R 2 =0.96; residual standard deviation (R.S.D.)=1.4%) between the fat and the water content of EBW. Body water was significantly correlated with SD 2 O. The body fat and energy content prediction equations from SD 2 O and BW were similar to those published in a thin-tailed ewe breed ( R 2 =0.92; R.S.D.=1.4 kg). The body fat equation prediction from body condition score and/or BW was significant ( R 2 =0.81; R.S.D.=1.9 kg) although not as precise as that using D 2 O.

Polyethylene glycol compared with ytterbium oxide as a total faecal output marker to predict organic matter intake of dairy ewes fed indoors or at pasture.

Several external markers can be used for estimating total faecal output in view of assessing ruminant intake at pasture. Among them, ytterbium (Yb) has been used for many years in various conditions. Polyethylene glycol (PEG) is a promising external marker because it can be rapidly determined using near-infrared spectroscopy (NIRS). The study consisted of 24 adult lactating dairy ewes over three periods (P1, P2 and P3), fed with three different diets: P1, total mixed ration (TMR); P2, Italian ryegrass (IRG); and P3, pasture. After an adaptation period, the ewes were administered a daily dose of ytterbium oxide (0.35 g/day) and PEG (20 g/day) for 2 weeks. During the last week, the daily organic matter intake (OMIOBS) was measured. Faecal samples were collected at milking time (0800 and 1600 h) to determine marker content, using only samples collected in the morning (PEGm) or by averaging samples (Yb, PEGma). Faecal marker content made it possible to assess total faecal output, either using the two recovery rates for PEG (0.98 or 0.87) or not. The OMIOBS was assessed on the basis of total faeces estimated with Yb (OMIYb) or PEG (OMIPEG), and the digestibility was calculated on the basis of feed analysis. With total TMR (P1), the OMIPEG, corrected with recovery rate (OMIPEGm98) or not corrected (OMIPEGm) was 2.40 kg/day and 2.50 kg/day, respectively, and was not different (P>0.05) from OMIOBS (2.51 kg/day), whereas OMIYb was lower (2.14 kg/day) (P<0.001). With IRG (P2), OMIPEGm98 (1.67 kg/day), OMIPEGm87 (1.51 kg/day) and OMIYb (1.59 kg/day) were not different (P>0.05) from OMIOBS (1.57 kg/day). With pasture (P3), the OMIPEGm (1.54 kg/day) and OMIPEGm98 (1.48 kg/day) were not different (P>0.05) from the OMI assessed from the biomass measurement (1.52 kg/day). The OMIYb (1.36 kg/day) was lower (P<0.05) but not different from OMIPEGm98 and OMIPEGm87. Spearmans rank correlation between OMIOBS and other OMIs (predicted with Yb or PEG P1 and P2) showed that it is possible to rank animals using PEG when there is a sufficiently wide range of OMIOBS (1.65 to 2.8 kg/day in P1) but not within a narrower range (1.47 to 1.72 kg/day in P2). In conclusion, the present study confirms that PEG is a valuable external faecal marker, easy to prepare (solution), administer and determine (NIRS). It can be used to assess intake with numerous animals at pasture, but only for groups, and not to quantitatively estimate individual OMI.

An assessment of Walk-over-Weighing to estimate short-term individual forage intake in sheep

The main limitation for determining feed efficiency of freely grazing ruminants is measurement of daily individual feed intake. This paper describes an investigation that assessed a method for estimating intake of forage based on changes in BW of ewes. A total of 24 dry and non-pregnant Romane ewes (12 hoggets, HOG; mean±SD 51.8±2.8 kg BW; body condition score (BCS) 2.6±0.2; and 12 adults, ADU; 60.4±8.5 kg BW; BCS 2.7±0.8) were selected for the study and moved from their rangeland system to a confined pen with controlled conditions and equipped with individual automatic feeders. The experiment lasted for 28 days (21 days adaptation and 7 days feed intake measurement). Ewes were fed hay and trained to use the electronic feeders (one feeding station per ewe) in which actual daily intake (H intake24) was measured. The pens were designed to maximize movement of trained ewes through an automated Walk-over-Weighing device, by using water and mineral salts as attractants. Total individual intake of hay measured in the automatic feeder at each meal (H intake) was compared with indirect estimates of feed intake determined using differences in the BW of the ewes (∆BW) before and 1 h following morning and afternoon feeding at fixed times. The BW, BCS, H intake, H intake24, as well as plasma non-esterified fatty acids (NEFA), glucose and insulin profiles were determined. The BW was higher in ADU v. HOG but BCS was not affected by parity. The H intake24 was affected by day of experiment as a consequence of reduced availability and intake of water on one day. Plasma glucose, NEFA and insulin were not affected by parity or day of experiment. The H Intake was and ∆BW tended to be higher in the morning in HOG, whereas H intake was and ∆BW tended to be higher in ADU at the afternoon meal. Irrespective of parity or feeding time, there was very strong correlation (r 2=0.93) between H intake and ∆BW. This relationship confirms that our indirect method of estimating individual forage intake was reliable within the strictly controlled conditions of the present experiment. The method appears suitable for use in short-term intensive group feeding situations, and has potential to be further developed for longer-term forage intake studies, with a view to developing a method for freely grazing ruminants.