Lymphocytes and MHC class II positive cells in the female rabbit reproductive tract before and after ovulation

In this study, we identified lymphocytes and MHC class II positive (MHC-II+) cells in the reproductive tract of female rabbits both before and after ovulation. CD43+ T cells were frequently present in the mucosa of the oviduct, cervix, and vagina, but far fewer positive cells were seen in the endometrium. The induction of ovulation did not change the cell density in these regions. KEN-5+ T cells and MHC-II+ cells were also frequently seen in the mucosa of the oviduct, cervix, and vagina both before and after ovulation. However, in the uterus, there were very few positive cells before ovulation, but the number increased dramatically after ovulation. Associated with the increase of KEN-5+ T cells, IL-2 mRNA expression in the uterus also increased after ovulation, suggesting that the uterus experienced an increase of T-cell activation. IgM- and IgA-positive B cells were not commonly seen in the reproductive tract and the induction of ovulation did not alter this. Our results suggest that the reproductive tract of female rabbits has the capacity to mount an immune response and that the immune cell distribution of the rabbit reproductive tract has some distinctive features compared with that found in other species.     

Effect of ovarian steroids on nitric oxide synthase in the rat uterus, cervix and vagina

The effects of estrogen (E2), progesterone (P) and E2 and P (E2 + P) were examined on nitric oxide synthase (NOS) activity in both cytosolic and particulate fractions isolated from the rat uterus, vagina, cervix and cerebral cortex. Additionally plasma nitrate + nitrite (NO3 + NO2) levels were measured in control and hormone treated rats. Cytosolic NOS was the predominant form being approximately 80% of the total in all four tissues. NOS activity in both fractions from all tissues was highly Ca-dependent (> 90%). Among the reproductive tract tissues, the highest activity was found in the cervix, which was nearly 5- and 2-fold higher than the uterus and vagina, respectively. NOS activity in the cerebral cortex was by far the highest being 5-fold higher than in the cervix. In contrast to the cortex, E2 treatment downregulated cytosolic NOS in all reproductive tract tissue, but this was statistically significant in only uterus. When compared with E2 treated rats, P increased cytosolic NOS in uterus, vagina, and particulate NOS in the cervix. The data do not give any indication whatsoever of differential effects of P in the uterus and cervix.     

Ultrasonography of the developing reproductive tract in ram lambs: effects of a GnRH agonist

In spring-born ram lambs, the testes (from 2 wk), prostate and vesicular glands (from 4 wk) were examined by ultrasonography every 2 wk up to 26 wk of age. Image analysis was done (numerical pixel values). Ram lambs were treated with a long acting formulation of a GnRH superagonist (Leuprolide acetate; 1.5 mg/kg) at 3 and 7 wk of age. In blood samples taken every 15 min for 8 h, mean serum LH, LH pulse amplitude, and basal and mean serum FSH concentrations were lower at 5 wk of age, and LH pulse frequency was lower at 15 wk of age in animals given Leuprolide acetate compared with those of the controls. There were no differences (P>0.05) in testis, prostate or vesicular gland development between treated and control animals. Testicular diameter of the left and right testes in transverse and longitudinal planes increased slowly to 8 wk of age, more rapidly to 18 wk of age, then more slowly to 26 wk of age (P<0.05). Numerical pixel values of testicular images decreased from 2 to 8 wk of age, increased to 22 wk of age and then plateaued. Width of the prostate increased from 4 to 26 wk of age, but length and width of the vesicular glands increased slowly to 8 wk of age, more rapidly to 18 wk of age and then plateaued (P<0.05). Numerical pixel values for the prostate declined from 4 to 8 wk and for the vesicular glands, declined from 4 to 10 wk of age; numerical pixel values increased to 12 wk and then decreased to a nadir at 18 wk, followed by a steady increase to 26 wk of age (P<0.05). We concluded that developmental patterns of numerical pixel values of the testes, prostate and vesicular glands in ram lambs reflect stages of development, but treatment with a GnRH superagonist at 3 and 7 weeks of age did not affect growth of testes, vesicular or prostate glands.     

Ultrasonographic imaging of the reproductive tract and surgical recovery of oocytes in Cebus apella (capuchin monkeys)

Two-dimensional real-time and Doppler ultrasonography are valuable non-invasive methods to assess reproductive anatomy and physiology. In adult, postpubertal female Cebus apella (capuchin monkeys), the objectives were to determine (1) uterine and ovarian dimensions, ovarian follicular dynamics, day of ovulation, and arterial blood flow of uterus and utero-ovarian ligament during the follicular phase of the menstrual cycle and (2) the number of oocytes aspirated from antral follicles at laparotomy. Based on two-dimensional, transabdominal B-mode ultrasonography, mean (+/- S.E.M.) length, height, width, and volume of the uterus were 17.9+/-0.4, 12.4+/-0.3, 13.6+/-0.3 mm, and 1.55+/-0.08 mL, respectively, and of the ovary were 13.4+/-0.2, 8.2+/-0.1, 7.7+/-0.1 mm, and 4.5+/-0.2 mL. Ovarian follicles were monitored for 6 days before ovulation, which occurred on day 9.3+/-0.5 (range, days 7-11; day 1=start of menses), with 10 of 12 ovulations in the right ovary. Diameter and volume of the preovulatory follicle were 10.1+/-0.2 mm and 0.55+/-0.03 mL (on the estimated day of ovulation) and of the CL were 8.1+/-0.4 mm and 0.3+/-0.05 mL. Resistivity and pulsatility indices were 0.86+/-0.02 and 2.15+/-0.11 for uterine arteries, and were 0.69+/-0.04 and 1.63+/-0.15 for the utero-ovarian ligament (UOL) artery; just prior to ovulation, both indices peaked (P<0.05) in the uterine artery ipsilateral to the side of ovulation, but both reached a nadir (P<0.05) in the UOL artery. In the absence of ovarian stimulation, 31 oocytes (diameter, 137+/-10 microm) were aspirated (average of 2 oocytes/(female attempt)) on days 5, 7, and 9. In conclusion, transabdominal ultrasonography facilitated assessment of reproductive anatomy and physiology in C. apella adult females. Resistance and pulsatility indices of uterine and UOL arteries changed near the time of ovulation. Dominant follicles were easiest to aspirate at 8-9 mm in diameter ( approximately day 9), with intact cumulus-oocyte complexes recovered from ovarian follicles 2-9 mm in diameter.     

Cervical patency during non-ovulatory and ovulatory estrus cycles in domestic cats

The cervix functions as a barrier to spermatozoa. Vaginal artificial insemination in cats is, therefore, likely to be successful only at the period of estrus when the cervix is open. This study aimed to define the period of cervical patency in cats in both non-ovulatory and ovulatory estrus cycles. A total of 15 reproductive cycles were studied in six cats during the estrous stage. Cervical patency was monitored with the cats under sedation, by infusing 2 mL of Iohexol contrast medium via a 3.5 French tomcat catheter into the cranial vagina during estrus. Day one of estrus was defined as the first day the cats showed estrous behavior. Non-ovulatory cycles were characterized by a serum progesterone concentration on days 11-15 that was below 5 nmol/L and a normal interestrus interval of 7-14 days. Ovulatory cycles were characterized by a serum progesterone concentration on days 11-15 that was above 5 nmol/L and an interestrus interval that exceeded 30 days. The cervix was considered to be open when the contrast medium was seen to enter the uterus, and to be closed when the contrast medium remained in the vagina. Blood samples were collected at each examination and were assayed for estradiol-17beta and progesterone concentrations. The cervix was open on the first day of standing estrus at a mean estradiol-17beta serum concentration of 87.4+/-21.8 pmol/L (range 14 to >or=180 pmol/L) and closed at an estradiol concentration of 47.1+/-12.4 pmol/L (range 4 to >or=180 pmol/L). In the ovulatory cycles the cervix was closed at a progesterone concentration of 9.8+/-4.4 nmol/L (range 0.6-28.4 nmol/L). There was no difference in the duration of cervical patency in non-ovulatory and ovulatory cycles (5.5+/-1.2 days and 5.2+/-0.5 days, respectively) (p>0.05). The higher overall mean concentrations of estradiol-17beta seen in the ovulatory cycles than in the non-ovulatory cycles, indicate that a high level of estradiol is necessary for induction of ovulation. Ovulation in 60% of unmated females in this study indicates that the techniques used for evaluation of cyclus stage and cervical opening have the potential to induce ovulation in the cat. This study demonstrates that cervical patency is not influenced by the occurrence of ovulation, but is due to individual variations between cats.     

Effects of abnormal ovarian cycles during pre-service period postpartum on subsequent reproductive performance of high-producing Holstein cows

The primary objective of this study was to investigate the effects of abnormal ovarian cycles during the pre-service postpartum period on subsequent reproductive performance of high-producing Holstein cows. The study was conducted in a commercial dairy farm with approximately 150 lactating cows, in a subtropical region of Japan. Animals were kept in free-stall barn, and fed a total mixed ration. Cows that calved from June 2001 to July 2002 were included in the study. Milk samples were collected twice weekly from 2 to 11 weeks postpartum, and progesterone concentrations in skim milk were determined by ELISA. After a voluntary waiting period of 40 days, cows detected in estrus were bred by artificial insemination (AI). Pregnancy was confirmed by palpation per rectum 40-70 days after AI. Out of a total of 91 cases, 39 (42.9%) had normal ovarian cycles (ovulation within 45 days after calving, followed by normal ovarian cycles), 32 (35.2%) had prolonged luteal phase (i.e. luteal activity for >20 days), and 12 (13.2%) had anovulation or delayed first ovulation (i.e. first ovulation did not occur until >45 days after calving). The remaining (8.8%) had other types of abnormalities. When compared with cows with a normal ovarian cycle, prolonged luteal phase cows had a lower 100 days AI submission rate, conception rate and pregnancy rate (84.2% versus 56.3%; P<0.05, 50% versus 16.7%; P<0.05 and 42.1% versus 9.4%; P<0.01, respectively), and longer intervals to first AI and to conception ( 67+/-6 days versus 98+/-7 days and 95+/-9 days versus 136+/-11 days; P<0.01 for each). Similarly, when compared with cows with normal ovarian cycles, those with anovulation had lower 100 days conception rate and pregnancy rate (50% versus 0%; P<0.05 and 42.1% versus 0%; P<0.01, respectively), and longer intervals to first AI and to conception ( 67+/-6 days versus 93+/-12 days; P<0.05 and 95+/-9 days versus 155+/-14 days; P<0.01, respectively). Survival analysis of the data for calving to conception interval showed that cows with prolonged luteal phase or anovulation were getting pregnant at a slower rate, and took longer to get pregnant than the cows with normal resumption of ovarian cyclicity postpartum. In conclusion, abnormal ovarian cycles during the pre-service period postpartum adversely affected reproductive performance, including AI submission rate, pregnancy rate, interval to first AI, and calving to conception interval in high-producing Holstein cows.     

Importance of using guarded techniques for the preparation of endometrial cytology smears in mares

Material for endometrial cytology can be collected by veterinarians using guarded or unguarded swabs, or digitally with a gloved hand, and is an important diagnostic tool in establishing the endometrial health of mares prior to breeding. The aim of this study was to determine whether the use of unguarded endometrial samples is a reliable indicator of the presence of neutrophils in the uterus. Duplicate endometrial smears were collected from 41 genitally normal, non-pregnant fertile mares by both double-guarded swabs (DGS) and in an unguarded manner by digital scraping (DS) of the endometrium. In 17 of the 41 mares, smears were also collected from the cranial vagina by DS. Cytological samples were collected from a further seven non-pregnant mares at different reproductive stages, and tissues (vestibule, vagina and cervix) from four reproductively normal mares were examined histologically after slaughter to detect the presence of neutrophils. Only 3/41 (7.3%) of the DGS endometrial smears had neutrophils present compared to 36/41 (87.8%) of the DS endometrial smears. The percentage of neutrophils in DGS endometrial smears ranged from 0 to 6% (mean = 0.41%), whereas those in the DS smears ranged from 2 to 90% (mean = 22.02%). Neutrophils were present in all vaginal smears (17/17, range=3-56% (mean = 22.18%)). There was a significant positive correlation (r = 0.84; P < 0.001) between the percentage of neutrophils in the vagina and in the DS endometrial smears. More neutrophils were found in the cervix, vagina and vestibule than in endometrial smears during the cycle (P<0.05). Neutrophils were also observed in tissue collected from the cervix, vagina and vestibule from reproductively normal mares at post-mortem. In conclusion, endometrial smears collected using unguarded techniques are very likely to be contaminated with neutrophils transferred from the vagina potentially leading to incorrect diagnosis of endometritis. When collecting samples for endometrial cytology it is important to use guarded techniques to ensure that only the endometrium is sampled to avoid contamination with cells carried over from other areas of the reproductive tract.     

Gamma-aminobutyric acid in the genital tract of the rat during the oestrous cycle

The highest values of gamma-aminobutyric acid (GABA) in the genital tract of the rat at different stages of the oestrous cycle were found in the oviduct (3.5-7 micrograms/mg protein) and the lowest in the ovary (50-100 ng/mg protein). The values for uterus and vagina ranged between 80 and 150 ng/mg protein. GABA (10-30 ng/microliter) was also found in fluid in the ovarian bursa. At 11:00 h, on the day of oestrus, GABA content increased in the ovaries but values in the oviducts were maximal at 11:00 h on the day of pro-oestrus. Variations in GABA content of the vagina were also found. Uterine cervix or uterine horn showed no changes during the oestrous cycle. The GABA content was not uniform throughout the oviduct: the highest values were found in the portion next to the ovary. At 10 days after removal of the right oviduct, GABA values in the ovary and ovarian bursa fluid decreased on the operated side. At 1 month after surgery, the values in ovary were normal but the values in ovarian bursa fluid were still low, suggesting that the source of ovarian GABA was not the oviduct. The variations observed in the present paper suggest an involvement of GABA in reproductive physiology.     

Anatomy and histology of the reproductive tract of the female Babirusa (Babyrousa celebensis)

The anatomy and histology of the female reproductive tract of the Indonesian wild pig Babyrousa celebensis was studied by means of reproductive tracts obtained from seven animals aged between two and 22 years of age. The ovary appeared to have the ability to ovulate up to four ova at one time. However, the combined ovarian output seemed to average 1.86 ova. Ovulation can take place at any time from puberty to old age (22 years). The opening to the uterine tube was indicated by a ‘flower-like' array of tall, broad epithelial ‘petals' arising from the luminal surface of the funnel. The mucosal surfaces of these structures were covered in a mixture of prominent ciliated cells and bulbous secretory cells. The uterine tube followed a tightly convoluted path to the tip of the uterine horn. The uterus was proportionately short. The anatomical construction of the uterus was similar to those of other suids in that the columnar endometrium was heavily folded, there was a rich supply of uterine glands in the lamina propria, and the uterus was provided with a good blood supply. The cervix was thick walled and had a spiral lumen.     

Effect of diet and GnRH administration on post-partum ovarian cyclicity in autumn-lambing ewes

Using autumn-lambing ewes, this study investigated (i) the effects of diet on gonadotrophin secretion and responsiveness of the hypothalamic-pituitary-ovarian axis to exogenous GnRH during the early post-partum period; and (ii) whether ovulation prior to completion of uterine involution results in an increased incidence of aberrant ovarian cycles. Thirty-two ewes rearing 1.9+/-0.12 lambs were equally allocated to two dietary treatments at lambing (22 October +/-0.2 day). Diets comprised ad libitum hay and 1.5 kg per ewe per day of one of two concentrates (11.5 MJ ME, 195 g CP per kg) containing 300 g kg(-1) cracked maize grain (M) or 300 g kg(-1) sugar beet pellets (S). Half of the ewes on each diet (G) received 25 i.v. injections of 250 ng GnRH in 2 ml 0.9% saline at 2 h intervals from days 12-14 post-partum while remaining ewes (N) were monitored for the resumption of spontaneous ovarian cyclicity. Blood samples were obtained from all ewes throughout the study (lambing to 18 December) for measurement of circulating hormone concentrations and the uteri and ovaries of all ewes were examined via laparoscopy on day 21 post-partum. There were no effects of dietary treatment on ewe daily live weight loss, lamb daily live weight gain or the immediate post-partum increase in circulating FSH concentrations. Diet did not affect insulin concentrations or LH pulse frequency on day 12 post-partum but LH pulse amplitude was lower in ewes fed concentrate M compared to concentrate S (1.4+/-0.10 versus 1.7+/-0.12 ng ml(-1), respectively, P<0.05) and this was associated with an increased interval to the resumption of spontaneous ovarian cycles (35+/-3.1 versus 26+/-2.1 day, respectively, P<0.05). Administration of exogenous GnRH increased (P<0.05) the proportion of ewes on both diets that ovulated within 20 days of parturition and advanced the onset of ovarian cyclicity in ewes fed concentrate M by 9.5 days (significance of interaction, P<0.05). Four ewes, all of which ovulated before day 22 post-partum, had extended luteal activity while in remaining ewes, duration of the first luteal phase was inversely related to the time of first ovulation (r(2)=0.16, P<0.05). Results demonstrate that (i) the onset of ovarian cyclicity is influenced by diet and can be advanced by administration of exogenous GnRH; and (ii) ovulation during the early post-partum period is associated with an increased incidence of extended luteal activity.     

Assessment of development of the testes and accessory glands by ultrasonography in bull calves and associated endocrine changes

The testes, prostate and vesicular glands of 10 bull calves were examined by ultrasonography every 2 wk from 2 to 46 wk of age, at which time the scrotal circumference (SC) of all the calves had reached pubertal size (28 cms). Computer-assisted image intensity analysis (numerical pixel values) was conducted. Blood samples were collected every other week from 2 to 46 wk of age. Testicular diameter increased in a linear manner from 2 to 46 wk of age, but the diameter measured in a transverse plane (caudal) was greater between 10 and 34 wk of age than when measured in a longitudinal (lateral) plane (P<0.05). Growth of the prostate and vesicular glands, based on dimensions, was linear, but vesicular gland length increased more rapidly after 32 wk of age (P<0.05). Image intensity of the vesicular glands and prostate declined from birth or 8 wk of age, respectively, to 14 wk of age, increased to 18 wk and then declined to a nadir at 30 wk, followed by a rapid increase to 34 wk of age for the vesicular glands and to 46 wk of age for the prostate (P<0.05). Image intensity of the testes showed an early increase to 6 to 8 wk of age and a subsequent increase from about 20 wk of age to 46 wk of age, with an inflection at 30 wk of age (P<0.05). There was a transient increase in mean serum concentrations of LH between 6 and 20 wk of age (P<0.05), and LH concentrations appeared to increase again after 36 wk of age (P>0.05). Mean serum concentrations of FSH declined with age (P<0.05). Mean serum concentrations of testosterone increased after 32 wk of age (P<0.05) In summary, numerical pixel values comprising the ultrasound images of the developing testes, prostate and vesicular glands revealed a complex development pattern that may reflect important details of developmental stages.     

Ovulation rate and distribution in the thoroughbred mare, as determined by ultrasonic scanning: the effect of age

Ovarian activity was monitored in 2181 oestrous cycles from 1136 thoroughbred brood mares to accurately determine overall ovulation rate, the relative distribution of ovulations between left and right ovary, whether mare age had an effect on these parameters and whether ovulation pattern in one cycle affected the pattern within the next. Ultrasonic scanning was used in preference to rectal palpation and slaughterhouse material due to its greater accuracy. Mare's ages ranged from 3 to 22 years. The results obtained indicated a similar incidence of ovulation occurring on the right (49.5% of all ovulations) or left ovary (50.5%). Multiple ovulations (MO) occurred in 22.4% of oestrous cycles. Significantly (P<0.01), more double ovulations (DO) were bilateral (57.2%) than unilateral (42.8%); 20.7% were unilateral right and 22.1% were unilateral left. There were 828 mares grouped into six age groups: A (3-5 years), B (6-8 years), C (9-11 years), D (12-14 years), E (15-18 years), F (18-22 years). The distribution of ovulation between the left and right ovary was not affected by age group, but significant association (P<0.01) was evident between age and MO. Group A had 15% of cycles resulting in MO compared to 19.4, 24.6, 25.8, 28.6 and 35.1% in groups B-F, respectively. A significant (P<0.01) association between MO in successive cycles was also demonstrated, with a greater chance (P<0.05) of successive single ovulations (SO) being on alternate ovaries. There was no significant repeatability of the classification of DO in one cycle compared to the next. We conclude that: (i) the mare is a symmetrical ovulator; (ii) 22.4% of cycles yield MO (ovulation rate 1.23); (iii) this MO incidence increased (P<0.01) with age; (iv) the exhibition of MO in one cycle was linked to an increased probability (P<0.01) of a MO occurring at the next one and (v) SO in one ovary was more likely (P<0.05) to be followed by a SO on the alternate ovary during the next cycle.     

Dominant activation of the hedgehog signaling pathway alters development of the female reproductive tract

The role of hedgehog (HH) signaling in reproductive tract development was studied in mice in which a dominant active allele of the signal transducer smoothened (SmoM2) was conditionally expressed in the Müllerian duct and ovary. Mutant females are infertile, primarily because they fail to ovulate. Levels of mRNA for targets of HH signaling, Gli1, Ptch1, and Hhip, were elevated in reproductive tracts of 24-day-old mutant mice, confirming overactivation of HH signaling. The tracts of mutant mice developed abnormally. The uterine luminal epithelium had a simple columnar morphology in control mice, but in mutants contained stratified squamous cells typical of the cervix and vagina. In mutant mice, the number of uterine glands were reduced and the oviducts were not coiled. Expression of genes within the Hox and Wnt families that regulate patterning of the reproductive tract were altered. Hoxa13, which is normally expressed primarily in the vagina and cervix, was expressed at 12-fold higher levels in the uterus of mutant mice compared with controls. Wnt5a, which is required for development of the cervix and vagina and postnatal differentiation of the uterus, was expressed at higher levels in the oviduct and uterus of mutant mice compared with controls. Mating mutant females with fertile or vasectomized males induced a severe inflammatory response in the tract. In summary, overactivation of HH signaling causes aberrant development of the reproductive tract. The phenotype observed could be mediated by ectopic expression of Hoxa13 in the uterus and elevated levels of Wnt5a in the oviducts and uterus.     

Effect of low colostrum intake on gastrointestinal development and uterine and cervical morphometrical architecture in the neonatal gilt

To assess the importance of natural variation in colostrum intake on piglet gastrointestinal and reproductive development, two equally sized female piglets from each of 27 litters were selected, one with low (average 226 g) and one with high (average 401 g) colostrum intake. At weaning (23 d of age), piglets were euthanised to perform macromorphological measurements on ileum, colon, cervix and uterus tissues, and to obtain tissue samples from the cervix and uterus for histology. Sections of uterine and cervical preparations were analysed using digital image analysis. Despite being selected for the same birth weight (average 1.1 kg, standard deviation 0.18 kg), piglets with low colostrum intake weighed 5.91 ± 0.17 kg and piglets with high colostrum intake weighed 6.96 ± 0.19 kg at weaning (P < 0.05). Most of the micro- and macroscopic measures such as length and weight of ileum and colon, cervix and uterus, luminal size of cervix and uterus, number of cervical crypts and uterine glands, were greater in gilts with high colostrum intake. The histological architecture of the uterus and cervix in gilts with high colostrum intake showed more complexity, reflecting more advanced development in these piglets. In conclusion, these data demonstrate that independent of birth weight, natural variation in colostrum intake is related to the overall development of neonatal piglets, affecting body growth, as well as growth and development of the gut and reproductive tract.     

Ultrasonographic image attributes of non-ovulatory follicles and follicles with different luteal outcomes in gonadotropin-releasing hormone (GnRH)-treated anestrous ewes

Ultrasonographic images are composed of multiple square picture elements called pixels. Quantitative changes in numerical pixel values (echotexture) determined by computer-assisted analysis of digital images reflect discrete changes in the microscopic structure and physiological status of ovarian antral follicles. The objective of the present study was to determine and compare the ultrasonographic attributes of non-ovulatory antral follicles that grew to an ostensibly ovulatory diameter (> or =5mm) and follicles with different luteal outcomes in response to gonadotropin-releasing hormone (GnRH) in anestrous Western White Face ewes (n=34). All animals received GnRH injections (250ng i.v. every 2h for 24h) followed by a bolus injection of 125microg of GnRH i.v. Ovarian images obtained by repeated transrectal ultrasonography were digitized and subjected to computerized analyses to determine the changes in follicular size and echotexture of the follicular antrum and wall. At the beginning of GnRH treatment, follicles that formed inadequate corpora lutea following ovulation (ICL; n=22) had higher (P<0.001) pixel intensity of the central and peripheral antrum compared with non-ovulatory follicles (n=40). Pixel intensity of the central follicular antrum was greater (P<0.01) in follicles that formed ICL compared with follicles that formed normal (full-lifespan) CL post-treatment (NCL; n=20) and mean pixel heterogeneity of the follicular wall was greater (P<0.05) in non-ovulatory follicles compared with follicles that gave rise to NCL. At the time of GnRH bolus injection (i.e., induction of a synchronous LH surge), the mean diameter of non-ovulatory follicles was greater (P<0.01) than that of all ovulating follicles, and pixel heterogeneity of the central follicular antrum was lowest (P<0.05) in non-ovulatory follicles. The mean diameter of luteinized unovulated follicles (n=9) tended to be greater (P<0.10) at 2.5 and 3 days after emergence, and pixel intensity of the follicular wall was lower (P<0.05) compared with non-luteinized follicles (n=8) at 1.5 and 2.5 days after emergence (beginning of the growth from approximately 3mm onwards). In conclusion, ovarian antral follicles with different outcomes after GnRH treatment (in seasonally anestrous ewes) had distinctive ultrasonographic characteristics.     

The effect of long non-reproductive periods on the genital health in captive female white rhinoceroses (Ceratotherium simum simum, C.s. cottoni)

White rhinoceroses suffer from a low reproductive rate in captivity. Intensive efforts to propagate specifically the northern white rhinoceros have been very limited. The dismal outlook for this subspecies in the wild makes successful ex situ breeding programs paramount. In this context, this study examined 48 southern and 6 northern white rhinoceroses using ultrasound and faecal hormone analysis to elucidate causes for female reproductive failure and to determine whether long non-reproductive periods have a detrimental impact on genital health. Results showed that 76% of the nulliparous females had intact hymenal membrane indicating these females had never been bred, at an age when their wild counterparts have delivered multiple offspring. Fifty-six percent of the studied population had various reproductive pathology. Cystic endometrial hyperplasia; leiomyomas of the cervix, uterus and ovary, adenoma; para-ovarian cysts and hydromucometra represent the scope of lesions identified. The stages of the lesions in nulliparous females correlated with age (r = 0.4, P < 0.05). Due to the severity of the lesions, 28% of the study population was considered post-reproductive. Therefore, the reproductive life span in some individuals was 10-20 years shorter than expected. However, in parous females the incidence of pathological lesions was significantly lower (P < 0.0001). Seventy-eight percent females studied had erratic or absent luteal activity. The hormone data corresponded with two ultrasonographic levels of ovarian activity, active and inactive, occurring within an age range of 3-19 years and 15-38 years, respectively. This suggests the lack of ovarian activity by reproductive mid-life in non-reproducing females. The accuracy of the ultrasound findings was validated by necropsy in nine animals showing a strong positive correlation (r2 = 0.9, p < 0.001). Our data suggests that the development of reproductive pathology and ovarian inactivity in white rhinoceros is an age-related consequence of long non-reproductive periods. This asymmetric ageing process of the genital organs can be prevented with the achievement of at least one pregnancy.     

Effect of subclinical uterine infection on cervical and uterine involution, estrous activity and fertility in postpartum buffaloes

Nili-Ravi buffaloes (n=29) that calved normally between August and November and did not develop any clinical reproductive disorder after calving were studied for the incidence of sub-clinical bacterial infection of the uterus and its effects on postpartum reproductive efficiency. The incidence of subclinical uterine infection was 24% (7/29). Involution of the cervix and uterus was slower (P < 0.01) in the infected group than in the normal group (45.6 vs 31.1 days and 46.3 vs 35.8 days), respectively. The mean diameters of cervix and gravid horn on Day 12 post partum and on completion of involution did not differ between buffaloes of the two groups. However, the rate of involution of the cervix and the gravid horn was lower in buffaloes of the infected group (2.2 vs. 2.7 mm/day and 2.6 vs. 3.2 mm/day). The mean interval to first post partum ovulation was similar in buffaloes in the infected (35.5 days) and the normal group (33.8 days). The life span of corpus luteum formed after first ovulation was shorter (11 days) in buffaloes of both groups than that of a normal estrous cycle (15 to 17 days). The incidence of silent ovulation was apparently higher in buffaloes of the infected group (83 vs. 60%) but the difference was not significant. For the first four months after calving, the mean interval to first postpartum estrus was longer in buffaloes of the infected group (73.0 vs. 47.7 days; P < 0.01). Similarly, the average service period was longer in buffaloes of the infected group (91.0 vs. 64.8 days; P < 0.05). The overall pregnancy rate for the first four months after calving did not differ between buffaloes of the two groups. We conclude that subclinical bacterial infection of the postpartum uterus delays the cervical and uterine involution which can, in turn, delay the occurrence of first postpartum estrus and prolong the service period in buffaloes.     

Differential ovarian expression of KiSS-1 and GPR-54 during the estrous cycle and photoperiod induced recrudescence in Siberian hamsters (Phodopus sungorus)

Kisspeptins, coded by the KiSS-1 gene, regulate aspects of the reproductive axis by stimulating GnRH release via the G protein coupled receptor, GPR54. Recent reports show that KiSS/GPR54 may be key mediators in photoperiod-controlled reproduction in seasonal breeders, and that KiSS-1/GPR54 are expressed in the hypothalamus, ovaries, placenta, and pancreas. This study examined the expression of KiSS-1/GPR54 mRNA and protein in ovaries of Siberian hamsters (Phodopus sungorus). Ovaries from cycling hamsters were collected during proestrus (P), estrus (E), diestrus I (DI), and diestrus II (DII). To examine KiSS-1/GPR54 during stimulated recrudescence, additional hamsters were maintained either in long day (LD 16L:8D, control) or short day (SD 8L:16D) for 14 weeks and then transferred to LD for 0-8 weeks. Staining of KiSS-1/GPR54 protein was detected by immunohistochemistry in steroidogenic cells of pre-antral and antral follicles, and corpora lutea. Immunostaining peaked in P and E, but decreased in the diestrus stages (P < 0.05). In recrudescing ovaries, KiSS-1/GPR54 immunostaining was low after 14 weeks of SD exposure (post-transfer [PT] week 0), and increased during the early weeks of recrudescence. Expression of KiSS-1/GPR54 mRNA was low with short day exposure, but increased during recrudescence and was higher at PT week 8 as compared to PT weeks 0 and 2 (P < 0.05). The elevated KiSS-1/GPR54 expression during P and E suggests a potential role in ovulation in Siberian hamsters. Transient increases in KiSS-1/GPR54 expression following LD stimulation are also suggestive of possible involvement in ovulation and/or restoration of ovarian function.     

“In vivo” pH values of the estrous reproductive tract of the gilt

The hydrogen ion concentration of the fluids of various portions of the reproductive tract was determined in 6 estrous gilts. Determinations were made with a pH cardiovascular catheter electrode placed in the lumen of the oviduct, uterus, cervix and vagina and a reference electrode placed in the peritoneal cavity. The mean pH values were 7.02, 6.98, 7.49, and 6.61 for the oviduct, uterus, cervix and vagina, respectively.