Feasibility of two screen media reduction interventions: Results from the SCREENS pilot trial.

BACKGROUND: Advancements in screen media devices has transformed the way families engage with screen media. Although these modern devices offer many opportunities, e.g. communication and research online, an in-depth understanding of how these devices affect our health, is lacking. Before a definite randomized controlled trial, the SCREENS pilot study was conducted to assess compliance to and feasibility of two interventions, a measurement protocol, and a survey-based recruitment strategy. Also, the potential of the interventions to impact leisure time spent non-sedentary in children six-to-ten years of age was explored. METHODS: Families (N = 12) were recruited through a population-based survey sent out in October of 2018 to adults (N = 1,675) in the Municipality of Middelfart, Denmark. Families were randomized to one of two two-week interventions; an Evening Restriction intervention (no screen media use after six pm) and a General Restrict intervention (limit entertainment-based screen media to three hours/week/person). Intervention compliance was assessed objectively by measuring household TV usage, smartphone and tablet activity via an application, and via screen media diaries. During baseline and follow-up, as part of larger protocol, family members wore two triaxial accelerometers for seven consecutive days. The potential of the interventions to impact non-sedentary time was explored based on means and standard errors (SEs). RESULTS: Despite almost 85% and 75% reductions in leisure screen media use 0% and 50% of families were compliant in the Evening Restrict group and General Restrict group, respectively, based on strict a priori criteria. Participant feedback indicated that the General Restrict intervention generally was feasibly. Compliance to the accelerometry wear protocol was high (median non-wear was <1 hour/week). Moreover, the recruitment strategy was implemented and was feasible. The General restrict intervention might increase children's non-sedentary time (mean (SE): 36.6 (23) min/day, N = 6). CONCLUSIONS: The General Restriction intervention, the accelerometer wear protocol and recruitment strategy, appeared feasible. TRIAL REGISTRATION: NCT03788525 at https://clinicaltrials.gov [Retrospectively registered; 27th of December, 2018].

Ovulation and ovulation rate in ewes under grazing conditions: factors affecting the response to short-term supplementation.

The relationships between ovulation rate and nutrition remain confused, probably because of uncontrolled variation in experimental conditions. To help resolve the problem, we analyzed data from 20 experiments conducted between 2002 and 2016, in Uruguay with grazing ewes. All experiments were carried out by a single laboratory under comparable conditions of experimental design and measured variables. The studies used a total of 3 720 ewes, of purebred Corriedale, Polwarth, or East Friesian x Polwarth genotypes. In all experiments, a control group grazed native pastures and extra nutrition was provided to the treatment groups using either improved pastures or supplements. Ovulation rate was measured by counting corpora lutea using laparoscopy or rectal ultrasound or by counting foetuses at ultrasound on day 45 of gestation. For statistical analysis, data were grouped according to nutritional treatment (control or supplemented) and, within these groups, type of supplement to provide energy or protein (protected or not from rumen degradation). Across all experiments, 92-99% of the ewes ovulated and the effects of diet, length of supplementation, and initial live weight and genotype are reported. Within diets, ovulation was most affected by overall energy intake during supplementation (P < 0.01). Ewes that grazed native pastures supplemented with protein supplements had higher ovulation rates (P < 0.05) than control ewes grazing only native pastures. The addition of tannins to the protein supplement, to protect it from degradation in the rumen, did not further increase the ovulation rate. In unsupplemented ewes that had access to legume pastures, ovulation rates did not increase when the legume pasture was rich in tannins although only ewes that grazed tanniniferous legumes had marginally higher ovulation rates than the control ewes (P < 0.05). When ewes grazing native pastures were supplemented with energy, their ovulation rate did not increase above those of nonsupplemented ewes. Live weight at the start of supplementation also affected ovulation rate. We conclude that ovulation was most affected by overall energy intake, whereas the factors that affected ovulation rate during short-tern nutritional supplementation were intake of protein from highly digested supplements or dietary protein protected from ruminal degradation.

Arcuate nucleus kisspeptin response to increased nutrition in rams.

Rams respond to acute nutritional supplementation by increasing the frequency of gonadotrophin-releasing hormone (GnRH) pulses. Kisspeptin neurons may mediate the effect of environmental cues on GnRH secretion, so we tested whether the ram response to nutrition involves activation of kisspeptin neurons in the arcuate nucleus (ARC), namely kisspeptin, neurokin B, dynorphin (KNDy) neurons. Rams were given extra lupin grain with their normal ration. Blood was sampled before feeding, and continued until animals were killed for collection of brain tissue at 2 or 11h after supplementation. In supplemented rams, LH pulse frequency increased after feeding, whereas control animals showed no change. Within the caudal ARC, there were more kisspeptin neurons in supplemented rams than in controls and a higher proportion of kisspeptin cells coexpressed Fos, regardless of the time the rams were killed. There were more Fos cells in the mid-ARC and mid-dorsomedial hypothalamus of the supplemented compared with control rams. No effect of nutrition was found on kisspeptin expression in the rostral or mid-ARC, or on GnRH expression in the preoptic area. Kisspeptin neurons in the caudal ARC appear to mediate the increase in GnRH and LH production due to acute nutritional supplementation, supporting the hypothesised role of the KNDy neurons as the pulse generator for GnRH.

Volatiles from Merino fleece evoke antennal and behavioural responses in the Australian sheep blow fly Lucilia cuprina.

To identify flystrike-related volatile compounds in wool from Merino sheep, the attractiveness of wool to Lucilia cuprina Wiedmann (Diptera: Calliphoridae) was examined. First, a selection of wool samples guided by previous knowledge of sheep lines, predicted to be more susceptible or more resistant to flystrike, was tested. The attractiveness of the 10 samples selected was not associated with field susceptibility: two samples from the more resistant line were identified as most attractive and two samples from the more susceptible line were identified as least attractive, based on the behavioural assays with gravid flies. Comparison of the headspace volatiles of these samples, using solid phase microextraction and gas chromatography-mass spectrometry-electroantennographic detection, revealed octanal and nonanal to be present in the attractive wool samples that elicited responses from the fly antenna. Furthermore, the two compounds were not present in wool that was least attractive to L. cuprina. In laboratory bioassays, octanal and nonanal evoked antennal and behavioural responses in gravid L. cuprina, thus confirming their potential role as semiochemicals responsible for attracting L. cuprina to Merino sheep.

Milk production and composition, and progeny performance in young ewes with high merit for rapid growth and muscle and fat accumulation.

In ewe lambs, acceleration of growth and accumulation of both muscle and fat leads to earlier sexual maturity and better reproductive performance. The next stage in the development of this theme is to test whether these aspects of growth in young ewes affect milk production in their first lactation and the growth of their first progeny. We studied 75 young Merino ewes that had known phenotypic values for depth of eye muscle (EMD) and fat (FAT), and known Australian Sheep Breeding Values for post-weaning weight (PWT) and depths of eye muscle (PEMD) and fat (PFAT). They lambed for the first time at 1 year of age. Their lambs were weighed weekly from birth to weaning at 10 weeks to determine live weight gain and weaning weight. Progeny birth weight was positively associated with live weight gain and weaning weight (P0.05). The PWT of the sire was positively associated with live weight gain (P0.05). The concentrations of fat, protein, lactose and total solids in the milk were not affected by the phenotype or genotype of the mothers or of the sires of the mothers, or by the sex of the progeny (P>0.05). We conclude that selection of young Merino ewes for better growth, and more rapid accumulation of muscle and fat, will lead to progeny that are heavier at birth, grow faster and are heavier at weaning. Moreover, milk production and composition do not seem to be affected by the genetic merit of the mother for post-weaning live weight or PEMD or PFAT. Therefore, Merino ewes can lamb at 1 year of age without affecting the production objectives of the Merino sheep industry.

Appraisal and standardization of curvilinear velocity (VCL) cut-off values for CASA analysis of Japanese quail (Coturnix japonica) sperm.

One of the basic steps in objective analysis of sperm motility is the subdivision of a motile sperm population into slow, medium and rapid categories based on their velocity. However, for CASA analysis of quail sperm, the velocity values for categorization of slow, medium and rapid sperm have not yet been standardized. To identify the cut-off values of "velocity curvilinear" (VCL) for quail sperm categorization, we captured and analysed 22,300 tracks of quail sperm using SCA® -CASA. The median and mean VCL values were 85 and 97 µm/s. To define the VCL cut-off values, we used two methods. In the first, we identified the upper (rapid sperm) and lower (slow sperm) cut-off values using: (i) median VCL ± 25% or ± 50% or ± 75% of median VCL value; (ii) first and third quartile values of VCL data (i.e. 25% cut-off setting); and (iii) 33% and 66% of VCL data. Among these settings, sperm categories and their corresponding motility characteristics recorded using the "25%" setting (i.e. slow ≤36 ≤ medium ≤154 ≤ rapid) were found the most realistic and coherent with male ranking by fertility. In the second method, we calculated heteroscedasticity in the total VCL data using PCA and the two-step clustering method. With this approach, the mean of the high and low clusters was 165 and 51 µm/s, respectively. Together, the mean from two methods suggested that, for SCA® -CASA categorization of quail sperm, sperm should be classed as "rapid" at VCL ≥160 µm/s and "slow" at VCL ≤45 µm/s.

In utero betamethasone affects 3ß-hydroxysteroid dehydrogenase and inhibin-α immunoexpression during testis development.

Prenatal glucocorticoids, commonly used in women at risk of preterm delivery, can predispose the newborn to disease in later life. Since male reproductive function is likely to reflect testis development during fetal life, we studied the effects of prenatal glucocorticoids on two key intra-testicular factors that play roles in cellular proliferation and differentiation, 3ß-hydroxysteroid dehydrogenase (3ß-HSD) and inhibin-α. Pregnant sheep (n=42) were treated with betamethasone (0.5 mg/kg) or saline (control) at 104, 111 and 118 days of gestation (DG). Testicular tissue was sampled from fetuses at 121 and 132DG, and from lambs at 45 and 90 postnatal days (PD). Within the betamethasone treated group, 3ß-HSD immunostaining area was greater at 121DG than at 90PD (P=0.04), but the intensity of immunostaining was higher at 90PD than at 121DG (P=0.04), 132DG (P=0.04) and 45PD (P=0.03). Control animals showed no changes in 3ß-HSD area or intensity of immunostaining. No significant differences were observed between treated and control animals in immunostaining area, but immunostaining was more intense in the treated group than in the control group at 90PD (P=0.03). For inhibin-α, the proportion of immunostaining area declined in treated offspring from 121DG to 45PD, in contrast to control values, but recovered fully by 90PD, concomitantly with the onset of spermatogenesis. In conclusion, prenatal betamethasone increased the postnatal testicular expression of inhibin-α but reduced the expression of 3ß-HSD. These effects could compromise androgen-mediated testicular development and therefore adult capacity for spermatogenesis.

Monitoring stress in captive and free-ranging African wild dogs (Lycaon pictus) using faecal glucocorticoid metabolites.

An understanding of stress physiology is important for species management because high levels of stress can hamper reproduction and affect an individual's ability to cope with threats to their survival, such as disease and human-wildlife conflict. A commonly used indicator of stress, faecal concentrations of cortisol metabolites (FCM), can be used to assess the impact of social, biological and environmental factors. Measurements of FCM are particularly valuable for endangered species that are logistically challenging to study and where non-invasive techniques are preferred. As the second most endangered canid in Africa, the African wild dog (Lycaon pictus) has been the focus of considerable conservation research, yet there is still little understanding of factors associated with stress, in either captive or free-ranging populations. The present study therefore aimed to determine whether stress levels differ between captive and free-ranging populations, and to detect social, biological and environmental factors that are stressful in these populations. Faecal samples were collected from 20 captive and 62 free-ranging animals. Within free-ranging populations, the sexes differed significantly, but there was no effect of social status, age or breeding period for either sex. Captive females had higher FCM concentrations than free-ranging females. In captive populations, FCM concentrations differed among zoos and with reproductive status in females, but were not related to age class or group-housing structure. In conclusion, FCM is a useful indicator of stress and should be considered an integrative aspect of management, for both in situ and ex situ African wild dog populations.

Twenty-four-hour profiles of metabolic and stress hormones in sheep selected for a calm or nervous temperament.

Even in the absence of stressors, temperament is associated with changes in the concentration of stress-responsive hormones and, possibly because of such changes, temperament can affect metabolism. We tested whether, in sheep bred for temperament for 14 generations, "nervous" females have greater concentrations of stress-responsive hormones in the absence of stressors than "calm" females, and whether these differences are associated with changes in the concentrations of metabolic hormones. In resting "calm" (n = 8) and "nervous" (n = 8) sheep, concentrations of cortisol, prolactin, leptin, and insulin were measured in blood plasma sampled via jugular catheter every 20 min for 24 h. The animals were individually penned, habituated to their housing and human handling over 7 wk, and fed before sampling began. Diurnal variation was evident for all hormones, but a 24-h cortisol pattern was detected in only 7 individuals. There was no effect of temperament on any aspect of concentrations of cortisol or prolactin, but "calm" animals had greater concentrations of insulin in the early afternoon than "nervous" animals (14.5 ± 1.1 vs 10.0 ± 1.6 µU/mL; P = 0.038), and a similar tendency was seen for leptin (P = 0.092). We conclude that selection for temperament affects the concentration of metabolic hormones in the absence of stressors, but this effect is independent of stress-responsive hormones.

Relationships among Puberty, Muscle and Fat, and Liveweight Gain during Mating in Young Female Sheep.

Greater depths of muscle are associated with better reproductive performance in ewe lambs, but, in adult ewes, reproductive performance also seems to vary with liveweight gain during the mating period. Therefore, in a large field study with Merino ewe lambs, we tested whether the relationships among eye muscle depth (EMD), fat depth (FAT) and reproductive performance depend on liveweight gain during the mating period. We selected lambs with a wide range in phenotypic values for depths of eye muscle (EMD) and fat (FAT) and assigned them to dietary treatments designed to achieve low (LOW, n = 244) or high (HIGH, n = 237) rates of liveweight gain during a 28-day mating period. The LOW treatment maintained live weight, whereas the HIGH treatment gained 179 ± 3.8 g/day (p < 0.001). From those ewe lambs that attained puberty, first oestrus was detected at live weight 37.8 ± 0.2 kg and age 232 days. The proportion of ewes that attained puberty increased with EMD (p < 0.01). Ewes from the HIGH treatment were more fertile (pregnant ewes per 100 ewes exposed to rams) and had a higher reproductive rate (foetuses in utero per 100 ewes exposed to rams; p < 0.001) than those from the LOW treatment. Fertility and reproductive rate were positively correlated with weight gain during mating as well as live weight at the start of mating, FAT and EMD (p < 0.05 to <0.001). We conclude that faster growth, due to either extra nutrition during mating or higher phenotypic potential for fat and muscle, will increase reproductive performance in ewe lambs mated at 8 months of age.

Reproductive hormonal patterns in pregnant, pseudopregnant and acyclic captive African wild dogs (Lycaon pictus).

African wild dogs are one of the most endangered canid species, with free-living populations declining as a consequence of habitat loss, disease and human conflict. Captive breeding is considered an important conservation strategy, but is hampered by a poor overall understanding of the reproductive biology of the species. To improve our basic knowledge, we studied hormone patterns in 15 female wild dogs using non-invasive faecal collections. By comparing longitudinal hormone profiles with behavioural and anatomical changes, females could be allocated among three reproductive classes: pregnant (n=1), pseudopregnant (n=9) and acyclic (n=4). We also monitored a single female in which contraception was induced with a deslorelin implant. Comparison of pseudopregnant and acyclic females showed that, in both classes, faecal oestradiol concentrations increased from anoestrus to pro-oestrus then declined into the oestrous and dioestrous phases. Progestagen concentrations rose steadily from anoestrus to the dioestrous phase in both pseudopregnant and acyclic females and, pseudopregnant females had significantly higher concentrations of progestagens than acyclic females in all phases of the oestrous cycle. Most females classed as pseudopregnant were found in female-only groups, suggesting that wild dogs are spontaneous ovulators. Furthermore, only one adult female did not ovulate, so suppression of reproduction in subordinates is likely to be behavioural rather than physiological.

Colostrum production in ewes: a review of regulation mechanisms and of energy supply.

In sheep production systems based on extensive grazing, neonatal mortality often reaches 15% to 20% of lambs born, and the mortality rate can be doubled in the case of multiple births. An important contributing factor is the nutrition of the mother because it affects the amount of colostrum available at birth. Ewes carrying multiple lambs have higher energy requirements than ewes carrying a single lamb and this problem is compounded by limitations to voluntary feed intake as the gravid uterus compresses the rumen. This combination of factors means that the nutritional requirements of the ewe carrying multiple lambs can rarely be met by the supply of pasture alone. This problem can overcome by supplementation with energy during the last week of pregnancy, a treatment that increases colostrum production and also reduces colostrum viscosity, making it easier for the neonatal lamb to suck. In addition, litter size and nutrition both accelerate the decline in concentration of circulating progesterone that, in turn, triggers the onsets of both birth and lactogenesis, and thus ensures the synchrony of these two events. Furthermore, the presence of colostrum in the gut of the lamb increases its ability to recognize its mother, and thus improves mother-young bonding. Most cereal grains that are rich in energy in the form of starch, when used as supplements in late pregnancy will increase colostrum production by 90% to 185% above control (unsupplemented) values. Variation among types of cereal grain in the response they induce may be due to differences in the amount of starch digested post-ruminally. As a percentage of grain dry matter intake, the amount of starch entering the lower digestive tract is 14% for maize, 8.5% for barley and 2% for oats. Supplements of high quality protein from legumes and oleiferous seeds can also increase colostrum production but they are less effective than cereal grains. In conclusion, short-term supplementation before parturition, particularly with energy-rich concentrates, can improve colostrum production, help meet the energy and immunological requirements for new-born lambs, and improve lamb survival.

Relationships among body composition, circulating concentrations of leptin and follistatin, and the onset of puberty and fertility in young female sheep.

The onset of puberty depends on the attainment of critical body mass, so should also be affected by increases in the rate of accumulation of muscle and adipose tissue. Adipose tissue and reproduction are linked by leptin. For muscle, a link has not yet been identified, although one possibility is follistatin. We assessed the relationships among circulating concentrations of follistatin and leptin and the rates of growth and accumulation of muscle and fat during pubertal development in female sheep. We used 326 animals with known phenotypic values for live weight (LW), depths of eye muscle (EMD) and fat (FAT), and known breeding values at post-weaning age for body mass (PWT) and depths of eye muscle (PEMD) and fat (PFAT). Leptin concentration was positively correlated with values for EMD, PEMD, FAT, PFAT, LW and PWT (P<0.001), whereas follistatin concentration was negatively correlated with values for EMD and PWT (P<0.001), and PEMD (P<0.01) and FAT (P<0.05). Leptin concentration was negatively related to age and positively related to live weight at first oestrus and the proportion of females that attained puberty (P≤0.05), and to fertility and reproductive rate (P<0.01). Follistatin concentration was negatively related to live weight at first oestrus and to fertility (P<0.01) and reproductive rate (P<0.05). There were positive correlations (P<0.001) between muscle accumulation and leptin concentration, and between muscle accumulation and reproductive performance. We conclude that leptin and follistatin are probably both involved in effects of accelerated accumulation of muscle and adipose tissues on the onset of puberty.

Nutrition, metabolic profiles and puberty in Brahman (Bos indicus) beef heifers.

The aim of the present study was to gain an improved understanding of the relationships between body weight (BW), body condition (BCS), and metabolic homeostasis, and the attainment of puberty in Brahman heifers in a subtropical environment. Brahman heifers (200±3kg BW; 2.00±0.0 BCS) were assigned to a moderate nutrition (MN, n=11) or improved nutrition (IN, n=11) treatment from 11 to 23 months-of-age. The heifers were monitored at regular intervals for circulating concentrations of GH, IGF-1, insulin, glucose and leptin, and ovarian follicular activity was recorded until the first ovulation. From approximately 16 months of age, heifers on IN had a greater (P<0.01) BW and greater (P<0.01) BCS than heifers on MN. Heifers on IN also had a generally improved metabolic homeostasis than heifers on MN which was reflected in greater circulating concentrations of insulin, leptin, IGF-1 and glucose in the former heifers. Heifers on IN attained puberty between 21 and 23 months of age and only one heifer on MN had reached puberty by 23 months. This study has shown that Brahman heifers on IN had a metabolic homeostasis that was supportive of reproductive maturation and puberty.

Follicle development, endocrine profiles and ovulation rate in adult Merino ewes: effects of early nutrition (pre- and post-natal) and supplementation with lupin grain.

In adult ewes, we tested whether ovarian function, including the response to short-term supplementation, was affected by the nutrition of their mothers during the pre-/post-natal period. A 2×2 factorial design was used with nutrition in early life (low or high) and a 6-day supplement (with or without) as factors. All ewes received three prostaglandin (PG) injections 7 days apart, and the supplement (lupin grain) was fed for 6 days from 2 days after the second until the third PG injection. We measured reproductive and metabolic hormones, studied follicle dynamics (ultrasonography), and evaluated granulosa cell numbers, aromatase activity and oestradiol (E2) concentrations in follicular fluid in healthy follicles at days 3 and 7 of supplementation. Ovulation rate was increased by 25% by exposure to high pre-/post-natal nutrition (1.5 vs 1.2; P<0.05), in association with a small decrease in FSH concentrations (P=0.06) and a small increase in insulin concentrations (P=0.07). The number of healthy antral follicles was not affected. Acute supplementation increased the number of granulosa cells (3.7±0.2 vs 3.0±0.2 million; P<0.05) in the largest follicle, and the circulating concentrations of E2 (4.6±0.3 vs 3.9±0.3 pmol/l; P<0.05) and glucose (3.4±0.03 vs 3.3±0.03 mmol/l; P<0.01). Both early life nutrition and acute supplementation appear to affect ovulation rate through changes in glucose-insulin homoeostasis that alter follicular responsiveness to FSH and therefore E2-FSH balance.

Ewe lambs with higher breeding values for growth achieve higher reproductive performance when mated at age 8 months.

We studied the relationships among growth, body composition and reproductive performance in ewe lambs with known phenotypic values for depth of eye muscle (EMD) and fat (FAT) and Australian Sheep Breeding Values for post-weaning live weight (PWT) and depth of eye muscle (PEMD) and fat (PFAT). To detect estrus, vasectomized rams were placed with 190 Merino ewe lambs when on average they were 157 days old. The vasectomized rams were replaced with entire rams when the ewe lambs were, on average, 226 days old. Lambs were weighed every week and blood was sampled on four occasions for assay of ghrelin, leptin and ß-hydroxybutyrate. Almost 90% of the lambs attained puberty during the experiment, at an average live weight of 41.4 kg and average age of 197 days. Ewe lambs with higher values for EMD (P < 0.001), FAT (P < 0.01), PWT (P < 0.001), PEMD (P < 0.05) and PFAT (P < 0.05) were more likely to achieve puberty by 251 days of age. Thirty-six percent of the lambs conceived and, at the estimated date of conception, the average live weight was 46.9 ± 0.6 kg and average age was 273 days. Fertility, fecundity and reproductive rate were positively related to PWT (P < 0.05) and thus live weight at the start of mating (P < 0.001). Reproductive performance was not correlated with blood concentrations of ghrelin, leptin or ß-hydroxybutyrate. Many ewe lambs attained puberty, as detected by vasectomized rams, but then failed to become pregnant after mating with entire rams. Nevertheless, we can conclude that in ewe lambs mated at 8 months of age, higher breeding values for growth, muscle and fat are positively correlated with reproductive performance, although the effects of breeding values and responses to live weight are highly variable.

In vitro initiation of the acrosome reaction in the emu (Dromaius novaehollandiae).

1. An assessment of the efficiency of the acrosome reaction (AR) provides an important predictor of the fertilizing potential of semen and for diagnosis of the causes of infertility. A standardized protocol was therefore developed for initiation of the acrosome reaction in emu spermatozoa in vitro, and the role of CaCl2 or perivitelline membrane (PVM) proteins in determining the outcome of the reaction was investigated. 2. The acrosome reaction (assessed by FITC-PNA) was successfully induced in live spermatozoa by incubation for 2 min in NaCl-TES medium supplemented with 5 mM CaCl2. The maximum response was 32% live acrosome-reacted spermatozoa (LAR) achieved after 10 min incubation. 3. Compared to the outcome with 5 mM CaCl2 or PVM protein alone, the response was significantly better with a combination of PVM protein and CaCl2. 4. A significant variation in the percentage of LAR spermatozoa among individual males was observed. No treatment affected the percentage of dead acrosome-reacted spermatozoa. 5. The results emphasize the important role played by both PVM proteins and Ca(2+) in the in vitro initiation of the acrosome reaction.

Apoptosis-related protein expression during pre- and post-natal testicular development after administration of glucocorticoid in utero in the sheep.

Pre-natal glucocorticoids are used in women at risk of preterm delivery to induce foetal lung maturation. However, glucocorticoids can produce negative outcomes for other tissues such as the reproductive system. We therefore tested the effects of pre-natal betamethasone on testicular morphology and apoptotic protein immune expression during pre- and post-natal development. Pregnant ewes (n = 42) bearing singleton male foetuses were randomly allocated to receive intramuscular injections of saline or betamethasone (0. 5 mg/kg) at 104, 111 and 118 days of gestation (DG). Testes were collected at 121 and 132 DG, and at 45 and 90 post-natal days (PD) and subjected to morphometric analysis (volume densities of sex cords and interstitial tissues; sex cord diameter). Immunohistochemistry (% stained area) was used to assess active caspase-3, Bax, Bcl-2 and cell-cycle proteins (PCNA). Compared with control values, betamethasone treatment decreased sex cord diameter at 121 DG, 45 and 90 PD, and sex cord volume at 90 PD. Active caspase-3 was decreased by betamethasone at 121 DG and 90 PD, but Bax was increased in all betamethasone groups. Bcl-2 and PCNA decreased in the betamethasone groups at 121 DG and 45 PD, but increased at 132 DG and 90 PD. We conclude that high levels of pre-natally administered glucocorticoid reduce foetal testicular development, perhaps via changes in the balance between pro- and anti-apoptotic proteins and cell-cycle proteins. These outcomes could compromise the future spermatogenic potential of male offspring.

Selection for superior growth advances the onset of puberty and increases reproductive performance in ewe lambs.

The reproductive efficiency of the entire sheep flock could be improved if ewe lambs go through puberty early and produce their first lamb at 1 year of age. The onset of puberty is linked to the attainment of critical body mass, and therefore we tested whether it would be influenced by genetic selection for growth rate or for rate of accumulation of muscle or fat. We studied 136 Merino ewe lambs with phenotypic values for depth of eye muscle (EMD) and fat (FAT) and Australian Sheep Breeding Values at post-weaning age (200 days) for live weight (PWT), eye muscle depth (PEMD) and fat depth (PFAT). First oestrus was detected with testosterone-treated wethers and then entire rams as the ewes progressed from 6 to 10 months of age. Blood concentrations of leptin and IGF-I were measured to test whether they were related to production traits and reproductive performance (puberty, fertility and reproductive rate). In total, 97% of the lambs reached first oestrus at average weight 39.4 ± 0.4 kg (mean ± s.e.m.) and age 219 days (range 163 to 301). Age at first oestrus decreased with increases in values for PWT (P < 0.001), and concentrations of IGF-I (P < 0.05) and leptin (P < 0.01). The proportion of ewe lambs that achieved puberty was positively related with increases in values for EMD (P < 0.01), FAT (P < 0.05) or PWT (P < 0.01), and 75% of the ewe lambs were pregnant at average weight 44.7 ± 0.5 kg and age 263 days (range 219 to 307). Ewe lambs that were heavier at the start of mating were more fertile (P < 0.001) and had a higher reproductive rate (P < 0.001). Fertility and reproductive rate were positively correlated with increases in values for EMD (P < 0.01), FAT (P < 0.05), PWT (P < 0.01) and leptin concentration (P < 0.01). Fertility, but not reproductive rate, increased as values for PFAT increased (P < 0.05). Leptin concentration increased with increases in values for EMD (P < 0.001), FAT (P < 0.001), PWT (P < 0.001), PEMD (P < 0.05) and PFAT (P < 0.05). Many of these relationships became non-significant when PWT or live weight was added to the statistical model. We conclude that selection for genetic potential for growth can accelerate the onset of puberty and increase fertility and reproductive rate of Merino ewe lambs. The metabolic hormones, IGF-I and leptin, might act as a physiological link between the growing tissues and the reproductive axis.

Interrelationships of nutrition, metabolic hormones and resumption of ovulation in multiparous suckled beef cows on subtropical pastures.

The effect of nutrition before and after calving on metabolic status and the resumption of ovulation postpartum was examined in multiparous sucked beef cows on subtropical pastures. At 6-7 months of gestation, Droughtmaster cows were randomly assigned on body weight (BW) and stage of gestation to two groups that received either standard subtropical pasture (SP, n = 7, 543 ± 12 kg BW) or improved pasture (IP, n = 7, 564 ± 12 kg BW). The two nutritional treatments were maintained after calving. Starting at 1 week after calving, cows were monitored for BW and body condition score (BCS, biweekly) and for circulating concentrations of insulin, glucose, IGF-1, GH and leptin (weekly). Ovarian follicular status was monitored weekly by trans-rectal ultrasonography. Fecal samples were obtained at 3-week intervals to ascertain percentage crude protein (%CP) and dry matter digestibility (DMD) of pastures. Crude protein and DMD were greater (P < 0.05) for cows on IP during the first 9 weeks after calving after which there were no differences between nutritional treatments. Cows on IP were heavier (P < 0.05) and had a greater (P < 0.01) BCS than cows on SP at 1 week after calving (585 ± 9 kg and 3.7 ± 0.2 BCS and 528 ± 21 kg and 2.3 ± 0.2 BCS, respectively). Cows on SP showed a gradual increase in BW and there were no differences in BW after approximately 7 weeks postpartum whilst BCS remained less for cows on SP. Plasma concentrations of insulin, glucose, IGF-1 and leptin were all greater (P < 0.01) for cows on IP compared with cows on SP, whilst GH did not differ. The diameter of the largest follicle did not differ between cows on IP and SP throughout the postpartum period. However, 7 of 7 cows on IP resumed ovulations between 12 and 15 weeks postpartum whilst only 1 of 7 cows on SP had resumed ovulation during the study. It is concluded from the findings that exposure of cows to IP and SP before and after calving resulted in two groups of cows with different metabolic homeostasis and that the greater circulating concentrations of insulin, glucose, IGF-1 and leptin promoted the earlier resumption of ovulation for cows on IP.