Two unanaesthetized female yellow baboons (Papio cynocephalus cynocephalus) were infused (i.v.) with [3H]oestradiol and two with [3H]progesterone, early in the follicular phases of their cycles. One month later, the two females infused with [3H]oestradiol were simultaneously infused with [14C]progesterone and [3H]dehydroepiandrosterone. All urine and faeces were collected for 96 h after infusion. The proportion of steroid excreted in faeces (versus urine) was 10.0% for oestradiol and 40% for progesterone. Peak excretion in urine occurred 4.5 h after infusion. Peak excretion in faeces occurred an average of 36.4 h after infusion, with remarkable consistency between steroids. Eighty per cent of faecal oestradiol and progesterone metabolites were excreted as free (rather than conjugated) steroids. Simply boiling (20 min) the dried faecal sample in 90% ethanol proved to be the most rapid and efficient means of extracting these steroid metabolites. High pressure liquid chromatography and immunoreactivity studies revealed that oestradiol was excreted in faeces as oestradiol (36%), oestrone (44%) and a conjugated metabolite that co-eluted with oestrone sulfate (20%). Progesterone was excreted as eight different free forms, only a minor portion of which was progesterone, and what appeared to be a conjugated metabolite that co-eluted with pregnanediol-glucuronide (20%). The free progesterone metabolites were identified by gas-chromatography-mass-spectrometry as epimers of 5-pregnane-3-diol and 5-pregnane-3-ol-one. These data suggest that currently available immunoassays for free oestradiol and oestrone should adequately characterize faecal oestrogen profiles in baboons. However, high variability in crossreactivities of various progesterone antisera to progesterone metabolites in baboons makes antiserum selection a more serious concern in attempts to quantify faecal progestogen dynamics.(ABSTRACT TRUNCATED AT 250 WORDS)
A study was conducted in captive baboons to determine (i) the impact of cereal dietary fibre on faecal progestogen excretion, and (ii) whether means of controlling dietary effects could be identified. Blood was collected on 3 days per week and faeces on 5 days per week from four unanesthetized cyclic female baboons, consecutively fed three diets of 5, 10 and 20% fibre for 90 days per diet. A 2 day lag time was detected before progesterone in the blood appeared in the faeces, regardless of diet (mean correlation was 0.62, P = 0.002). Increased dietary fibre had a negative effect on progestogen excretion (P < 0.004). Correspondence between blood and faecal progestogens was consistently greatest and the effect of dietary fibre least when faecal progestogens were expressed g-1 dry faeces. Several means of indexing faecal steroid excretion rates were examined including dehydroepiandrosterone (DHEA) and a number of byproducts of cholesterol metabolism. The cholesterol metabolite, cholestanone, was positively correlated with dietary fibre (r = 0.27; P < 0.04). Multiplying faecal progestogen concentration by the cholestanone g-1 dry faeces concentration increased the correlation between serum and cholestanone-indexed faecal progestogens (r = 0.78, P = 0.0001) compared with nonindexed progestogens (r = 0.71, P = 0.0001). We conclude that expressing faecal progestogens g-1 dry faeces may be sufficient and the most cost-effective method for controlling for most dietary effects when the objective is monitoring longitudinal endocrine status in baboons. However, it may be appropriate to express faecal progestogens by cholestanone concentrations when increased precision is needed to overcome the effects of profound variations in dietary fibre.
of Columbia (0. E. W., J. L. B., M. B., M.A.B., K.A.C., J.G. H.), and the Oklahoma City Zoological Park, Oklahoma City, Oklahoma (J. G.)Under the mandate of a Species Survival Plan (SSP), reproductive status was assessed in 128 cheetahs maintained in 18 different institutions in North America. A mobile laboratory research team evaluated cheetahs using anesthesia, serial blood sampling, electroejaculation (males), and laparoscopy (females). Biomaterials were also collected for parallel studies of genetics, nutrition, and health. There was no mortality, and cheetahs were capable of reproducing naturally after these intense manipulatory examinations. No marked differences were observed in reproductive or endocrine characteristics between proven and unproven breeders. However, males consistently produced teratospermic ejaculates, and cheetah sperm were compromised in conspecific or heterologous in vitro fertilization systems. Structurally abnormal sperm were found to be filtered by the oocyte's zona pellucida. More than 80% of the females were anatomically sound, but morphological and endocrine evidence suggested that -50% or more of the population may have had inactive ovaries at the time of the examination. Males ranging in age from 15 to 182 months produced spermic ejaculates, but motile sperm numberdejaculate and circulating testosterone concentrations were highest in males 60 to 120 months old. Parovarian cysts were observed in 51.5% of female cheetahs, but comparisons between proven and unproven subpopulations revealed that this abnormality likely had no influence on fertility. Fresh luteal tissue was not observed in any nonpregnant or nonlactating female, strongly suggesting that the cheetah is an induced ovulator. Overall survey results were discussed in the context of the etiology of reproductive inefficiency, especially with respect to the potential importance of biological versus management factors. Four high priority research areas in cheetah reproductive biology were identified: 1) continuous monitoring of ejaculate quality in the extant population, while studying the impact of pleiomorphisms on fertility; 2) determining the potential relationship between libido and androgen production (excretion) in males; 3) confirming the extent of cyclic, or acyclic, ovarian activity in females; and 4) continued development of assisted reproductive techniques for enhancing management. In summary, a multidisciplinary, multi-institutional survey coordinated through the SSP is both possible and useful for generating a physiological and health database beneficial to driving further research and management initiatives. 0 1993 Wiley-Liss, Inc.
Immature cumulus-oocyte complexes (COCs) were recovered from freshly excised domestic cat ovaries and graded at a magnification of x40 for the condition of the cumulus oophorus of the oocyte cytoplasm. Grade I and II COCs were those with a uniformly dark cytoplasm and a readily identifiable, eccentrically located germinal vesicle. Grade I COCs had five or more cumulus oophorus cell layers, whereas grade II complements had less than five cell layers. Grade III and IV COCs were those undergoing progressive stages of oocyte cytoplasmic deterioration indicated by transparency or mosaic fragmentation and partial-to-complete loss of cumulus oophorus cells. In Expt 1, 699 oocytes were cultured for maturation and fertilization in vitro. More (P < 0.05) oocytes from grade I COCs matured (59.3%) and fertilized (29.7%) than from all other grades. Maturation and fertilization success did not differ (P > 0.05) for grade II (32.4, 11.6%, respectively) and grade III (21.9, 5.1%) oocytes, but these values were superior (P < 0.05) to those of grade IV (5.1, 1.4%). In Expt 2, 1040 COCs were graded, cultured for maturation and then inseminated. Of grade I oocytes, 24.4% developed into blastocysts compared with only 5.3% of grade II oocytes (P < 0.05). In general, oocytes from grade III and IV COCs were incapable of cleaving or growing in vitro. Of the 1739 COCs collected for both experiments, 12.3% met grade I criteria, the only category that provided consistent maturation, fertilization and development to blastocyst stage in vitro. In summary, a highly heterogeneous population of cumulus-oocyte complexes can be separated in the cat on the basis of grossly apparent morphological characteristics that, in turn, reflect functional differences in the ability of oocytes to mature, fertilize and develop in vitro.
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