Effect of follicular fluid supplementation on in vitro maturation of porcine oocytes [abstract]

Abstract only availableThe reproductive process that occurs in the female is controlled by a series of critical events. One such event that must occur is the "maturation" of the oocyte prior to its ovulation, so that it is capable of being fertilized. This maturation process occurs naturally in vivo and has been mimicked in vitro. Currently, there are two methods of determining oocyte maturation after being subjected to an oocyte in vitro maturation (IVM) protocol. The first method involves fixing and staining the oocytes to identify the developmental stage of each oocyte. The second method is based on visually determining whether each oocyte has extruded a polar body, which indicates that the oocyte has resumed meiosis and reached metaphase II. Follicular fluid (FF) is believed to play an important role in the in vivo maturation of oocytes. Our hypothesis was: 1) that the addition of FF to the IVM medium would increase the number of oocytes reaching metaphase II, and 2) that the success rate was dependent on the percentage of FF in the culture media. To test this hypothesis, we utilized porcine oocytes and porcine follicular fluid (pFF). Porcine ovaries were collected from a nearby slaughter facility and transported to the lab where the oocytes were aspirated from follicles on the ovaries of prepuberal gilts. Oocyte culture media consisted of: 1) TCM 199 as a control medium, 2) TCM 199 + 10% pFF, 3) TCM 199 + 25% pFF, or 4) TCM 199 + 50% pFF. Oocytes were randomly allocated to one of the four culture media and incubated at 37.5°C for approximately 46 hours. After incubation the cumulus complex was removed using hyaluronidase, and oocytes were examined for the presence of an extruded polar body. The control medium incubation resulted in 54.25% of oocytes reaching metaphase II (i.e. they had an extruded polar body). Incubation of oocytes in TCM 199 + 10% pFF resulted in a 57.96% maturation rate; incubation in TCM 199 + 25% pFF resulted in a 54.61% maturation rate and incubation in TCM 199 + 50% pFF resulted in a 48.10% maturation rate. Although there was no significant difference among the treatments, we suggest that these data provide evidence that the addition of pFF to the IVM culture of porcine oocytes may be beneficial, but not when added in excess of 25%.CAFNR On Campus Research Internshi

Sheep pregnancy checking by ultrasonic sound (1993)

Ultrasonic sound waves of about 2 million cycles per second can pass through living tissue. Used to determine sheep pregnancy, these sound waves are painless and have no harmful after-effects to the ewe or fetus

Low protein intake during the preconception period in beef heifers affects offspring and maternal behaviour

Maternal low protein diets prior to conception alter embryo and fetal development and are associated with detrimental outcomes in the offspring in many species. The aim of this study in beef cattle was to investigate the effect of preconception dietary protein upon maternal and offspring behaviour at birth concomitant with the associated hormonal profile. Sixty days prior to conception, nulliparous yearling heifers (n = 85) were fed either a High (PreH: 18%; n = 43) or Low (PreL: 10%; n = 42) crude protein diet, followed by a control diet throughout gestation. After calving, each cow-calf pair was penned individually, accelerometers fitted, and each pen observed continuously via video recordings. Cows fed on the low protein diet during preconception showed an increase in standing time (P [less than] 0.01); while calves born to heifers receiving the PreL diet showed an increase in suckling time (P = 0.04). These calves were also heavier at birth than calves from PreH mothers (P [less than] 0.01). In conclusion, low maternal dietary protein prior to conception in beef heifers modifies both offspring feeding behaviour and birth weight, and cow’s standing times

Sex differences in metabolic and adipose tissue responses to juvenile-onset obesity in sheep

Sex is a major factor determining adipose tissue distribution and the subsequent adverse effects of obesity-related disease including type 2 diabetes. The role of gender on juvenile obesity and the accompanying metabolic and inflammatory responses is not well established. Using an ovine model of juvenile onset obesity induced by reduced physical activity, we examined the effect of gender on metabolic, circulatory, and related inflammatory and energy-sensing profiles of the major adipose tissue depots. Despite a similar increase in fat mass with obesity between genders, males demonstrated a higher storage capacity of lipids within perirenal-abdominal adipocytes and exhibited raised insulin. In contrast, obese females became hypercortisolemic, a response that was positively correlated with central fat mass. Analysis of gene expression in perirenal-abdominal adipose tissue demonstrated the stimulation of inflammatory markers in males, but not females, with obesity. Obese females displayed increased expression of genes involved in the glucocorticoid axis and energy sensing in perirenal-abdominal, but not omental, adipose tissue, indicating a depot-specific mechanism that may be protective from the adverse effects of metabolic dysfunction and inflammation. In conclusion, young males are at a greater risk than females to the onset of comorbidities associated with juvenile-onset obesity. These sex-specific differences in cortisol and adipose tissue could explain the earlier onset of the metabolic-related diseases in males compared with females after obesity

Liver lipid content and inflammometabolic indices in peripartal dairy cows are altered in response to prepartal energy intake and postpartal intramammary inflammatory challenge.

This study evaluated the effect of feeding a control diet (CON) or a moderate energy diet (overfed, OVE) during the dry period (∼45d) and a postpartum intramammary lipopolysaccharide (LPS) challenge on blood metabolic and inflammatory indices, milk production, and hepatic gene expression. A subset of cows (n=9/diet) in CON (1.34 Mcal/kg of dry matter) and OVE (1.62 Mcal/kg of dry matter) received an intramammary LPS challenge (200 μg; CON-LPS, OVE-LPS, respectively). Liver biopsies were harvested at -14 d from calving, and postpartum at 2.5h post-LPS on d 7, 14, and 30. Prepartum, the OVE group was in more positive energy balance (EB) and had greater body condition score compared with CON. In contrast, during wk 1 postpartum and before the LPS challenge, the OVE group was in greater negative EB than CON. Prepartal diet did not affect postpartal milk production or dry matter intake. At 2h postchallenge on d 7, we observed an increase in serum nonesterified fatty acids (NEFA) and bilirubin and a decrease in hydroxybutyrate, regardless of diet. That was coupled with greater haptoglobin in OVE-LPS compared with CON-LPS. In addition, OVE-LPS cows versus CON nonchallenged, OVE nonchallenged, and CON-LPS had greater liver triacylglycerol (TAG) concentration 2.5h postchallenge on d 7. The concentration of TAG in liver of OVE-LPS remained elevated by 30d postpartum. The liver TAG concentration in OVE-LPS compared with CON-LPS cows was associated with greater serum concentration of NEFA and reactive oxygen metabolites on d 10 and 14 postpartum. Cows in OVE-LPS also had greater concentrations of ceruloplasmin, cholesterol, and vitamin E from d 10 through 21. Among 28 genes associated with fatty acid oxidation, inflammation, oxidative stress, and gluconeogenesis, only SAA3 (which encodes an acute phase protein) was greater in CON-LPS compared with OVE-LPS at 2.5h postchallenge. Expression of HP, which encodes another acute phase protein, was greater in OVE-LPS than in CON-LPS at 14 and 30 d postpartum. Several inflammation-related genes (TNF, IRAK1, NFKB1, ANGPTL4) showed markedly decreased expression between 7 and 14 d, after which expression remained unchanged. No differences were observed in several genes of the growth-hormone/insulin-like growth factor-1 axis, except for SOCS2, expression of which decreased markedly between 7 and 14 d in OVE-LPS but not in CON-LPS. These data suggest that overfeeding a moderate-energy diet prepartum alters the response of the cow to an intramammary challenge after calving and may predispose it to sustained liver lipidosis

SEASONAL VARIATION OF NUTRITIONAL HORMONES IN CAPTIVE FEMALE MOOSE

The health status of animals may be inferred from the patterns of hormonal concentrations and other chemical characteristics in blood samples. Baseline endocrine data representing the nutritional and reproductive condition of moose are currently unknown. In this study, we examined the seasonal patterns of 3 nutritional hormones (leptin, ghrelin, insulin-like growth factor-1) in 3 captive, non-pregnant female moose (Alces alces) fed a maintenance diet from November to August. Plasma concentrations for leptin, ghrelin, and IGF-1 averaged 1.36 ± 0.81 ng/mL, 0.229 ± 0.110 ng/mL, and 114.0 ± 30.5 ng/mL, respectively; only ghrelin displayed a seasonal change. Plasma ghrelin concentration was significantly elevated (P < 0.001) during winter months suggesting it may be sensitive to seasonal changes and indicative of nutritional status

Nutritional skewing of conceptus sex in sheep: effects of a maternal diet enriched in rumen-protected polyunsaturated fatty acids (PUFA)

<p>Abstract</p> <p>Background</p> <p>Evolutionary theory suggests that in polygynous mammalian species females in better body condition should produce more sons than daughters. Few controlled studies have however tested this hypothesis and controversy exists as to whether body condition score or maternal diet is in fact the determining factor of offspring sex. Here, we examined whether maternal diet, specifically increased n-6 polyunsaturated fatty acid (PUFA) intake, of ewes with a constant body condition score around the time of conception influenced sex ratio.</p> <p>Methods</p> <p>Ewes (n = 44) maintained in similar body condition throughout the study were assigned either a control (C) diet or one (F) enriched in rumen-protected PUFA, but otherwise essentially equivalent, from four weeks prior to breeding until d13 post-estrus. On d13, conceptuses were recovered, measured, cultured to assess their capacity for interferon-tau (IFNT) production and their sex determined. The experiment was repeated with all ewes being fed the F diet to remove any effects of parity order on sex ratio. Maternal body condition score (BCS), plasma hormone and metabolite concentrations were also assessed throughout the study and related to diet.</p> <p>Results</p> <p>In total 129 conceptuses were recovered. Ewes on the F diet produced significantly more male than female conceptuses (proportion male = 0.69; deviation from expected ratio of 0.5, P < 0.001). Conceptus IFNT production was unaffected by diet (P > 0.1), but positively correlated with maternal body condition score (P < 0.05), and was higher (P < 0.05) in female than male conceptuses after 4 h culture. Maternal plasma hormone and metabolite concentrations, especially progesterone and fatty acid, were also modulated by diet.</p> <p>Conclusion</p> <p>These results provide evidence that maternal diet, in the form of increased amounts of rumen-protected PUFA fed around conception, rather than maternal body condition, can skew the sex ratio towards males. These observations may have implications to the livestock industry and animal management policies when offspring of one sex may be preferred over the other.</p

Nutritional Skewing of Conceptus Sex in Sheep: Effects of a Maternal Diet Enriched in Rumen-Protected Polyunsaturated Fatty acids (PUFA)

doi:10.1186/1477-7827-6-21Evolutionary theory suggests that in polygynous mammalian species females in better body condition should produce more sons than daughters. Few controlled studies have however tested this hypothesis and controversy exists as to whether body condition score or maternal diet is in fact the determining factor of offspring sex. Here, we examined whether maternal diet, specifically increased n-6 polyunsaturated fatty acid(PUFA) intake, of ewes with a constant body condition score around the time of conception influenced sex ratio.The research was supported by USDA/CSREES/NRI Grant 2001-35203- 10693 (to RMR) and a Life Sciences Molecular Biology Fellowship, University of Missouri (partial salary support for MPG)

Effect of pre- and postnatal growth and post-weaning activity on glucose metabolism in the offspring

Maternal caloric restriction during late gestation reduces birth weight, but whether long-term adverse metabolic outcomes of intra-uterine growth retardation (IUGR) are dependent on either accelerated postnatal growth or exposure to an obesogenic environment after weaning is not established. We induced IUGR in twin-pregnant sheep using a 40% maternal caloric restriction commencing from 110 days of gestation until term (∼147 days), compared with mothers fed to 100% of requirements. Offspring were reared either as singletons to accelerate postnatal growth or as twins to achieve standard growth. To promote an adverse phenotype in young adulthood, after weaning, offspring were reared under a low-activity obesogenic environment with the exception of a subgroup of IUGR offspring, reared as twins, maintained in a standard activity environment. We assessed glucose tolerance together with leptin and cortisol responses to feeding in young adulthood when the hypothalamus was sampled for assessment of genes regulating appetite control, energy and endocrine sensitivity. Caloric restriction reduced maternal plasma glucose, raised non-esterified fatty acids, and changed the metabolomic profile, but had no effect on insulin, leptin, or cortisol. IUGR offspring whose postnatal growth was enhanced and were obese showed insulin and leptin resistance plus raised cortisol. This was accompanied by increased hypothalamic gene expression for energy and glucocorticoid sensitivity. These long-term adaptations were reduced but not normalized in IUGR offspring whose postnatal growth was not accelerated and remained lean in a standard post-weaning environment. IUGR results in an adverse metabolic phenotype, especially when postnatal growth is enhanced and offspring progress to juvenile-onset obesity

Tissue cell stress response to obesity and its interaction with late gestational diet

Intra-uterine growth restriction in late pregnancy can contribute to adverse long term metabolic health in the offspring. We utilised an animal (sheep) model of maternal dietary manipulation in late pregnancy, combined with exposure of the offspring to a low activity, obesogenic environment after weaning, to characterise the effects on glucose homeostasis. Dizygotic twin-pregnant sheep were either fed to 60% of requirements (nutrient restriction (R)) or fed ad libitum (~ 140% of requirements (A)) from 110 days gestation until term (~147d). After weaning (~3 months of age), their offspring were kept in either a standard (in order to remain lean) or low activity, obesogenic environment. R mothers gained less weight and produced smaller offspring. As adults, obese offspring were heavier and fatter with reduced glucose tolerance, irrespective of maternal diet. Molecular markers of stress and autophagy in liver and adipose tissue were increased with obesity, with gene expression of hepatic Grp78 and of omental Atf6, Grp78 and Edem1 only being increased in R offspring. In conclusion, the adverse effect of juvenile onset obesity on insulin responsive tissues can be amplified by previous exposure to a suboptimal nutritional environment in utero, thereby contributing to earlier onset of insulin resistance