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Headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for the determination of 5alpha-androst-2-en-17-one and -17beta-ol in the female Asian elephant: application for reproductive monitoring and prediction of parturition.

Asian elephants are not self-sustaining in captivity. The main reasons for this phenomenon are a low birth rate, an aging population, and poor calf-rearing. Therefore, it is essential that reproductive rates had to be improved and there is need for rapid quantitative measures to monitor reproductive functions focussing on estrous detection and the prediction of the period of parturition. The objective of this study was to develop a method which combines headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for analyses of 5alpha-androst-2-en-17beta-ol and -17-one to prognose estrous and to predict the period of parturition. SPME was carried out with a CTC Combi Pal system. The course of the luteal phase-specific substance 5alpha-androst-2-en-17beta-ol and -17-one followed a cyclic pattern in which the follicular and luteal phases could be clearly distinguished (mean estrous cycle length, 15+/-1.4 weeks). Based on daily urine samples, estrous prognosis might be possibly based on the initial 5alpha-androst-2-en-17beta-o1 increase at the end of the follicular phase. Parturition prognosis was performed in three elephant cows based on the 5alpha-androst-2-en-17beta-o1 drop to baseline levels 5-4 days prior parturition. Experiments revealed that 5alpha-androst-3alpha-ol-17-one and probably 5alpha-androst-3alpha-ol-17beta-ol are generated from sulfate conjugates by a thermal process.

Androstane-3,17-diol↗

Human myometrial gene expression before and during parturition.

Identification of temporal and spatial changes in myometrial gene expression during parturition may further the understanding of the coordinated regulation of myometrial contractions during parturition. The objective of this study was to compare the gene expression profiles of human fundal myometrium from pregnant women before and after the onset of labor using a functional genomics approach, and to further characterize the spatial and temporal expression patterns of three genes believed to be important in parturition. Fundal myometrial mRNA was isolated from five women in labor and five women not in labor, and analyzed using human UniGEM-V microarrays with 9182 cDNA elements. Real-time polymerase chain reaction using myometrial RNA from pregnant women in labor or not in labor was used to examine mRNA levels for three of the genes; namely, prostaglandin-endoperoxide synthase 2 (PTGS2), calgranulin B (S100A9), and oxytocin receptor (OXTR). The spatial expression pattern of these genes throughout the pregnant uterus before and after labor was also determined. Immunolocalization of cyclooxygenase-2 (also known as PTGS2) and S100A9 within the uterine cervix and myometrium were analyzed by immunohistochemistry. Few genes were differentially expressed in fundal myometrial tissues at term with the onset of labor. However, there appears to be a subset of genes important in the parturition cascade. The cellular properties of S100A9, its spatial localization, and dramatic increase in cervix and myometrium of women in labor suggest that this protein may be very important in the initiation or propagation of human labor.

Calgranulin B↗

Regulation of progesterone levels during pregnancy and parturition by signal transducer and activator of transcription 5 and 20alpha-hydroxysteroid dehydrogenase.

The two highly related signal transducers and activators of transcription (Stats), Stat5a and Stat5b, are major mediators of prolactin signaling in both the mammary gland and in the ovary. Deficiencies in Stat5b, or in both Stat5a and Stat5b, result in loss of pregnancy during midgestation and are correlated with an increase in ovarian 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) and a decrease in serum progesterone, which normally declines only immediately before parturition. To determine the relative contribution of 20alpha-HSD to progesterone metabolism and Stat5 function during pregnancy and parturition, we created a 20alpha-HSD-deficient strain of mice by gene disruption. Mice deficient for 20alpha-HSD sustain high progesterone levels and display a delay in parturition of several days demonstrating that 20alpha-HSD regulates parturition downstream of the prostaglandin F2alpha receptor in an essential and nonredundant manner. Moreover, 20alpha-HSD deficiency partially corrected the abortion of pregnancies associated with Stat5b deficiency, supporting the concept that prolactin activation of Stat5b is important in suppressing 20alpha-HSD gene expression and thereby allowing the maintenance of progesterone levels that are required to sustain pregnancy.

20-alpha-Hydroxysteroid Dehydrogenase↗

Genetic analysis of litter size, parturition length, and birth assistance requirements in primiparous sows using a joint linear-threshold animal model.

The aim of this study was to investigate whether selection for number of live born piglets has led to prolonged parturition and increased requirement for birth assistance, resulting in increased numbers of stillborn piglets. Data were collected from 6,718 primiparous Norwegian Landrace sows farrowing between 2001 and 2003. The need for birth assistance was recorded as a binary response. Physical intervention in the birth of piglets and/or hormonal treatment by the farmer was recorded as birth assistance. The duration of the parturition was analyzed as a binary trait (<4 h and >4 h). The statistical model used for analysis included contemporary groups of herd-year, litter breed, season of farrowing, parity in which the sow was born, a regression on the age of sow at farrowing, an additive genetic effect, and a service sire effect. A full Bayesian approach via Gibbs sampling was adopted to estimate the genetic relationships between these four traits. A total chain length of 100,000 iterations was run. The first 10,000 samples were discarded as burn-in, and the remaining 90,000 iterations were retained without thinning for post-Gibbs analysis. The highest direct heritability was estimated for the number of live-born piglets (h2 = 0.07), followed by the duration of farrowing (h2 = 0.05), the need for birth assistance (h2 = 0.05), and the number of stillborn piglets (h2 = 0.04). The genetic correlations revealed that the number of live and stillborn piglets was uncorrelated; however, the number of live piglets born had a moderate genetic correlation to the need for birth assistance (rg = 0.24 +/- 0.01) and duration of farrowing (rg = -0.20 +/- 0.01), whereas the number of stillborn piglets was highly correlated to the need for birth assistance (rg = 0.74 +/- 0.01) and the duration of parturition (rg = 0.66 +/- 0.01). The duration of farrowing and the need for birth assistance were genetically highly correlated (rg = 0.89 +/- 0.00). For all traits, the service sire variance was approximately one quarter in magnitude compared with its respective genetic variance. The results showed that selection for the number of live born piglets is not expected to influence the number of stillborn piglets. Increasing the number of live piglets born through selection should have a slight negative effect on farrowing duration and a minor increase in the need for birth assistance. Sows with a high genetic potential for birth assistance and prolonged parturition were more likely to give birth to greater numbers of stillborn piglets.

Animals↗

Effects of body condition score at parturition and postpartum supplemental fat on metabolite and hormone concentrations of beef cows and their suckling calves.

To determine the effects of BCS at parturition and postpartum lipid supplementation on blood metabolite and hormone concentrations, 3-yr-old Angus x Gelbvieh beef cows, which were nutritionally managed to achieve a BCS of 4 +/- 0.07 (479.3 +/- 36.3 kg of BW) or 6 +/- 0.07 (579.6 +/- 53.1 kg of BW) at parturition, were used in a 2-yr experiment (n = 36/yr). Beginning at 3 d postpartum, cows within each BCS were assigned randomly to be fed hay and a low-fat control supplement or lipid supplements with either cracked high-linoleate or high-oleate safflower seeds until d 61 of lactation. The diets were formulated to be isonitrogenous and isocaloric, and the safflower seed supplements were formulated to achieve 5% DMI as fat. On d 31 and 61 of lactation, blood samples were collected preprandially and then hourly postprandially (at 0, 1, 2, 3, and 4 h). Serum insulin (P = 0.27) and glucose (P = 0.64) were not affected by BCS at parturition. The mean concentrations of plasma NEFA (P = 0.08) and beta-hydroxybutyrate (P = 0.08) tended to be greater, and serum IGF-I was greater (P < 0.001) in BCS 6 than BCS 4 cows. Conversely, serum GH was greater (P = 0.003) for BCS 4 cows, indicating that regulation of IGF by GH may have been uncoupled in BCS 4 cows. The postpartum diet did not affect NEFA (P = 0.94), glucose (P = 0.15), IGF-I (P = 0.33), or GH (P = 0.62) concentrations. Oleate-supplemented cows had greater (P = 0.03) serum insulin concentrations, whereas control cows had greater (P = 0.01) plasma beta-hydroxybutyrate concentrations. Concentrations of NEFA (P = 0.05) and glucose (P < 0.001) were greater, and beta-hydroxybutyrate tended (P = 0.07), to be greater at d 3, whereas serum IGF-I was greater (P = 0.003) at d 6 of lactation. Similar concentrations of NEFA, glucose, GH, and IGF-I indicate that the nutritional status of beef cows during early lactation was not influenced by lipid supplementation. However, perturbations of the somatotropic axis in BCS 4 cows indicate that the influence of energy balance and BCS of the cow at parturition on postpartum performance should be considered when making managerial decisions.

Animal Feed↗

Putative serotonin reuptake inhibitor-induced spawning and parturition in freshwater bivalves is inhibited by mammalian 5-HT2 receptor antagonists.

Selective serotonin reuptake inhibitors (SSRIs) can mimic the physiological actions of serotonin, and in bivalve molluscs they induce zebra mussel spawning and fingernail clam parturition. We have elucidated further the pharmacology of SSRI-induced spawning and part-urition by blocking these reproductive processes with two mammalian 5-HT(2) receptor antagonists, cyproheptadine and mianserin. These two antagonists were potent inhibitors of both spawning and parturition induced by the SSRIs fluvoxamine, fluoxetine, and zimelidine. In zebra mussels, both cyproheptadine and mianserin significantly blocked spawning induced by fluvoxamine and by zimelidine. In the fingernail clams Sphaerium spp., both cyproheptadine and mianserin blocked fluvoxamine-induced parturition. A possible mechanism of action for SSRI-induced spawning and parturition in bivalves is suggested.

Animals↗

Mammary gland immunology around parturition. Influence of stress, nutrition and genetics.

Adequate immune functions are essential for the defence against udder infections. Detailed knowledge about the immune response and important defence factors is essential in order to find new ways for the prevention and treatment of udder infections leading to mastitis. Work should be concentrated on ways of minimising the negative influence on immune functions and/or ways of stimulating these functions, especially during periods of immune suppression. A depression of important immune functions has been reported around parturition and there is a higher prevalence of clinical mastitis and other diseases during this period. Immunosuppression is often associated with high levels of glucocorticoids in blood, a common finding around parturition and during stressful conditions. A number of stressors are present around calving, e.g. parturition, onset of lactation and changes in feeding and management regimes. Adequate management including feeding strategies and routines are important for the immune functions. Metabolic stress as well as deficiencies in vitamins and minerals around parturition and during the first month of lactation can have a negative influence on the immune functions and thereby increase the risks for udder infections and mastitis. There seem to be a genetic variability in certain immune functions among periparturient cows. This might indicate a possibility to find markers for genetic selection of individuals with a well-developed immune system without negative effects on milk productivity.

Animals↗

Effects of hormones on prolactin binding to normal mammary glands and pregnancy-dependent mammary tumors in GR/A female mice near parturition.

Pregnancy-dependent mammary tumors in GR/A female mice are different from normal mammary glands in hormone responsiveness and prolactin binding near parturition. The binding was modified by several hormones. Thus, prolactin binding under the hormonal conditions favorable for the progression of tumors was compared between normal and neoplastic mammary glands. Slices of the glands were incubated in 1 ml Medium 199 containing 125I-labeled ovine prolactin and the radioactivity incorporated was counted as the index of binding. Prolactin binding to either normal or neoplastic glands on day 19 of pregnancy was low and not affected by ovariectomy on day 15, and by daily injections of progesterone alone or in combination with estradiol benzoate between days 14 and 19 of pregnancy. One day after parturition (litter was removed immediately after parturition), the binding to tumors was raised significantly, while, the binding to normal glands remained low. Injections of progesterone alone or in combination with human placental lactogen twice a day from 1 day before parturition induced little changes in the binding to either normal or neoplastic glands. These results suggest no alteration of mammary response for prolactin binding to exogenous hormones in the process of transformation of the glands from normal to neoplastic states, and low predictability of membrane prolactin receptors for tissue responsiveness to the hormone.

Animals↗

Radioimmunoassay of plasma and pituitary oxytocin in pregnant rats during various stages of pregnancy and parturition.

Radioimmunoassayable plasma and pituitary oxytocin (ROT) was measured during pregnancy and parturition. The highest mean plasma ROT level was seen in maternal rats on day 20 of pregnancy followed by a non significant decrease on day 21 reaching the basal level just before delivery. Plasma ROT level declined continuously up to the time of delivery of the first pup and then gradually increased until the completion of parturition. The highest maternal mean pituitary ROT level was found on day 21 pregnancy. Just before delivery of the first pup, the mean pituitary ROT level significantly declined by 22%. Pituitary ROT level declined steadily during the delivery of the first five pups. This pattern of reduction was reversed and ROT levels increased between the delivery of the fifth pup and the completion of labor. The increase of maternal pituitary and plasma ROT levels during delivery of the pups suggest that maternal ROT may play a role in normal parturition. Fetal hypothalamic-pituitary ROT does not seem to play any part in parturition.

Animals↗

Timing of parturition in two species of viviparous lizard: influences of beta-adrenergic stimulation and temperature upon uterine responses to arginine vasotocin (AVT).

The southern snow skink Niveoscincus microlepidotus is a viviparous alpine lizard with biennial reproduction, in which embryos are fully developed before winter but parturition is delayed until spring. We aimed to determine whether, in this species, in vitro uterine preparations are responsive to arginine vasotocin (AVT) and prostaglandin (PGF(2alpha)) in autumn and spring, and whether pre-treatment with the beta-adrenergic agonist isoproterenol decreases the effectiveness of AVT in stimulating uterine contractions. Using the spotted snow skink (Niveoscincus ocellatus), an annually breeding species, we aimed to determine influences of temperature and the beta-adrenergic system upon the response to AVT in vivo. In both N. microlepidotus and N. ocellatus females are more responsive to AVT than to PGF(2alpha), and that the response to AVT is decreased, but not prevented, by beta-adrenergic stimulation. In N. microlepidotus, uteri are equally responsive in both seasons to the hormones administered. In N. ocellatus environmental conditions, specifically, temperature, modulate the response to AVT in vivo with the time to parturition increasing as temperature decreases. We conclude that in these viviparous squamates the endocrine cascade leading to parturition is modulated by the beta-adrenergic system, and that this may reflect the mechanism by which the timing of parturition is tied to suitable environmental conditions.

Adrenergic beta-Agonists↗

Changes in cervical compliance at parturition independent of uterine activity.

A new animal model has been developed to measure intrinsic changes of cerivcal compliance during spontaneous parturition or exposure to hormonal manipulation. Intermittent measurements of cervical compliance and continous measurements of uterine and cervical pressure were made during the last month of gestation of nine pregnant ewes. Cervical compliance increased abruptly and dramatically during spontanous and dexamethasone-induced parturition. Maternal progesterone supplementation at parturition inhibited uterine contractions but not the increase in cervical compliance, demonstrating the independence of the two events. The cervix was found to contract rhythmically and vigorously with a gradual decrease in activity as parturition approached.

Animals↗

Circulating hyaluronic acid in nonpregnant, pregnant, and postpartum guinea pigs: elevated levels observed at parturition.

OBJECTIVE: Dilatation of the uterine cervix at parturition is associated with an increase in cervical hyaluronic acid content. The objective is to test the hypothesis that circulating hyaluronic acid is increased at parturition. STUDY DESIGN: Serum hyaluronic acid levels from nonpregnant (n = 5), pregnant (n = 13), and postpartum (n = 4) adult Hartley guinea pigs were determined with a radiometric assay that utilizes iodine 125-labeled hyaluronic acid-binding protein. Results were analyzed for statistical significance with Student's paired t test and regression analysis. RESULTS: The serum hyaluronic acid level in nonpregnant animals was 238 +/- 88 ng/ml (mean +/- SEM). During pregnancy, serum hyaluronic acid levels were 127 +/- 12 and 126 +/- 34 ng/ml at 25 and 50 to 63 days' gestation, respectively. At parturition, hyaluronic acid levels increased fivefold to 765 +/- 111 ng/ml (p less than 0.001). Hyaluronic acid levels returned to antepartum values 2 days post partum (153 +/- 27 ng/ml). There was no significant difference between arterial and venous levels. CONCLUSION: Circulating hyaluronic acid levels increase significantly at parturition in the guinea pig.

Animals↗

Fever response in the guinea pig before and after parturition.

The febrile response to an intramuscular injection of bacterial endotoxin (E. coli 4 microgram/kg) was tested in guinea pigs at the end of pregnancy in the time period extending from 8 days before until 3 days after parturition. In comparison to non-pregnant female controls both fever height and fever index were reduced in mother guinea pigs one week before parturition. This response was gradually reduced and reached its minimum on the last day before parturition. Immediately after parturition the fever response was still suppressed in mother animals as well as in newborns. Several hours after birth the fever response increased again in both groups of animals. The onset time and duration of fever were, however, shorter than in controls. The full fever responsivity was not reached until several days postpartum. Apparently the guinea pig develops an active antipyresis during the last phase of pregnancy. This resembles the suppression of fever in ewes at term of pregnancy where endogenous arginine-vasopressin has been proposed as an antipyretic agent. The vasopressinergic neuronal systems have therefore been localized by immunohistochemical methods in the brains of the guinea pigs whose responses to bacterial endotoxin were studied. These studies, which are described in detail in a following paper, support the involvement of vasopressin in natural antipyresis in the guinea pig.

Animals↗

Effect of constant light on parturition and postpartum reproduction in the rat.

Light-dark (LD) cyclicity contributes to a number of reproductive events in the rat, including estrus cyclicity, and the timing of parturition and postpartum estrus. Constant light (LL) disrupts the female's cyclicity, and fertility is typically diminished. To ascertain whether constant light similarly diminishes the fertility of postpartum females, LL and LD animals were compared over three successive pregnancies on several reproductive measures including parturition, postpartum estrus and spontaneous ovulation, the timing of estrus relative to parturition, and the number, body weights, and viability of offspring. While the 24 h pattern of parturition differed for LL and LD groups, postpartum estrus and ovulation occurred reliably in both groups. The LD group showed greater variation in the birth-estrus interval, the result of a delayed estrus in those females giving birth late in the light phase. The likelihood of spontaneous ovulation and overall successful reproduction was high for both groups. Thus, unlike the cycling female, the fertility of the postpartum female is relatively unaffected by constant light. As such, these findings suggest that LD cyclicity is not critical for postpartum ovulation and estrus.

Animals↗

Induction of parturition in swine with prostaglandin analogs and oxytocin.

Two trials were conducted to increase the effectiveness of parturition induction with PG analogs. In trial 1, 39 sows were treated with the analog Luprostiol (Pronilen, Vemie/Kempen). Of these, 19 received an additional injection of 30 i.u. oxytocin 24 h later. In trial 2, 126 sows were divided into four groups, three of which were treated with the analog Alfaprostol (Vetem/Milano), while one served as control. Two of the PG-treated groups received an additional oxytocin injection either 24 or 20 h later, respectively. The response to either of the PG analogs resembled that observed during earlier studies although the percentage of sows responding was somewhat lower (70% and 77%, respectively). Oxytocin increased the number of respondents to 84 and 89%, respectively, and parturition commenced promptly. In trial 1, parturition started within 2.1+/-1.6 (0 to 5) h after oxytocin, in trial 2 within 1.57+/-1.06 (0.5 to 3) h when the interval between prostaglandin and oxytocin was 24 h, and 1.81+/-0.84 (0.5 to 3) h when it was 20 h (x+/-SD). Parturition and viability, birth weight and weaning weight of piglets were normal. The proportion of sows in trial 2 that showed increased body temperature during the first 3 days post partum was reduced from 22.9% in the control group to 8.6, 14.3 and 7.1% in the three groups receiving just PG or PG and oxytocin after 24 or 20 h, respectively.

Journal Article↗

Clenbuterol (planipart(trade mark)) for the postponement of parturition in cattle.

Clenbuterol is a highly specific, long-acting (four to eight hours) beta-two sympathomimetic which causes bronchodilation and tocolysis (myometrial paralysis). The tocolytic effect was explored as a means to control parturition and reduce dystocia. Forty-six heifers were injected i.m. with either Clenbuterol or saline placebo in a randomly controlled experiment. Animals were treated when a cervical dilation of five centimeters or more was detected by vaginal examination. Length of first, second and third stages of parturition, ease of parturition, maternal pelvic area and calf viability were compared between treatment groups. Treatment with Clenbuterol increased (P<0.025; 119 vs 468 mins) the time heifers were in Stage I. However, the lengths of Stages II and III, pelvic area at birth and calf viability were not influenced by treatment. Diameter of the cervix at treatment was negatively related to the length of Stage I delay. Pelvic area also significantly affected the length of Stage II. Clenbuterol effectively delays Stage I of parturition with no adverse effects on the fetus or dam.

Journal Article↗

Release of PGF(2alpha) during parturition and the postpartum period in the ewe.

Peripheral plasma levels of 15-keto-13, 14-dihydro-PGF(2alpha) were determined from one week before parturition until five weeks postpartum in six ewes. Starting at approximately 1 nmol/l one week before parturition, the prostaglandin metabolite levels rose successively during the week preceding parturition and peaked at 70 to 125 nmol/l during parturition. During the first three days postpartum the concentrations of the 15-keto-13, 14-dihydro-PGF(2alpha) decreased to 0.5 to 1 nmol/l. These levels were maintained for three weeks and low levels of 50 to 100 pmol/l were reached around four weeks postpartum. The ewe, as well as other ruminants, has a sustained release of elevated concentrations of PGF(2alpha) postpartum. It seems likely that PGF(2alpha) plays a role in uterine involution.

Journal Article↗

Bacterial contamination of the uterus after parturition and its effect on the reproductive performance of cows on large-scale dairy farms.

The bacteriological status of the uteri of 150 cows was examined on 14 large-scale dairy farms 10 to 20 d after parturition and then twice again in 2-wk intervals during uterine involution. The degree of bacterial contamination and the proportion of the infected uterus as well as the composition of the isolated flora were determined. The antibiotic sensitivity of the important bacterium strains was also studied. The relationship between the reproductive data and bacterial contamination of the uteri was analyzed, and the influence of certain environmental and genetic factors on the rate of infection was examined. The proportion of cows having moderate or more serious uterine infection remained above 30 % at the end of involution on more than 35 % of the farms. Infections were caused mainly by streptococci, E. coli and corynebacteria. The bacterium strains showed broad resistance against commericially available antibiotics. There were significant differences in the length of time from the parturition to the first insemination and conception among cows at the various farms. There was a significant correlation between these differences and the bacteriological status of the uteri. Reproductive data were the lowest in the group of cows infected with Corynebacterium pyogenes. The rate of the infected uteri was considerably higher after the third parturition and in cows producing less than 20 l of milk per day. No connection was found between the bacteriological status of the uteri and the breed, type of housing, and season of the parturitions.

Journal Article↗