Marc Coryn

Risk factors for post partum ovarian dysfunction in high producing dairy cows in Belgium: A field study

An epidemiological study of risk factors for postpartal ovarian disturbances was carried out on 334 high-yielding dairy cows in 6 well-managed Belgian herds. Ovarian activity was closely monitored using progesterone profiles, based on twice weekly RIA-analysis for progesterone in milk fat, starting at 10 d after calving and continuing until the confirmation of a new pregnancy. Attention was focused on abnormal cyclicity during the preservice, postpartum period; cows were divided into 6 different categories. Three of these categories (normal profile, delayed cyclicity, and prolonged luteal phase) were of major importance and were analyzed using a multiple variable logistic regression model. Season of calving (stable vs pasture, odds ratio (OR)=5.7), an extended length of the previous dry period (> 77 vs < or = 63 d, OR=2.9), problem calvings (OR=3.6), abnormal vaginal discharge (OR=4.5), health problems during the first month of lactation (clinical disease, OR=5.4; ketosis, OR=11.3), and clinical parameters illustrating the appearance of a severe negative energy balance significantly increased the risk for delayed cyclicity before service. Parity (> or = 4 vs 1, OR=2.5), problem calvings (OR=2.9), occurrence of puerperal disturbances (OR ranged from 3.5 to 11.0), health problems during the first month of lactation (OR=3.1), and an early resumption of ovarian cyclicity after calving (< 19 d vs > 32 d, OR=2.8) increased the risk for prolonged luteal cycles before service.

Immunohistochemical detection of estrogen receptors in the canine uterus and their relation to sex steroid hormone levels.

Cyclic changes in estrogen receptor expression in the uterine tissue of 60 female dogs were evaluated, using an immunohistochemical technique on formalin-fixed paraffin-embedded sections. The expression of estrogen receptors in the uterine horns, body and cervix was quantified by means of an immunohistochemical score. A negative correlation was found between staining scores in the uterine horns and serum progesterone levels. Generally, staining scores in the uterine horns were highest during proestrus, declined during estrus and were lowest during early metestrus. During anestrus high staining scores for estrogen receptors were observed, indicating sensitivity for estrogens in a sexual quiescence stage. Compared with the uterine horns, high staining scores were found in the uterine body and cervix during estrus and metestrus. No positive staining for estrogen receptors was detected in 1 pregnant uterus. Fluctuations in estrogen receptors were more pronounced in endometrial stroma cells than in epithelial cells of the uterine horns. The importance of stromal cells in the sexual cyclicity of the canine uterus should not be underestimated when studying uterine endocrinology and pathology.

Cystic endometrial hyperplasia- pyometra complex in the bitch: should the two entities be disconnected?

The uteri of 26 clinically healthy bitches and 42 bitches with a clinical suspicion of pyometra were examined histologically using a computerized image analysis system. Histologic lesions were characterised mainly by thickening or atrophy of the endometrium and by varying degrees of cystic changes of the glands. These lesions were observed in most of the clinically healthy bitches as well as in all of the clinically ill animals. In most of the ill bitches a variable degree of inflammation also was found. Some bitches with clinical signs indicative for pyometra had no inflammatory reaction in the uterus. These bitches were misdiagnosed as suffering from pyometra, confirming the difficulty of diagnosing pyometra by simple clinical examination. Determination of sex hormone serum levels revealed that all dogs in both groups were either in metestrus or in anestrus. Based on the results of this study the cystic endometrial hyperplasia-pyometra complex can be divided in two entities: a cystic endometrial hyperplasia-mucometra complex and an endometritis-pyometra complex. Both entities bear many similarities with each other, except for the inflammatory reaction in the endometritis-pyometra complex. It is concluded from this study that the latter complex probably does not necessarily follow the former, but that both can arise de novo.

Insulin resistance: the link between metabolic disorders and cystic ovarian disease in high yielding dairy cows?

A preliminary investigation was performed to examine whether insulin resistance is a factor in the pathogenesis of cystic ovarian disease (COD) in high-yielding dairy cows. In total 30 cows, of which 15 were diagnosed as suffering from COD based on the anamnesis and clinical examination, and the other 15 served as matched controls, were subjected to an intravenous glucose tolerance test (IVGTT). The aim of the study was to investigate whether insulin activity was altered in COD cows. Differences in glucose clearance between the COD cows and their controls were analyzed comparing the fractional turnover rate (k), the glucose half-time (T1/2), and the area under the curve (AUC) 60 and 120 min after infusion. Differences in insulin response were analyzed comparing the insulin increment, the insulin peak concentration, and the AUC 60 and 120 min after glucose infusion. Although insulin resistance, attended by a secondary hyperinsulinemia, is stated to directly contribute to the ovarian abnormalities that characterize the polycystic ovary syndrome (PCOS) in human medicine, this was not observed in COD cows. On the contrary, COD cows appeared to have a low insulin response following an intravenous glucose load as compared with their matched controls. This was illustrated by significantly lower insulin increments (P = 0.04) and lower insulin peak concentrations (P = 0.04). As COD cows had a significantly lower insulin response to a standard glucose load, it was concluded that insulin could be a factor in the pathogenesis of COD in dairy cows.

Estrogen-α and progesterone receptor expression in cystic endometrial hyperplasia and pyometra in the bitch

Estrogen-alpha receptor (ER) and progesterone receptor (PR) were examined immunohistochemically in uteri of normal bitches, in uteri of bitches with cystic endometrial hyperplasia-mucometra (CEH-M) and in uteri of bitches with endometritis-pyometra (E-P), under exogenous progesterone treatment. In the CEH-M group, the ER- and PR-scores of all uterine cell types were higher than the ER- and PR-scores of normal uteri, although these differences were not always statistically significant. The ER-scores of E-P group were significantly lower than the ER-scores of the normal uteri and CEH-M group. The PR-scores of the E-P group tended to be higher than the PR-scores of the normal uteri, except for the surface epithelium, although these differences were not statistically significant. Exogenous progesterone treated bitches with CEH-M or E-P showed reduced ER- and PR-scores in the different uterine cell types, compared with the corresponding nontreated CEH-M or E-P group. The differences in ER and PR expression between CEH-M and E-P suggest different factors in the pathogenesis of both entities. Although, these changes in ER and PR expression do not seem to be directly involved in the pathogenesis of CEH-M and E-P. It is suggested that for CEH-M and progestin induced CEH-M a hormone dependent pathway is responsible. For P, the trigger may be bacterial infection.

Immunolocalization of progesterone receptors in the canine ovary and their relation to sex steroid hormone concentrations

The aim of the present study was to describe the normal cellular distribution of progesterone receptors in the canine ovary at different stages of the oestrous cycle. Samples of both ovaries were obtained from 75 healthy adult bitches of various breeds and ages, including five pregnant bitches and three bitches that had just delivered. The presence of progesterone receptors was visualized by immunohistochemistry on paraffin wax sections using a monoclonal antibody. Nuclear staining for progesterone receptors was observed in the surface epithelium, cortical tubules, rete ovarii, follicle cells, thecal cells, luteal cells, granulosa cell cords and ovarian stroma. The staining intensity for progesterone receptors in the follicle cells increased with the stage of follicle development, indicating an intrafollicular role of progesterone in the mechanism of ovulation and luteinization. The stronger staining intensities for progesterone receptors in thecal cells compared with follicle cells may be explained by the fact that thecal cells mediate some effects of steroid hormones on the follicle cells in secondary and tertiary follicles. Little correlation was found between the expression of progesterone receptors in follicle cells and oestradiol, progesterone or testosterone concentrations. This finding indicates a different regulating mechanism for progesterone receptors in canine ovarian follicles compared with other tissues of the genital tract. During pregnancy all groups of ovarian cells had lower staining intensity scores than during the oestrous cycle, although the sex steroid hormone concentrations in pregnant bitches were similar to those in non-pregnant bitches during the luteal phase of the oestrous cycle. The lower expression of progesterone receptors during pregnancy may be due to higher tissue concentrations of progesterone that are not reflected in the serum because of haemodilution and increased metabolism and clearance during pregnancy.

EFFECTS OF LONG-TERM TREATMENT WITH BOVINE SOMATOTROPIN ON FOLLICULAR DYNAMICS AND SUBSEQUENT OOCYTE AND BLASTOCYST YIELD IN AN OPU-IVF PROGRAM

Fourteen Holstein-Friesian heifers between 15 and 22 months of age with normal reproductive tracts were used in an experimental set up to investigate the effect of a long term rBST-treatment on the follicular population prior to transvaginal ovum pick-up (OPU). The estrous cycle of the animals was synchronized by means of a double injection of 2 ml cloprostenol 11 days apart. The heifers were divided in 2 equal groups (n = 7) in which the animals had the same average body weight, one group receiving a weekly subcutaneous injection of 640 mg recombinantly derived bovine somatotropin (rBST) on Mondays (rBST-treatment group) and a control group, being injected with 10 ml of saline. Heifers in both groups were submitted to OPU twice a week on Mondays and Thursdays using a 5 Mhz transducer and a disposable, 55 mm long, 20-g short bevelled needle at a vacuum pressure corresponding to approximately 13 ml water/min. The experimental period lasted for 10 weeks (April to June), each animal receiving a total of 10 injections and being submitted to OPU for 20 times. Oocytes were subsequently matured and cultured in a separate drop per cow following conventional IVF procedures. A blood sample was taken on heparin immediately after each OPU session, for determination of blood progesterone concentrations to assess the influence of treatment and OPU procedure on the cows estrous cycle. Although results show a significant increase in the total number of follicles and medium sized follicles in the rBST-treated group, no statistically significant different number of retrieved oocytes between the rBST-treated and nontreated group could be detected. The average number of retrieved oocytes per session per cow was comparable, being 6.4 for the treated and 6.0 for the control group. Additionally, the average number of blastocysts per cow per session did not differ significantly between groups, being 1.41 in the rBST-treated group and 1.53 in the control group. The number of cultured oocytes which developed to the blastocyst stage was 22% in the rBST-treated group, which was not significantly different from 25% in the control group. Repeated OPU appeared to induce a certain degree of acyclicity in both treated and nontreated animals.

Clinical, morphological and endocrinological aspects of cryptorchidism in the horse

The authors analysed clinical, histological and hormonal data obtained from 205 cryptorchid horses. The majority of the unilaterally and bilaterally retained testes were located in the inguinal canal; however, the ratio of inguinal vs abdominal retention appeared to decrease with advancing age. In unilateral cryptorchidism, a pronounced preference was noted for left abdominal retention, whereas for inguinal cryptorchids, the retained testes occurred equally on both sides. Right inguinal retention was found to decrease with advancing age. Histology of cryptorchid testes revealed apparently normal Leydig cells and arrested spermatogenesis. Plasma testosterone concentrations were similar in normal stallions and unilateral cryptorchids, even in those which had the scrotal testis removed. Plasma oestradiol-17beta levels were lower in unilateral cryptorchids than in stallions.

Changes in sex hormone receptors during administration of progesterone to prevent estrus in the bitch.

The development of lesions and the changes in sex hormone receptors were studied in the uteri of bitches under progesterone treatment. Twelve weeks after the onset of treatment, there was atrophy of the endometrium and increased thickness of the myometrium, without cystic dilatation of endometrial glands. This was accompanied by a dramatic reduction in estrogen-alpha and progesterone receptors in all cell types of the uterine wall. By 24 weeks after the onset of treatment, however, the endometrium was thickened due to the development of cysts of endometrial glands, while the myometrium thickness had returned to normal. The estrogen-alpha and progesterone receptors in most cell types of the uterine wall were again within the normal range. These results clarify and reconcile some apparent contradictions in the literature. They show that sex hormone receptors in most cell types of the uterine wall, especially endometrial gland cells and stromal cells, escape progestin (down) regulation after prolonged exogenous administration of progesterone.

The control of superovulation in the bovine with a PMSG-antiserum

Abstract A PMSG antiserum prepared in turkeys has been injected on the first day of standing heat in 20 superovulated cows (2000–3000 I.U. PMSG on D 10 , 2×500 mcg Estrumate on D 12 ). The treatment did not reduce the individual variability in superovulatory response but resulted in a higher ovulation (15,7 CL vs 9,4), a decreased number of unruptured follicles (2,8 vs 6,5), a better fertilisation (80% vs 60%) and a shorter external heat with less inseminations to be performed.