Behavioural development in the neonatal lamb: effect of maternal and birth-related factors

The survival of the neonate relies on the integrated expression of appropriate behaviours from both the mother and young. In precocious species, like the sheep, the speed with which the lamb stands and seeks the udder is related to lamb survival. In this study the effects of birth difficulty, lamb birth weight, ewe loss or gain in backfat from conception to delivery, ewe parity, litter size, and lamb sex on neonatal lamb behavioural development were investigated in over 500 lambs of two breeds (Scottish Blackface and Suffolk). Lambs that required assistance to be delivered were significantly slower to perform all neonatal behaviours than unassisted lambs (P<0.001), and were less active over the first 3 days after delivery (P<0.05). There were no effects of lamb birthweight that were not accounted for by the increased likelihood of requiring assistance in heavier birth weight lambs. Ewes that mobilised less body fat during their pregnancy produced lambs that stood and sucked quickly (P<0.001), and were more active over the first 3 days of life. Lambs born to first parity ewes were slower to stand and suck than lambs born to experienced ewes (P<0.001). There was an improvement in time taken by lambs to stand, seek the udder and to suck with each increase in ewe parity. Litter size had an additional retarding influence on the behaviour of multiple-born lambs that could not be accounted for by birthweight. In the Suffolk breed male lambs were slower to stand and suck than female lambs, this effect was not seen in Blackface lambs. These data demonstrate that lambs that require assistance at birth, even if they survive the birth process, lambs born to ewes that lose a lot of condition over pregnancy or first parity ewes, triplet lambs and, at least in some breeds, male lambs are slower to progress through the sequence of neonatal behaviours. These lambs are, therefore, at greater risk of not surviving to weaning.     

Maternal serum progesterone concentration during pregnancy and lamb birth weight at parturition in Javanese Thin-Tail ewes with different litter sizes

Pregnant ewes, (N=38), with similar body weight and age (19, 13 and 6 carrying 1, 2 and 3 fetuses, respectively) were used to study the relationship between maternal serum progesterone concentration during pregnancy and lamb birth weight at parturition. Average maternal serum progesterone concentration in the ewes carrying 1, 2, and 3 fetuses was: 5.3±0.3, 6.2±0.7 and 6.6±0.5 ng/ml, during weeks 0 to 7 of gestation; 16.9±1.4, 25.3±1.5, and 26.8±2.5 ng/ml, during weeks 8 to 20; and 13.2±1.0, 18.7±1.0, and 19.8±1.7 ng/ml, during the entire gestation period respectively. Total lamb birth weight in ewes carrying 1, 2, and 3 fetuses was 1.9±0.1, 3.2±0.2, and 4.2±0.4 kg, respectively. In the respective litter sizes, ewes with higher mean serum progesterone concentration during the whole pregnancy gave birth to lambs with higher birth weight (r²=0.76, 0.42 and 0.46, for ewes carrying 1, 2 and 3 fetuses, respectively). The results of the study indicated that prenatal growth could probably be improved by increasing endogenous secretion of maternal progesterone during pregnancy.     

Low-intensity contraction activates the alpha1-isoform of 5'-AMP-activated protein kinase in rat skeletal muscle

Skeletal muscle expresses two catalytic subunits, alpha1 and alpha2, of the 5'-AMP-activated protein kinase (AMPK), which has been implicated in contraction-stimulated glucose transport and fatty acid oxidation. Muscle contraction activates the alpha2-containing AMPK complex (AMPKalpha2), but this activation may occur with or without activation of the alpha1-containing AMPK complex (AMPKalpha1), suggesting that AMPKalpha2 is the major isoform responsible for contraction-induced metabolic events in skeletal muscle. We report for the first time that AMPKalpha1, but not AMPKalpha2, can be activated in contracting skeletal muscle. Rat epitrochlearis muscles were isolated and incubated in Krebs-Ringer bicarbonate buffer containing pyruvate. In muscles stimulated to contract at a frequency of 1 and 2 Hz during the last 2 min of incubation, AMPKalpha1 activity increased twofold and AMPKalpha2 activity remained unchanged. Muscle stimulation did not change the muscle AMP concentration or the AMP-to-ATP ratio. AMPK activation was associated with increased phosphorylation of Thr(172) of the alpha-subunit, the primary activation site. Muscle stimulation increased the phosphorylation of acetyl-CoA carboxylase (ACC), a downstream target of AMPK, and the rate of 3-O-methyl-d-glucose transport. In contrast, increasing the frequency (>or=5 Hz) or duration (>or=5 min) of contraction activated AMPKalpha1 and AMPKalpha2 and increased AMP concentration and the AMP/ATP ratio. These results suggest that 1) AMPKalpha1 is the predominant isoform activated by AMP-independent phosphorylation in low-intensity contracting muscle, 2) AMPKalpha2 is activated by an AMP-dependent mechanism in high-intensity contracting muscle, and 3) activation of each isoform enhances glucose transport and ACC phosphorylation in skeletal muscle.     

Effects of induction of parturition in ewes with dexamethasone or oestrogen on concentrations of immunoglobulins in colostrum, and absorption of immunoglobulins by lambs

Immunoglobulins were measured in colostral whey from untreated ewes or ewes treated on days 142-145 of gestation with 15 mg oestradiol benzoate (ODB) or 10, 17.5 or 25 mg dexamethasone m-sulfobenzoate (DEX) to induce parturition. DEX at all levels reduced the concentrations of immunoglobulins in colostrum, but ODB had no effect. In a second experiment, similar treatments of ODB and DEX were applied on day 143 of gestation. Lambs were removed from their dams at birth and fed a standard volume of pooled colostrum taken from untreated ewes. They were injected with an antigen (Brucella abortus) at 12 h and 4 weeks of age. Concentrations of immunoglobulins in blood plasma at 24 h after birth of lambs from DEX-treated ewes were lower than for lambs from untreated ewes or from those treated with ODB. Response to the antigen, monitored to 12 weeks of age, was unaffected by either drug. The impairment of transfer of immunoglobulins from ewe to lamb offers a possible explanation for the higher mortality among older lambs found previously after treatment of ewes with DEX.     

Real-time ultrasonography for determining pregnancy status and viable fetal numbers in ewes

The reproductive tracts of 26 estrus synchronized, bred ewes were scanned with a portable 5.0 MHz real-time ultrasound unit within 1 to 6 d postbreeding. Intrarectal scanning was performed on alternate days until Days 28 to 30 and twice weekly until Days 50 of gestation. Transabdominal uterine scans were conducted twice weekly from Days 25 to 65 and continued weekly until parturition. A total of 24 ewes (92%) became pregnant. A nonpregnant ewe was recognized 100% of the time by both methods of ultrasonic screening. Correct identification of a gravid ewe as pregnant was 100% from Days 51 to 150 of gestation using transabdominal real-time ultrasonography. There was a significant association (P < 0.005) between the number of lambs born and the number of fetuses observed using transabdominal real-time ultrasonography after Day 25 of gestation. Accurate differentiation of fetal numbers by transabdominal scanning was 100.0% for ewes carrying one lamb and 97.3% for ewes carrying two lambs at Days 51 to 75 of gestation. Fetal attrition was documented in one ewe at Day 49 of gestation. Hydrops allantois was diagnosed in another ewe at 110 d of gestation. A total of 37 lambs were born to 23 ewes in the project flock. No congenital abnormalities were noted in any of the lambs. Transabdominal real-time ultrasonography is a safe, rapid, accurate and practical method for assessing pregnancy status, fetal number and fetal viability in sheep.     

AMP-activated protein kinase is required for the lipid-lowering effect of metformin in insulin-resistant human HepG2 cells

The antidiabetic drug metformin stimulates AMP-activated protein kinase (AMPK) activity in the liver and in skeletal muscle. To better understand the role of AMPK in the regulation of hepatic lipids, we studied the effect of metformin on AMPK and its downstream effector, acetyl-CoA carboxylase (ACC), as well as on lipid content in cultured human hepatoma HepG2 cells. Metformin increased Thr-172 phosphorylation of the alpha subunit of AMPK in a dose- and time-dependent manner. In parallel, phosphorylation of ACC at Ser-79 was increased, which was consistent with decreasing ACC activity. Intracellular triacylglycerol and cholesterol contents were also decreased. These effects of metformin were mimicked or completely abrogated by adenoviral-mediated expression of a constitutively active AMPKalpha or a kinase-inactive AMPKalpha, respectively. An insulin-resistant state was induced by exposing cells to 30 mm glucose as indicated by decreased phosphorylation of Akt and its downstream effector, glycogen synthase kinase 3alpha/beta. Under these conditions, the phosphorylation of AMPK and ACC was also decreased, and the level of hepatocellular triacylglycerols increased. The inhibition of AMPK and the accumulation of lipids caused by high glucose concentrations were prevented either by metformin or by expressing the constitutively active AMPKalpha. The kinase-inactive AMPKalpha increased lipid content and blocked the ability of metformin to decrease lipid accumulation caused by high glucose concentrations. Taken together, these results indicate that AMPKalpha negatively regulates ACC activity and hepatic lipid content. Inhibition of AMPK may contribute to lipid accumulation induced by high concentrations of glucose associated with insulin resistance. Metformin lowers hepatic lipid content by activating AMPK, thereby mediating beneficial effects in hyperglycemia and insulin resistance.     

Effects of exogenous progesterone on gestation length, foetal survival and colostrum yield in ewes

Twin bearing mature ewes (n=40) were treated with exogenous progesterone (100mg daily in oil) or vehicle (oil control) from Day 143 of gestation until lambing to investigate the effects on gestation length, foetal survival and colostrum yield and composition. Compared to control ewes, progesterone treated ewes had increased (P<0.05) serum progesterone concentrations (by 4.3 ng/ml) before lambing and in the first day post-partum (by 10 ng/ml). Progesterone treatment increased gestation length (150.4+/-0.6 days versus 147.8+/-0.6 days, P<0.05) and colostrum yield at 1h after lambing (P<0.05) but the colostrum had a lower concentration of IgG (P=0.02). In the first 24h after lambing, total colostrum and IgG yields were not different between groups. Four (20%) of the progesterone treated ewes produced either one or two dead lambs, while one ewe died on day 155 without initiating the birth process. We conclude that the daily administration of 100mg progesterone resulted in extended gestation length and reduced lamb survival but did not lower colostrum yield.     

The effect of prenatal photoperiodic history on the postnatal endocrine status of female lambs

Postnatal photoperiodic experience plays a pivotal role in determining the timing of ovarian activity in female lambs. This study examines whether a photoperiodic history gained while in utero is able to influence this timing.Pregnant Soay ewes were maintained in either long days (n = 7, 18 h light : 6 h dark; group PLD) or short days (n = 12, y h light : 18 h dark; group PSD) from 25 days of gestation. At birth, female lambs (n = 8 per group) were transferred to long days for 10 weeks, and then placed under short days until the end of the experiment at 38 weeks of age. Blood samples were collected from lambs on the day of birth and three times weekly for the duration of the study and the resulting plasma assayed for progesterone and prolactin.Although both gestational photoperiods produced, at best, abbreviated periods of ovarian activity, lambs born to ewes which experienced long days during gestation (group PLD) exhibited elevated plasma progesterone concentrations significantly earlier (P < 0.05) than lambs born to ewes exposed to short days during gestation (group PSD) (mean ± SEM, 193 ± 17 versus 244 ± 14 days for PLD and PSD groups, respectively. Plasma prolactin concentrations in newborn lambs born between late December and early April were not affected by the ambient photoperiod, but reflected the artificial daylength experienced by their mothers during gestation. Lambs born to ewes maintained under long days during gestation (group PLD) had significantly higher prolactin concentrations on the day of birth than lambs born to ewes maintained under short days during gestation (group PSD) (45 ± 5.4 ng/ml versus 7 ± 3.7 ng/ml respectively, P < 0.001). The mean birth weight, rate of live weight gain and live body weight of lambs at the end of the experiment did not vary significantly between treatment groups. These results suggest that the ovine foetus is sensitive to photoperiodic information prior to birth, and develops a photoperiodic history which, under the present experimental conditions, modulates the subsequent endocrine status of the neonatal lamb.     

Feeding pregnant ewes a high-salt diet or saltbush suppresses their offspring's postnatal renin activity

If ewes consumed a high-salt diet or saltbush during the last 3 months of pregnancy and for 3 weeks after birth, we expected the renin activity of their lamb to be suppressed at birth and at 3 weeks of age. We also expected an increase in the concentration of cations other than sodium in the ewe's milk and an increase in the plasma Na concentration of the lamb at birth. To test these hypotheses, Merino ewes were fed a high-salt diet (14% NaCl) in an animal house and compared to control ewes eating a control diet (2% NaCl). In addition, we compared ewes grazing saltbush (about 13% salt in diet) to ewes grazing pasture from day 60 of pregnancy to 3 weeks after birth. Lambs born to ewes consuming saltbush had 85% lower (P < 0.001) renin activity than offspring from ewes consuming pasture at 3 weeks of age. Similarly, lambs born to ewes consuming a high-salt diet had 20% lower renin activity at birth and 3 weeks (P = 0.07). Feeding ewes a high-salt diet or saltbush altered the mineral composition of the milk; the largest change was a 10% increase in K levels (P 0.05). Consuming a high-salt diet or saltbush lowered the plasma Na of ewes at 130 days of gestation (by 3-5 mmol/l; P < 0.001), but only lambs from ewes fed the high-salt diet had a lower plasma Na at birth (P < 0.05). Suppression of the renin activity of lambs could lead to permanent physiological changes in salt balance in later life.     

Reducing the level of nutrition of twin-bearing ewes during mid to late pregnancy produces leaner prime lambs at slaughter

The Australian prime lamb industry is seeking to improve lean meat yield (LMY) as a means to increasing efficiency and profitability across the whole value chain. The LMY of prime lambs is affected by genetics and on-farm nutrition from birth to slaughter and is the total muscle weight relative to the total carcass weight. Under the production conditions of south eastern Australia, many ewe flocks experience a moderate reduction in nutrition in mid to late pregnancy due to a decrease in pasture availability and quality. Correcting nutritional deficits throughout gestation requires the feeding of supplements. This enables the pregnant ewe to meet condition score (CS) targets at lambing. However, limited resources on farm often mean it is difficult to effectively manage nutritional supplementation of the pregnant ewe flock. The impact of reduced ewe nutrition in mid to late pregnancy on the body composition of finishing lambs and subsequent carcass composition remains unknown. This study investigated the effect of moderately reducing ewe nutrition in mid to late gestation on the body composition of finishing lambs and carcass composition at slaughter on a commercial scale. Multiple born lambs to CS2.5 target ewes were lighter at birth and weaning, had lower feedlot entry and exit weights with lower pre-slaughter and carcass weights compared with CS3.0 and CS3.5 target ewes. These lambs also had significantly lower eye muscle and fat depth when measured by ultrasound prior to slaughter and carcass subcutaneous fat depth measured 110 mm from the spine along the 12th rib (GR 12th) and at the C-site (C-fat). Although carcasses were ~5% lighter, results showed that male progeny born to ewes with reduced nutrition from day 50 gestation to a target CS2.5 at lambing had a higher percentage of lean tissue mass as measured by dual energy X-ray absorptiometry and a lower percentage of fat during finishing and at slaughter, with the multiple born progeny from CS3.0 and CS3.5 target ewes being similar. These data suggest lambs produced from multiple bearing ewes that have had a moderate reduction in nutrition during pregnancy are less mature. This effect was also independent of lamb finishing system. The 5% reduction in carcass weight observed in this study would have commercially relevant consequences for prime lamb producers, despite a small gain in LMY.     

Breed, litter and parity effects on placental weight and placentome number, and consequences for the neonatal behaviour of the lamb

Lamb survival is impaired in low birth weight lambs, and those that are slow to stand and suck. Many of the factors that influence lamb vigour, such as parity, litter size, and breed, may exert their effects, at least partially, before birth by influencing placenta development. Our hypothesis was that retarded lamb behavioural development was due to differences in placentation in these animals. Data were collected from Blackface and Suffolk lambs in the first 2 h after birth and placentas were collected when delivered. Suffolk lambs, which were behaviourally slower and had lower rectal temperatures than Blackface lambs, were associated with larger but less efficient placentas (placental efficiency defined as foetal weight supported per g placenta) with fewer foetal cotyledons than Blackface placentas. Triplet lambs were significantly slower than twin or single lambs to suck and had lower rectal temperatures. Although placenta efficiency increased with litter size, placenta and cotyledon weight, and cotyledon number increased with twinning but not thereafter. It seemed likely that triplet lambs suffered some placental insufficiency in comparison to other litter sizes. Lambs born to first parity mothers were slower to stand and reach the udder than lambs of more experienced ewes, and first parity ewes also had smaller and less efficient placentas although cotyledon number was not affected. Male lambs tended to be slower than female lambs for most behaviours, although rectal temperatures were not affected. The sire of the lamb also influenced lamb behaviour and rectal temperature. Both lamb sex and lamb sire influenced the average weight of placental cotyledons, thus some of the sire effect on the behaviour and birth weight of his progeny might be mediated through placental development. Lamb neonatal vigour was correlated with placental efficiency suggesting that lamb behaviour immediately after birth is related to placental development and function.     

Late foetal life nutrient restriction and sire genotype affect postnatal performance of lambs

This experiment investigates the effects of maternal nutrient restriction in late gestation on the offsprings' postnatal metabolism and performance. Forty purebred Shropshire twin lambs born to ewes fed either a high-nutrition diet (H) (according to standard) or a low-nutrition (L) diet (50% during the last 6 weeks of gestation) were studied from birth until 145 days of age. In each feeding group, two different sires were represented, 'growth' (G) and 'meat' (M), having different breeding indices for the lean : fat ratio. Post partum all ewes were fed the same diet. Lambs born to L-ewes had significantly lower birth weights and pre-weaning growth rates. This was especially pronounced in L-lambs born to the M-ram, which also had markedly lower pre-weaning glucose concentrations than the other three groups of lambs. L-lambs converted milk to live weight with an increased efficiency in week 3 of life. Their glucose concentrations and growth rates were both increased. Plasma glucose concentrations in LM-lambs became similar to those observed in H-lambs post-weaning. However, LM-lambs continued to be lighter than the other groups throughout the experimental period and were unable to compensate for the reduced weight at birth despite having the highest daily fractional growth rates. LG-lambs had the highest plasma glucose concentrations of all four groups of lambs, and they indeed reached body weights comparable to those of the H-lambs by 145 days of age. The increased growth rate post-weaning in L-lambs was not reflected in fat deposition, as L-lambs had lower fat deposition than H-lambs. This may relate to the lower plasma insulin levels found in the L-lambs than in the H-lambs. In conclusion, a 50% reduction of maternal nutrient supply in the last 6 weeks of gestation reduces the birth weight and pre-weaning growth of the offspring due to lower milk intake. Growth rates can be restored when an adequate post-weaning diet is provided, but the prenatal nutrition may programme postnatal metabolism differentially depending on genotype, thus affecting the ability of the ad libitum-fed lamb to achieve a given body weight by a certain age.     

Circulating LH, FSH, prolactin, testosterone, delta 4-androstenedione, dehydroepiandrosterone sulfate and cortisol levels in the fetus in late gestation and in newborn male and female lambs

Gonadotropins, prolactin (PRL), testosterone (T), delta 4-androstenedione, dehydroepiandrosterone sulfate and cortisol (F) levels were determined from 14 days before birth to term in 3 female and 3 male ovine fetuses with a chronically implanted venous catheter, and in the same animals from birth to 72 h of age. In both sexes, plasma gonadotropins and androgens were low throughout the period of study while plasma F increased with gestational age. After birth, plasma gonadotropins and PRL tended to increase progressively with time while PRL concentrations were significantly higher in female than in male lambs. F and T concentrations decreased significantly within the first 12 and 6 h of postnatal life. Higher T values were again observed at 36 h in male lambs. These data indicate that the fetal hypothalamic-pituitary-gonadal axis is relatively quiescent in the last 14 days of gestation but is activated within the first 72 h after birth.     

Low pasture allowance until late gestation in ewes: behavioural and physiological changes in ewes and lambs from lambing to weaning

Low pasture allowance during gestation affects ewes’ BW at parturition, the bond with their lamb, lamb development, and thus also may affect their responses to weaning. The objectives were to determine if native pasture allowance from before conception until late pregnancy affects ewe–lamb behaviours at lambing, ewes’ milk yield, lambs’ BW, and the behavioural and physiological changes of ewes and lambs at weaning. From 23 days before conception until 122 days of pregnancy, 24 ewes grazed on two different native pasture allowances: high (10 to 12 kg of dry matter (DM)/100 kg of BW per day; HPA treatment; n=12) or low (5 to 8 kg of DM/100 kg of BW per day; LPA treatment; n=12). Thereafter, all ewes grazed on Festuca arundinacea and received rice bran and crude glycerine. Ewes’ body condition score (BCS) and BW were recorded during pregnancy and postpartum periods. Milk yield was determined on days 32, 41 and 54 after lambing. Lambs’ BW was recorded from birth until 72 days after lambing. Latency from parturition until the ewe licked her lamb, maternal behaviour score (a test that evaluates maternal attachment to the lamb) and latency for lamb to stand up and suckle were determined. The behaviour of the lambs and ewes was recorded before and after weaning (at 65 days). The ewes’ serum total protein, albumin and globulin concentrations were measured before and after weaning. The HPA ewes presented greater BW (P<0.005) and BCS (P<0.005) than the LPA ewes during pregnancy and postpartum (P<0.04), and had a greater milk yield than the LPA ewes (P<0.03). Treatments did not influence any behaviour at lambing, lambs’ BW, neither the ewes’ behavioural and physiological changes at weaning. HPA lambs paced and vocalized more than LPA lambs (P<0.0001). The variation of albumin concentration before and after weaning was greater in the HPA lambs than in the LPA lambs (P<0.0001). In conclusion, although ewes’ BW, BCS and milk production were affected by pasture allowance until late pregnancy, this did not affect the behaviours that lead to the establishment of the mother–young bond, nor the ewes’ behavioural responses at weaning. Lambs reared by ewes that grazed on low pasture allowance during pregnancy presented fewer behavioural changes and a lower decrease of albumin concentration after weaning. Lambs’ BW was not affected by the feeding received by their mothers.     

Role of AMPKalpha2 in basal, training-, and AICAR-induced GLUT4, hexokinase II, and mitochondrial protein expression in mouse muscle

We investigated the role of AMPKalpha2in basal, exercise training-, and AICAR-induced protein expression of GLUT4, hexokinase II (HKII), mitochondrial markers, and AMPK subunits. This was conducted in red (RG) and white gastrocnemius (WG) muscle from wild-type (WT) and alpha2-knockout (KO) mice after 28 days of activity wheel running or daily AICAR injection. Additional experiments were conducted to measure acute activation of AMPK by exercise and AICAR. At basal, mitochondrial markers were reduced by approximately 20% in alpha2-KO muscles compared with WT. In both muscle types, AMPKalpha2 activity was increased in response to both stimuli, whereas AMPKalpha1 activity was increased only in response to exercise. Furthermore, AMPK signaling was estimated to be 60-70% lower in alpha2-KO compared with WT muscles. In WG, AICAR treatment increased HKII, GLUT4, cytochrome c, COX-1, and CS, and the alpha2-KO abolished the AICAR-induced increases, whereas no AICAR responses were observed in RG. Exercise training increased GLUT4, HKII, COX-1, CS, and HAD protein in WG, but the alpha2-KO did not affect training-induced increases. Furthermore, AMPKalpha1, -alpha2, -beta1, -beta2, and -gamma3 subunits were reduced in RG, but not in WG, by 30-60% in response to exercise training. In conclusion, the alpha2-KO was associated with an approximately 20% reduction in mitochondrial markers in both muscle types and abolished AICAR-induced increases in protein expression in WG. However, the alpha2-KO did not reduce training-induced increases in HKII, GLUT4, COX-1, HAD, or CS protein in WG, suggesting that AMPKalpha2 may not be essential for metabolic adaptations of skeletal muscles to exercise training.     

Litter-size-dependent intrauterine growth restriction in sheep

Regulation of foetal development in sheep depends on interactions between the intrinsic capacity of the foetus for growth and the maternal environment. Lambs born in multi-foetus litters have relatively small placentae with fewer cotelydons, and lower birth weights. Litter-size-dependent intrauterine growth restriction (IUGR) is evident at mid gestation when metabolic needs of the conceptus are moderate, and overnutrition of ewes with multiple foetuses does not promote growth of their foetuses to the size of singletons. Those observations suggest that placental and conceptus growth in multi-foetus pregnancies is reprogrammed at mid gestation by an as yet undefined mechanism to attenuate foetal growth. This may protect the foetus from severe nutritional insult during late gestation, when its daily growth rate is at a maximum. In that way, lambs born in large litters with relatively lower birth weights may not experience the long-term physiological insults that can be observed in small lambs born to undernourished ewes.     

Effect of maternal nutrient restriction in early to mid gestation and thyrotrophin-releasing hormone on lamb survival following Caesarean section delivery near to term

We investigated the influence of restricted maternal nutrition between 28 and 77 days gestation on survival and thermoregulatory adaptation following Caesarean section delivery near to term. This study was designed to examine the hypothesis that adaptation after birth would be compromised in those lambs born to nutrient restricted ewes. We further hypothesised that this would be due in part to inadequate hypothalamic-pituitary-thyroidal function. Lambs born to nutrient restricted ewes were untreated (RU) or treated with thyrotrophin-releasing hormone (TRH; RT) immediately prior to umbilical cord clamping. Single bearing ewes consumed either 6.60 MJ x day(-1) (controls, n = 4) or 3.00 MJ x day(-1) (nutrient restricted, n = 15) from 28-77 days gestation, after which all ewes consumed 7.20 MJ x day(-1). All lambs born to control ewes commenced continuous breathing and began to shiver following Caesarean section delivery and survived to 6 h after birth. Only 4 out of 9 RU lambs established continuous breathing and survived to 6 h after birth compared with all RT lambs. Six hours after birth, RT lambs possessed perirenal brown adipose tissue with a higher thermogenic activity than 6 h old RU or control lambs. Lamb birth weight was similar in all groups. In conclusion, near-term lambs born to ewes nutrient restricted in early to mid gestation are at increased risk of death following Caesarean section delivery. Survival after birth can be significantly enhanced if TRH is administered to the lambs immediately before delivery.     

Effects of postnatal dexamethasone on blood-brain barrier permeability and brain water content in newborn lambs

We showed that antenatal corticosteroids reduced blood-brain barrier permeability in fetuses at 60 and 80%, but not 90% of gestation, and decreased brain water content in fetuses. Our objective was to examine the effects of postnatal corticosteroids on regional blood-brain barrier permeability and brain water content in newborn lambs. Three dexamethasone treatment groups were studied in 3- to 5-day-old lambs. A 0.01 mg/kg dose was selected to estimate the amount of dexamethasone that might have reached fetuses via antenatal treatment of ewes in our previous studies. The other doses (0.25 and 0.5 mg/kg) were chosen to approximate those used clinically to treat infants with bronchopulmonary dysplasia. Lambs were randomly assigned to receive four intramuscular injections of dexamethasone or placebo given 12 h apart on days 3 and 4 of age. Blood-brain barrier function was measured with the blood-to-brain transfer constant (K(i)) to alpha-aminoisobutyric acid, brain plasma volume was measured with polyethylene glycol for the calculation of K(i,) and brain water was measured by wet-to-dry tissue weights. Postnatal treatment with corticosteroids did not reduce barrier permeability in newborn lambs. Brain blood volume was higher in the 0.25 and 0.5 mg/kg dose dexamethasone groups than in the placebo group. Brain water content did not differ among the groups. We conclude that postnatal treatment with corticosteroids did not reduce regional blood-brain barrier permeability or brain water content but increased the brain plasma volume in newborn lambs. These findings are consistent with our previous work indicating that barrier permeability is responsive to corticosteroids at 60 and 80% of gestation and brain water regulation at 60% of gestation, but not in near-term fetuses or newborn lambs.     

Influence of gestational overfeeding on myocardial proinflammatory mediators in fetal sheep heart

Maternal overnutrition is associated with predisposition of offspring to cardiovascular disease in later life. Since maternal overnutrition may promote fetal and placental inflammatory responses, we hypothesized that maternal overnutrition/obesity increases expression of fetal cardiac proinflammatory mediators and alter cardiac morphometry. Multiparous ewes were fed either 150% of National Research Council (NRC) nutrient recommendations (overfed) or 100% of NRC requirement (control) from 60 days prior to mating to gestation Day 75 (D75), when ewes were euthanized. An additional cohort of overfed and control ewes were necropsied on D135. Cardiac morphometry, histology, mRNA and protein expression of toll-like receptor 4, iNOS, IL-1a, IL-1b, IL-6, IL-18, CD-14, CD-68, M-CSF and protein levels of phosphorylated I-κB and nuclear factor κB (NF-κB) were examined. Immunohistochemistry was performed to assess neutrophil and monocyte infiltration. Crown rump length, left and right ventricular free wall weights as well as left and right ventricular wall thickness were significantly increased in D75 fetuses of overfed mothers. Hematoxylin and eosin staining revealed irregular myofiber orientation and increased interstitial space in fetal ventricular tissues born to overfed mothers. Oil red O staining exhibited marked lipid droplet accumulation in the overfed fetuses. Overfeeding significantly enhanced TLR4, IL-1a, IL-1b IL-6 expression, promoted phosphorylation of IκB, decreased cytoplasmic NF-κB levels and increased neutrophil and monocyte infiltration. Collectively, these data suggest that maternal overfeeding prior to and throughout gestation leads to inflammation in the fetal heart and alters fetal cardiac morphometry.     

Effect of plane of nutrition of 1- and 2-year-old ewes in early and mid-pregnancy on ewe reproduction and offspring performance up to weaning

The objective of the present study was to determine the effect of plane of nutrition in early pregnancy (EP) and mid-pregnancy (MP), on the productive performance of 1- and 2-year-old ewes and their offspring. Over 2 successive years, between days 0 and 39 after synchronized mating (EP), 1- (n = 117) and 2- (n = 52) year-old ewes were allowed 60% (low, L-EP), 100% (medium, M-EP) or 200% (high, H-EP) of requirements for maintenance (M). Between days 40 and 90 (MP), 1-year-old ewes were allowed 140% (M-MP) or 200% (H-MP), while 2-year-old ewes were allowed 80% (M-MP) or 140% (H-MP) of their M requirement. After day 90, all ewes were fed to meet requirements for late pregnancy. Increasing the plane of nutrition between days 0 and 39 resulted in increases in live weight (LW) (P < 0.001) and body condition score (BCS) (P < 0.001) during the EP period (H-EP > M-EP > L-EP), differences that in 1-year-old ewes were sustained to lambing (P < 0.05). On day 42 of gestation H-EP ewes had lower plasma progesterone concentrations than L-EP or M-EP ewes in 1- (P < 0.01) and 2- (P < 0.001) year olds. This was concomitant with diet H-EP tending to reduce the number of lambs born per ewe in both age groups (P = 0.06 and 0.07, respectively). Foetuses from 1-year-old L-EP ewes had smaller cranial (P < 0.01) and abdominal (P < 0.05) diameters at day 53 of gestation, with H-EP lambs tending to be heaviest at birth (P = 0.07). Similar findings were recorded for 2-year-old ewes. One-year-old ewes offered diet L-EP presented negative maternal behaviours more frequently (P < 0.05), while the incidence of lamb mortality at 6 weeks tended to be greater for L-EP lambs (P = 0.07). In MP, 1-year-old ewes offered diet M-MP were associated with foetuses with bigger abdominal diameters at day 78 (P < 0.05). However, there were no differences in lamb weight or size at term (P > 0.05). These ewes exhibited more positive maternal behaviours (e.g. increased grooming frequency and duration; P < 0.05) than ewes offered diet H-MP, and their offspring were more successful in suckling (P < 0.05). Results suggest that in young ewes, a temporary nutrient restriction in EP resulted in increased prolificacy. However, ewes and their offspring were lighter at birth and ewe maternal behaviour was poorer, resulting in increased lamb mortality. In MP, a medium plane of nutrition offered to 1-year-old ewes led to improved maternal and offspring behaviour.