Biochemical and hormonal characterization of follicles from follicular and luteal phase ovaries of goat and sheep.

Follicles from goat and sheep ovaries were characterized for their biochemical and hormonal parameters to investigate the effect of developmental stage of follicles on ovarian steroidogenesis. The follicles were isolated mechanically from follicular and luteal phase ovaries and divided in 6 morphologically different groups (small, medium and large follicular and small, medium and large luteal). Follicles were characterized for their contents of protein, DNA, estradiol-17 beta and progesterone and the activity of 3 beta-hydroxysteroid dehydrogenase. There was a progressive increase in the contents of all these biomolecules and activity of the enzyme as size of follicles increased in both the follicular and luteal phase ovaries. Follicles from follicular phase ovaries exhibited higher estradiol-17 beta content than those shown by luteal phase follicles. The reverse pattern was obtained for progesterone content. The results provide the basic data on biochemical and hormonal entities at different stages of follicular development in small ruminants which may be useful for in vitro studies on regulation of follicular development and steroidogenesis.     

Concentrations of progesterone,testosterone and estradiol-17β in the serum during the estrous cycle of Asian elephants (Elephas maximus)

The levels of progesterone, testosterone and estradiol-17β in serum samples from two female Asian elephants were measured for the period of 32 months from February 1987 to September 1989. Serum samples were collected weekly from unanesthetized elephants. Each elephant showed eight ovarian cycles in 32 months. Ovarian cycles, characterized by changes in concentrations of serum progesterone, averaged 16.8 ± 0.6 (mean ± SEM. n = 14) weeks in length. The changes in concentrations of testosterone in the serum showed a similar pattern to those of progesterone with a striking increase noted during the luteal phase. The highest levels of serum estradiol-17β were noted when progesterone levels showed low basal values. These results suggest that estradiol-17β may be an index of follicular maturation during the estrous cycle in Asian elephants, and that the ovaries of Asian elephants may produce testosterone in the luteal phase.     

Ultrasonographic and laparoscopic evaluation of the reproductive tract of the captive female African lion (Panthera leo)

The use of transabdominal ultrasonography to assess the oestrous cycle has not been previously described in the African lion (Panthera leo). Twelve sexually mature lionesses and five female cubs had their reproductive organs assessed by transabdominal ultrasound. Ovarian findings were compared to laparoscopic findings while performing laparoscopic ovariectomy or salpingectomy. Vaginal cytology was performed and serum progesterone levels were determined. By combining all data the oestrous cycle stage of each lion was determined. One lion was far pregnant and was not operated on. In adults a uterine body could be seen ultrasonographically in 67% of lions while mural structures could be distinguished in 44% of lions. Five uterine horns could be seen in 3 lions. In 12 adults 10 ovaries were found of which eight had discernable follicles or luteal structures. During laparoscopy 12 active ovaries were seen with luteal structures seen in 11 ovaries and follicles in 2 ovaries. Using laparoscopy as the gold standard, ultrasonography had a sensitivity of 66% and specificity of 83% to detect ovarian reproductive activity. Two uterine cysts and a cluster of periovarian cysts were seen in three different lions. Three lions were pregnant, two were in oestrus, three in a luteal phase (dioestrus), and four were in anoestrus. Transabdominal ultrasound in combination with serum progesterone levels and vaginal cytology can be used to assess ovarian cyclical activity with reasonable accuracy in captive bred lions.     

Gonadotropin releasing hormone treatment of dairy cows with ovarian cysts. I. Gross ovarian morphology and endocrinology

Dairy cows diagnosed as having ovarian cysts were assigned to receive either sterile water or 100 mug GnRH (5 cows/group). Immediately prior to treatment and three days post-treatment, ovaries were observed via paralumbar laparotomy, photographed and visible structures and ovarian size recorded. Nine to thirteen days post-treatment, ovaries were removed. Blood plasma was collected for hormone determinations prior to each surgery, 1.5 and 3.0 hours and 1, 5 and 9 days post-treatment. Although concentrations were similar between groups prior to treatment, concentrations of progesterone were higher and LH and estradiol-17beta lower for GnRH treated cows than control cows, immediately prior to ovariectomy. A layer of luteal tissue approximately 5 mm thick was present around the periphery of the cystic structure at ovariectomy in 4 of 5 GnRH treated cows, but in only one control cow. The thickness of the luteal layer around the periphery of the ovarian cysts was correlated -.82, .78 and -.63 with estradiol-17beta, progesterone and LH, respectively. In summary, response to GnRH treatment in cows with ovarian cysts appears to be characterized in most cases by luteinization of the cystic structures.     

Serum LH,progesterone and estradiol-17β levels throughout the ovarian cycle,during the early stage of pregnancy and after the parturition and the abortion in the common marmoset,Callithrix jacchus

In the ovarian cycle of common marmosets, serum progesterone began to increase at two to three days after estradiol-17β or LH surge, attained a peak of 25–70 ng/ml and then declined to a level of under 2 ng/ml before the ensuing rise in estradiol-17β and LH. Serum estradiol-17β increased to 700–5,500 pg/ml during the luteal phase, synchronizing with progesterone. It is suggested that the corpus luteum secreted estradiol-17β as well as progesterone. The cycle length as determined from the interval between successive LH surges was approximately 28 days. During the luteal phase, the levels of progesterone and estradiol-17β were higher than in Old World monkeys and women, but marmosets were not accompanied by any clinical symptoms due to excessive progesterone and estradiol-17β. This suggests that such unresponsiveness to progesterone and estradiol-17β in marmosets reflects the small amount of estradiol-17β receptor and presumably also the lower function of the post receptor system. Recovery of the post-partum ovarian cycle in two marmosets differed from that observed in Old World monkeys and women. The first LH surge was found on the ninth and tenth day after parturition and the first ovulation led to the next pregnancy. This suggests that the suckling stimulus of newborns in the common marmoset does not cause any delay in recovery of the ovarian cycle. In three cases of abortion, the recovery of the ovarian cycle was almost the same as that in the case of normal parturition: the first LH surge appeared on the 10th, 14th, and 34th day after abortion.     

Morphological changes of the ovary and hormonal changes through the ovarian cycle of the common marmoset (Callithrix jacchus)

Laparoscopic observations of morphological changes of the ovary during the ovarian cycle in conjunction with radioimmunoassay of serum progesterone and estradiol-17β was investigated as a method of monitoring the ovarian cycle in the common marmoset. In the common marmoset, plural follicles first appeared in each ovary five days prior to ovulation. At three to four days prior to ovulation one or two follicles developed into translucent blisters on the surface of the ovary. As the follicles filled with follicular fluid, they became larger and clearer until one to two days prior to ovulation, at which time they formed well defined, transparent bubbles protruding from the surface of the ovary. After ovulation, the ovulation point could be detected at the center of the follicle, sometimes surrounded by a corpus of engorged blood vessels. Ovulations of the plural follicles were not simultaneous, and due to the time lag ovulations took at least 12 to 20 hrs in four out of seven animals examined. After two to five days of ovulation the corpus hemorragicum, a bright red protrusion made of tissue and blood disrupted by ovulation, was found. Subsequently, the color of the formatted corpus luteum changed from dark-red to yellow then to yellow white. While the corpus luteum remained reddish in color serum progesterone was maintained at as high levels as in the luteal phase. There was no mature follicle or corpus luteum in subordinate female ovaries.     

Progesterone receptor is not required for progesterone action in the rat corpus luteum of pregnancy

In this study, we investigated whether progesterone exerts a local action regulating the function of the corpus luteum of pregnancy in rats. The luteal activities of the enzymes 3beta-hydroxysteroid dehydrogenase (3beta-HSD), involved in progesterone biosynthesis, and 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD), that catabolizes progesterone and reduces progesterone secretion by the corpus luteum, were evaluated after intrabursal ovarian administration of progesterone in pregnant rats that had received a luteolytic dose of prostaglandin F2alpha (PGF2alpha). Luteal 3beta-HSD activity decreased and 20alpha-HSD activity increased after PGF2alpha treatment (100 microg x 2 intraperitoneally on Day 19 of pregnancy at 12:00 p.m. and 4:00 p.m.) when compared with controls sacrificed at 8:00 p.m. on Day 20 of pregnancy. This effect of PGF2alpha on the luteal 3beta-HSD and 20alpha-HSD activities was abolished in animals that also received an intraovarian dose of progesterone (3 microg/ovary on Day 19 of pregnancy at 8:00-9:00 a.m.). In a second functional study, luteal cells obtained from 19-day pregnant rats responded to the synthetic progestin promegestone (R5020) in a dose-dependent manner, with an increase in the progesterone output. In addition, the glucocorticoid agent hydrocortisone did not affect progesterone accumulation in the same luteal cell culture. We also examined by immunocytochemistry the expression of progesterone receptors (PR) in the corpora lutea during pregnancy and demonstrated the absence of PR in this endocrine gland in all the days of pregnancy studied. In the same pregnant rats, positive staining for PR was observed in cells within the uteroplacental unit, such as cells of the decidua basalis and trophoblast giant cells of the junctional zone. In addition, positive PR staining was observed in the ovarian granulosa and theca cells of growing follicles, but not in corpora lutea of ovaries obtained from cycling rats at proestrus. In summary, this report provides further evidence of a local action of progesterone regulating luteal function in the rat despite the absence of a classic PR.     

Female reproductive patterns in nonhibernating bats

The major reproductive events in the oestrous cycles of nonhibernating mega- and microchiropteran species are reviewed. However, special attention is given to the reproductive biology of the temperate North American species, Tadarida brasiliensis mexicana (Molossidae), the Mexican free-tailed bat, which expresses dextral uterine and ovarian dominance. Only the larger right ovary is capable of producing an ovulatory follicle and the left has long been considered atrophic. In order to elucidate the normal oestrous cycle and define the structural and functional characteristics of the ovaries of this nonhibernating bat several analytical technique were utilized. These included light microscopic analysis, histochemical localization of delta 5-3 beta-hydroxysteroid dehydrogenase (HSD) activity, and radioimmunoassay of seasonal plasma progesterone levels. Interstitial tissue was found in both ovaries, but the left was almost entirely an interstitial organ. Histochemical analysis demonstrated that the gonads have seasonally varying amounts of delta 5-2 beta-HSD localized either in the thecal cells of the Graafian follicle or in the interstitium. The corpus luteum persisted throughout gestation, reaching maximum development just prior to parturition. Circulating progesterone levels correlated directly with luteal gland size and peaked at 106 ng/ml when the corpus luteum was largest.     

Monitoring the reproductive status of female gorillas (Gorilla gorilla gorilla) by measuring the steroid hormones in fecal samples

The reproductive status of female gorillas (Gorilla gorilla gorilla) was estimated by measuring the sex steroid hormones in fecal samples instead of in blood samples. Fecal samples from female gorillas were used to examine the reliability of this non-invasive assay system, which included the extraction method for estradiol-17β (E₂) and progesterone (P) from fecal samples. Fecal samples from three female gorillas were collected daily for about 55 days, and fecal E₂ and P were assayed to clarify the fluctuation patterns of these steroids in the feces. Fecal sampling from one female was repeated for another 50-day period (starting 75 days after the end of the first observation period) and assayed to confirm if the menstrual cycle of this subject was ovulatory. Although fecal E₂ concentration measurements were quantitative by using this assay system, fecal P concentration measurements were semi-quantitative. Relative amounts of fecal P in fecal samples were estimated by using the values of B/B₀ (bound/total binding in the radioimmunoassay system). Two of the four fluctuation patterns of fecal hormones observed throughout the menstrual cycle for the three female gorillas were typical for normal ovulatory cycles. In the subject observed for two periods, one pattern was typical and the other atypical. The results show that this non-invasive method is simple and practical for monitoring the reproductive status of great apes as well as Old World monkeys.     

Luteal function in cows following destruction of ovarian follicles at midcycle

Luteal function was studied in the absence of non-ovulatory ovarian follicles to determine if these follicles are involved in luteal regression in cattle. After at least one estrous cycle, cows were assigned randomly to treatment (n=5) or control (n=5). All cows were laparotomized on day 10 postestrus (Estrus = day 0). During laparotomy of treated cows, all visible follicles on both ovaries were destroyed by electrocautery, and follicular growth was prevented by ovarian x-irradiation. In controls, laparotomy and ovarian manipulation were as in treated cows but follicles were not destroyed and ovaries were not irradiated. On day 22 postestrus, ovaries of 4 treated cows contained no visible follicles and concentrations of estradiol-17beta in jugular plasma (0.4 +/- 0.1 pg/ml) were less (P<0.05) than in controls (3.2 +/- 0.4 pg/ml). Daily mean concentrations of LH from surgery to day 22 postestrus in treated cows did not differ from controls. On day 22 postestrus, progesterone in jugular plasma and weights of corpora lutea in treated cows were greater (P<0.05) than in controls. Between days 12 and 18 postestrus, concentrations of estradiol-17beta and PGF(2)alpha in utero-ovarian venous plasma of controls increased prior to detectable declines in concentrations of progesterone. Therefore, non-ovulatory ovarian follicles present during mid to late diestrus are necessary for luteal regression in non-pregnant cattle.     

Fecal steroid metabolites and reproductive monitoring in a female Tsushima leopard cat (Prionailurus bengalensis euptilurus)

Although the Tsushima leopard cat (Prionailurus bengalensis euptilurus) is one of the most endangered mammals in Japan, its reproductive physiology and endocrinology have been not elucidated. The objective was to establish the non-invasive monitoring of reproductive endocrinology in a female Tsushima leopard cat and to identify the types of fecal reproductive steroid metabolites in this species. Fecal concentrations of estrogen and progestin were determined by enzyme immunoassays, from 60 d before to 60 d after the last copulation, during three pregnancies. Fecal estrogen metabolite concentrations were increased before/around the mating period and after mid-pregnancy. Fecal progestin metabolite concentrations increased after the last copulation and remained high during pregnancy. The gestation period was 65.0 ± 0.6 d (mean ± SD). Fecal extracts were separated by high-performance liquid chromatography for identification of fecal metabolites. Fecal estrogens were identified as estradiol-17β and estrone. Fecal progestins during pregnancy contained 5α-reduced pregnanes: 5α-pregnan-3α-ol-20-one, 5α-pregnan-3β-ol-20-one and 5α-pregnan-3,20-dione, and nonmetabolized progesterone was barely detected in feces. In conclusion, measurement of fecal estrogen and progestin metabolites was effective for noninvasive reproductive monitoring in the Tsushima leopard cat. An immunoassay for fecal estradiol-17β concentrations seemed useful to monitor follicular activity, whereas an immunoassay with high cross reactivity for 5α-reduced pregnanes was useful to monitor ovarian luteal activity and pregnancy.     

Ultrasonographic and laparoscopic evaluation of the reproductive tract in older captive female cheetahs (Acinonyx jubatus)

The study uniquely described the clinical value of transabdominal ultrasonography for monitoring features characterizing the estrous cycle in female cheetahs (Acinonyx jubatus). The reproductive tracts of 21 female, nulliparous, and relatively aged (median: 11 and interquartile range: 9.25–14 years) captive cheetahs resident on two sites in Namibia were assessed by transabdominal ultrasound. Subsequently, the ovarian findings on ultrasound were compared with direct visualization while performing laparoscopic sterilization. A combination of these observations supported by concurrent sampling for vaginal cytology and serum progesterone concentrations defined the estrous status of individual animals. At one site, six cheetahs had been implanted with the GnRH agonist, deslorelin as a contraceptive at least once within the preceding 11 years. On ultrasound, 31 uterine horns and 35 ovaries with discernible structures on 28 (86%) were visualized in the 21 cheetahs. The uterine body was difficult to visualize because of its intrapelvic location. Eleven of 19 uteri (58%) visualized showed endometrial edema suggestive of estrogenization. The uteri of four cheetahs (19%) showed evidence of mild cystic endometrial hyperplasia. Paraovarian cysts were seen on ultrasound (n = 21) and laparoscopy (n = 26) in 16 (76.2%) and 18 (85.7%) cheetahs, respectively. Ovarian volumes obtained from ultrasonographically determined dimensions predicted cyclic activity. Laparoscopy showed that 19 ovaries had discernible follicular structures. In the study population, 10 (47.6%) cheetahs were in proestrus or estrus; none in the luteal phase; and 11 (52.4%) in anestrus. Transabdominal ultrasound, in combination with serum progesterone concentrations and vaginal cytology, was used with acceptable accuracy to assess cyclic ovarian activity in captive cheetahs. A considerable proportion of this aged population showed ovarian activity and the prevalence of paraovarian cysts was notable. A history of prior deslorelin treatment was not associated with either reproductive activity or uterine pathology.     

Ovarian function in Nelore (Bos taurus indicus) cows after post-ovulation hormonal treatments

Maternal recognition of pregnancy in the cow requires successful signaling by the conceptus to block luteolysis. Conceptus growth and function depend on an optimal uterine environment, regulated by luteal progesterone. The objective of this study was to test strategies to optimize luteal function, as well as prevent a dominant follicle from initiating luteolysis. Nelore (Bos taurus indicus) beef cows (n=40) were submitted to a GnRH/PGF(2alpha)/GnRH protocol. Cows that ovulated from a dominant ovarian follicle (ovulation=Day 0) were allocated to receive: no additional treatment (G(C); n=7); 3000IU of hCG on Day 5 (G(hCG); n=5); 5mg of estradiol-17beta on Day 12 (G(E2); n=6); or 3000IU of hCG on Day 5 and 5mg of estradiol-17beta on Day 12 (G(hCG/E2); n=5). Ultrasonographic imaging of the ovaries, assessment of plasma progesterone concentration, and detection of estrus were done daily from Day 5 to the day of subsequent ovulation. Treatment with hCG induced an accessory CL, increased CL volume, and plasma progesterone concentration throughout the luteal phase (P<0.01). Estradiol-17beta induced atresia and recruitment of a new wave of follicular growth; it eliminated a potentially estrogen-active, growing ovarian follicle within the critical period for maternal recognition of pregnancy, but it also hastened luteolysis (Days 16 or 17 vs. Days 18 or 19 in non-treated cows). In conclusion, the approaches tested enhanced luteal function (hCG) and altered ovarian follicular dynamics (estradiol-17beta), but were unable to extend the life-span of the CL in Nelore cows.     

Patterns of estrogen and progesterone receptors in monkey endometrium during the normal menstrual cycle

Total concentrations of estradiol-17β (E₂) and progesterone (P) receptors (R) were measured in the endometrium of rhesus monkeys ($\text{Macaca mulatta}$) during the normal menstrual cycle. The endometrium was collected at abdominal fundal hysterotomy on days 8, 12, 15, 18 and 24 of the menstrual cycle. Visual inspection of the ovaries and measurement of E₂, P, follicle stimulating hormone (FSH) and luteinizing hormone (LH) provided assuredness of normal ovarian function. Exchange procedures were used in order to measure the total concentrations of E₂R and PR in nuclear and cytosol fractions. The pattern of estrogen receptor showed a slight increase in the cytosol and nuclear concentrations at the preovulatory interval. Later, the total E₂R concentration was decreased when P increased during the luteal phase. Cytosol PR synthesis was parallel to the serum E₂ increase during the late follicular phase. Secretion of P by the corpus luteum was accompanied by a rapid nuclear translocation and concomitant decrease in cytoplasmic PR. Thereafter the total PR concentration declined during the second half of the luteal phase. These findings in monkey endometrium are similar to those reported for human endometrium during the normal menstrual cycle and further establish the utility of these surrogate primates in investigations indicative of human endometrial function.     

Clinical analysis of reproductive failure among female buffaloes (Bubalus bubalis) under village management in Andhra Pradesh

Analysis of the clinical data of 20, 439 infertile graded Murrah female buffaloes (Bubalus bubalis ) of small and marginal farmers in Andhra Pradesh, examined over a period of two years, revealed that 2.50, 73.77 and 23.73 percent of the animals had reproductive disorders which were of anatomical, functional and non-specific infectious nature, respectively. Highest frequencies were of infantile tracts, ovarian quiescence and endometritis in each group, which together constituted 79.13 percent. The average gross incidence of various conditions encountered in order of frequency was: ovarian quiescence (56.36%), endometritis (20.68%), silent ovulations (5.3%), anovular heats (4.35%), sub-oestrus (3.48%), luteal persistency (3.37%), genital infantilism (2.09%), vaginitis (o.99%), salpingitis (0.98%), cystic ovarian degeneration (0.91%), bursal adhesions (0.75%), cervicitis (0.3%), hypoplasia of the ovaries and tracts (0.24%), paraovarian cysts (0.15%) and hydrosalpinx (0.03%). The incidence of quiescent ovaries followed seasonal trend with significant differences between seasons of the year both in heifers and cows. Between herds and between years, no marked difference in the incidence of different reproductive disorders was observed. It was estimated that 2.93 percent of the animals had disorders which rendered them unfit for breeding and, hence, sterile. Freemartinism was recorded in four cases. The comparative clinical features of freemartinism and cystic ovarian dgeneration in buffalo cows were described. The factors predisposing for higher incidence of endometritis in the species were also discussed.     

Asymmetric secretion of principal ovarian venous steroids in the primate luteal phase

We have characterized the degree of asymmetry of ovarian steroid secretion in the luteal phase of the menstrual cycle in rhesus and cynomolgus monkeys. Femoral blood levels of FSH, LH, progesterone, estradiol and 17-hydroxyprogesterone were determined. In addition, laparotomies were performed in the early, mid or late luteal phase to facilitate localization of the corpus luteum and collection of ovarian venous blood. We conclude that: 1) the ovary bearing the active corpus luteum contributes virtually all of the progesterone entering peripheral circulation in the luteal phase; 2) the ipsilateral ovary secretes more 17-hydroxyprogesterone than the contralateral one, although both are active in the luteal phase; and 3) the asymmetrical secretion of estradiol was manifest only in the early and mid-luteal phase, with ovarian symmetry being reestablished in the late luteal phase.     

Serum estradiol-17beta concentrations during spontaneous silent estrus and after prostaglandin treatment in the mare

Serum estradiol-17beta concentrations were determined during silent estrus in the mare. Relationships between serum estradiol-17beta concentration, corpus luteum regression, follicular development, ovulation, prostaglandin treatment and behavioral estrus were investigated. The expression of behavioral estrus was found to be related to the patterns of progesterone and estradiol-17beta secretion during the periovulatory period. When compared to normal estrous cycles, silent estrus was accompanied by a significantly lower maximum serum estradiol-17beta concentration (47.8 vs 34.6 pg/ml), a significantly longer interval from maximum estradiol-17beta concentration to ovulation (1.7 vs 4.0 days), and a significantly shorter interval from corpus luteum regression to ovulation (5.3 vs 2.8 days). Silent estrus following prostaglandin treatment was related to a significantly shorter interval from prostaglandin treatment to ovulation (3.6 +/- 0.4 days) than from normal corpus luteum regression to ovulation (5.3 +/- 0.3 days). Silent estrus appeared to be related to changes in follicular estradiol-17beta secretion and to the pattern of its secretion as related to regression of the corpus luteum. There appeared to be not only less estradiol-17beta present, but also less time available after luteal regression for it to interact with the central nervous system to elicit the changes necessary to cause behavioral estrus. There fore, unusual relationships between luteal function and folliculogenesis can result in one type of silent estrus. Significant correlations (P<0.05) were found between follicle size and serum estradiol-17beta concentration whenever behavioral estrus occurred [follicle diameter in mm = 0.96 (serum estradiol-17beta in pg/ml) + 6.08 and 0.73 (serum estradiol-17beta + 13.32 for control and normal estrus following prostaglandin treatment groups, respectively]. During silent estrus, however, no significant correlations between follicle size and serum estradiol-17beta concentration were observed.     

Cryptosporidium sp. and Giardia sp. infections in mountain gorillas (Gorilla gorilla beringei) of the Bwindi Impenetrable National Park, Uganda

For conservation purposes and because of growing ecotourism, some mountain gorilla (Gorilla gorilla beringei) populations have been habituated to humans. Fecal specimens (n = 100) of nonhabituated and human-habituated gorillas (5 populations; 6 age classes) were tested for Cryptosporidium sp. oocysts and Giardia sp. cysts by conventional staining and immunofluorescent antibody (IFA). Cryptosporidium sp. infections (prevalence 11%) were not restricted to very young gorillas but were observed in 3-yr-old to >12-yr-old gorillas; most of the infections (73%) occurred in human-habituated gorillas. The prevalence of Giardia sp. infections was 2%; 1 nonhabituated gorilla was concomitantly infected. Oocysts of Cryptosporidium sp. in the gorilla stools were morphologically, morphometrically, and immunologically undistinguishable from a bovine isolate of Cryptosporidium parvum oocysts. Mean concentration of Cryptosporidium sp. oocysts and Giardia sp. cysts in gorilla stools was 9.39x10(4)/g, and 2.49x10(4)/g, respectively. There was no apparent relationship between oocyst concentration and gorilla age, sex, or habituation status. Most Cryptosporidium sp. infections found in gorillas with closest proximity to people may be a result of the habituation process and ecotourism. This study constitutes the first report of Cryptosporidium sp. infections in the family Pongidae, in the free-ranging great apes, and in the species of gorilla.     

Temporal association of serum progesterone concentrations and vaginal cytology in walruses (Odobenus rosmarus)

Concentrations of serum estradiol-17β and progesterone were monitored in six female walruses using an enzyme immunoassay. Progesterone concentrations increased from March to May in females aged 6 y or older, and subsequently declined (October). No significant elevation of estradiol-17β concentration was detected before an elevation of progesterone concentration. Vaginal smears from four females were examined with Papanicolaou staining. In all females, most epithelial cells were basophilic intermediate-superficial cells; no color change from basophilic to eosinophilic of the cells was detected. Meanwhile, the percentage of anucleate cells in vaginal smears reached its highest value before the elevation of progesterone concentration, followed by an increase in the percentage of leukocytes. We inferred that the change in populations of anucleate cells and leukocytes in vaginal smears reflected ovarian status and CL formation in female walruses.     

Possible role of cyclooxygenase II in the acquisition of ovarian luteal function in rodents

The development of the corpus luteum (CL), which involves angiogenesis, is essential for the establishment of early pregnancy. We investigated the roles of the prostaglandin synthases cyclooxygenase (COX) I and COX-II in angiogenesis and progesterone production in the newly formed CL, using inhibitors of the COX enzymes and the gonadotropin-induced pseudopregnant rat as a model. Injection of indomethacin, a nonselective COX inhibitor, on the day of ovulation and the following day decreased serum levels of progesterone, as did injection of the selective COX-II inhibitor NS-398. In contrast, a selective COX-I inhibitor, SC-560, had no effect on serum progesterone concentrations. None of the inhibitors had any effect on the weight of the superovulated ovaries or on the synthesis of progesterone by cultured luteal cells. To determine whether changes in angiogenesis are responsible for the decrease in progesterone synthesis, we measured hemoglobin and CD34 levels in luteinized ovaries following injection of COX inhibitors and measured the relative frequency of cells positive for platelet-endothelial cell adhesion molecule as a specific marker for endothelial cells. All of these parameters were reduced by the COX-II inhibitors, suggesting that changes in the vasculature are responsible for the decrease in serum progesterone. Histological examination of ovarian corrosion casts indicated that NS-398 inhibited the establishment of luteal capillary vessels following the injection of hCG. The results are consistent with the hypothesis that the activity of COX-II is associated with the formation of functional CL via its stimulation of angiogenesis.