Polyunsaturated fatty acids in male and female reproduction

In Westernized societies, average consumption of n-6 polyunsaturated fatty acids (PUFAs) far exceeds nutritional requirements. The ratio of n-6 to n-3 PUFAs is generally >10:1 whereas on a primitive human diet it was closer to 1:1. Diets fed to intensively farmed livestock have followed a similar trend. Both n-6 and n-3 PUFAs can influence reproductive processes through a variety of mechanisms. They provide the precursors for prostaglandin synthesis and can modulate the expression patterns of many key enzymes involved in both prostaglandin and steroid metabolism. They are essential components of all cell membranes. The proportions of different PUFAs in tissues of the reproductive tract reflect dietary consumption. PUFA supplements (particularly n-3 PUFAs in fish oil) are promoted for general health reasons. Fish oils may also benefit fertility in cattle and reduce the risk of preterm labor in women, but in both cases current evidence to support this is inconclusive. Gamma-linolenic acid containing oils can alter the types of prostaglandins produced by cells in vitro, but published data to support claims relating to effects on reproductive health are lacking. Spermatozoa require a high PUFA content to provide the plasma membrane with the fluidity essential at fertilization. However, this makes spermatozoa particularly vulnerable to attack by reactive oxygen species, and lifestyle factors promoting oxidative stress have clear associations with reduced fertility. Adequately powered trials that control for the ratios of different PUFAs consumed are required to determine the extent to which this aspect of our diets does influence our fertility.     

Regulatory effect of steroid hormones and fetal tissues on expression of oxytocin receptor in the endometrium of late pregnant ewes.

Oxytocin receptors play an important role in the establishment of pregnancy and parturition in ruminants. Previous studies in cyclic and early pregnant ewes have indicated that receptor concentrations are regulated by steroid hormones and fetal secretory products. This study investigated the effect of oestradiol and progesterone, or co-culture with placenta or corpus luteum on oxytocin receptor expression. Endometrial explants from late pregnant ewes were cultured for up to 96 h in various treatment combinations. After culture, tissues were subjected to in situ hybridization and autoradiography with 125I-labelled oxytocin receptor antagonist to localize and measure the expression of oxytocin receptor mRNA and protein. Results were quantified as absorbance units from autoradiographs. Oxytocin receptors were confined to the endometrial luminal epithelium and both mRNA and 125I-labelled oxytocin receptor antagonist binding were upregulated spontaneously in basic serum-free medium. Upregulation occurred earlier in the presence of oestradiol (0.1 mumol l-1) but the final receptor concentration was similar to that found in the basic medium. Continuous progesterone treatment (1 mumol l-1) and co-culture with corpus luteum both delayed the increase in oxytocin receptor mRNA, but a short initial (4 h) period in progesterone-free basic medium resulted in loss of the inhibitory effect. Co-culture with placental tissues had no effect. In conclusion, oxytocin receptor expression in the luminal epithelium increased immediately on removal from the maternal environment. This occurred regardless of treatment and did not require the presence of steroid hormones, but could be accelerated or delayed by oestradiol and progesterone, respectively. There may be an additional inhibitory factor present in the corpus luteum.     

The impact of chronic environmental stressors on growing pigs, Sus scrofa (Part 1): stress physiology, production and play behaviour

Commercially farmed animals are frequently housed in conditions that impose a number of concurrent environmental stressors. For pigs housed indoors, elevated levels of mechanical noise, atmospheric ammonia and low light intensities are commonplace. This experiment examined the effects on growing pigs of chronic exposure to combinations of commercially relevant levels of these potential stressors. Four-week-old hybrid female pigs (n = 224) were housed under experimentally manipulated conditions of nominally either <5 or 20 ppm atmospheric concentration of ammonia (24 h), a light intensity of 40 lux or 200 lux (12 h) and mechanical noise at either ⩽60 or 80 dB(A) (24 h) for 15 weeks in a fully factorial arrangement (2³) of treatments. The response of pigs to these environmental factors was assessed using a suite of physiological, production and behavioural measures. These included indicators of hypothalamic–pituitary–adrenal (HPA) axis activation such as salivary cortisol and adrenal morphometry, as well as body weight, food conversion efficiency and general health scores. Play behaviour was recorded as it is thought to be inversely related to stress. Chronic exposure to ammonia produced the strongest effect, shown by lower concentrations of salivary cortisol and larger adrenal cortices in the pigs reared under 20 ppm ammonia, which may have been indicative of a period of HPA activation leading to a downregulation of cortisol production. The pigs in the ammoniated rooms also performed less play behaviour than pigs in non-ammoniated rooms. There was evidence for an interaction between high noise and ammonia on the health scores of pigs and for brighter light to ameliorate the effect of ammonia on salivary cortisol. However, there was no measurable impact of these potential stressors on the productivity of the pigs or any of the other physiological parameters measured. We conclude that there should be little concern in terms of performance about the physical stressors tested here, within current European Union legal limits. However, 20 ppm ammonia may have had an adverse influence on the well-being of growing pigs. In this study, all other aspects of the pigs’ husbandry were optimal; therefore, it is possible that under less favourable conditions, more pronounced effects of ammonia, noise and dim light would be observed.     

Mortality in Holstein-Friesian calves and replacement heifers, in relation to body weight and IGF-I concentration, on 19 farms in England

The incidence of mortality and culling in Holstein-Friesian heifers from birth through first calving was determined on 19 dairy farms selected from across southern England. The outcome of 1097 calvings was determined. Size (BW, heart girth, crown-rump length and height at withers) and insulin-like growth factor-I concentration of live heifer calves were measured at a mean age of 26 ± 0.7 days (n = 506). Associations between the heifer-level variables and mortality were determined using clustered binary logistic regression. Perinatal mortality (stillbirths and mortality within the first 24 h of birth) of male and female calves was 7.9%. This figure was significantly higher in cases where calving assistance was required (19.1% v. 5.6%, P < 0.001) and in twin births (18.5% v. 7.0%, P < 0.05), and was lower in pluriparous v. primiparous dams (5.6% v. 12.1%, P < 0.01). On average, 6.8% of heifers died or were culled between 1 day and 6 months of age. Low BW at 1 month was associated with reduced subsequent survival up to 6 months. Between 6 months and first calving, a further 7.7% of heifers either died (42%) or were culled (58%); accidents and infectious disease accounted for the majority of calf deaths between 6 and 15 months, whereas infertility (16/450 animals served, 3.5%) was the main reason for culling following the start of the first breeding period. In total, 11 heifers (2.2%) were culled as freemartins; eight at birth and three around service. Overall, 14.5% of liveborn potential replacement heifers died or were culled before first calving.     

Metabolic profiles and progesterone cycles in first lactation dairy cows

This study investigated the ovarian function, metabolic profiles and fertility in first lactation Holstein-Friesian dairy cows (mean 305 day milk yield: 7417+/-191kg, n=37). Reproductive profiles obtained from milk progesterone analysis were categorized into normal (n=17) and four abnormal profiles (delayed ovulation, DOV1, n=9; DOV2, n=2; persistent corpus luteum, PCL1, n=6; PCL2, n=4; 1: immediately post-calving, 2: subsequent cycles). Fifty-five percent of cows had abnormal profiles with half of these being categorized as DOV1. Fertility of DOV1 and DOV2 cows was reduced whereas PCL1 and PCL2 cows had similar reproductive competence to normal profile cows. DOV1 animals had higher milk energy values, lower energy balances, lower dry matter intakes (DMI) and greater body weight and body condition score (BCS) losses post-calving than normal profile animals. DOV1 animals also had lower insulin-like growth factor-I (IGF-I) and higher betahydroxybutyrate (BHB) concentrations and tended to have the lower insulin and glucose concentrations in the pre-service period than normal profile cows. All PCL animals had vulval discharges postpartum. Despite this, the DMI, body weight and BCS changes, IGF-I concentrations and fertility of PCL1 animals was similar to normal profile cows. In conclusion, the high prevalence of delayed ovulation post-calving (DOV1) in primiparous high yielding cows lasted long enough (71+/-8.3 days) to have a detrimental impact on fertility and was associated with significant physiological changes. This study did not establish any detrimental effects of PCL profiles on fertility or production parameters.     

The effect of a diet containing excess quickly degradable nitrogen (QDN) on reproductive and metabolic hormonal profiles of lactating dairy cows

The objective of this experiment was to examine the effects of an excess intake of quickly degradable nitrogen (QDN) on metabolic and reproductive parameters in lactating dairy cows. Twenty-two lactating dairy cows were fed a total mixed ration once daily. The control diet was a typical ration for high producing cows in the UK (CP = 17.5%, ME = 11.8 MJ/kg DM). The cows were randomly divided into two groups, control diet (control; n = 12) and excess QDN diet (QDN; n = 10). The QDN group was fed an additional 250 g of urea per cow per day, from 10 days before insemination (day 0) until the end of the experiment, 17 days after the second insemination. Ten days before insemination, a synchronized oestrus was induced and the cows inseminated twice, 48 and 72 h after synchronization, with commercial frozen semen from a single sire. Ovaries were scanned using B-mode ultrasonography 10 days before insemination and then daily from 3 days before insemination. Eighteen of the cows (9 control and 9 QDN) were sampled more intensively to determine the pulsatile pattern of secretion of luteinizing hormone (LH) and growth hormone (GH). Cows were slaughtered 17 days after insemination, the reproductive tracts recovered and flushed to retrieve embryos. The excess QDN diet resulted in elevated (P < 0.05) plasma urea concentrations 3 days after starting urea feeding and these were maintained until the end of the experiment. However, the excess QDN diet did not significantly affect daily milk production or plasma concentrations of insulin and IGF-I. The QDN treatment did not significantly affect pulsatile patterns of secretion of LH and GH or the number of small (< 0.5 cm diameter) and medium to large follicles (> 0.5 cm diameter). Twenty cows ovulated following synchronization (control 11/12; QDN 9/10). There were no significant differences between the control and the QDN groups in the peak concentrations of oestradiol during the follicular phase or in the post-ovulatory pattern of plasma and milk progesterone secretion. Embryos and/or foetal membranes were recovered from 10 cows (5 control and 5 QDN). The results of the current study show that feeding excess QDN, as urea, for 27 days commencing 10 days before insemination had no effect on reproductive or metabolic hormonal parameters. Ovulation and the formation and function of the post-ovulatory corpus luteum were also unaffected by excess QDN. These data suggest that the harmful effects of excess intakes of QDN on fertility occur after 17 days following ovulation.     

Regulation of insulin-like growth factor binding protein-6 expression in the reproductive tract throughout the estrous cycle and during the development of the placenta in the ewe

Insulin-like growth factors (IGF-I and IGF-II) are essential for normal uterine development and have been particularly implicated in fetal and placental growth. A family of six IGF binding proteins enhance or attenuate IGF-stimulated cell proliferation. In this study we have used in situ hybridization to map the distribution of IGFBP-6, one of the lesser known of the IGFBPs, in sections of the uterus collected from cyclic, anestrous, and ovariectomized nonpregnant ewes and from the uterus and placenta of early pregnant (13-55 days) and unilaterally pregnant ewes. In nonpregnant ewes IGFBP-6 mRNA (measured as arbitrary optical density units from autoradiographs) was abundant in the periepithelium and caruncles, with lower levels in the endometrial stroma and myometrium. In most regions IGFBP-6 mRNA showed cyclic variations with concentrations maximal around ovulation and the early luteal phase. In addition, 16 out of 25 ewes expressed IGFBP-6 mRNA in their endometrial glands between estrus and Day 2. Measurements of IGFBP-6 mRNA were high in anestrous ewes (equivalent values to ovulation) but low in ovariectomized ewes (equivalent values to mid to late luteal phase). In pregnant ewes IGFBP-6 mRNA was found in similar regions to those recorded during the cycle. In the periepithelium and caruncular stroma IGFBP-6 mRNA levels were higher during early pregnancy than in the midluteal phase. In the unilateral pregnant ewes there was no difference in IGFBP-6 mRNA measured between pregnant and nonpregnant horns. In conclusion, IGFBP-6 mRNA is differentially regulated during the estrous cycle and pregnancy and may be functionally important in modulating IGF activity in the uterus and placenta by virtue of its strong affinity and ability to regulate IGF-II mediated actions.     

Cloning and gene map assignment of the Xiphophorus DNA ligase 1 gene

Fishes represent the stem vertebrate condition and have maintained several gene arrangements common to mammalian genomes throughout the 450 Myr of divergence from a common ancestor. One such syntenic arrangement includes the GPI-PEPD enzyme association on Xiphophorus linkage group IV and human chromosome 19. Previously we assigned the Xiphophorus homologue of the human ERCC2 gene to linkage group U5 in tight association with the CKM locus. CKM is also tightly linked to the ERCC2 locus on human chromosome 19, leading to speculation that human chromosome 19 may have arisen by fusion of two ancestral linkage groups which have been maintained in fishes. To investigate this hypothesis further, we isolated and sequenced Xiphophorus fish genomic regions exhibiting considerable sequence similarity to the human DNA ligase 1 amino acid sequence. Comparison of the fish DNA ligase sequence with those of other species suggests several modes of amino acid conservation in this gene. A 2.2-kb restriction fragment containing part of an X. maculatus DNA ligase 1 exon was used in backcross hybrid mapping with 12 enzyme or RFLP loci. Significant linkage was observed between the nucleoside phosphorylase (NP2) and the DNA ligase (LIG1) loci on Xiphophorus linkage group VI. This assignment suggests that the association of four DNA repair-related genes on human chromosome 19 may be the result of chance chromosomal rearrangements.