Normal Parturition Research Articles

Determination of ceftiofur derivatives in serum, endometrial tissue, and lochia in puerperal dairy cows after subcutaneous administration of ceftiofur crystalline free acid

Puerperal uterine infections are often associated with decreased reproductive performance in dairy cows. Routine treatment protocols include the systemic administration of antibiotics. Antibiotic drugs, however, should be administered daily over at least 5 d. The objective of this study was to determine concentrations of ceftiofur derivatives in serum, endometrial tissue, and lochia after subcutaneous administration of ceftiofur crystalline free acid in 6 clinically healthy puerperal dairy cows with normal parturition. Samples were taken immediately before treatment, 2h after, and then every 24h over a 7-d period. Concentrations of ceftiofur derivatives were quantified using an HPLC assay. In serum and endometrial tissue, ceftiofur derivatives could be detected above the reported minimum drug concentrations required to inhibit relevant pathogens such as Escherichia coli and Arcanobacterium pyogenes over a 7-d period. Concentrations of desfuroylceftiofuracetamide at 5 d after administration of ceftiofur crystalline free acid were 1.21±0.61 μg/mL in serum, 0.86±0.61 μg/mg in endometrial tissue, and 0.96±1.15 μg/mL in lochia. In lochia, mean concentrations of ceftiofur derivatives also remained above the minimal inhibitory concentration of relevant pathogens, but showed greater variations between cows.

Coxibs strike back

Coxibs strike back

Peripartal plasma concentrations of 15-ketodihydro-PGF 2α, cortisol, progesterone and 17-β-estradiol in Martina Franca jennies

The transition from intra- to extrauterine environment represents a very delicate phase, in which the successful coordination of maturation is strictly connected with several hormonal changes during the last weeks of gestation and at parturition. While the peripartal endocrinology in the mare has been deeply investigated, the peripartal hormonal changes in the jenny need further evaluation. The aim of this study is to evaluate the mean 15-ketodihydro-PGF 2α (PGFM), cortisol (C), progesterone (P4), and 17β-estradiol (E2) levels during the peripartal period in this species. Ten Martina Franca jennies, with normal gestational length and parturition, were enrolled. From each jenny, blood was collected twice a day from 10 d before to 7 d after parturition and from the plasma obtained PGFM, C, P4 and E2 were analyzed by RIA. Higher, constant PGFM concentrations were observed in the pre-foaling days compared to the decreasing levels detected the days after delivery, as previously observed in the mare. During the whole period of observation no significant differences in plasma C levels were detected. In contrast to the mare, P4 has always been detectable and the highest level found at −2.5 days was significantly different compared to samples obtained between −10 and −4.5 days and between 1.5 and 7 days after foaling. Finally, E2 showed higher concentrations before foaling, with the highest values between −3 and −1.5 days, decreasing only one day before foaling. A positive correlation was found between PGFM and P4, during the last 4 days of gestation, while a positive correlation between PGFM and E2 was observed during the prepartum. Despite some similarities with the mare exist, differences have been found in P4 and E2 profiles, underlining once more the differences in the physiology of this two species.

Developmental dioxin exposure of either parent is associated with an increased risk of preterm birth in adult mice

We have previously described diminished uterine progesterone response and increased uterine sensitivity to inflammation in adult female mice with a history of developmental exposure to TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin). Since parturition in mammals is an inflammatory process mediated in part by a decline in progesterone action, toxicant-mediated disruption of progesterone receptor (PR) expression at the maternal–fetal interface would likely impact the timing of birth. Therefore, in the current study, we examined pregnancy outcomes in adult female mice with a similar in utero exposure to TCDD. We also examined the impact of in utero TCDD exposure of male mice on pregnancy outcomes in unexposed females since the placenta, a largely paternally derived organ, plays a major role in the timing of normal parturition via inflammatory signaling. Our studies indicate that developmental exposure of either parent to TCDD is associated with preterm birth in a subsequent adult pregnancy due to altered PR expression and placental inflammation.

Acquisition of Clostridium difficile by piglets

Clostridium difficile is recognized as an important cause of nosocomial diarrhoea in humans especially in association with administration of antibiotics. In pigs, C. difficile can cause neonatal enteritis and can be isolated from faeces from both diseased and healthy animals. The presented prospective study describes how soon C. difficile can be isolated from newborn piglets after normal parturition and how C. difficile spreads within a pig farm. Six sows, their farrowing crates and their litters at one farm were sampled until C. difficile was found in all piglets. Within 48h after birth, all 71 piglets became positive for C. difficile (two piglets were already positive within 1h post partum), all sows became positive within 113h after parturition and the farrowing crates were found intermittently positive. C. difficile could also be detected in air samples and in samples of teats of the sows. All isolates belonged to PCR ribotype 078. Twenty-one C. difficile ribotype 078 isolates, found at the farm, were further analyzed by MLVA (multiple-locus variable-number tandem repeat analysis) and belonged to one clonal complex, except one isolate.To be sure that piglets were not born already infected with C. difficile ribotype 078, 38 caesarean derived piglets were sampled immediately after surgery. All piglets tested negative at delivery and stayed negative for C. difficile ribotype 078 during the 21 days in which they were kept in sterile incubators.This study shows that C. difficile ribotype 078 spreads easily between sows, piglets and the environment. Vertical transmission of C. difficile ribotype 078 was not found and is very unlikely to occur.

Assessment of feline fetal viability by conceptus echobiometry and triplex Doppler ultrasonography of uterine and umbilical arteries

The aims of the present study were to: (1) evaluate blood flow in the uterine (UA) and umbilical arteries (Uma) in the pregnant queen, by measuring the resistive index (RI) and pulsatility index (PI); (2) to note the presence or absence of the early diastolic notch and diastolic flow in the UA and Uma flow waveforms, respectively; and (3) perform conceptus echobiometry for fetal growth assessment during pregnancy. Eight healthy pregnant domestic Brazilian Shorthair queens were examined from Days 10 to 50 after mating (mating = Day 0). Triplex Doppler and B-mode ultrasonography were used to assess blood flow and conceptus echobiometry. All pregnancies ended with a normal parturition and birth of live kittens. Prior to parturition, all conceptus dimensions increased significantly, whereas RI and PI peaked between Days 33 and 43 followed by a decrease ( P < 0.05). The PI least on Day 50. The RI and PI of Uma decreased ( P < 0.05) during two periods in the fetal development, i.e. from Days 22 to 40 (0.79 ± 0.01 and 1.64 ± 0.04), and from Days 41 to 50 (0.75 ± 0.01 and 1.39 ± 0.05), representing the increased Uma perfusion. Both the complete disappearance of the early diastolic notch in the UA, and the appearance of diastolic flow in the Uma occurred on Day 42 ± 1. It was concluded fetal echobiometry, UA and Uma perfusion, were important end points to assess fetal viability in queens. Furthermore, the current reference values provided a baseline for monitoring normal and abnormal pregnancies in queens.

A Novel Biased Allosteric Compound Inhibitor of Parturition Selectively Impedes the Prostaglandin F2α-mediated Rho/ROCK Signaling Pathway

The prostaglandin F2alpha (PGF2alpha) receptor (FP) is a key regulator of parturition and a target for pharmacological management of preterm labor. However, an incomplete understanding of signaling pathways regulating myometrial contraction hinders the development of improved therapeutics. Here we used a peptidomimetic inhibitor of parturition in mice, PDC113.824, whose structure was based on the NH(2)-terminal region of the second extracellular loop of FP receptor, to gain mechanistic insight underlying FP receptor-mediated cell responses in the context of parturition. We show that PDC113.824 not only delayed normal parturition in mice but also that it inhibited both PGF2alpha- and lipopolysaccharide-induced preterm labor. PDC113.824 inhibited PGF2alpha-mediated, G(alpha)(12)-dependent activation of the Rho/ROCK signaling pathways, actin remodeling, and contraction of human myometrial cells likely by acting as a non-competitive, allosteric modulator of PGF2alpha binding. In contrast to its negative allosteric modulating effects on Rho/ROCK signaling, PDC113.824 acted as a positive allosteric modulator on PGF2alpha-mediated protein kinase C and ERK1/2 signaling. This bias in receptor-dependent signaling was explained by an increase in FP receptor coupling to G(alpha)(q), at the expense of coupling to G(alpha)(12). Our findings regarding the allosteric and biased nature of PDC113.824 offer new mechanistic insights into FP receptor signaling relevant to parturition and suggest novel therapeutic opportunities for the development of new tocolytic drugs.

Use of the progesterone (P4) receptor antagonist aglepristone to characterize the role of P4 withdrawal for parturition and placental release in cows

In late pregnant cows, progesterone (P(4)) is mainly of luteal origin. However, the trophoblast may provide high local P(4) concentrations in the uterus. To test for the importance of a complete P(4) withdrawal for parturition-related processes and placental release, the P(4) receptor (PGR) blocker aglepristone (Ap) was administered to three cows on days 270 and 271 of pregnancy. A complete opening of the cervix was observed 46.5±7.3 h after the start of treatment. However, expulsion of the calves was impaired obviously because of insufficient myometrial activity, and placental membranes were retained for at least 10 days. Measurement of P(4) concentrations indicated that PGR blockage induced luteolysis. To investigate the role of P(4) withdrawal for the prepartal tissue remodeling of the placentomes, the caruncular epithelium was evaluated by morphometry, and the percentage of trophoblast giant cells (TGCs) relative to the total number of trophoblast cells were assessed. Caruncular epithelium in Ap-treated cows (D272+Ap) was immature (30.5±3.3%) and not different from untreated controls (elected cesarean section (CS) on day 272; D272-CS; 31.5±1.4%), whereas it was significantly reduced at normal term (D280.5±1.3; 21.0±6.1%; P=0.011). Correspondingly, the percentage of TGCs were 20.1±1.4 in D272+Ap, 22.1±4.8 in D272-CS, and 9.8±3.9 at term (P=0.001). No effect was detected on placental estrogen synthesis. The results showed that in late pregnant cows, P(4) withdrawal only induces a limited spectrum of the processes related to normal parturition and is not a crucial factor for the prepartal tissue remodeling in placentomes and the timely release of the placenta.

Interleukin-6 Is an Essential Determinant of On-Time Parturition in the Mouse

IL-6 abundance in amniotic fluid and uterine tissues increases in late gestation or with infection-associated preterm labor. A role in regulation of labor onset is suggested by observations that IL-6 increases expression of genes controlling prostaglandin synthesis and signaling in isolated uterine cells, but whether IL-6 is essential for normal parturition is unknown. To evaluate the physiological role of IL-6 in parturition in mice, we investigated the effect of Il6 null mutation on the timing of parturition and expression of genes associated with uterine activation. Il6 null mutant mice delivered 24 h later than wild-type mice, although circulating progesterone fell similarly in both genotypes during the prepartal period. Il6 null mutant mice were also refractory to low doses of lipopolysaccharide sufficient to induce preterm delivery in wild-type mice. The characteristic late-gestation elevation in uterine expression of Oxtr mRNA encoding oxytocin receptor, and peripartal increases in Ptgfr and Ptgs2 mRNAs regulating prostaglandin synthesis and signaling were delayed by 24 h in Il6 null mutant mice. Conversely, Ptger4 mRNA encoding the prostaglandin E receptor-4 was abnormally elevated in late-gestation in Il6 null mutant mice. Administration of recombinant IL-6 from d 11.5 postcoitum until term restored the normal timing of delivery and normalized Ptger4 mRNA expression in late gestation. We conclude that IL-6 has a key role in controlling the progression of events culminating in parturition and that it acts downstream of luteolysis in the uterus to regulate genes involved in the prostaglandin-mediated uterine activation cascade.

The Matrix Metalloproteases and Endothelin‐1 in Infection‐Associated Preterm Birth

Preterm birth (PTB) is clinically defined as any delivery which occurs before the completion of 37 weeks of gestation, and is currently the most important problem in obstetrics. In the United States, PTB accounts for 12-13% of all live births, and, with the exception of fetuses suffering from anomalies, is the primary cause of perinatal mortality. While the risk factors for PTB are numerous, the single most common cause is intrauterine infection. As there is currently no FDA-approved therapy for infection-associated PTB, understanding the pathogenesis of preterm labor (PTL) and delivery should be given high priority. The matrix metalloproteinases (MMPs) are a family of enzymes that have been implicated in normal parturition as well as infection-triggered rupture of membranes and preterm birth. Several lines of evidence also suggest a role for endothelin-1 (ET-1) in infection-associated preterm delivery. This paper focuses on the evidence that the MMPs and ET-1 act in the same molecular pathway in preterm birth.

217 EFFECT OF FREQUENCY OF MILKING ON POSTPARTUM EVENTS IN SISTANI CATTLE

Prolonged postpartum anestrus is a major concern in Bos indicus breeds. Sistani cattle is an Iranian Bos indicus beef breed with more than 5000 years of history in the northwest part of Iran, Sistan. This breed is well adapted to the harsh, arid environment, which includes 120 days of hot winds. Small holders in the Sistan region use Sistani cattle for both meat and milk. The purpose of this study was to investigate the effect of frequency of milking on the postpartum events in this breed. Healthy Sistani cattle (n = 41) with 3.3 ± 0.3 parity and normal parturition were randomly assigned to 3 groups after parturition and were monitored for 3 months. Cows in Groups 1 and 2 were milked once (n = 15) or twice (n = 14) per day, respectively. The control (n = 12) cows were not milked. All cows were allowed to be suckled by their calves once per day. Throughout the 3 months of monitoring after calving, blood samples were collected weekly and assayed for progesterone using RIA. Standing estrus was detected every day at 0700, 1400, and 2100 h. The first rise in progesterone after calving was considered when it was above 1.5 ng/mL. Data were analyzed using GLM procedure in SAS (SAS Institute, Cary, NC). There was no significant difference in the variables measured among groups (P &gt; 0.05); therefore, the data were pooled (Table 1). The number of estrus, the day of the first estrus, and the first rise in progesterone after calving were 2.9 ± 1.42, 31.2 ± 1.77, and 38.8 ± 2.20, respectively (Table 1). Average number of service per conception (1.6 ± 0.12) and days open (94.4 ± 7.63) in Sistani cattle may be considered to be superior reproductive indices to those reported in the majority of Bos indicus breeds. In conclusion, once-daily or twice-daily milking did not influence postpartum reproductive indices of Sistani cattle, indicating that twice-daily milking may be advised to enhance the profitability of small holders. In Sistani cattle, ovarian activity and estrous behavior resumed fairly soon after partition, allowing for earlier conception in this breeds. Table 1.Postpartum reproductive indices of Sistani cattle with varying frequencies of milking This research was funded by the Research Deputy of the University of Tehran and Animal Science Research Institute.

Canine placenta: a source of prepartal prostaglandins during normal and antiprogestin-induced parturition

Expression of cyclooxygenase 2 (COX2, now known as PTGS2), prostaglandin E2 synthase (PTGES, PGES), and prostaglandin F2alpha synthase (PGFS), of the respective receptors PTGFR (FP), PTGER2 (EP2), and PTGER4 (EP4) and of the progesterone receptor (PGR, PR) was assessed by real-time PCR, immunohistochemistry (IHC), or in situ hybridization (ISH) in utero/placental tissue samples collected from three to five bitches on days 8-12 (pre-implantation), 18-25 (post-implantation), and 35-40 (mid-gestation) of pregnancy and during the prepartal luteolysis. Additionally, ten mid-pregnant bitches were treated with the antiprogestin aglepristone (10 mg/kg bw (2x/24 h)); ovariohysterectomy was 24 and 72 h after the second treatment. Plasma progesterone and 15-ketodihydro-PGF2alpha (PGFM) concentrations were determined by RIA. Expression of the PGR was highest before implantation and primarily located to the endometrium; expression in the placenta was restricted to the decidual cells. PTGS2 was constantly low expressed until mid-gestation; a strong upregulation occurred at prepartal luteolysis concomitant with an increase in PGFM. PGFS was upregulated after implantation and significantly elevated through early and mid-gestation. PTGES showed a gradual increase and a strong prepartal upregulation. PTGFR, PTGER2, and PTGER4 were downregulated after implantation; a gradual upregulation of PTGFR and PTGER2 occurred towards parturition. ISH and IHC co-localized PGFS, PTGFR, PTGES, and PTGS2 in the trophoblast and endometrium. The changes following application of aglepristone were in the same direction as those observed from mid-gestation to prepartal luteolysis. These data suggest that the prepartal increase of PGF2alpha results from a strong upregulation of PTGS2 in the fetal trophoblast with the withdrawal of progesterone having a signalling function and the decidual cells playing a key role in the underlying cell-to-cell crosstalk.

Decreased adherence and spontaneous separation of fetal membrane layers – amnion and choriodecidua – a possible part of the normal weakening process

IntroductionThe fetal membrane (FM) layers, amnion and choriodecidua, are frequently noted to have varying degrees of separation following delivery. FM layers normally separate prior to rupture during in vitro biomechanical testing. We hypothesized that the adherence between amnion and choriodecidua decreases prior to delivery resulting in separation of the FM layers and facilitating FM rupture. MethodsFM from 232 consecutively delivered patients were examined to determine the extent of spontaneous separation of the FM layers at delivery. Percent separation was determined by the weight of separated FM tissue divided by the total FM weight. Separately, the adherence between intact FM layers was determined. FM adherence was tested following term vaginal delivery (13), term unlabored cesarean section (10), and preterm delivery (6). ResultsSubjects enrolled in the two studies had similar demographic and clinical characteristics. FM separation was present in 92.1% of membranes. Only 4.3% of FM delivered following spontaneous rupture of the fetal membranes (SROM) had no detectable separation. 64.7% of FM had greater than 10% separation. FM from term vaginal deliveries had significantly more separation and were less adherent than FM of term unlabored, elective cesarean section (39.0±34.4% vs 22.5±30.9%, p=.046 and 0.041±0.018N/cm vs 0.048±0.019N/cm, p<.005). Preterm FM had less separation and were more adherent than term FM (9.95±17.7% vs 37.5±34.4% and 0.070±0.040N/cm vs 0.044±0.020N/cm; both p<.001). ConclusionsSeparation of the amnion from choriodecidua at delivery is almost universal. Increased separation is associated with decreased adherence as measured in vitro. Increased separation and decreased adherence are seen both with increasing gestation and with labor suggesting both biochemical and mechanical etiologies. The data are consistent with the hypothesis that FM layer separation is part of the FM weakening process during normal parturition.

Avaliação física e dos níveis séricos de cortisol de bezerros neonatos da raça Nelore, nascidos de partos normais e auxiliados

O presente trabalho visou avaliar a vitalidade e os níveis séricos de cortisol de bezerros neonatos, nascidos de partos eutócicos e auxiliados, bem como determinar os índices de partos distócicos em vacas receptoras de embriões nelores. Cerca de 310 bezerros foram avaliados. Observou-se a ocorrência de 38 partos distócicos (38/310) e de oito abortos (8/310). Os bezerros nascidos de partos distócicos mais duradouros, entre quatro e seis horas, apresentaram os maiores valores médios de frequência respiratória e dos níveis séricos de cortisol. A temperatura retal foi mais elevada em animais nascidos de partos laboriosos do que naqueles nascidos de partos normais. As taxas de morbidade e mortalidade em animais neonatos nelores foram de 77,93% e 3,44%, respectivamente. O emprego do escore APGAR, ao nascimento, demonstrou que 90% dos bezerros apresentavam-se deprimidos, independente do tipo de nascimento.

Progesterone withdrawal as the lactogenic trigger in the rat

Lactogenesis, as indicated by the appearance of mammary lactose, follows ovariectomy and/or hysterectomy and the fall in the concentration of progesterone in the plasma preceding normal parturition in the pregnant rat. It is suppressed by administered progesterone in all cases, and by prolactin after hysterectomy. The natural lactogenic signal may occur 30 hr. before parturition, due to conversion of progesterone to 20α-hydroxypregn-4-en-3-one in the ovaries. Changes in the concentration of corticosterone, oestrogen and prolactin in the plasma either do not occur or do not constitute a lactogenic signal.

Reproduction and maternal behavior in insulin-regulated aminopeptidase (IRAP) knockout mice

During human pregnancy, a circulating form of insulin-regulated aminopeptidase (IRAP EC 3.4.11.3), often termed oxytocinase or placental leucine aminopeptidase (PLAP), is present in plasma. It is proposed that circulating IRAP plays an important role in regulating the circulating levels of oxytocin and/or vasopressin during pregnancy. We assessed the reproductive and maternal profile of global IRAP knock out mice. No differences in the reproductive profile were observed, with normal gestational period, litter size and parturition recorded. However, western blot analysis of pregnant mouse serum, failed to detect IRAP, a result which was confirmed by fluorimetric IRAP enzyme assay. A review of the literature revealed that the presence of IRAP in the maternal circulation during pregnancy has been only reported in humans. Moreover, the sequence, Phe 154 Ala 155, identified as the cleavage site for the release of soluble IRAP, is restricted to members of the homindae family. Therefore the absence of IRAP from the circulation in mice, and other species during pregnancy, is due to the inability of a secretase to cleave placental IRAP to produce a soluble form of the enzyme. Given the expression of IRAP in areas of the brain associated with oxytocin modulated maternal behavior, we also investigated whether the IRAP global knockout mice had improved maternal responses. Using standard tests to assess maternal behavior, including pup retrieval, feeding and nurturing, no differences between knock out and wild type dams were observed. In conclusion, the physiological significance of circulating IRAP during human pregnancy cannot be addressed by investigations on mice.

Oxytocin Plasma Levels in Cows with Normal Parturition or Dystocia and with Placental Retention*)

Plasma oxytocin level was determined by RIA in eight cows with normal parturition or with dystocia and retained foetal membranes. Both concentration and profile of oxytocin measured around parturition in cows with these disturbances were markedly changed, compared to the cows with the normal course of parturition. Oxytocin was released in the greatest amounts during physiological parturition or during parturition with dystocia in response to the Ferguson reflex only. In cows with dystocia lengthen of the period of the low oxytocin level was observed, though cervix was dilated. Then, it seems that some cases of placental retention are related to the low oxytocin level.

Acid–Base Changes in Canine Neonates Following Normal Birth or Dystocia

There are limited data concerning blood gas parameters in neonatal dogs. Knowledge of the normal physiology may facilitate effective therapeutic intervention and potentially reduce neonatal mortality. This study examined acid-base parameters in pups born at normal parturition (n = 27) compared with those born after obstetrical assistance or caesarean operation (n = 13) and those born following oxytocin (OXY) administration for treatment of uterine inertia (n = 11). Pups were subjected to an objective scoring method of neonatal health adapted from use in humans (the Apgar score) at birth and again at 5 and 60 min after birth. Venous blood samples were collected at 5 and 60 min after birth for evaluation of blood gas parameters. At birth, all pups had low Apgar scores and a mixed acidosis. The base excess was lowest for pups delivered after OXY administration. The Apgar score improved for all pups after 5 min of birth and there was an improvement in carbon dioxide tension, base excess and venous blood pH at 1 h, although in all pups a metabolic acidosis persisted. These data provide an important insight into neonatal physiology and the variability of blood gas parameters in pups born at normal and abnormal parturition and provide the basis for clinical decision making following dystocia.

Peripartum Haemodynamic Status of Bitches with Normal Birth or Dystocia

There has been limited investigation of parturition in the bitch and there is little information published on clinical and obstetrical examination other than opinion and anecdote. While there are substantial data on haemodynamic and vascular changes during normal parturition in humans, little is known about the physiological events in the dog. This study was aimed at maternal haemodynamic changes occurring during normal parturition and to investigate how these were modified in bitches with dystocia (DYST) treated either medically or via assisted delivery and caesarean operation. Three groups of 10 bitches were investigated; those with normal parturition, those with DYST corrected by manipulative assistance or caesarean operation and those with uterine inertia treated by oxytocin administration. Heart rate, systolic and diastolic blood pressure, electrocardiogram and blood glucose concentration were measured pre-partum, intra-partum, immediately after parturition and 1 h later. Heart rate was high at all times throughout the study and the majority of bitches had normal sinus rhythm. Blood pressure was generally within the normal range, and although systolic and diastolic blood pressure was highest during the intra-partum period and sometimes during the immediate post-partum period, there were no significant differences between groups. All bitches had blood glucose concentrations within the normal range throughout the study although pre-partum concentrations were statistically lower than many of the other time periods. The study provides useful physiological data that will facilitate monitoring and clinical management of bitches throughout normal parturition and DYST.

Normal parturition and management of dystocia in dogs and cats

THIS article describes the normal stages of parturition and discusses how to recognise and manage dystocia in dogs and cats. In each case, it is important to have a good...