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Author (up) Brown, J.L.; Wasser, S.K.; Wildt, D.E.; Graham, L.H.
Title Comparative Aspects of Steroid Hormone Metabolism and Ovarian Activity in Felids, Measured Noninvasively in Feces Type Journal Article
Year 1994 Publication Biol Reprod Abbreviated Journal
Volume 51 Issue 4 Pages 776-786
Keywords Animal; Carbon; Radioisotopes; Carnivora; Cats; Chromatography; High; Pressure; Liquid; Comparative Study; Estradiol; metabolism; Estrone; feces; chemistry; Female; Ovary; physiology; Pregnancy; Progesterone; Pseudopregnancy; Support; Non-U.S.Gov't; browse; non; government; gov't; us; 170
Abstract Noninvasive fecal assays were used to study steroid metabolism and ovarian activity in several felid species. Using the domestic cat (Felis catus) as model, the excretory products of injected [14C]estradiol (E2) and [14C]progesterone (P4) were determined. Within 2 days, 97.0 +/- 0.6% and 96.7 +/- 0.5% of recovered E2 and P4 radioactivity, respectively, was found in feces. E2 was excreted as unconjugated estradiol and estrone (40%) and as a non-enzyme- hydrolyzable conjugate (60%). P4 was excreted primarily as non-enzyme- hydrolyzable, conjugated metabolites (78%) and as unconjugated pregnenolone epimers. A simple method for extracting fecal steroid metabolites optimized extraction efficiencies of the E2 and P4 excretion products (90.1 +/- 0.8% and 87.2 +/- 1.4%, respectively). Analysis of HPLC fractions of extracted fecal samples from the radiolabel-injected domestic cats revealed that E2 immunoreactivity coincided primarily with the unconjugated metabolized [14C]E2 peak, whereas progestogen immunoreactivity coincided with a single conjugated epimer and multiple unconjugated pregnenolone epimers. After HPLC separation, similar immunoreactive E2 and P4 metabolite profiles were observed in the leopard cat (F. bengalensis), cheetah (Acinonyx jubatus), clouded leopard (Neofelis nebulosa), and snow leopard (Panthera uncia). Longitudinal analyses demonstrated that changes in fecal E2 and P4 metabolite concentrations reflected natural or artificially induced ovarian activity. For example, severalfold increases in E2 excretion were associated with overt estrus or exogenous gonadotropin treatment, and elevated fecal P4 metabolite concentrations occurred during pregnant and nonpregnant (pseudopregnant) luteal phases. Although overall concentrations were similar, the duration of elevated fecal P4 metabolites during pseudopregnancy was approximately half that observed during pregnancy. In summary, steroid metabolism mechanisms appear to be conserved among these physically diverse, taxonomically related species. Results indicate that this hormone-monitoring approach will be extremely useful for elucidating the hormonal regulatory mechanism associated with the reproductive cycle, pregnancy, and parturition of intractable and endangered felid species.
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ISSN 0006-3363 ISBN Medium
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Notes Document Type: eng Approved no
Call Number SLN @ rana @ 251 Serial 198
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Author (up) Brown, J.L.; Wasser, S.K.; Wildt, D.E.; Graham, L.H.
Title Steroid Metabolism and the Effectiveness of Fecal Assays for Assessing Reproductive Status in Felids Type Journal Article
Year 1994 Publication Biology of Reproduction Abbreviated Journal
Volume 50 Issue suppl 1 Pages 185
Keywords fecal-analysis; breeding; reproduction; medicine; medical; zoo; zoos; veterinary; snow-leopard; snow leopard; browse; fecal; analysis; 1420
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Notes Twenty-seventh Annual Meeting of the Society for the Study of Reproduction, Ann Arbor, Michigan, USA, July 24-27, 1994 Call Number: QP251 .B56 Approved no
Call Number SLN @ rana @ 203 Serial 197
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Author (up) Johnston, L.A.; Donoghue, A.M.; O'Brien, S.J.; Wildt, D.E.
Title Rescue and maturation in vitro of follicular oocytes collected from nondomestic felid species Type Journal Article
Year 1991 Publication Biol Reprod Abbreviated Journal
Volume 45 Issue 6 Pages 898-906
Keywords Animal; Carnivora; anatomy; histology; physiology; Cells; Cultured; Female; Fertilization; Vitro; inVitro; Male; Oocytes; cytology; growth; development; ovarian; Follicle; Spermatozoa; Support; Non-U.S.Gov't; U.S.Gov't; P.H.S.; browse; non; us; gov't; government; 230
Abstract The potential for rescuing immature oocytes from the ovaries of females of rare felid species which die or undergo medical ovariohysterectomy was evaluated. Ovaries were recovered from 13 species representing 35 individuals in good-to-poor health. Although the majority of females were 10 yr of age or older and in fair-to-poor health, a total of 846 oocytes were recovered of which 608 (71.9%) were classified as fair-to- excellent quality. One hundred of these oocytes were used for initial maturation classification and as parthogenetic controls. Overall, of the 508 fair-to-excellent quality oocytes placed in culture, 164 (32.3%) matured to metaphase II in vitro. For species in which 3 or more individuals yielded oocytes, mean oocyte maturation rates were as follows: 36.2%, tiger; 27.9% leopard; and 8.3%, cheetah. In vitro insemination of oocytes resulted in fertilization (2 polar bodies, 2 pronuclei, or cleavage) rates of 9.1% to 28.6% (leopard) using homologous fresh spermatozoa and 4.0% (lion) to 40.0% (puma) using homologous frozen-thawed spermatozoa. Inseminations using heterologous (domestic cat) spermatozoa also resulted in fertilized oocytes in the tiger, leopard, snow leopard, puma, serval, and Geoffroy's cat (range in fertilization rate, 5.0% for leopard to 46.2% for puma). Cleaved embryos resulted from the insemination of leopard oocytes with homologous sperm (n = 1 embryo) and puma oocytes with domestic cat sperm (n = 3 embryos). These results demonstrate that immature ovarian oocytes from rare felid species can be stimulated to mature in vitro despite an excision-to-culture interval as long as 36 h.(ABSTRACT TRUNCATED AT 250 WORDS)
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ISSN 0006-3363 ISBN Medium
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Notes Document Type: eng Approved no
Call Number SLN @ rana @ 172 Serial 503
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Author (up) Roth, T.L.; Armstrong, D.L.; Barrie, M.T.; Wildt, D.E.
Title Seasonal effects on ovarian responsiveness to exogenous gonadotrophins and successful artificial insemination in the snow leopard (Uncia uncia) Type Journal Article
Year 1997 Publication Reprod Fertil Dev Abbreviated Journal
Volume 9 Issue 3 Pages 285-295
Keywords Animal; Carnivora; anatomy; histology; Blood; physiology; Estradiol; Female; Gonadotropins; administration; dosage; pharmacology; Chorionic; Equine; Human; Insemination; artificial; methods; veterinary; Laparoscopy; Male; Oocytes; cytology; Ovary; drug; effects; Ovulation; Induction; Pregnancy; Progesterone; Seasons; Support; Non-U.S.Gov't; browse; non; us; gov't; government; 400
Abstract Ovaries of the seasonally-breeding snow leopard (Uncia uncia) were examined to determine whether they were responsive to exogenous gonadotrophins throughout the year. The potential of laparoscopic artificial insemination (AI) also was assessed for producing offspring. During the non-breeding, pre-breeding, breeding and post-breeding seasons, females (n = 20) were treated with a standardized, dual- hormone regimen given intramuscularly (600 I.U. of equine chorionic gonadotrophin followed 80-84 h later with 300 I.U. of human chorionic gonadotrophin (hCG)). Laparoscopy was performed 45-50 h after administration of hCG, and all ovarian structures were described. Females with fresh corpora lutea (CL) were inseminated, and anovulatory females were subjected to follicular aspiration to examine oocyte quality. Snow leopards responded to exogenous gonadotrophins throughout the year. Mean number of total ovarian structures (distinct follicles mature in appearance plus CL) did not differ (P > or = 0.05) with season, but the proportion of CL: total ovarian structures was greater (P < 0.01) for the breeding season compared with all other seasons. The proportion of females ovulating was greater (P < 0.05) during the breeding and post-breeding seasons than during the pre-breeding and non- breeding seasons respectively. No Grade-1 quality oocytes were recovered from follicles of anovulatory females. Serum concentrations of oestradiol-17 beta appeared elevated in all females, and neither oestradiol-17 beta concentrations nor progesterone concentrations differed (P > or = 0.05) among seasons. Of 15 females artificially inseminated, the only one that was inseminated in the non-breeding season became pregnant and delivered a single cub. This is the first successful pregnancy resulting from AI in this endangered species.
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ISSN 1031-3613 ISBN Medium
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Notes Document Type: eng Approved no
Call Number SLN @ rana @ 325 Serial 830
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Author (up) Roth, T.L.; Howard, J.G.; Donoghue, A.M.; Swanson, W.F.; Wildt, D.E.
Title Function and culture requirements of snow leopard (Panthera uncia) spermatozoa in vitro Type Journal Article
Year 1994 Publication J Reprod Fertil Abbreviated Journal
Volume 101 Issue 3 Pages 563-569
Keywords Animal; Carnivora; physiology; Cell; survival; Cells; Cultured; Comparative; study; Culture; Media; Female; Male; Pentoxifylline; Sperm; Motility; Ovum; Interactions; Spermatozoa; cytology; Support; Non-U.S.Gov't; U.S.Gov't; P.H.S.; browse; non; us; gov't; government; 190
Abstract Electroejaculates from eight snow leopards were used to determine how the motility of spermatozoa was influenced by (i) type of media (Ham's F10, PBS, human tubal fluid or RPMI-1640); (ii) holding temperature (23 degrees C versus 37 degrees C); (iii) washing of spermatozoa and (iv) a sperm metabolic enhancer, pentoxifylline. The duration of sperm motility was assessed by evaluating samples in each treatment every hour for 6 h and a sperm motility index (a value combining percentage sperm motility and rate of forward progression) calculated. Spermatozoa from the Ham's F10, PBS and PBS plus pentoxifylline treatments were also co-incubated with zona-intact, domestic cat eggs that were fixed and evaluated for spermatozoa bound to the zona pellucida, penetrating the outer and inner layers of the zona pellucida and within the perivitelline space. During the 6 h co-incubation, the sperm motility index in PBS with pentoxifylline was greater (P < 0.05) than in PBS alone which, in turn, was greater (P < 0.05) than in the other three test media. Washing the spermatozoa enhanced (P < 0.05) motility in both PBS and PBS plus pentoxifylline relative to unwashed samples, but there was no effect (P > 0.05) of holding temperature. Pentoxifylline supplementation enhanced (P < 0.05) the proportion of cat eggs with bound, but not penetrated, snow leopard spermatozoa in the inner layer of the zona pellucida, and there were no spermatozoa in the perivitelline space.(ABSTRACT TRUNCATED AT 250 WORDS)
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ISSN 0022-4251 ISBN Medium
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Notes Document Type: eng Approved no
Call Number SLN @ rana @ 253 Serial 828
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Author (up) Roth, T.L.; Swanson, W.F.; Wildt, D.E.
Title Snow leopard (Panthera unica) sperm longevity in vitro is not influenced by protein or energy source supplements but is affected by buffer source Type Journal Article
Year 1995 Publication Theriogenology Abbreviated Journal
Volume 43 Issue 1 Pages 309
Keywords genetics; Sperm; captivity; medical; medicine; veterinary; zoo; breeding; browse; 1400
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Notes Annual Conference of the International Embryo Transfer Society, Calgary, Alberta, Canada, January 8-10, 1995 Document Type: English Call Number: QP251 .T44 Approved no
Call Number SLN @ rana @ 280 Serial 829
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Author (up) Roth, T.L.; Swanson, W.F.; Wildt, D.E.; Collins, D.; Burton, M.; Garell, D.M.
Title Snow leopard (Panthera uncia) spermatozoa are sensitive to alkaline pH, but motility in vitro is not influenced by protein or energy supplements Type Miscellaneous
Year 1996 Publication Journal of Andrology Abbreviated Journal
Volume 17 Issue Pages 558-566
Keywords Felid,comparative biology,sperm culture medium,sperm function,capacitation,sodium bicarbonate.
Abstract To better understand the biology of snow leopard spermatozoa and to facilitate developing assisted reproduction, a series of studies was conducted to: 1) identify the component(s) of complex culture media responsible for the detrimental effect on sperm survival in vitro, 2) optimize medium for supporting sperm viability, and 3) evaluate sperm capacitation in vitro. Constituents of complex media were added systematically to phosphate-buffered saline (PBS) to isolate the factor(s) influencing snow leopard sperm motility in vitro. Sperm capacitation was also assessed following incubation in PBS with bovine serum albumin (BSA), fetal calf serum (FCS), or heparin. For maintaining sperm motility, there was no benefit (P ? 0.05) to supplementing PBS with low (5%) or high (20%) concentrations of snow leopard serum (SLS) versus FCS or BSA. Likewise, adding supplemental energy substrates (pyruvate, glucose, lactate, or glutamine) did not enhance or hinder (P ? 0.05) sperm motility. However, motility rapidly decreased (P < 0.05) with the addition of NaHCO3 to PBS or Ham's F10 nutrient mixture. Surprisingly, Ham's F10 with no buffering component or with both NaHCO3 and N-Z-hydroxyethylpiperazine-N'-2- ethanesulfonic acid (HEPES) maintained sperm motility at levels similar (P ? 0.05) to PBS. Although sperm motility in all treatments decreased with time, there was a strong inverse relationship (P < 0.01; r = 0.90) between motility and sample pH at 6 hours. Spermatozoa incubated in PBS containing FCS, BSA, or heparin did not undergo the acrosome reaction when exposed to calcium ionophore. In summary, alkaline pH has a profound detrimental effect on snow leopard sperm motility, and capacitation does not occur under conditions that normally promote this event in other felid species. These results clearly demonstrate a high degree of interspecific variation among felids in fundamental sperm function, and they provide evidence for the necessity of basic research when developing assisted reproduction in little-studied nondomestic species.
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Notes Approved no
Call Number SLN @ rana @ 897 Serial 831
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