Underfed Animals Research Articles

The effects of follicle stimulating hormone (FSH)-induced controlled ovarian hyperstimulation and nutrition on implantation-related gene expression in caruncular tissues of non-pregnant sheep

Disturbances at the conceptus-maternal interface can have detrimental effects on pregnancy outcome. Additionally, changes in body condition and exogenously administered gonadotropins could affect ovarian and uterine function, including cell proliferation and ovulation rates, and alter endometrial receptivity. In ruminants, endometrial caruncles maintain placental function via interaction with fetal chorionic cotyledons. Here, the effects of feeding regimens on the expression of selected genes known to be involved in uterine receptivity were investigated in the caruncles of control and FSH-superovulated ewes. Sheep were grouped according to their diet: control fed (CF), overfed (OF) or underfed (UF), and were either superovulated with FSH (SOV) or untreated (CON, naturally cycling) (n = 3–5/group). Caruncular samples for the assessment of the transcript levels of 11 target genes were collected at either the early (day 5) or mid-luteal (day 10) phases of the luteal lifespan, resulting in 12 groups of animals. The day of the estrous cycle affected the expression of ITGAV, ITGB3, FGF10 and IGFBP3 mRNA. There was lower expression of MUC1, and higher expression of FGF10, ITGB3 and FN1, on day 10 in CF_SOV animals. Compared with CF, expression of integrins (ITGB3, ITGA5 and ITGA4) was higher in OF and UF, and higher transcript levels of HGF and IGFBP3 in UF animals on day 10. Expression of ITGA5, ITGB1, -3, -5 and MUC1 was greater in OF_SOV than CF_SOV at day 10. In conclusion, it appears that imbalanced nutrition, by altering the expression of genes responsible for intercellular communication, cell adhesion, and encoding for growth factors, could affect the uterine responsiveness to exogenously applied hormonal stimulation and, likely, uterine receptivity.

46 How far we could go reducing crude protein with the use of supplemental amino acids

Abstract Feeding growing pigs with diets providing the required amount of essential and non-essential amino acids (AA) reduces energy expenditure and minimizes N excretion. Low protein diets can be obtained by supplementing feeds with crystalline AA. Numerous experiments have evaluated the ideal dietary AA concentration at different growth stages, but reducing dietary protein with the use of supplemental AA is limited by the inaccuracy of the principles used to estimate AA requirements. One of these principles states that growing animals need AA for maintenance and growth. Maintenance requirements are related to BW whereas the efficiency of AA utilization (e.g., 72% for Lys) and body protein AA composition are constant (e.g., 7% for Lys). These parameters are, however, affected by AA restriction, meal frequency, energy supply, genetics, etc. Even when controlling these factors, individual pigs respond differently to the same AA supply. Yet pigs are raised in groups and fed with a unique feed for long periods. Individual pigs within a given population differ in terms of BW, ADG, health status, etc., and consequently, differ in the amount of AA they need at a given time. Therefore, when feeding a group of pigs, the concept of maintenance and growth requirements may not be appropriate. In this situation, nutrient requirements should be seen as the optimal balance between the proportion of animals that needs to be overfed and underfed. Given that for most AA, underfed animals exhibit reduced performance, whereas overfed animals exhibit near-optimal performance, optimal growth is obtained when nutrients are provided to satisfy the requirements of the most demanding animals. There is therefore a trade-off between performance and dietary protein reduction. The inaccuracy of the principles used to estimate AA requirements, both for individual animals and populations, limits how far we can go reducing dietary protein with the use of supplemental AA.

Developmental programming of somatic growth, behavior and endocannabinoid metabolism by variation of early postnatal nutrition in a cross-fostering mouse model

BackgroundNutrient deprivation during early development has been associated with the predisposition to metabolic disorders in adulthood. Considering its interaction with metabolism, appetite and behavior, the endocannabinoid (eCB) system represents a promising target of developmental programming.MethodsBy cross-fostering and variation of litter size, early postnatal nutrition of CB6F1-hybrid mice was controlled during the lactation period (3, 6, or 10 pups/mother). After weaning and redistribution at P21, all pups received standard chow ad libitum. Gene expression analyses (liver, visceral fat, hypothalamus) were performed at P50, eCB concentrations were determined in liver and visceral fat. Locomotor activity and social behavior were analyzed by means of computer-assisted videotracking.ResultsBody growth was permanently altered, with differences for length, weight, body mass index and fat mass persisting beyond P100 (all 3>6>10,p<0.01). This was paralleled by differences in hepatic IGF-I expression (p<0.01). Distinct gene expression patterns for key enzymes of the eCB system were observed in fat (eCB-synthesis: 3>6>10 (DAGLα p<0.05; NAPE-PLD p = 0.05)) and liver (eCB-degradation: 3>6>10 (FAAH p<0.05; MGL p<0.01)). Concentrations of endocannabinoids AEA and 2-AG in liver and visceral fat were largely comparable, except for a borderline significance for higher AEA (liver, p = 0.049) in formerly overfed mice and, vice versa, tendencies (p<0.1) towards lower AEA (fat) and 2-AG (liver) in formerly underfed animals. In the arcuate nucleus, formerly underfed mice tended to express more eCB-receptor transcripts (CB1R p<0.05; CB2R p = 0.08) than their overfed fellows. Open-field social behavior testing revealed significant group differences, with formerly underfed mice turning out to be the most sociable animals (p<0.01). Locomotor activity did not differ.ConclusionOur data indicate a developmental plasticity of somatic growth, behavior and parameters of the eCB system, with long-lasting impact of early postnatal nutrition. Developmental programming of the eCB system in metabolically active tissues, as shown here for liver and fat, may play a role in the formation of the adult cardiometabolic risk profile following perinatal malnutrition in humans.

Mitotic index and morphological characteristics of ovarian small follicles from goats submitted to nutritionally unbalanced regimens.

The objective of this study was to assess the influence of nutritional regimens such as adequate feeding, restricted feeding, and underfeeding-refeeding on the follicle growth and development from caprine ovaries. Goats were divided into three different groups (n = 5 per group). For 24 weeks, goats received elephant grass plus concentrate to provide 1.5 (n = 5) and 0.72 (n = 10) times the energy requirements for maintenance of live weight. Underfed goats were subsequently refed for 6 weeks with the diet of the nourished group (1.5 times the energetic requirements of maintenance). Follicular morphology and morphometry, as well as granulosa cells mitotic index were assessed. Ovarian follicles were classified as small or large preantral follicles, or as small or large antral follicles. Ovarian volume was smaller in animals from both underfed and refed groups than in those animals from fed group. Although no difference in the total number of normal follicles was observed among the nutritional groups, underfed animals presented higher percentages of atretic preantral and small antral follicles when compared with fed animals. Large antral follicles from underfed and refed goats presented a lower mitotic index when compared with fed ones. In conclusion, ovaries from goats challenged with prolonged undernutrition will be functionally compromised, which is characterized by atresia of preantral and small antral follicles and decreased mitotic index of large antral follicles. Refeeding those animals will not recover ovarian function to a same level experienced by goats fed a diet with adequate energy requirements.

Periconceptional undernutrition modifies endocrine profiles and hepatic gene expression in sheep.

This study investigated whether a 22-day period of undernutrition (half maintenance) could affect maternal endocrine responses and liver gene expression during early pregnancy (day 7). Thirty-five ewes were fed 1.5 (n=15) or 0.5 (n=20) their maintenance requirements and slaughtered on day 7 of the oestrus cycle or pregnancy (oestrus=day 0). Insulin, IGF, leptin and non-esterified fatty acids (NEFA) were determined on days -14, 0 and 7. Transcripts of the IGF family and adipokines receptors were determined in the liver by real-time RT-PCR. Underfed animals presented lower body weight and body condition, greater plasma concentration of NEFA, and lower plasma concentrations of leptin, insulin and IGF1 compared to adequately fed animals. Underfed ewes presented greater hepatic expression of IGFBP2 than well-fed ewes, but tended to have lesser expression of IGFBP5. While no effect of undernutrition on IGFBP4 and ADIPOR2 mRNA expressions was observed, they were increased by pregnancy in underfed animals. This study shows that undernutrition modifies endocrine profiles and hepatic gene expression of IGFBP2 and 5. The pregnancy status increased hepatic gene expression of IGFBP4 and ADIPOR2 mRNA in undernourished ewes.

Does the Fat Tailed Damara Ovine Breed Have a Distinct Lipid Metabolism Leading to a High Concentration of Branched Chain Fatty Acids in Tissues?

Fat tailed sheep breeds are known for their adaptation to nutritional stress, among other harsh production conditions. Damara sheep, native to Southern Africa, have recently been exported to other areas of the world, particularly Australia, aiming to produce lamb in semi-arid regions. Damaras have a unique hanging fat tail, a fat depot able to be mobilized under nutritional stress. In this article we perform an in-depth characterization of the fatty acid profiles of the fat tail in underfed and control Damara rams. Profiles were very similar between experimental groups, with the exception of palmitic acid (16:0) that was lower (P = 0.014) in underfed animals. However, the most striking result was the very high proportions of non-terminal branched chain fatty acids found in the fat tail adipose tissue, as well as the gastrocnemius muscle of Damara rams. The muscle of Dorper and Merino rams used in the same experiment did not present non-terminal branched chain fatty acids, suggesting that Damara rams have a unique lipid metabolism. Herein, we interpret this trait relating it to a higher ability of Damara sheep to digest fibrous fodder and to putative differences in the propionate metabolism by comparison to other sheep breeds.

Thermoregulation of nutrient-restricted hair ewes subjected to heat stress during late pregnancy

In summers with severe hot environmental temperatures, thermoregulation is expected to be critical during the last third of gestation of hair ewes, and this effect can be overstated by suboptimal feeding conditions. Twenty-four multiparous hair sheep ewes with approximately 100d of pregnancy were randomly assigned to two treatments to evaluate the effects of nutritional restriction (NR) during late gestation on some serum metabolites, physiological variables, and thyroid hormone concentrations under intense heat load (maximum temperature 42–45°C). Treatments were as follows: (1) control, ewes fed free choice wheat straw plus a concentrate at a rate of 500g/d, and (2) nutritionally-restricted (NR) ewes, which were offered wheat straw only ad libitum. Body weight of NR ewes dropped 5.1kg (P<0.05), while control ewes gained 4.7kg during the study period. Rectal temperature was not affected (P>0.05) by feeding regime either in the morning or afternoon hours. Respiration rate was greater (P<0.05) in control ewes (20 and 40breaths/min higher in the day and night, respectively) on d 130 and 145 of pregnancy compared to NR ewes. Skin temperatures during the morning and afternoon were affected (P<0.05) by feeding regime at d 130 and 145 of pregnancy. In the morning, NR ewes presented greater (P<0.05) head and rump temperature at day 145 of gestation, and lower (P<0.05) udder temperatures at d 130 and 145 than control ewes. In the afternoon, skin temperatures of NR ewes were higher (P<0.05) in head and right flank on d 130 and 145 of pregnancy, and in udder at day 145 compared to control ewes. Serum glucose was higher (P<0.05) in NR ewes than control animals at day 145 of pregnancy. Serum cholesterol, triglyceride and thyroid hormones were not affected by nutritional restriction. Overall, it was found that nutritionally restricted ewes were less affected by intense heat loads than well-fed ewes, apparently due to the lower metabolic heat produced by this underfed animals. Also, it was apparent that the lower respiration rate of NR ewes was compensated by a greater body surface temperature.

Early Metabolic Programming of Puberty Onset: Impact of Changes in Postnatal Feeding and Rearing Conditions on the Timing of Puberty and Development of the Hypothalamic Kisspeptin System

Kiss1 neurons have recently emerged as a putative conduit for the metabolic gating of reproduction, with leptin being a regulator of hypothalamic Kiss1 expression. Early perturbations of the nutritional status are known to predispose to different metabolic disorders later in life and to alter the timing of puberty; however, the potential underlying mechanisms remain poorly defined. Here we report how changes in the pattern of postnatal feeding affect the onset of puberty and evaluate key hormonal and neuropeptide [Kiss1/kisspeptin (Kp)] alterations linked to these early nutritional manipulations. Female rats were raised in litters of different sizes: small (four pups per dam: overfeeding), normal (12 pups per dam), and large litters (20 pups per litter: underfeeding). Postnatal overfeeding resulted in persistently increased body weight and earlier age of vaginal opening, as an external sign of puberty, together with higher levels of leptin and hypothalamic Kiss1 mRNA. Conversely, postnatal underfeeding caused a persistent reduction in body weight, lower ovarian and uterus weights, and delayed vaginal opening, changes that were paralleled by a decrease in leptin and Kiss1 mRNA levels. Kisspeptin-52 immunoreactivity (Kp-IR) in the hypothalamus displayed similar patterns, with lower numbers of Kp-IR neurons in the arcuate nucleus of postnatally underfed animals, and a trend for increased Kp-positive fibers in the periventricular area of early overfed rats. Yet, gonadotropin responses to Kp at puberty were similar in all groups, except for enhanced responsiveness to low doses of Kp-10 in postnatally underfed rats. In conclusion, our data document that the timing of puberty is sensitive to both overfeeding and subnutrition during early (postnatal) periods and suggest that alterations in hypothalamic expression of Kiss1/kisspeptin may underlie at least part of such programming phenomenon.

Early pregnancy alters the metabolic responses to restricted nutrition in sheep

This study investigated whether a 27-day period of nutrition at half-maintenance during early pregnancy (up to Day 14) could alter maternal endocrine responses. Forty-six ewes were fed all or half of their maintenance requirements and slaughtered on Day 14 of the oestrous cycle or pregnancy. We used real time RT-PCR to study gene expression of growth hormone receptor (GHR) and leptin in adipose tissue and GHR, GHR1A and of the insulin-like growth factor I (IGF-I) in the liver. Blood profiles of metabolites and metabolic hormones were also determined. Throughout the experiment, underfed animals presented lower body weight and body condition, greater plasma concentrations of non-esterified fatty acids (NEFA), and lower plasma concentrations of leptin, compared to adequately fed animals. Undernutrition affected the patterns of gene expression in adipose and hepatic tissues, and the responses differed between pregnant and non-pregnant ewes. In adequately fed ewes, pregnancy up-regulated leptin mRNA expression in adipose tissue, a response that was impaired in underfed ewes. The hepatic expression of IGF-I mRNA was increased by pregnancy in underfed animals while no effect was observed in adequately fed ewes. It remains to be determined whether the changes in the endocrine milieu are paralleled by modifications in uterine gene expression that could alter the environment of the embryo during early pregnancy.

Desensitization of gonadotropin responses to kisspeptin in the female rat: analyses of LH and FSH secretion at different developmental and metabolic states

Kisspeptins have emerged as potent elicitors of gonadotropin secretion and, therefore, putative targets for pharmacological intervention. In this context, desensitization of gonadotropin responses to continuous administration of kisspeptins has begun to be characterized, but information so far available is mostly restricted to LH responses in males, whereas the similar phenomenon in females, of obvious therapeutic interest, remains virtually unexplored. We report herein LH and FSH responses to continuous intracerebral administration of kisspeptin in female rats at different developmental and metabolic states. Infusion of kisspeptin-10 to adult female rats induced a transient elevation in serum LH concentrations, followed by a precipitous drop and normalization of LH levels thereafter. Elevation of LH after kisspeptin infusion was prolonged in underfed animals; a phenomenon mimicked by leptin administration. Conversely, FSH levels were persistently heightened along continuous kisspeptin infusion, but duration of this response was shortened by undernutrition. In pubertal females, LH and FSH levels remained elevated at the end of a 7-day infusion of kisspeptin; responses whose magnitude was augmented by subnutrition but not mimicked by leptin. In all settings, terminal gonadotropin-releasing hormone responses were fully preserved, suggesting that eventual desensitization must occur upstream from the pituitary. In summary, our current data document the pharmacological consequences of continuous administration of kisspeptin to female rats, with remarkable differences being detected between LH and FSH responses, in different developmental and metabolic states. These observations of potential pharmacological interest might help also to delineate the physiological roles of kisspeptins in the dynamic regulation of gonadotropin secretion in the female.

Compensating for a bad start: catch‐up growth in juvenile lizards (Amphibolurus muricatus, agamidae)

In most natural environments, food availability varies unpredictably through space and time, and growth rates of individual organisms respond accordingly. However, growth rates are not necessarily a simple function of current nutritional conditions: growth rates can be affected by earlier nutritional experience as well as current circumstances. Thus, even a brief period of dietary restriction early in life might influence growth rates later on: either reducing them (if early experience sets subsequent rates, as in the "silver spoon" effect) or increasing them (if underfed individuals can compensate by growing more rapidly to cancel out the early decrement). Alternatively, later growth may be unaffected by earlier rates of growth. We experimentally manipulated food supply (and thus, growth rates) of hatchling lizards (Amphibolurus muricatus) for 1 month post-hatching, then maintained both high-food and low-food animals under identical nutritional conditions in outdoor enclosures for another 6 months. Low food abundance early in life significantly reduced juvenile growth, but these previously underfed animals exploited the subsequent (common garden) conditions to grow much faster than their larger (initially better-fed) siblings. Thus, the two groups were indistinguishable in body size at 6 months of age. Intriguingly, the compensatory growth occurred in winter, a period that is generally unsuitable for rapid growth in ectotherms.

Prolonged underfeeding of sheep increases myostatin and myogenic regulatory factor Myf-5 in skeletal muscle while IGF-I and myogenin are repressed.

The IGF axis is nutritionally sensitive in vivo and IGFs stimulate myoblast proliferation and differentiation in vitro, while myostatin inhibits these processes in vitro. We hypothesised that underfeeding would reversibly inhibit the myogenic activity of satellite cells in vivo together with decreased IGF-I and increased myostatin in muscle. Satellite cell activity was measured indirectly from the expression of proliferating cell nuclear antigen (PCNA) and the myogenic regulatory factors (MRFs), MyoD, Myf-5 and myogenin. Young sheep were underfed (30% of maintenance) and some killed after 1, 4, 12, 17, 21 and 22 weeks. Remaining underfed animals were then re-fed a control ration of pellets and killed after 2 days, and 1, 6 and 30 weeks. Expression of PCNA and MRFs decreased during the first week of underfeeding. This coincided with reduced IGF-I and myostatin mRNA, and processed myostatin. Subsequently, Myf-5, MyoD, myostatin mRNA and processed myostatin increased, suggesting that satellite cells may have become progressively quiescent. Long-term underfeeding caused muscle necrosis in some animals and IGF-I and MRF expression was increased in these, indicating the activation of satellite cells for muscle repair. Re-feeding initiated rapid muscle growth and increased expression of PCNA, IGF-I and the MRFs concurrently with decreased myostatin proteins. In conclusion, these data indicate that IGF-I and myostatin may work in a coordinated manner to regulate the proliferation, differentiation and quiescence of satellite cells in vivo.

Opioid effects on glucose and eicosanoid metabolism in isolated uterus of ovariectomized and non-ovariectomized restricted diet rats

The effect of a 25-day restricted diet (50% of the normal food intake) on uterine glucose metabolism of ovariectomized (25 days) and non-ovariectomized rats, was studied. Underfeeding reduces 14CO2 production from U14C-glucose in intact animal. However, in spayed rats, results are the opposite. In intact rats receiving a low food intake, the effect of the addition to the KRB medium of various agonist opioids, was studied. Dinorphin A did not bring about any change. On the other hand, β endorphin increased glucose metabolism. Also, the addition of Dago and Dadle increased 14CO2 production, while their corresponding specific blockers, β-FNA and Naltrindole, reversed it. Ovariectomized rats subjected to food restriction are not affected by opioid agonists. In vitro morphine, like endogenous opioids, increased 14CO2 in intact restricted diet rats. Arachidonic acid metabolism in these rats show that underfeeding brings about a decrease in PGF2α and PGE2, but the addition of morphine does not alter this situation, for which eicosanoids metabolites are not related to the effect of morphine. The morphine effect was not altered by naloxone. The subcutaneous injection of morphine increased glucose metabolism in intact underfed animals, while naloxone reduced 14CO2 in spayed rats subjected to underfeeding. It can be concluded that uteri from ovariectomized rats receiving a restricted diet are influenced by a mechanism of upregulation related to endogenous opioids. These likely originate in other tissues, and so prevent us from seeing the morphine effect.

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 31 kg. Animals of groups M and LH were slaughtered at similar BW (50 kg). 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.

Effect of Restricted Food Intake on Production, Catabolism, and Effects of IGF-I and Cyclic Nucleotides in Cultured Ovarian Tissue of Domestic Nutria (Myocastor coypus)

The aims of these in vitro experiments were to examine the effects of short-term food restriction on ovarian secretory activity and the role of IGF-I and cAMP- and cGMP-dependent intracellular mechanisms in the control of ovarian function in domestic nutria. Slices of ovary from sexually mature animals kept under conditions of normal and restricted (12 of standard ration) feeding were cultured with or without IGF-I (50 ng/ml), cAMP analogues (dbcAMP and Rp-cAMPS), and cGMP analogues (8-pCPT-cGMP and Rp-8-Br-PET-cGMPS; all at 100 nM). In nonovarian cells dbcAMP activates and Rp-cAMPS inhibits protein kinase A, while 8-p-CPT-cGMP activates and RP-8-Br-PET-cGMPS inhibits protein kinase G and cGMP-gated ion channels. IGF-I release and catabolism, as well as the release of progesterone (P), estradiol (E), and cAMP by the cultures, were evaluated using RIA. IGF-I did not affect cAMP release, while each of the cAMP and cGMP analogues inhibited IGF-I release in both control and experimental groups. Fasting did not affect cAMP or IGF-I release. It partially prevented the effect of Rp-cAMPS, but not of other cyclic nucleotides, on IGF-I release and inhibited IGF-I catabolism. The Rp-cAMPS and Rp-8-Br-PET-cGMPS also inhibited IGF-I catabolism and the effects were greater with tissue from food-restricted than control animals. Ovaries from the underfed nutria secreted significantly more P and less E than those from normally fed animals. IGF-I and both cAMP analogues, given alone, did not affect P release whereas a combination of IGF-I and Rp-cAMPS increased P output in control, but not in the experimental group. The 8-pCPT-cGMP had no effect P release. Rp-8-Br-PET-cGMPS, given alone or in combination with IGF-I, dramatically increased P secretion by tissue from control but not underfed animals. Estradiol secretion by tissue from underfed animals was stimulated by IGF-I, dbcAMP, Rp-cAMPS, 8-pCPT-cGMP, and Rp-8-Br-PET-cGMPS as well as by combinations of IGF-I and Rp-cAMPS or Rp-8-Br-PET-cGMPS; these effects were not seen with control tissue. The results demonstrate that: (1) ovaries of domestic nutria secrete IGF-I, P, E, and cAMP; (2) cAMP and cGMP can influence IGF-I release and catabolism; (3) the cyclic nucleotides may have an IGF-I-mediated effect on P and E output; (4) IGF-I and cyclic nucleotides can prevent the effect of undernutrition on E, but not on P release; (5) effects of cAMP and cGMP on P and E are probably not mediated by protein kinase A, protein kinase G, or cGMP-gated ion channels; and (6) food restriction can influence ovarian IGF-I catabolism, P, and E release and modulate the effects of cyclic nucleotides and IGF-I on steroidogenesis. It is concluded that ovarian secretory activity may be regulated separately by nutrition and the cyclic nucleotide-IGF-I system, and there may be functional interrelationships between these mechanisms.

Effects of Morphine and Naloxone on Glucose Metabolism in Uterine Strips from Ovariectomized and Non-Ovariectomized Restricted Diet Rats

The effect of underfeeding over glucose metabolism in uteri isolated from ovariectomized and non-ovariectomized rats subjected to a restricted diet for 25 days (50% of the normal food intake), was studied. Underfeeding decreases 14CO2 formation from U14 C-glucose in intact animal uteri. While in ovariectomized rats (25 days), the effect is the opposite. The addition of morphine 10−6 M to the medium does not affect rats fed ad libitum. However, 14CO2 levels increase significantly in intact animals receiving a restricted diet. In ovariectomized rats morphine does not show any activity, regardless of the type of diet rats were subjected to. None of the rat groups seems to be sensitive to naloxone 10−6 M. The s.c. injection of morphine (4 mg.kg−1) increases glucose metabolism only in intact rats provided with a restricted diet, while naloxone (2.5 mg.kg−1) produces a decrease of 14CO2 in ovariectomized underfed animals. To conclude, morphine either ‘in vivo’ or ‘in vitro’ is active only in uteri from intact rats subjected to underfeeding. Naloxone produces a decrease in 14CO2 production, particularly when it is s.c. injected to ovariectomized rats undergoing a dietary restriction. Since the uterus does not react to naloxone, the effect of the opiod blocker may be the result of endogenous opioids originated in other tissues.

Effects of underfeeding and refeeding on weight and cellularity of splanchnic organs in ewes.

We assessed the effects of a long and severe period of underfeeding, followed by a rapid refeeding with a high-concentrate diet, on weight, protein mass, and cellularity of the splanchnic organs in adult ewes. Twenty-four ewes, allocated to four groups of six, were fed a forage diet (50% regrowth of natural grassland hay and 50% wheat straw) either at maintenance (groups M and MO) or at 40% maintenance (groups U and UO) for 78 d. Groups M and U were then slaughtered, and groups MO and UO were subsequently overfed a high-concentrate diet (52% hay, 20% barley, 16% rapeseed meal, 4% fish meal, and 8% Megalac) at 236% maintenance for 26 d before being slaughtered. During the experiment, feed was adjusted to maintain feed supply at a constant percentage of animal requirements. After slaughter, fresh weight, dry weight, and protein mass of the reticulorumen, omasum, abomasum, small intestine, large intestine, and liver were measured. Cellularity was assessed from nucleic acids and protein contents for both ruminal mucosa and muscular-serosa layers, jejunum, and liver. The concentrations of ubiquitin and cathepsin D mRNA were measured in ruminal mucosa and muscular-serosa layers and in jejunum. Underfeeding decreased protein mass of splanchnic organs, especially in liver (-29%) and reticulorumen (-39%). Refeeding previously underfed animals increased protein mass of liver (+102%) and small intestine (+59%). No carry-over effect of the previous level of intake (UO vs. MO) was observed on the protein mass of splanchnic tissues after 26 d of refeeding. Variations in liver mass were mainly due to hypertrophy, as determined by the protein:DNA ratio, whereas variations in small intestinal mass were mainly due to hyperplasia, as determined by the amount of DNA. By contrast, changes in rumen mass associated with increasing ME intake seemed to be related to hypertrophy in the muscular-serosal component and hyperplasia in the epithelial component. The concentrations of ubiquitin and cathepsin D mRNA in the rumen and jejunum were not modified by feeding level, demonstrating that the expression of these genes for proteolytic enzymes was unchanged under these conditions.

Digestive and metabolic adaptations of ruminants to undernutrition, and consequences on reproduction.

In response to undernutrition, short- (days) and medium-term (weeks) adaptations are more pronounced for splanchnic organs than for other tissues. For the latter, the long-term response involves a sequential mobilization (fat > muscle > bone) with relative priorities differing among anatomical sites. Among chemical components, the body lipids are extensively used (up to 80%) in underfed animals, while the range of protein utilization is limited (up to 15-20%). The decrease in energy expenditure during undernutrition is mostly due to a short- and medium-term decrease in feeding activity, and in the mass and activity of splanchnic tissues. In the long-term, energy expenditure and tissue masses decrease concomitantly, but there does not appear to be a significant decrease in expenditure per unit tissue weight. Nitrogen losses decrease in response to decline in nitrogen fluxes and also due to sparing by renal activities and sometimes by urea recycling. However, ruminants do not seem to be able to compensate for a low level of intake (below maintenance) by an increase in digestive efficiency. Numerous hormones (insulin, growth hormone, insulin-like growth factor I, catecholamines, thyroid hormones, cortisol, leptin, etc.) are involved in the changes during undernutrition of nutrient fluxes between tissues, either through control of synthesis and/or degradation in peripheral tissues (adipose tissue and muscle), or through hepatic conversions of substrates (gluconeogenesis, ureagenesis and ketogenesis), in order to maintain the constancy of the internal environment (homeostasis) and/or to sustain productive functions (teleophoresis). However, reproductive process may be blocked in underfed animals.

Effects of ovarian steroids on glucose metabolism in uterus horns isolated from underfed rats. Influence of castration.

The effect of a restricted diet (50% of the normal intake) during 25 days, on the glucose metabolism was explored in uterus horn strips isolated from intact and ovariectomized rats. In intact underfed rats, the formation of 14CO2 from U14C-glucose was significantly lower than in their controls. In castrated rats, the formation of 14CO2 increased after underfeeding. The return to ad libitum feeding increased glucose metabolism in both groups. Total food deprivation for 4 days diminished 14CO2 formation both in non-ovariectomized rats and in ovariectomized ones as measured at the end of this fasting period. This diminution was greater in intact rats. The increase of glucose metabolism provoked by the administration of s.c. estradiol, progesterone or their combination was more important in underfed intact animals. In intact rats, blood levels of endogenous estradiol and progesterone decreased as a result of underfeeding. In spayed ones, their low concentration increased in relation to their controls.

Nutrient metabolism and utilization in the liver

The role of ruminant liver in modifying the profile of absorbed nutrients before presented to peripheral tissues is illustrated with literature data on hepatic nutrient exchanges. These data are mainly from experiments with multicatheterized dairy cows enabling estimation of blood flow rates and nutrient balances across the liver. In cow liver there is net uptake of propionate, butyrate, valerate, long-chain fatty acids, lactate, amino acids, NH 4 + and O 2, while there is net release of acetate, ketone bodies, glucose, urea and CO 2. Approximately 55% of urea synthesized in the liver is recycled to portal drained viscera. More than 40% of amino acid net absorption is removed by the liver resulting in an amino acid deficit for milk protein synthesis. This could indicate an underestimation of amino acid absorption. Estimated negative carbon balances across the liver can be related to incomplete data concerning lipid exchange and also to carbon use in gluconeogenesis as hepatic uptake of gluconeogenic carbon is less than hepatic release of glucose-carbon. Within a range of glucose flux rates from 3 to 15 mol/d milk yield is linearly related to glucose flux rate. This relationship and published relationships between feed intake and hepatic glucose output are used to calculate regression estimates of basal glucose requirement in cows (2 mol/d) as well as glucose requirement for milk synthesis (0.4 mol/kg milk). Glucose synthesis in the liver is under both metabolic (stimulation by substrate availability; inhibition by glucose itself) and hormonal control (stimulation by glucagon and probably by somatotropin; inhibition by insulin). In early lactation with increased lipid mobilization and fatty acid oxidation in the liver a high rate of gluconeogenesis is ensured by regulation of metabolite transport across mitochondrial membranes. However, in severely underfed animals with excessive lipid mobilization and ketogenesis glucose synthesis is depressed. Reference is given to published mathematical models of liver metabolism in lactating cows. Simulation results obtained with such models have brought further attention to the need for more quantitative knowledge, both regarding hepatic nutrient exchanges (in particular lipids and proteins) and also inter-organ nutrient metabolism (eg. regulation of amino acid catabolism). It is concluded that future feed evaluation systems for ruminants should be based on dynamic, mechanistic whole animal models simulating feed intake, digestion and absorption as well as nutrient metabolism and utilization.