Equine amnionitis and fetal loss: mare abortion following experimental exposure to Processionary caterpillars (Ochrogaster lunifer)

Reasons for performing study: In Australia, there have been recent reports of unusual abortions in mid‐ to late‐gestation mares. These were clinically distinct from other recognised causes of pregnancy loss and the term ‘equine amnionitis and fetal loss’ (EAFL) was adopted to describe this syndrome. Initial investigations concluded that possible causal factors included the presence on affected stud farms of Processionary caterpillars (Ochrogaster lunifer). Objectives: To determine if exposure of pregnant mares to Processionary caterpillars or their shed exoskeletons can induce EAFL. Methods: Processionary caterpillars and their shed exoskeletons were collected and stored frozen. Mid‐gestation mares were dosed with a slurry of caterpillars or shed exoskeleton by nasogastric intubation. Their clinical responses and times to abortion were recorded. All aborted fetuses were autopsied and samples taken for bacteriological and virological culture and histopathology. Results: Intubating mares in mid‐pregnancy with preparations of either whole Processionary caterpillars or shed caterpillar exoskeletons induced abortion with few impending clinical signs. The gross pathological and bacteriological findings of the aborted fetuses were similar to those observed in field cases of EAFL. Potential relevance: Possible exposure to Processionary caterpillars should be considered when examining cases of fetal loss in the mare. The present results provide a starting point to further explore the aetiology and pathogenesis of EAFL.     

Validation of the BioPRYN enzyme‐linked immunosorbent assay for detection of pregnancy‐specific protein‐B (PSPB) and diagnosis of pregnancy in American bison (Bison bison)

This study assessed the accuracy of the commercial BioPRYN^® ELISA for the detection of pregnancy‐specific protein‐B (PSPB) using a single blood sample to determine pregnancy status in American bison (Bison bison). A total of 49 bison cows were used in the study, and sampled at two time‐points during the gestation period, fall and spring, correlating with early‐ to mid‐term gestation (average 62.9 days post‐mating) and mid‐ to late‐term gestation (average 229.2 days post‐mating), respectively. Sensitivity of the test during early‐ to mid‐term gestation sampling period (fall) was 87.1%, while specificity was 100%; sensitivity of the test during late‐term gestation sampling period (spring) was 96.3%, while specificity remained at 100%. In total, the test showed a total sensitivity of 91.4%, specificity of 100% and total accuracy of 93.8%, similar to domestic cattle. Use of the single‐sample BioPRYN^® ELISA in American Bison for pregnancy diagnosis is economical and practical, minimizing animal handling time, frequency and subsequent stress while providing accurate results for pregnancy diagnosis at 62 days post‐mating. This method should be considered over more traditional pregnancy diagnosis methods for use in managed bison herds.     

The nutritional status of pregnant and non-pregnant mares grazing South East Queensland pastures

It has been reported that the increasing nutritional demands from mid to late pregnancy of grazing mares may not be met when these stages of gestation coincide with pastures being affected by frost. It was established in this study that grass/legume pastures could support the nutritional requirements of brood mares by providing digestible energy intakes of 68.0 and 91.7 MJ/day and digestible nitrogen intake of 91.2 and 138 g/day during mid and late pregnancy, respectively.     

Heart rate and heart rate variability in the pregnant mare and its foetus

Abortion and preterm birth of foals are major reasons for reproductive losses in the horse. Risk pregnancies require close supervision so that adequate treatment can be initiated in time. The aim of this study was to determine normal values in heart rate (HR) and heart rate variability (HRV) of the pregnant mare compared to her foetus and to detect physiological changes during ongoing gestation. In mares, the RR interval decreased from 1480±29 ms on day 270 of pregnancy to 1190±58 ms on day 330 of pregnancy (p<0.05). In contrast, foetal RR interval increased during the same time period from 611±23 ms on day 270 of gestation to 756±25 ms on day 330 of gestation (p<0.05). Concomitantly, maternal HR increased and foetal HR decreased. No further changes in RR interval occurred during the last 10 days before foaling, neither in the mare nor the foetus. In the last hours preceding parturition, maternal RR interval was lower than at all times earlier in pregnancy (average of 1037±13 ms) but did not change during this time. Maternal HRV did not change during gestation. Marked changes in HRV occurred only during the last minutes of foaling. Then, all HRV variables increased significantly (standard deviation of beat-to-beat interval: p=0.01, root mean square of successive beat-to-beat differences: p<0.01). The cardiovascular system of pregnant mares adapted to the demands of ongoing pregnancy with an increase in HR. We have no evidence that in healthy mares, pregnancy is a major stressor.     

The Relationship Between the Positive Identification of the Embryo Proper in Equine Pregnancies Aged 18-28 Days and its Future Viability: A Field Study

Early embryonic loss (EEL) negatively affects the reproductive efficiency of equine reproduction. A sign of future EEL is when the embryo proper (EP) fails to develop within the embryonic vesicle after 30 days of gestation. The earlier the identification of impending EEL, the earlier the mare can be rebred to allow a second chance of pregnancy. The objectives of this study were to determine the percentage of embryonic vesicles with a visible EP at 18-28 days of gestation and to study the association between the presence/absence of the EP at different days of gestation and the future viability of the pregnancy. A total of 1,256 pregnancies were identified and followed by transrectal B-mode ultrasonography 12-45 days post ovulation in mares of the same Thoroughbred farm. The identification of the EP was attempted once during days 18-28. Pregnancy reconfirmation was performed on days 35-45. The percentage of vesicles with an EP increased gradually from days 18 (2.8%) to 21 (86.9%) (P < .05). From day 20 onward, the EEL rate of mares with vesicles without an EP was significantly higher (P < .05) than that of vesicles with a positive identification of an EP. In conclusion, the EP of the equine vesicle can be identified reliably with B-mode ultrasonography in the majority of mares (>71%) on day 20 of gestation. The lack of a positive identification of an EP from day 20 onward is associated with a higher EEL rate.     

Mid‐gestation pregnancy is not disrupted by a 5‐day gastrointestinal mucosal cytoprotectant oral regimen of misoprostol

Reasons for performing study: To investigate effects of a 5‐day oral misoprostol regimen recommended for use in horses as a gastrointestinal mucosal cytoprotectant during colic on mid‐gestation pregnancies. Objectives: To monitor cervical tone, ultrasonographic characteristics of the uterus, cervix and conceptus, as well as serum progesterone and oestrone sulphate concentrations, and observations of general health, behaviour and comfort of mid‐gestation mares given a 5‐day course of misoprostol or control treatment. Methods: Eleven light horse and pony mares with known breeding dates were administered 5 µg/kg bwt misoprostol orally, twice daily for 5 days. General health and pregnancy status were monitored daily during treatment via general physical examination, as well as palpation and ultrasonography per rectum of the uterus, cervix and conceptus. Jugular serum was obtained during and for 5 days following treatment for assay of progesterone and oestrone sulphate concentrations. Additionally, daily 12 h video samples of the mares were obtained to evaluate behaviour and comfort. Results: All findings, including cervical tone, ultrasonographic characteristics of the uterus, cervix and conceptus, as well as progesterone and oestrone sulphate concentrations, and observations of general health, behaviour and comfort, were similar during misoprostol and control treatment. Conclusions: Treatment of pregnant mares with a gastrointestinal mucosal cytoprotectant regimen of oral misoprostol for 5 days did not disrupt pregnancy, nor adversely affect the general health and comfort of these mares. Additional investigation of treatment at earlier and later stages of gestation, for longer‐term treatment, as well as evaluating neonates for developmental disturbances, would add further information on safety of misoprostol during gestation. Potential relevance: These results provide some assurance of safety of a 5‐day gastrointestinal mucosal cytopretectant regimen of oral misprostol in mid‐gestation pregnant mares.     

Transabdominal Ultrasonography for Pregnancy Diagnosis and Estimation of Gestational Age in Dromedary Camels

The objective of this study was to evaluate the use of transabdominal ultrasonography as a method for pregnancy diagnosis and establish some foetal biometrics guidelines of pregnancy staging. Transabdominal ultrasonography was performed weekly on six pregnant camels from the 4th to the 19th week and at 2‐week intervals from the 20th to the 52nd week of gestation. Six areas were selected for examination: caudal abdominal (CAA), inside the thigh, above the base of the udder; middle abdominal (MIA), from the base of the udder to the umbilicus; and cranial abdominal (CRA), from the umbilicus to the xiphoid cartilage. Several foetal parameters including orbital diameter (ORD), biparietal diameter (BPD), abdominal diameter (ABD), chest depth (CHD) and ruminal diameter (RUD) were measured using standardized scan plans. The relationships between the gestational age and the foetal parameters were evaluated. From the 6th to the 12th week, the conceptus was always observed through the left CAA approach. Between the 13th and 27th week, the foetus was entirely visualized by the MIA examination. From the 28th to the 52nd week, the conceptus was mainly detected through the left CRA approach. The foetal parameters were measured frequently during mid‐ and late gestation. All regression and correlation coefficients were highly significant (p < 0.0001). In conclusion, transabdominal ultrasonography was shown to be a reliable technique for pregnancy diagnosis and estimation of gestational age in dromedary camels.     

Factors influencing equine chorionic gonadotrophin production in the mare

Reasons for performing study: An opportunity to monitor equine chorionic gonadotrophin (eCG) production during 61 pregnancies in 25 Thoroughbred mares mated to the same Thoroughbred stallion was utilised in order to further knowledge regarding factors involved in the production of this hormone. Objectives: To examine the effects of maternal body condition, exercise and parity on eCG production. Methods: In the first experiment, maiden mares were fed either a moderate (n = 9) or an excessive (n = 10) food intake throughout gestation. In the second experiment, 5 mares were exercised daily during pregnancy and eCG production rates were compared to 5 nonexercised mares. In the third experiment, eCG profiles were compared in 9 mares during 3 successive pregnancies. Equine chorionic gonadotrophin secretion was assessed as area under the curve (AUC), peak serum concentration, timing of the peak and the rate of decline. In addition, a mean eCG profile of 61 pregnancies was created to provide means and ranges for the above parameters. Results: In Experiment 1, eCG production was significantly higher in moderately rather than excessively fed mares in terms of AUC and peak eCG concentrations. In Experiment 2, the mean AUC did not differ between exercised and nonexercised animals but mean eCG concentrations were significantly higher in nonexercised mares between Days 60 and 90 of gestation. In Experiment 3, eCG became undetectable significantly earlier in gestation in the third parity. The mean eCG profile of 61 pregnancies showed a peak of 64.5 ± 3.7 iu/ml at 62.4 ± 1.0 days after ovulation and was undetectable by 134.1 ± 1.7 days. Peak eCG levels reduced by 50% 22.6 ± 1.13 days. Conclusions: Some of the factors examined clearly influenced eCG production rate, the secretion of this hormone and its rate of disappearance from the blood. Potential relevance: The results provide insights into some factors that govern the production of the placental gonadotrophin, eCG.     

Changes in urinary and plasma oestrone sulphate concentrations after induction of foetal death in mares at 45 days of gestation

Foetal death was induced in 10 Standardbred mares at day 45 of gestation by injecting 20 to 45 ml of hypertonic (24% W/V) saline into the conceptus at surgery. Ten mares underwent sham treatment and acted as controls. Blood and urine samples were collected every other day between days 30 and 45 post ovulation and at 0, 3 and 6 h relative to the infusion of saline in the treated mares, or sham treatment in control mares. Blood and urine samples were then collected daily between days 46 and 55 post ovulation. Urine oestrone sulphate (E1S) concentrations, measured by radioimmunoassay, increased between day 34 and day 36 of gestation in treated and control groups. In mares in which foetal death was induced, urine E1S concentrations declined post-operatively and were significantly (p less than .05) lower than controls by day 50. In plasma, E1S concentrations showed a major increase between days 36 and 40 in both groups. This was followed by a rapid decline after treatment in saline-injected mares, so that by day 48 plasma E1S concentrations in treated mares were significantly (P less than .05) lower than the controls. The results show that urinary and plasma E1S concentrations rise rapidly during early pregnancy, and are associated with a viable foetus after day 45 of pregnancy.     

Factors Affecting Plasma Pregnancy-associated Glycoprotein 1 Concentrations Throughout Gestation in High-producing Dairy Cows

This study was designed to establish the factors, if any, which could affect plasma pregnancy-associated glycoprotein-1 (PAG-1) expression in a study population of 87 pregnant, high-producing dairy cows. The factors examined were: semen providing breed (Holstein-Friesian vs Limousin), outcome of gestation (male vs female newborn, and singleton vs twin pregnancies), lactation number, milk production at pregnancy diagnosis, plasma progesterone concentration, season of gestation (warm period, March–November vs cool period, December–February), and day of gestation (40, 90, 120, 150, 180 and 210). Pregnancy was diagnosed by transrectal ultrasound on day 40 post-insemination and by palpation per rectum on days 90, 120, 150, 180 and 210. Blood samples were collected from each animal immediately before each pregnancy diagnosis. The relative contributions of the different factors on PAG-1 concentrations were evaluated by GLM repeated measures analysis of variance. No significant effects of the herd, foetal sex, milk production, lactation number and plasma progesterone concentrations were observed. In contrast, twin pregnancy, the use of Limousin semen and conception during the cool period were correlated with significantly increased plasma PAG-1 concentrations throughout gestation. Our data indicate that both cow well-being during early placental development, determined in our conditions by reduced heat stress when conception occurred in the cool season, and crossbreed pregnancies lead to improved PAG-1 production throughout the gestation period.     

Anomalous Pregnancies during Late Embryonic/Early Foetal Period in High Producing Dairy Cows

This study analyses anomalous cases of gestation ending in pregnancy loss during the early foetal period and their effect on progesterone and plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations. Data derived from a large-scale ultrasound pregnancy diagnosis programme in high producing dairy cows. Over a 3-year period (2004–2007), a very low incidence (0.5%: 15 of 3094) of anomalous pregnancies was recorded. The results revealed that the following anomalies were detected on days 35–41 of gestation in cows carrying singletons with one single corpus luteum: embryo death in eight cows (0.3%); and embryo in the uterine horn contralateral to the corpus luteum in seven cows (0.2%). All these animals suffered pregnancy loss during the early foetal period. In cows carrying dead embryos, no signs of conceptus degeneration were observed on pregnancy diagnosis. Amnion size (approximately 25 mm diameter) and uterine horn fluid contents were estimated to be similar to those of the normal pregnant cows in this period. In the contralateral gestations, live embryos were observed in all ultrasound checks before pregnancy loss. Uterine fluid contents increased in the two cows in which gestation continued for more than a week. In the cases of embryo death but not in those of contralateral gestation, a drop in PAG-1 levels was noted prior to pregnancy loss. Two cows carrying dead embryos increased with time allantoic fluid contents. The PAG-1 values increased with time in one cow bearing a dead embryo (from 2.31 to 6.79 ng/ml) and in two of the contralateral gestations (from 1.66 to 2.33 ng/ml and from 0.39 to 6.79 ng/ml, respectively). Results of this study indicate that the foetal membranes continue to undergo some activity following embryo death, and that contralateral pregnancy may determine failure of the gestation process.     

Videoendoscopic evaluation of the mare's uterus: III. Findings in the pregnant mare.

During a 5-year period 65 Pony and 20 Thoroughbred pregnant mares were subjected to videoendoscopic hysteroscopy from 10 to 266 days of gestation. The aims of these examinations were to 1) observe foetal and placental development in vivo (60 Pony and 10 Thoroughbred mares); 2) eliminate one of unicornuate twin conceptuses (9 Thoroughbred mares); 3) recover embryonic foetal and placental tissues non-surgically for experimental purposes (47 Pony mares); 4) induce focal separation of the placenta in late gestation as an experimental model of placentitis (5 pony mares and 1 Thoroughbred mare). It was possible to view the embryo and all constituent extra-embryonic membranes of the conceptus between Days 10 and 87 of gestation without having to perforate the allantochorion. This allowed study of physiological processes such as the coordinated uterine contractions responsible for conceptus motility between Days 7 and 17, active foetal movements, which began as early as Day 34, and invasion of chorionic girdle cells into the endometrium, which occurred between Days 34 and 38. From Day 90, vision of the foetus was reduced or prevented by the increased thickness of the allantochorion. Transendoscopic recovery of the conceptus was successful in all of 10 mares under 30 days of gestation, whereas only 10 of 18 attempts between Days 30 and 45 produced the conceptus without resorting to uterine lavage after initial rupture of the allantochorion. All 9 attempts to eliminate one of unilateral twin conceptuses were unsuccessful and the technique was abandoned as clinically unsuitable. Nevertheless, 9 of 22 (41%) single conceptuses remained viable after one or more hysteroscopic examinations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Proteins and Enzymes in Uterine Lavage Fluid of Postpartum and Nonparturient Mares

Uterine lavage fluids from postpartum and nonparturient mares were compared to determine when the normal secretory capacity of the postpartum uterus is restored. Lavage fluids were obtained from cyclic nonparturient mares on the second, fourth or fifth day of oestrus, and 3, 8, or 14 days after ovulation (seven mares/sampling day). Twelve intact postpartum mares were sampled 1 to 28 days postpartum (group A: 1, 6, 12 and 20; group B: 2, 8, 14 and 24; group C: 4, 10, 16 and 28 days postpartum; four mares/group). Three ovariectomized (OVX) postpartum mares were sampled as mares in group C. Samples were analysed for neutrophils, bacteria, total protein concentration, proteolytic and antiproteolytic activities and for various lysosomal enzyme activities. In nonparturient mares, activities of acid phosphatase, β‐glucuronidase (B‐Gase), and N‐acetyl‐β‐D‐glucosaminidase (NAGase) in uterine lavage fluids were significantly higher in mid‐ and late‐dioestrus than in mid‐ to late‐oestrus (p < 0.05). Lysozyme concentration, trypsin‐inhibitor capacity (TIC), and plasmin activity were below the detection limit in nonparturient mares. One to four days postpartum, total protein, acid phosphatase, B‐Gase, and NAGase were high but declined rapidly thereafter. Lysozyme and plasmin activities were high 1 to 6 days postpartum. TIC peaked around day 6 postpartum. On day 16 postpartum, acid phosphatase, B‐Gase, and NAGase, being progesterone‐dependent, tended to be higher in intact mares than in OVX ones (p < 0.1). Total protein and lysozyme concentrations, TIC, and B‐Gase (p < 0.01) and acid phosphatase (p < 0.05) activities were significantly higher in parturient mares during postpartum oestrus than in oestrous nonparturient mares. High total protein concentration and TIC, and detectable lysozyme and plasmin activities during postpartum oestrus were associated with uterine inflammation. During dioestrus, differences between postpartum and nonparturient mares were not statistically significant and suggested that the endometrium of postpartum mares had resumed its normal secretory capacity by this time.     

Immunohistochemical Localization of Oestrogen Receptors α and β, Progesterone Receptor and Aromatase in the Equine Placenta

The functions of placental oestrogens during equine pregnancy are still unclear. Yet, they may act predominantly as local regulators of growth and differentiation in the microplacentomes. Thus, expression patterns of oestrogen receptors (ERs) α and β were investigated in the microcotyledonary placenta from pregnant mares at 110, 121, 179, 199 and 309 days of gestation by immunohistochemistry. In microplacentomes, both the ER isoforms were detected in trophoblast (T) cells, chorionic villous stroma (FS), microcaruncular epithelium (ME) and microcaruncular stroma (MS). Proportions of positive cells were 38–91% (T), 11–41% (FS), 55–89% (ME), 17–51% (MS) for ERα and 66–76% (T), 21–37% (FS), 41–68% (ME) and 24–55% (MS) for ERβ. Between days 110 and 199, proportions of cells positive for progesterone receptor (PR) varied between 19% and 62% (T), 3% and 50% (CS), 15% and 46% (ME), and 4% and 33% (MS). At day 309, PR was virtually absent in T, CS and ME (percentages < 0.1), whereas in MS 14.3% of cells were still positive. The expression of ERs and PR in equine microplacentomes gives evidence for a role of placental steroids as regulators of placental growth, differentiation and function. The detection of ERα, ERβ and PR in foetal and maternal vascular tissue suggests that placental steroids are also involved in the control of placental angiogenesis and /or vascular functions. The co-localization of ERs with aromatase in T suggests auto- or intracrine functions of oestrogens in this cell type.     

Transabdominal ultrasonographic determination of fetal gender in the horse during mid-gestation

Gender determination of the equine fetus using transabdominal ultrasonography was studied in 20 mares. One group of 10 research mares was scanned repeatedly every 2 weeks from 100 days gestation to parturition, while the second group of 10 client mares was subjected to echography once during mid-gestation. In males, the penis and/or prepuce was observed on 71 occasions from 102 days to 258 days gestation. On cross-sectional views, the male external genitalia had a round shape with parallel linear echogenic foci up to approximately 140 days gestation and then appeared triangular. Fetal testes were oval in shape in frontal view and had an homogeneous ultrasonographic appearance. Females were diagnosed on 23 occasions from 118 days to 227 days gestation based on the presence of the mammary glands and teats. Fetal ovaries appeared homogeneous with a characteristic circular echo from 100 days to 134 days gestation. Gender identifications (n = 98) based on the presence of the penis and/or prepuce in males and mammary glands and teats or fetal gonads in females were all correct, in agreement with the sex of the foals at birth. The optimal window of time was defined in both sexes as 100 to 220 days gestation. Thereafter, it was increasingly difficult to identify the anatomical structures cited above. Fetal sex was mainly determined using the transabdominal approach (87/98). However, the transrectal approach was useful in cases in which fetuses were either in posterior presentation or located very high in the mares abdomen. Good quality diagnostic scanners used typically in equine reproduction and equipped with a 5.0 MHz probe can be used for this procedure up to 160 days gestation, after which a 3.5 MHz transducer is often necessary due to increasing fetal size.     

Types of colic and frequency of postcolic abortion in pregnant mares: 105 cases (1984-1988)

The records of 105 pregnant mares and 105 nonpregnant horses with colic admitted to an equine hospital were reviewed. The 2 groups had similar types of colic and short-term survivability. Of the 105 pregnant mares, 31 were treated medically and 74 required surgical intervention. Thirty-three of the 105 mares died or were euthanatized. Thirteen (18%) of the 72 remaining mares aborted. Of 4 mares with severe medical cases, 2 died, 1 aborted, and 1 aborted and died. Of 27 horses with medical cases that required less intensive treatment, none died and 2 aborted. Of the 74 horses that required surgery, 45 survived to termination of pregnancy (foaling or abortion); 36 of these mares (80%) had a live foal. The type of surgical lesion had no effect on pregnancy outcome. Stage of gestation at initial examination, duration of anesthesia, or intraoperative hypoxia or hypotension had no effect on pregnancy outcome. However, when hypoxia occurred during colic surgery in the last 60 days of pregnancy, the mares either aborted or delivered severely compromised foals that did not survive.     

The effects of an advanced uterine environment on embryonic survival in the mare

Reasons for performing the study: During embryo transfer (ET) the equine embryo can tolerate a wide degree of negative asynchrony but positive asynchrony of >2 days usually results in embryonic death. There is still confusion over whether this is due to the inability of the embryo to induce luteostasis or to an inappropriate uterine environment. Objectives: To assess embryo survival and development in an advanced uterine environment. Hypothesis: Embryo–uterine asynchrony, not the embryo's inability to induce luteostasis, is responsible for embryonic death in recipient mares with a >2 days chronologically advanced uterus. Methods: Experiment 1: Thirteen Day 7 embryos were transferred to the uteri of recipient mares with luteal prolongation, occasioned by manual crushing of their own conceptus, such that donor–recipient asynchrony was between +13 and +49 days. Experiment 2: Day 7 embryos were transferred to recipient mares carrying their own conceptus at Days 18 (n = 2), 15 (n = 2), 14 (n = 4), 12 (n = 4) or 11 (n = 4) of gestation. In addition, Day 8 embryos were transferred to 4 pregnant recipient mares on Day 11 of gestation. Results: No pregnancies resulted following transfer of Day 7 embryos to recipients in prolonged dioestrus with asynchronies between +13 and +49 days. However, the use of early pregnant mares as recipients resulted in 5/20 (25%) twin pregnancies, 4 of which came from the transfer of a Day 8 embryo to a Day 11 recipient. All transferred embryos showed retarded growth, with death occurring in 4/5 (80%). Conclusions and potential relevance: The results emphasise the importance of an appropriate uterine environment for embryo growth and the inability of equine embryos to survive transfer to a uterus >2 days advanced even when luteostasis is achieved. It is possible that in normal, non‐ET equine pregnancy, embryo–uterine asynchrony may account for some cases of embryonic death.     

Flutamide Influences Placental Aldo–Keto Reductase Family 1 Member C1 (AKR1C1) Expression in Pigs

Aldo–keto reductase family 1 member C1 (AKR1C1) catalyses the conversion of progesterone into inactive 20α‐dihydroxyprogesterone. It is suggested that AKR1C1 expression in the placenta prevents from the cytotoxic effect of progesterone on foetuses during late pregnancy. The aim of the study was to determine whether the anti‐androgen flutamide administered during late pregnancy (83–89 days of gestation) or before parturition (101–107 days of gestation) influences AKR1C1 expression in the porcine placenta. AKR1C1 mRNA and protein levels were measured using real‐time PCR and western blotting, respectively. Immunolocalization of AKR1C1 within placentas was also performed. Flutamide significantly increased AKR1C1 mRNA (p = 0.008) and protein (p = 0.019) expression only during the pre‐parturient period in pigs. AKR1C1 protein was immunolocalized in the epithelial and stromal cells of foetal and maternal part of placenta at both stages of gestation. Following flutamide treatment, the intensity of staining was higher (p = 0.045) on day 108 of gestation. In conclusion, porcine placental AKR1C1 expression seems to be regulated by an androgen signalling pathway and may be involved in foetal survival by preventing the detrimental effect of progesterone.     

Pulmonary Maturation in Canine Foetuses From Early Pregnancy to Parturition

Development of the foetal respiratory system includes both pulmonary growth and maturation. In human medicine, a higher incidence of respiratory distress is reported in newborn males. This study aimed to identify different phases of canine foetal lung maturation throughout pregnancy, to determine the stage of pregnancy in which surfactant production begins and to compare pulmonary development of male and female foetuses. Pregnant bitches (34) were subjected to elective ovariohysterectomy and allocated into four groups, according to the stage of pregnancy: 30–40 days of pregnancy (n = 10), 41–50 days (n = 10), 51–60 days (n = 10) and bitches in the first stage of parturition (n = 4). Foetal lungs were histologically processed and evaluated by optical microscopy. The pseudoglandular phase was identified between the 35th day and 46th day of gestation; the onset of canalicular and saccular periods was observed, respectively, from the 48th day and 60th day of pregnancy. Lungs from foetuses at term were in the saccular phase; thus, the development into the alveolar period occurs in the neonatal period. The histological analyses revealed that respiratory tract development is centrifugal, from upper to lower airways. Therefore, it is possible to identify distinct development periods in different portions of the same organ. In conclusion, the saccular phase of lung development begins around 57 and 60 days of pregnancy, the period in which surfactant production is believed to occur. Male and female foetuses present similar pulmonary development from early pregnancy until parturition.     

High rate of transplacental infection and transmission of Neospora caninum following experimental challenge of cattle at day 210 of gestation

In order to investigate the pathogenesis of neosporosis following a primary infection in late pregnancy, cattle were subcutaneously challenged with 5 × 10⁸Neospora caninum (NC1 isolate) tachyzoites at day 210 of gestation and serial necropsies were then carried out at 14, 28, 42 and 56 days post-infection (dpi). No abortions occurred and all the foetuses were viable at the time of euthanasia. There was a high rate of vertical transmission, as parasites were detected by immunohistochemical labelling and PCR in all the foetuses from 28 dpi. Focal necrotic lesions were observed in the placentomes of the placenta from 28 dpi and showed resolution during later time points, denoted by infiltration of inflammatory cells at 42 dpi and fibrosis at 56 dpi. Foetuses at 28 and 42 dpi showed scarce and isolated lesions which are unlikely to represent a threat to foetal viability. No lesions were observed in the foetuses at 14 or 56 dpi suggesting control of the infection and resolution of the lesions by maternal and foetal immune responses. Once infection was established, it could not be cleared from the host and vertical transmission of the parasite occurred in all infected hosts. Parasite was detected in the placenta at 28 dpi, while in previous experimental infections of cattle at day 70 and 140 of gestation using the same challenge model, it was already present at day 14 post infection. This suggests that a change in the maternal immune response plays a crucial role in limiting the initial infection during the last term of pregnancy.