Ovulation,Pregnancy Rate and Early Embryonic Development in Vernal Transitional Mares Treated with Equine‐ or Porcine‐FSH

The objective of this study was to compare the efficacy of purified equine‐ and porcine‐FSH treatment regimes in mares in early vernal transition. Mares (n = 22) kept under ambient light were examined ultrasonographically per‐rectum, starting January 30th. They were assigned to one of two treatment groups using a sequential alternating treatment design when a follicle ≥ 25 mm was detected. In the eFSH group, mares were treated twice daily with equine‐FSH, and in the pFSH group mares were treated twice daily with porcine‐FSH; treatments were continued until follicle(s) ≥ 35 mm, and 24 h later hCG was administered. Oestrous mares were inseminated with fresh semen and examined for pregnancy on days 11–20 post‐ovulation. In the eFSH group, 11/11 (100%) mares developed follicle(s) ≥ 35 mm, 8/11 (73%) ovulated and 6/8 (75%) conceived. In the pFSH group, 5/11 (45%) developed follicle(s) ≥ 35 mm, 4/11 (36%) ovulated and 3/4 (75%) conceived. Treatment with eFSH resulted in a greater ovarian stimulation; higher number of pre‐ovulatory‐sized follicles, higher number of ovulations and higher number of embryos (p < 0.05). Following ovulation, serum progesterone concentrations were correlated with the number of CLs and supported early embryonic development; maternal recognition of pregnancy occurred in all pregnant mares. We concluded that eFSH can be used to effectively induce follicular growth and ovulation in vernal transitional mares; however, if bred, diagnosis and management of twins’ pregnancies would be required prior to day 16 because of the increased risk of multiple embryos per pregnancy. Conversely, the current pFSH treatment regime cannot be recommended.     

Corpus luteum Function and Pregnancy Outcome in Buffaloes during the Transition Period from Breeding to Non‐Breeding Season

The aim in this study was to investigate corpus luteum function and embryonic loss in buffaloes mated by artificial inseminations (AI) during the transitional period from breeding to non‐breeding season. The study was carried out using 288 multiparous Italian Mediterranean Buffalo cows at 110 ± 4 days in milk. The buffaloes were mated by AI after synchronization of ovulation by the Ovsynch‐TAI protocol 25 days after AI buffaloes underwent trans‐rectal ultrasonography to assess embryonic development. Pregnancy diagnosis was confirmed on Days 45 and 70 after AI by rectal palpation. Buffaloes pregnant on Day 25 but not on Day 45 were considered to have undergone late embryonic mortality (LEM), whilst buffaloes pregnant on Day 45 but not on Day 70 were considered to have undergone foetal mortality (FM). Corpus luteum size and blood flow were determined by real‐time B‐mode/colour‐Doppler on day 10 after AI in 122 buffaloes. The resistive index (RI) and pulsatility index (PI) were recorded at the time. Milk samples were collected on Days 10, 20 and 25 after AI in all inseminated buffaloes for the assay of whey P4 concentrations. Data were analysed by anova . Pregnancy rate on Day 25 after AI was 48.6% (140/288) and declined to 35.4% (102/288) and 30.6% (88/288) by Day 45 and Day 70 respectively. The incidences of LEM and FM were respectively 27.1% (38/140) and 13.7% (14/102). Pregnant buffaloes had greater (p < 0.01) whey concentrations of P4 from Day 20 onwards than buffaloes which showed LEM, whilst P4 in buffaloes that showed FM did not differ from the other two groups on Day 10 and Day 20. Corpus luteum blood flow on Day 10 after AI showed higher RI (p < 0.05) and PI (p = 0.07) values in buffaloes that subsequently were not pregnant on Day 25 compared with pregnant buffaloes. Buffaloes that were not pregnant on Day 45 also had a higher (p = 0.02) RI value on Day 10 than pregnant buffaloes, whilst PI values on Day 10 did not differ for the two groups of buffaloes. It was concluded that blood flow to the corpus luteum on Day 10 after AI influences corpus luteum function as judged by P4 secretion and also embryonic development and attachment in buffaloes.     

Endometrial Phospholipase A2 Activity During the Oestrous Cycle and Early Pregnancy in Mares

The aim of this study was to determine phospholipase A2 (PLA2) kinetics and activity in the mare’s endometrium during the oestrous cycle and early pregnancy. Phospholipase A2 is responsible for the liberation of arachidonic acid from phospholipids, which is the first limiting step in prostaglandins synthesis. Phospholipase A2 activity was measured using an assay based on the liberation of oleic acid from 1‐palmitoyl‐2‐[¹⁴C] oleoyl phosphatidylcholine. The enzyme was shown to be calcium dependent, to have an optimum pH of 8 and an apparent Michaelis constant of 127 μm . Enzyme activity was low in the endometrium of early luteal phase tissue but increased significantly (p < 0.001) during the late luteal phase (5.39 ± 0.16; 3.48 ± 0.33, 6.85 ± 0.59, and 9.96 thinsp;± thinsp;1.23 thinsp;nmol oleic acid released/mg protein at oestrus, and Days 3, 8 and 14 after ovulation, respectively). The mean PLA2 activity in endometrial tissue from pregnant mares (4.23 ± 0.74) was significantly lower (p < 0.01) than from cyclic animals during late dioestrus (9.96 ± 1.23). The results indicate that PLA2 activity in equine endometrium changes with the stage of the oestrous cycle and thus may be influenced by systemic hormone concentrations. The inhibitory effects of conceptus products on secretion of prostaglandin during early pregnancy were associated with a competitive inhibitor that decreased endometrial PLA2 activity.     

Corpus Luteum Development and Function after Supplementation of Long‐Acting Progesterone During the Early Luteal Phase in Beef Cattle

Strategic supplementation of P4 may be used to increase conception rates in cattle, but timing of supplementation in relation to ovulation, mass of supplementary P4 and formulation of the P4‐containing supplement has not been determined for beef cattle. Effects of supplementation of long‐acting progesterone (P4) on Days 2 or 3 post‐ovulation on development, function and regression of corpus luteum (CL) were studied in beef cattle. Cows were synchronized with an oestradiol/P4‐based protocol and treated with 150 or 300 mg of long‐acting P4 on Day 2 or 3 post‐ovulation (6–7 cows/group). Colour‐doppler ultrasound scanning and blood sample collection were performed from Day 2–21.5. Plasma P4 concentrations were greater (p < 0.05) from Day 2.5–5.5 in the Day 2‐treated groups and from Day 3.5–5.5 in the Day 3‐treated cows than in the control group. CL area and blood flow during Day 2–8.5 did not differ (p > 0.05) among groups, suggesting no effect of P4 treatment on luteal development. The frequency of cows that began luteolysis before Day 15 was greater (p < 0.04) in cows treated with 300 mg than in the controls, but there were no differences between non‐treated and 150 mg‐treated cows. The interval from pre‐treatment ovulation to functional and structural luteolysis was shorter (p < 0.01) in the combined P4‐treated groups than in the control cows. In conclusion, was showed for the first time that long‐acting P4 supplementation on Day 2 or 3 post‐ovulation increases P4 concentrations for ≥3 day, has no effect on luteal development, but anticipates the beginning of luteolysis in beef cattle.     

Postnatal Hypothyroidism Does Not Affect Prepubertal Testis Development in Boars

Young boars were treated with propiothiouracil to induce hypothyroidism to examine its effects on postnatal testicular development. Treatments with 0.1% 4‐propyl‐2‐thiouracil (PTU) in drinking water started after weaning, at 3 weeks of age and all boars were severely hypothyroid at 6 weeks of age as determined by measuring T3 and T4 in blood plasma. Boars were castrated at different ages up to 20 weeks and their testes used for histological and immunohistochemical analyses. Although small but significant reduction in testis weight was observed from 8 to 12 weeks of age, this was not accompanied by significant difference in testicular volume. By 20 weeks of age, at the beginning of puberty, the differences in testis weights between control and treated groups of boars disappeared suggesting there is no lasting effect of hypothyroidism on postnatal development of boar testis. Immunohistochemical staining was used to determine the presence of molecular markers in both Sertoli and Leydig cells. Again, there were no differences between testes from control and treated boars in the pattern or intensity of immunostaining using antibodies against 3β‐hydroxysteroid dehydrogenase, antimullerian hormone or proliferating cell nuclear antigen (PCNA). Immunostaining with antibodies against PCNA showed interesting results as it was observed that Sertoli cells still express this marker of proliferating cells at 14 weeks of age, later than previously suggested cessation of Sertoli cell proliferation. This study suggests that hypothyroidism in boars does not have similar effects on postnatal testis development as reported in some other species.     

Effect of Administration of Oxytocin During Diestrus on Corpus Luteum Function and Endometrial Oxytocin Receptor Concentration in Cycling Mares

The objectives of this study were to (1) compare the effect of twice versus once daily administration of oxytocin on days 7-14 after ovulation on the duration of corpus luteum (CL) function and (2) determine the effect of oxytocin treatment on endometrial oxytocin receptor concentration in mares. In experiment 1, mares were randomly assigned to three groups on day 7: (1) untreated control group (n = 7), (2) twice daily oxytocin treatment group (n = 7), and (3) once daily oxytocin treatment group (n = 8). Oxytocin-treated mares received 60 U of oxytocin intramuscularly (IM) the respective number of times each day on days 7 through 14. One of seven control mares (14%), five of seven (71%) twice daily oxytocin-treated mares, and five of eight (63%) once daily oxytocin-treated mares had prolonged CL function. There was no significant difference in the proportion of mares with prolonged CL function between the two oxytocin-treated groups, and collectively, oxytocin treatment increased (P < .05) the proportion of mares with prolonged CL function compared with no treatment. In experiment 2, mares were randomly assigned to two groups (n = 5/group): (1) saline-treated control mares, and (2) oxytocin-treated mares. Beginning on day 7, control mares received 3 mL of sterile saline IM twice daily, and oxytocin-treated mares received 60 U of oxytocin IM twice daily through day 14. On day 15, endometrial oxytocin-binding capacity was determined (as a measure of oxytocin receptor concentration), and there was no difference (P > .1) between control and oxytocin-treated mares (1,465.7 ± 108 and 1,382.8 ± 108 fmol/mg protein [mean ± standard error of mean], respectively).     

The Pattern of Embryonic Fixation and its Relationship to Pregnancy Loss in Thoroughbred Mares

Ultrasonographic pregnancy records of 195 mares from six Thoroughbred stud farms, over a period of 7 years were retrospectively analysed to assess the effect of various factors on embryonic vesicle (EV) fixation pattern and pregnancy outcome. Of the total of 746 pregnancies analysed, significantly (p < 0.01) more EV fixations were evident in the right uterine horn than in the left (53.35% vs 46.65% respectively). There was no significant effect of either, the side of ovulation, or age of the mare, on the side of EV fixation. However, EV fixation, was significantly (p < 0.001) more likely to occur in the right uterine horn in maiden and barren mares (65.75% vs 57.45% respectively). The age and reproductive status of the mare as well as foal heat breeding failed to demonstrate a consistent effect on pregnancy loss relative to the side of EV fixation. In lactating and foal heat bred mares, EVs were significantly (p < 0.0001) more frequently established in the contralateral horn to the one from which the mare delivered her most recent foal. In lactating mares, significantly (p < 0.05) higher embryonic and pregnancy losses were observed in the ipsilateral horn. In conclusion, (a) side of EV fixation was (i) independent of the side of ovulation and mare age (ii) significantly (p < 0.001) affected by reproductive status, (b) neither age of mare nor reproductive status had any effect on pregnancy loss rates relative to the side of EV fixation and (c) in lactating mares the EV had a greater chance of fixation and survival in the horn contralateral to the one from which the mare delivered her most recent foal.     

Age-Dependent Effect of Foal Heat Breeding on Pregnancy and Embryo Mortality Rates in Thoroughbred Mares

The aim of the present study was to evaluate the correlation of age and heat cycle to determine reproductive efficiency in young and aged Thoroughbred mares bred on foal heat (FH) or on second heat (SH) after foaling. Embryo mortality (EmbM) was determined every time a mare was found open after a positive pregnancy diagnosis. Parturition to breeding interval, pregnancy rate (PregR) and EmbM rate were the dependent variables and the treatments were breeding on the FH or on SH. The cutoff age to obtain above-average probability for the EmbM was 10 years old. PregR in mares bred on FH was lower compared with SH (P < .01); however, it was neither affected by the age of mares (P > .05) nor by the age group of mares (P > .05). Regarding FH and SH, there was a difference in PregR in young mares (P < .01), unlike in aged mares (P > .05). EmbM rate was not different between mares bred on FH or SH (P > .05) although it was affected by age of mares (P < .01). EmbM was higher in oldest than young mares (P < .01). Aged mares bred on FH had a significantly higher EmbM rate compared with the young group also bred on FH (P < .01). In conclusion, the reproductive efficiency of Thoroughbred mares bred on FH is dependent of the age. Aged mares (≥10 years old) should be bred at their SH to reduce EmbM and improve reproductive performance.     

Metabolic Profile of Broodmares During Late Pregnancy and Early Post‐Partum

Pregnancy represents a specific physiological status characterized by continuous adjustments that affect maternal metabolism of all nutrients. In the last trimester of pregnancy, mare's nutrient requirements greatly increase and most pregnancy‐associated diseases are likely to occur. Therefore, we aimed to assess the metabolic profile of broodmares focusing on the last 3 months of pregnancy and the early post‐partum. Fifteen pregnant mares (Group A) were monitored from 263 ± 3 days of pregnancy until 21 days after foaling. Seven non‐pregnant mares (Group B) were used as the control group. Blood samples were collected weekly by jugular venipuncture throughout the experimental period, and additional blood samples were collected within 24 ± 12 h of foaling. Obtained sera were analyzed for urea (Ur), creatinine (Cre), total protein (TP), total, direct and indirect bilirubin (tB, dB, iB), triglycerides (TG), total cholesterol (tChol) and β‐hydroxybutyrate (BHB). During pregnancy, Ur (p = 0.015) and dB (p = 0.028) were higher in Group A than Group B. Serum Cre, tB, iB and TG (p < 0.001) decreased after foaling; furthermore, Group A exhibited lower tChol (p < 0.001) and higher BHB (p < 0.001) than Group B during the study. There were significant interactions of time × peripartum on Ur (p = 0.007), Cre (p < 0.001), tB (p = 0.018), TG (p < 0.001) and tChol (p < 0.001). These results can be used in the interpretation of biochemical studies in late gestation mares which develop systemic disease. The improved understanding of the metabolic profile during the peripartum might assist in monitoring the health status of the broodmares to promote foetal growth and well‐being.     

Expression and Hormonal Regulation of E‐Cadherin in Canine Uterus During Early Pregnancy

E‐cadherin, a Ca^{2 +} ‐dependent cell adhesion molecule, is necessary for endometrial receptivity to blastocyst implantation. The aim of this study was to investigate the differential expression of E‐cadherin in canine uterus during early pregnancy and its regulation under different conditions by in situ hybridization. E‐cadherin mRNA expression was at a low level in the glandular epithelium on days 6, 12 and 17 of pregnancy. On days 20 and 23 of pregnancy, E‐cadherin mRNA was highly expressed in the glandular epithelium surrounding the embryo, but not in the luminal epithelium and declined in villi and placenta on day 28 of pregnancy. During oestrous cycle, a moderate level of E‐cadherin mRNA expression was found in the luminal and glandular epithelium of canine uteri at oestrus stage. The same expression was also found at anoestrus stage. Progesterone slightly induced the expression of E‐cadherin mRNA in the luminal and glandular epithelium of ovariectomized canine uterus. These results suggest that E‐cadherin expression is closely related to canine implantation and can be up‐regulated by progesterone.     

Relationship Between Fertility at Foal Heat and Blood Biochemistry Parameters Monitored During the Peripartum Period in Thoroughbred Mares

This study aimed to examine fertility at foal heat and its relevance to body condition score (BCS) and blood nutritional metabolites in Thoroughbred mares. Thoroughbred mares foaled from 2006 to 2009 were included and classified into two groups: group C (conception; n = 34), which included mares that conceived during foal heat (within 3 weeks after foaling), and group NC (nonconception; n = 39), which included mares that did not conceive despite mating during their foal heat. BCS and blood samples were obtained 1 month before the expected foaling date and 1, 2, 3, and 4 weeks after foaling. Total protein (TP), albumin (Alb), blood urea nitrogen (BUN), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), total cholesterol (T-Cho), triglyceride (TG), nonesterified fatty acid (NEFA), calcium (Ca), inorganic phosphate (iP), and magnesium (Mg) levels were measured using an automatic clinical chemistry analyzer. Repeated measures analysis of variance (ANOVA) and Student t-test were used to examine the differences between the two groups. No significant differences were observed between the two groups in any of the above-mentioned parameters at 1 month before the expected foaling date (Student t-test). Furthermore, no significant differences were observed between the two groups in serum levels of TP, Alb, AST, GGT, T-Cho, NEFA, Ca, and Mg and BCS at postpartum periods (repeated measures ANOVA). Serum TG, BUN, and iP levels remained lower in group NC than in group C after foaling (P < .05, repeated measures ANOVA). Although the mechanism by which these nutritional factors affect a decline in reproductive performance remains unclear, our results suggest that blood biochemical tests can detect potential imbalances in nutrition and metabolism, even if there is no difference in BCS.     

Blood Lymphocyte Subpopulations, Neutrophil Phagocytosis and Proteinogram During Late Pregnancy and Postpartum in Mares

The aim of this study was to evaluate peripheral blood lymphocyte subpopulations, neutrophil phagocytic capacity and proteinogram characteristics in mares, during the last trimester of pregnancy and in postpartum. Measurement of phagocytosis and quantification of T‐lymphocyte subsets were done by flow cytometry. Quantification of T‐lymphocyte subsets was performed with monoclonal antibodies specific for CD2, CD3, CD4 and CD8 cell markers. Natural killer and B‐cell counts were estimated mathematically. Serum proteinogram was obtained by electrophoresis. No significant differences were observed between gestation and postpartum on CD4⁺, CD8⁺ and NK⁺ lymphocyte subsets, CD4 : CD8 ratio and phagocytosis. The percentage of cells expressing CD3 (64.2 ± 1.8) and CD2 (68.4 ± 1.7) (Mean ± SEM) was reduced during gestation vs postpartum (69.7 ± 1.5 and 73.8 ± 1.4 respectively) (p < 0.05). During pregnancy, CD19⁺ (31.6 ± 1.7) was higher than in postpartum (26.2 ± 1.4) (p < 0.05). Total T cells (2911 ± 227 cells/μl), T helper cells (2144 ± 169 cells/μl) and T‐cytotoxic cells (767 ± 68 cells/μl) were depressed in pregnancy, when compared with postpartum (4093 ± 337 cells/μl; 3004 ± 276 cells/μl; 1089 ± 94 cells/μl respectively) (p < 0.01). Total white blood cell count was reduced during pregnancy (8815 ± 427 cells/μl) with respect to postpartum (10742 ± 446 cells/μl) (p < 0.01), while neutrophil count did not change. Total proteins, albumin, α₁,α₂,β₁, β₂, γ globulins and albumin : globulin did not differ. Our results suggest that the physiological immune depression occurring in mares, during gestation might be due to T‐helper and T‐cytotoxic lymphocytes reduction.     

Inhibition of Nitric Oxide Synthase with L-NAME Does Not Increase Lactate Production at Rest or During Short-term High-intensity Exercise in Thoroughbred Horses

The present study was carried out to determine whether inhibition of nitric oxide (NO) synthase promotes anaerobic metabolism in exercising horses, resulting in a significantly increased blood lactate concentration. N -nitro-L-arginine methyl ester (L-NAME) is a potent inhibitor of NO synthase that has been tested in horses and other species. Two sets of experiments, namely placebo (saline control) and L-NAME (20 mg/kg, IV) studies, were carried out on seven healthy, sound, exercise-trained, Thoroughbred horses in random order, 6 to 7 days apart. In both experiments, an incremental exercise protocol was used and data were obtained at rest, during submaximal exercise performed at 8 m/s on a 4.5% uphill grade, and during galloping at 14 m/s on a 4.5% uphill grade – a workload that not only elicited maximal heart rate and induced exercise-induced pulmonary haemorrhage, but also could not be sustained for more than 90 s. Measurements were also made in the recovery period. Mixed-venous blood samples, obtained at matched intervals in the two sets of experiments, were analysed in triplicate for determining the lactate concentration. Following administration of L-NAME, significant bradycardia occurred at rest (27±1 vs 37±2 beats/min in the placebo trials; p<0.0001) as well as during submaximal exercise (183±4 vs 200±4 beats/min in the placebo trials; p<0.001), but the heart rate increased during galloping at 14 m/s on a 4.5% uphill grade to reach values observed in the placebo trials (215±2 beats/min) and significant differences were not found. At rest, the mixed-venous blood lactate concentration was similar in the two experiments. With exercise, the mixed-venous blood lactate concentration increased progressively as work intensity increased in both trials, but significant differences were not found between the placebo and the L-NAME experiments during submaximal exercise, near-maximal exercise or recovery. These experiments demonstrated that inhibition of NO synthase in Thoroughbred horses does not promote enhanced anaerobic metabolism at rest or during short-term incremental exercise leading to galloping at maximal heart rate.     

Treatment of Persistent Mating‐Induced Endometritis in Mares with the Non‐Steroid Anti‐Inflammatory Drug Vedaprofen

Recently, successful treatment of mares with a history of persistent mating‐induced endometritis (PMIE) with dexamethasone has been reported. As systemic treatment of horses with glucocorticoids should be handled with caution, we tested the hypothesis that treatment with the non‐steroid anti‐inflammatory drug (NSAID) vedaprofen, an inhibitor of cyclooxygenase‐2, may have comparative, positive effects on fertility. Barren mares with a history of repeated PMIE were treated with vedaprofen (n = 8; initially 2 mg/kg bodyweight followed by 1 mg/kg orally twice daily) from 1 day before the first insemination to 1 day after ovulation or left untreated (n = 9). All mares received oxytocin (20 I.E. s.c.) thrice daily. Uterine swabs were collected for bacteriology and cytology. The day after ovulation, fluid accumulation was detected in three control mares and four treated mares (n.s.). The percentage of neutrophils in uterine cytology was significantly increased in comparison to the day before ovulation irrespective of treatment. Pregnancy was confirmed in two of nine mares in the control group and seven of eight mares in the treatment group (p < 0.05). NSAIDs may positively affect fertility in mares with a history of PMIE.     

Neospora caninum Infection Does Not Affect the Fertility of Dairy Cows in Herds with High Incidence of Neospora‐associated Abortions

This present study, was designed to establish whether Neospora caninum infection affects the fertility of high‐producing dairy cows, and was based on yearly serological screening for neosporosis and on the confirmation of N. caninum infection in aborted foetuses. Assessment was made of 7518 artificial inseminations (AI) (applied to parous cows) performed in three herds. Of these inseminations, 2540 (33.8%) resulted in pregnancy; 34% of which corresponded to seronegative cows (2226 of 6556 AI performed in seronegative cows) and 32.6% to seropositive animals (314 of 962). Abortion occurred in 97 (30.1%) of the 314 pregnancies recorded in dams seropositive for N. caninum, while only 93 (4.2%) of the 2226 pregnancies in seronegative animals ended in abortion. Logistic regression analysis indicated no significant effects of N. caninum antibody titre, days in milk and milk production at insemination on fertility. Factors found to affect fertility were herd, season of insemination, lactation and insemination number, semen‐providing bull and AI technician. Our results indicate that N. caninum infection does not affect the fertility of high‐producing dairy cows.     

Equine Endometrial Gland Density and Endometrial Thickness Vary among Sampling Sites in Thoroughbred Mares

The secretions of the equine endometrial glands are essential for the survival, growth, and development of the conceptus in early pregnancy, and endometrial gland density is directly related to successful pregnancy outcome. Endometrial biopsy is routinely used to assess the reproductive potential of broodmares. Some previous studies have shown that equine endometrial glands are uniformly distributed throughout the uterus; however, other work has shown variation of the endometrial architecture between biopsy sites, suggesting that a single biopsy is not representative of the entire endometrium. The aims of this study were to assess and compare the endometrial gland density and thickness at four sampling sites in the uterus (the central segment of each uterine horn, the uterine horn-body junction, and the caudal portion of the uterine body). Endometrial samples from five nulliparous Thoroughbred mares in diestrus were obtained at necropsy and used for subsequent histomorphometric analysis. The caudal uterine body had a significantly lower endometrial gland density and endometrial thickness than the other sites. This may result in nutrient deprivation and reduced survival of embryos or fetuses in this region of the uterus. The endometrial gland density and endometrial thickness did not significantly differ between the other regions sampled, indicating that they are similarly suitable for embryonic implantation and fetal development. Our results suggest that the endometrial structure of the caudal uterine body of the mare is not representative of the endometrial morphology at other sites. Thus, the caudal uterine body is not a suitable site for routine endometrial biopsy.     

Faecal Oestrogens and Progesterone Metabolites in Mares of Different Breeds During the Last Trimester of Pregnancy

Non‐invasive pregnancy diagnosis in mares by measuring faecal oestrogens has been performed over years with great accuracy. However, results have indicated breed‐related differences in the amount of excreted steroids during late pregnancy. Therefore faecal samples were collected during the last 4 months of pregnancy of Thoroughbred (n=10), New Forest pony (n=9), Shetland pony (n=10) and Iceland pony mares (n=11). Concentrations of oestrogens, 20α‐hydroxy‐ and 20‐oxopregnanes were measured using enzyme immunoassays. Breed differences concerning both levels (though significant only in case of oestrogens) and time course of measured steroids were observed. There was a highly significant time effect (p < 0.00001) and an interaction between time and breeds (p < 0.02) for all steroids measured, suggesting that the time effect differs for different breeds. Oestrogen concentrations showed a decrease towards parturition, whereas in 20α‐hydroxy‐ and 20‐oxopregnane levels a pronounced increase was found 2 and 1 months, respectively, before parturition. A breed effect was only significant (p=0.001) when comparing oestrogen concentrations and was mainly due to Iceland ponies, which had the lowest concentrations especially during the last 2 months of pregnancy. An almost significant (p=0.06) breed effect was found for 20‐oxopregnanes. In Iceland mares an additional increase in faecal pregnane content was already observed earlier, reaching maximum levels before the 60th day ante‐partum (a.p.), followed by a decrease until the 30th day a.p. The ratio of 20‐oxopregnanes to oestrogens in the samples was significantly higher (p < 0.006) in Iceland ponies in comparison with any other breed throughout all months before parturition. The breed differences observed in the amounts of oestrogens and/or progestagens present during late pregnancy may demonstrate micro‐evolutionary changes in the endocrine system of a species.     

Effect of a Single Injection of Long-acting Progesterone on the First Ovulation in Early and Late Spring Transitional Mares

Since 1966, exogenous progestins have been used in equine practice for pregnancy maintenance, estrous suppression, and control of erratic sexual behavior. This study was designed to investigate the use of a new compounded controlled-release progesterone preparation (BioRelease P4 LA 300) in early and late spring transitional mares. In the first experiment, the pharmacodynamic properties of the preparation were studied in five geldings. In the second experiment, the use of a single intramuscular injection (600 mg) was tested in 68 embryo-recipient mares maintained under natural photoperiod in the Southern Hemisphere. Experiment 1 demonstrated elevated serum concentrations of progesterone (>1 ng/mL) for 7.6 ± 2.2 days. In experiment 2, there was no effect of treatment in mares that were treated on September 18, independent of their follicular status at day of treatment (10 to 15 mm; 20 to 25 mm, respectively). When mares with a follicular size of 20 to 25 mm were treated on October 14, significantly more progestin-treated mares (10/12; 83%) ovulated between 10 and 24 days after treatment than untreated controls (3/12; 25%) (P < .05). Additionally, there was a trend in mares treated on October 14 for a shorter treatment to ovulation interval (mean ± SD, 18.6 ± 8.7 days) compared with untreated controls (mean ± SD, 26.7 ± 14.7 days) (P = .07). Administration of one single injection of long-acting progesterone is a simple and effective method of controlling the first ovulation of the season in late transitional mares.     

交感神经对妊娠早期小鼠子宫内膜发育的影响

为了研究交感神经对小鼠妊娠早期(妊娠第1～9天)子宫内膜发育的影响,本试验采用交感神经阻断剂6-羟多巴胺(6-OHDA,100 mg·kg~(-1)体质量)腹腔注射成年雌性昆明小白鼠,损毁外周交感神经.利用放射免疫分析法、组织学和免疫组织化学技术,检测了小鼠妊娠早期血清17β-雌二醇水平、子宫腺发育和子宫内膜细胞增殖的变化.结果显示:(1)在妊娠第1天(E1)、E5和E7 6-OHDA处理组血清中17β-雌二醇的含量分别降低了63.4%(P<0.01)、35.7%(P<0.05)和25.5%(P<0.05),而在E3和E9分别升高了79.3%(P<0.05)和156.7%(P<0.01);(2)6-OHDA处理降低了妊娠早期子宫腺的比例,与同期对照组相比分别下降了37.5%、38.6%、41.8%、61.4%和58.1%(P<0.01);(3)6-OHDA处理降低了妊娠早期子宫内膜细胞的增殖,使E1、E3、E5、E7和E9子宫内膜增殖细胞分别减少了27.2%、30.2%、40.7%、27.2%和27.7%(P<0.01).上述结果表明,交感神经对雌激素的分泌以及妊娠早期子宫组织结构的重建具有重要的调控作用,进而调节动物早期胚胎的着床和发育.     

Expression of hLAMP‐1‐Positive Particles During Early Heart Development in the Chick

Heart development requires coordinated activity of various factors, the disturbance of which can lead to congenital heart defects. Heart lectin‐associated matrix protein‐1 (hLAMP‐1) is a matrix protein expressed within Hensen's node at Hamburger–Hamilton (HH) stage 4, in the lateral mesoderm by HH stages 5–6 and enhanced within the left pre‐cardiac field at HH stage 7. At HH stages 15–16, hLAMP‐1 expression is observed in the atrioventricular canal and the outflow tract. Also, the role of hLAMP‐1 in induction of mesenchyme formation in chick heart has been well documented. To further elucidate the role of this molecule in heart development, we examined its expression patterns during HH stages 8–14 in the chick. In this regard, we immunostained sections of the heart during HH stages 8–14 with antibodies specific to hLAMP‐1. Our results showed prominent expression of hLAMP‐1‐positive particles in the extracellular matrix associated with the pre‐cardiac mesoderm, the endoderm, ectoderm as well as neuroectoderm at HH stages 8–9. After formation of the linear heart tube at HH stage 10, the expression of hLAMP‐1‐stained particles disappears in those regions of original contact between the endoderm and heart forming fields due to rupture of the dorsal mesocardium while their expression becomes confined to the arterial and venous poles of the heart tube. This expression pattern is maintained until HH stage 14. This expression pattern suggests that hLAMP‐1 may be involved in the formation of the endocardial tube.