Biomedical subjects

J G Howard

Publications and source records attributed to J G Howard.

At least 37 records · Page 2Linked to original sources

PubMed1992

Sperm capacitation in the domestic cat (Felis catus) and leopard cat (Felis bengalensis) as studied with a salt-stored zona pellucida penetration assay.

The ability of domestic cat or leopard cat spermatozoa to penetrate zonae pellucidae (ZP) of salt-stored, domestic cat oocytes was examined as an assay for sperm capacitation. Ovarian oocytes were recovered after ovariectomy and matured in vitro for 18-36 h. Following removal of cumulus cells, the oocytes were used fresh, or stored (4 degrees C, 0.5-24 weeks) in a HEPES-buffered hypertonic salt solution. Electroejaculated, washed sperm (2-4 x 10(6) sperm/ml) were preincubated for 1.0 h (38 degrees C, 5% CO2 in air) and then co-incubated (2 x 10(5) sperm/ml) with fresh or stored oocytes for 6.0 h. Gametes were incubated in a protein-free, modified Tyrode's solution (TLP-PVA) or in the same medium containing 4.0 mg/ml bovine serum albumin (BSA; TALP-PVA). Treatments were compared for percentage ZP penetration (defined as sperm heads reaching more than halfway through the ZP) as an index of sperm capacitation. In both the domestic cat and leopard cat, there was no difference (P greater than 0.05) in sperm penetration of fresh ZP (domestic cat, 42.5 +/- 5.4%; leopard cat, 38.6 +/- 2.8%) or stored ZP (domestic cat, 32.4 +/- 4.2%; leopard cat, 27.6 +/- 2.3%). Sperm incubated in protein-free medium (TLP-PVA) were less capable (P less than 0.05) of ZP penetration (domestic cat, 14.6 +/- 5.9%; leopard cat, 7.9 +/- 3.0%) than sperm incubated in medium TALP-PVA containing BSA (domestic cat, 60.3 +/- 5.9%; leopard cat, 58.4 +/- 3.0%).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Correlation of sperm viability with gamete interaction and fertilization in vitro in the cheetah (Acinonyx jubatus).

Sperm-oocyte interaction in vitro was studied in the cheetah, a species known to produce poor quality ejaculates and to experience low rates of fertility. Twelve female cheetahs were injected (i.m.) with eCG followed by hCG 84 h later. Twenty-four to 26 h post hCG, each was subjected to laparoscopic oocyte aspiration. A sperm motility index (SMI) was calculated for each of 9 cheetah sperm donors that produced ejaculates averaging 41.3 +/- 22.9 x 10(6) motile sperm and 28.4 +/- 4.9% structurally normal sperm. Each ejaculate was used to inseminate cheetah oocytes from 1 or 2 females and salt-stored, domestic cat oocytes. The presence of ovarian follicles (greater than or equal to 1.5 mm in diameter) showed that all females responded to exogenous gonadotropins (range, 11-35 follicles/female). A total of 277 cheetah oocytes was collected from 292 follicles (94.9% recovery; 23.1 +/- 2.2 oocytes/female). Of these, 250 (90.3%) qualified as mature and 27 (9.7%) as degenerate. Of the 214 mature oocytes inseminated, 56 (26.2%) were fertilized, and 37 (17.3%) cleaved to the 2-cell stage in vitro; but the incidence of in vitro fertilization (IVF) varied from 0 to 73.3% (p less than 0.001) among individual males. When oocytes from individual cheetahs (n = 5) were separated into two groups and inseminated with sperm from a male with an SMI greater than 0 after 6 h coincubation versus an SMI = 0 at this time, the mean fertilization rates were 28/44 (63.6%) and 0/37 (0%), respectively (p less than 0.05). Of the 117 domestic cat oocytes coincubated with cheetah sperm, 96.6% contained 1 or more cheetah sperm in the outer half of the zona pellucida (ZP). Although the mean number of cheetah sperm penetrating the outer ZP of the cat oocyte was similar (p greater than 0.05) among all males, there was a positive correlation between the number of sperm reaching the inner half of the ZP and fertilization rate in vitro (r = 0.82; p less than 0.05). Compared to IVF efficiency in the domestic cat and tiger as reported in earlier studies, IVF efficiency in the cheetah is low. Because oocytes from 11 of 12 cheetahs were fertilized in vitro, there is no evidence that the female gamete is incompetent. Although sperm pleiomorphisms may contribute to poor reproductive performance, examination of the data on the basis of individual sperm donors reveals that effective gamete interaction in the cheetah is dictated, in part, by sperm motility.

Animals↗

PubMed1992

The effect of pre-ovulatory anaesthesia on ovulation in laparoscopically inseminated domestic cats.

Laparoscopic intrauterine artificial insemination (AI) of electroejaculated spermatozoa was used to compare embryo development and conception rates in domestic cats inseminated either before or after ovulation. Females were given a single (100 iu) injection of pregnant mares' serum gonadotrophin (PMSG) followed by either 75 or 100 iu human chorionic gonadotrophin (hCG) 80 h later. Cats were anaesthetized (injectable ketamine HCl/acepromazine plus gaseous halothane) 25-50 h after administration of hCG for laparoscopic assessment of ovarian activity and for transabdominal AI into the proximal aspect of the uterine lumen. At the time of AI, 23 cats were pre-ovulatory (25-33 h after hCG injection) and 30 were post-ovulatory (31-50 h after hCG injection). Pre-ovulatory females produced 10.5 +/- 1.1 follicles and no corpora lutea compared with 1.9 +/- 0.5 follicles and 7.5 +/- 0.9 corpora lutea for the post-ovulatory group (P < 0.05). Six days later, the ovaries of nine pre-ovulatory and 12 post-ovulatory females were re-examined and the reproductive tracts flushed. On this day, pre-ovulatory cats produced fewer corpora lutea (2.8 +/- 1.5; P < 0.05) and embryos (0.4 +/- 0.3; P < 0.05) than post-ovulatory females (18.9 +/- 3.3 corpora lutea; 4.6 +/- 1.2 embryos). Two of the 14 cats (14.3%) inseminated before ovulation and not flushed became pregnant compared with 9 of 18 cats (50.0%) inseminated after ovulation and up to 41 h after hCG injection (P < 0.05). These results indicate that ovulation in cats is compromised by pre-ovulatory ketamine HCl/acepromazine/halothane or laparoscopy or by both and that electroejaculated spermatozoa deposited by laparoscopy in utero, after ovulation, result in a relatively high incidence of pregnancy. Because ovulation usually occurs 25-27 h after injection of hCG, the lifespan for fertilization of the ovulated ovum appears to be at least 14 h in vivo in cats.

Anesthesia, Inhalation↗

PubMed1991

Impact of season on seminal characteristics and endocrine status of adult free-ranging African buffalo (Syncerus caffer).

Pituitary, gonadal and adrenal activity were compared in free-living, adult African buffalo bulls during the breeding and nonbreeding seasons. Frequent blood samples were collected for 2 h from anaesthetized bulls treated intravenously with saline, gonadotrophin-releasing hormone (GnRH, 200 micrograms), human chorionic gonadotrophin (hCG, 10,000 i.u.) or adrenocorticotrophic hormone (ACTH, 1.5 mg). Electroejaculates also were collected from anaesthetized bulls during the breeding and nonbreeding seasons. Pretreatment testosterone concentrations among bulls varied more during the breeding (0.17-23.0 ng/ml) than the nonbreeding (0.15-2.21 ng/ml) season. The variation within the breeding season was attributed to 8 of 25 bulls producing higher (P less than 0.05) serum testosterone (High-T; 16.28 +/- 2.03 ng/ml) and testicular LH receptor (1.53 +/- 0.22 fmol/mg testis) concentrations compared with their seasonal counterparts (Low-T; 0.95 +/- 0.26 ng/ml; 0.38 +/- 0.04 fmol/mg) or with all bulls during the nonbreeding season (0.90 +/- 0.27 ng/ml; 0.31 +/- 0.04 fmol/mg). The magnitude of GnRH- and hCG-induced increases in serum testosterone was similar (P greater than 0.05) between Low-T bulls and bulls during the nonbreeding season. In the High-T animals treated with GnRH or hCG, serum testosterone did not increase, suggesting that secretion was already maximal. Peak serum LH concentrations after GnRH were greater (P less than 0.05) in bulls during the nonbreeding than the breeding season; FSH responses were similar (P greater than 0.05). ACTH treatment did not increase serum cortisol concentrations above the 2-fold increase measured in bulls treated with saline, hCG and GnRH (P greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

Comparative semen cryopreservation in ferrets (Mustela putorius furo) and pregnancies after laparoscopic intrauterine insemination with frozen-thawed spermatozoa.

A study was conducted to determine an optimum technique for semen cryopreservation and the biological competence of frozen-thawed ferret spermatozoa. Fifty-two fresh electroejaculates from 4 males were evaluated for sperm percentage motility, forward progressive motility, motility index (SMI) and acrosomal integrity. To determine the optimum temperature for maintaining sperm motility in vitro and the influence of glycerol on sperm motility, seminal aliquants were diluted in TEST diluent (containing either 0 or 4% glycerol) and maintained at 25 degrees or 37 degrees C. For cryopreservation, semen was diluted in each of 3 cryodiluents (TEST, PDV, BF5F), cooled for 30 min at 5 degrees C and pelleted on solid CO2 or frozen in 0.25 ml straws (20 degrees C/min to -100 degrees C). Following thawing, SMI and acrosomal integrity were determined. Ten females with maximum vulval swelling were given 90 i.u. human chorionic gonadotrophin and laparoscopically inseminated in utero with spermatozoa previously frozen using the optimum diluent and freeze-thaw method. The maintenance temperature of 25 degrees C was superior (P less than 0.05) to 37 degrees C for sustaining sperm motility, and glycerol did not influence (P greater than 0.05) motility for up to 11 h of culture. After thawing, motile spermatozoa were recovered in all treatment groups, but sperm motility and normal acrosomal ratings were highest using the PDV diluent, the pelleting method and thawing at 37 degrees C (P less than 0.05). Seven of the 10 ferrets (70%) inseminated with spermatozoa frozen by this approach became pregnant and produced 31 kits (mean litter size 4.4; range 1-9 kits).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

Seasonal variation in pituitary-gonadal function in free-ranging impala (Aepyceros melampus).

Blood, testicular biopsies and electroejaculates were collected from adult male impala, free-ranging in the Kruger National Park (Republic of South Africa), during the breeding (rut; April-May) and nonbreeding (September-October) seasons. Blood samples were collected at 5-min intervals for 120 min from anaesthetized males (n = 7 impala/group) treated intravenously with saline, gonadotrophin-releasing hormone (GnRH: 1 microgram/kg body weight) or human chorionic gonadotrophin (hCG: 10 or 30 iu/kg). Semen was collected from six more animals during the breeding season and 12 animals during the nonbreeding season using a standardized electroejaculation protocol. Ejaculates obtained during the nonbreeding season were of inferior quality to those collected during the breeding season, and were characterized by lower sperm concentrations, poorer sperm motility and more morphologically abnormal sperm forms. Within season, there were no differences in testosterone secretion between the two hCG doses, and these responses were similar to those observed after GnRH, but during the rut, testosterone secretion stimulated by both GnRH and hCG was approximately nine times greater than during the nonbreeding season. This seasonal increase in testosterone production was associated with a doubling in testicular volume and concentrations of luteinizing hormone (LH) receptors. Although concentrations of testicular follicle-stimulating hormone (FSH) receptors were similar between seasons, receptor content increased during rut as a result of increased testicular volume. In contrast to testosterone secretion, basal LH and FSH secretions were unaffected by season and GnRH-induced gonadotrophin secretion was reduced during rut.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

Human menopausal gonadotropin induces ovulation in sheep, but embryo recovery after prostaglandin F2alpha synchronization is compromised by premature luteal regression.

Using pregnant mares' serum gonadotropin (PMSG) and follicle stimulating hormone (FSH-P) as conventional gonadotropins, human menopausal gonadotropin (hMG) was tested for its comparative ability to induce multiple ovulations in sheep. Estrous cycles were synchronized using either prostaglandin F2alpha (PGF2alpha) or progestogen (MAP)-impregnated pessaries. During the mid-luteal phase, control ewes received serial saline injections, whereas test females (which also served as embryo donors) received either a single PMSG injection (1200 IU) or serial injections of FSH-P (total, 21 mg) or hMG (total, 1350 IU) over 3.5 d. These sheep were naturally mated and artificially inseminated (AI) in utero. Number of CL and transferable-quality embryos 5 d after AI was greater (P<0.05) in FSH-P-and hMG-treated donors than in PMSG-treated ewes. The lower number of transferable-quality embryos produced by PMSG-treated donors was attributed to a reduced (P<0.05) fertilization rate compared with that of the other treatment groups. There were no differences (P>0.05) in daily circulating estradiol-17beta and progesterone concentrations among the gonadotropin treatment groups. Gonadotropin-treated ewes demonstrated estrus approximately 24 h earlier than control ewes and, therefore, exhibited an accelerated estradiol-17beta surge and rise in circulating progesterone. Progesterone production in gonadotropin-treated ewes was also more variable than in the controls; this was due, in part, to premature luteal regression which occurred in 4 of 10 PMSG-, 3 of 10 FSH-P- and 6 of 10 hMG-treated ewes also given PGF2alpha. Ewes with prematurely regressing CL experienced transient luteal tissue development within 4 d of ovulation and produced no embryos. Overall results 1) demonstrate that serial administration of hMG induces multiple ovulations in sheep comparable to FSH-P, and 2) suggest that PGF2alpha treatment during ovulation induction adversely affects newly formed luteal tissue compromising subsequent embryo recovery.

Journal Article↗

PubMed1990

Oocyte recovery, maturation and fertilization in vitro in the puma (Felis concolor).

Eight female pumas were treated i.m. with 1000 (N = 5) or 2000 (N = 3) i.u. PMSG followed 84 h later by 800 i.u. hCG. Eggs were recovered 24-26 h after hCG from ovarian follicles by using laparoscopy and transabdominal aspiration. Mature eggs were inseminated in vitro 4-6 h later whereas immature eggs were cultured for 24 h and then inseminated. Electroejaculates from 3 pumas were diluted with mKRB before insemination to evaluate the influence of sperm concentration on fertilization. Seven of 8 pumas responded with follicle development, and 140 eggs were recovered from 145 follicles (96.6%; 77 mature, 43 immature, 20 degenerate eggs; mean +/- s.e.m., 20.0 +/- 5.9 eggs/female). Overall fertilization rate was 43.5% (total eggs fertilized = 40) despite using inseminates containing 82-99% pleiomorphic spermatozoa. Of the 36 immature oocytes matured in vitro and inseminated, 12 were fertilized even though 50% of the inseminating spermatozoa contained an acrosomal defect. Fertilization rate of mature oocytes collected from follicles appeared unrelated (P greater than 0.05) to PMSG dose or number of spermatozoa/inseminate. This study demonstrates that a high proportion of follicular eggs can be recovered laparoscopically from adult pumas treated with PMSG and hCG. These gametes are capable of being fertilized in vitro (immediately or after maturation in vitro) even with low quality semen with a high incidence of sperm pleiomorphisms.

Animals↗

PubMed1990

Reproductive anatomy, manipulation of ovarian activity and non-surgical embryo recovery in suni (Neotragus moschatus zuluensis).

Marked disparity in the uterine horn dimensions and relative degrees of caruncle development in suni suggested that exclusive or predominant dextral implantation occurs in association with bilateral ovulatory activity. Daily urinary measurements of pregnanediol-3 alpha-glucuronide revealed an oestrous cycle of approximately 21 days in length. Ovarian activity was controlled for synchronization of oestrus by using progestagen-impregnated intravaginal sponges and multiple ovulations were induced by using exogenous gonadotrophin therapy. An effective transcervical uterine catheterization technique was developed for the non-surgical collection of embryos. The efficiency of embryo recovery performed 5 days after sponge removal was 50.0%.

Animals↗

PubMed1989

Effect of cryoprotective diluent and method of freeze-thawing on survival and acrosomal integrity of ram spermatozoa.

A multifactorial study analyzed the effects of freezing method, cryoprotective diluent, semen to diluent ratio, and thawing velocity on post-thaw motility, progressive status, and acrosomal integrity of ram spermatozoa. Although semen to diluent ratio (1:3 vs 1:6, v/v) had no effect (P greater than 0.05), overall post-thaw spermatozoal viability was highly dependent on freezing method and cryoprotectant. Improved results were obtained by freezing semen in 0.5-ml French straws compared to dry ice pelleting. Manually freezing straws 5 cm above liquid nitrogen (LN2) was comparable to cooling straws in an automated, programmable LN2 unit. Of the two cryoprotective diluents tested, BF5F (containing the surfactant component sodium and triethanolamine lauryl sulfate) yielded approximately 50% fewer (P less than 0.05) spermatozoa with loose acrosomal caps compared to TEST. Thawing straws in a water bath at a higher velocity (60 degrees C for 8 sec) had no effect (P greater than 0.05) on spermatozoal motility, progressive status ratings, or acrosomal integrity when compared to a lower rate (37 degrees C for 20 sec). For the TEST group, thawing pellets in a dry, glass culture tube promoted (P less than 0.05) percentage sperm motility at 3 and 6 hr post-thawing, but for BF5F diluted semen this approach decreased the % of spermatozoa with normal apical ridges. The results suggest that the poor fertility rates often experienced using thawed ram semen likely result not only from reduced sperm motility, but also from compromised ultrastructural integrity. This damage is expressed by an increased loosening of the acrosomal cap, a factor which appears insensitive to freezing method but markedly influenced by the cryoprotective properties of the diluents tested.

Acrosome↗

PubMed1989

Semen characteristics and testosterone profiles in ferrets kept in a long-day photoperiod, and the influence of hCG timing and sperm dilution medium on pregnancy rate after laparoscopic insemination.

Five domestic ferrets previously maintained for 12 weeks under a 16L:8D photoperiod were electroejaculated weekly for 15-65 weeks while continuing to be exposed to the prolonged light cycle. Two ferrets sustained spermatogenesis for 20 and 26 weeks, while sperm production in the remaining males either was sporadic or decreased, remained depressed and then increased to peak levels observed in other males. Regardless of the temporal spermatogenesis patterns within males, the number of electroejaculated spermatozoa with residual cytoplasmic droplets or abnormal acrosomes increased in all ferrets over time. Diluted ejaculates meeting artificial insemination criteria were deposited intravaginally or by transabdominal laparoscopy into the uterine horns of females treated 0 or 24 h earlier with 90 i.u. hCG. Vaginal insemination was ineffective (0 pregnancies in 10 attempts), but 17/24 ferrets (70.8%) inseminated laparoscopically became pregnant and delivered live young (mean litter size, 5.2 kits). Number of motile spermatozoa deposited in utero (1.6-10.0 x 10(6) cells), presence of glycerol in the sperm dilution medium (0 versus 4%) and time of hCG administration (0 versus 24 h before insemination) had no effect on pregnancy results or litter size.

Animals↗

PubMed1989

Reproductive biology of the domestic cat with special reference to endocrinology, sperm function and in-vitro fertilization.

Explore the source record for details and available documents.

Animals↗

PubMed1988

A comparative analysis of ejaculate and hormonal characteristics of the captive male cheetah, tiger, leopard, and puma.

Male cheetahs, tigers, leopards, and pumas maintained under the same conditions were anesthetized and 1) serially bled before, during, and after electroejaculation (EE); 2) serially bled only (AO); or 3) serially bled before and after receiving adrenocorticotropin hormone (ACTH). Ejaculates from leopards contained higher (p less than 0.05) sperm concentrations than cheetahs and pumas but lower (p less than 0.05) sperm motility ratings than all other species. Tigers produced a larger seminal volume and the greatest number of motile sperm/ejaculate (p less than 0.05). The percentage of morphologically abnormal spermatozoa was greater (p less than 0.05) in cheetahs (64.6%), leopards (79.5%), and pumas (73.5%) than in tigers (37.5%). The most prevalent spermatozoal deformities included a tightly coiled or bent flagellum, a deranged midpiece, or a residual cytoplasmic droplet. Mean baseline serum cortisol concentrations in leopards were 2- and 4-fold greater (p less than 0.05) than in tigers and cheetahs, respectively. Basal cortisol concentrations in pumas were similar to those of tigers, but irrespective of treatment increased 2-fold (p less than 0.01) during the bleeding period. An acute rise and fall in cortisol attributable to EE was observed only in cheetahs. In tigers and leopards, mean peak cortisol concentrations after ACTH were similar to maximal values observed after EE. However, peak cortisol levels in cheetahs and pumas after ACTH were greater (p less than 0.01) than the concentrations measured after EE, indicating that these manipulatory procedures were not eliciting a maximal adrenal response. In the EE groups, luteinizing hormone (LH) and testosterone levels in cheetahs were lower (p less than 0.05) than in other species, whereas levels of both hormones were comparable (p greater than 0.05) in tigers, leopards, and pumas. Elevated cortisol levels in cheetahs and pumas had no discernible effect on LH/testosterone patterns; however, the results were equivocal in tigers, and, among leopards, testosterone concentrations consistently declined over time. In this study, using a standardized approach, we identify different ejaculate and endocrine characteristics of captive cheetahs, tigers, leopards, and pumas. The data extend earlier observations and demonstrate that some, but not all, Felidae species ejaculate high numbers of pleiomorphic spermatozoa. However, inter-species differences in sperm integrity do not appear related to inter-species variations in cortisol, LH, or testosterone. The observation of continuously declining testosterone concentrations only in leopards after AO, EE, or ACTH treatment suggests that rising and/or elevated cortiso

Acinonyx↗

PubMed1988

Comparison of embryo recovery, embryo quality, oestradiol-17 beta and progesterone profiles in domestic cats (Felis catus) at natural or induced oestrus.

Domestic cats experiencing a natural or FSH-induced oestrus were studied. Mated cats produced fewer (P less than 0.01) unfertilized oocytes and more (P less than 0.01) morulato blastocyst-stage embryos of better quality after a natural oestrus than after FSH treatment. Serum oestradiol-17 beta concentrations were lower (P less than 0.05) and progesterone levels rose earlier (P less than 0.05) in the induced oestrus compared to the natural oestrus group. Morula/blastocyst-stage embryos from both groups transferred to 15FSH/hCG-treated recipients produced 3 pregnancies and 2 live-born litters (1 from a natural oestrus donor and 1 from an FSH-treated donor). These results indicate that fertilization rates and embryo quality in domestic cats appear to be compromised by the FSH treatment, probably because of altered oestradiol-17 beta and progesterone concentrations.

Animals↗

PubMed1987

Similarity in ejaculate-endocrine characteristics in captive versus free-ranging cheetahs of two subspecies.

Ejaculate-endocrine characteristics were measured in 23 captive cheetahs (Acinonyx jubatus jubatus) in North American zoos and in 8 free-ranging cheetahs (A.j. raineyi) in eastern Africa (Tanzania). A standardized electroejaculation protocol was used, and numbers of motile spermatozoa were similar (p greater than 0.05) between groups. Of the spermatozoa collected by electroejaculation, 70.6 +/- 3.3% and 75.9 +/- 4.4% were morphologically abnormal in the captive "North American" and in the free-ranging, eastern African populations, respectively. Adrenal activity, as measured by an acute, temporal rise and fall in serum cortisol levels during and after electroejaculation, was no different (p greater than 0.05) between groups. Although serum luteinizing hormone (LH) levels were less (p less than 0.05) in the free-ranging than in the captive animals, serum testosterone concentrations were similar. The data indicate that the comparatively poor reproductive performance of cheetahs maintained in zoological parks is not attributable to a captivity-induced response afflicting the male. Furthermore, there is no evidence that ejaculate/endocrine characteristics differ between the two subspecies. Because adrenal/gonadal activity and the number of pleiomorphic spermatozoa are similar between the test groups, the results suggest that spermatozoal diversity originates as a result of the extreme genetic monomorphism observed universally in the species.

Acinonyx↗

PubMed1986

Reproductive physiology of the clouded leopard: I. Electroejaculates contain high proportions of pleiomorphic spermatozoa throughout the year.

Ejaculates were analyzed from clouded leopards (Neofelis nebulosa) subjected to a regimented anesthesia/electroejaculation protocol. Group I males (n = 4), maintained individually in an environment with natural fluctuations in photoperiod, were electroejaculated on the same day at monthly intervals (January-December). Group II clouded leopards (n = 8), maintained in random zoo populations throughout the U.S., were evaluated on a single occasion. Phase contrast and electron microscopy indicated a high proportion of structurally abnormal spermatozoa in seminal fluid (Group I range, 14.8-78.9%; Group II range, 32.3-93.0%), the predominant deformity being a tightly coiled flagellum. Semen quality, including spermatozoal concentration and the incidence of abnormal sperm forms, varied (p less than 0.05) among males. Evaluating the numbers of motile spermatozoa/ejaculate (MS/E) among individual males from Group I on a monthly basis suggested a seasonal influence; gradually increasing MS/E values with peaks in June and July were observed in three of four animals. A simultaneous analysis of international breeding records for captive female clouded leopards demonstrated that 46.2% of parturitions occurred in March and April, indicating that most estrual periods occurred from late December through February. These data suggest that a physiological asymmetry may exist in peak reproductive performance between the male and female clouded leopard, perhaps as a result of differing adaptations to the captive environment. Motile spermatozoa can be recovered throughout the year using electroejaculation and, when used over time, a standardized procedure can determine a hierarchy of seminal quality among males of unknown reproductive potential. The relatively high proportions of structurally abnormal spermatozoa in the ejaculates of the clouded leopard may be related to a low degree of genetic variation within the species and/or hyperadrenal activity in captive populations.

Aging↗

PubMed1986

Reproductive physiology of the clouded leopard: II. A circannual analysis of adrenal-pituitary-testicular relationships during electroejaculation or after an adrenocorticotropin hormone challenge.

A circannual analysis was made of serum cortisol, luteinizing hormone (LH), and testosterone concentrations in the male clouded leopard (Neofelis nebulosa). Group I males (n = 4), maintained in a standardized environment, were bled serially during a regimented anesthesia/electroejaculation episode occurring monthly (beginning in January, ending in December). Additional sampling intervals were conducted under anesthesia only (control, n = 8), anesthesia plus a single adrenocorticotropin hormone challenge (ACTH, Cortrosyn, n = 4), or anesthesia plus a single 25 micrograms injection of gonadotropin-releasing hormone (GnRH, Gonadorelin, n = 4). Group II males (n = 6) from various zoological collections were sampled serially under the same semen collection conditions on one random occasion within the year. Serum cortisol levels were 2 times greater than values measured in comparable studies involving other felid species. Cortisol concentrations were similar during electroejaculation and control (anesthesia only) episodes, and mean levels did not rise as a result of semen collection. Adrenocorticotropin caused an immediate rise in cortisol to levels at least 1.5 times greater than electroejaculated or control counterparts. Mean concentrations of basal cortisol in individual males gradually increased as the year progressed, possibly as a consequence of repeated psychogenic stress. Between seasons, there were no differences in mean LH; however, testosterone levels were greater (p less than 0.05) in the winter compared to all other seasons. There were no differences (p greater than 0.05) between individual males in secretory patterns or mean concentrations of cortisol, LH, or testosterone. Within males, distinct temporal fluctuations were observed in both LH and testosterone during the approximately 80-min sampling interval. Neither LH nor testosterone profiles appeared affected by cortisol patterns during electroejaculation or after an ACTH challenge. A bolus of GnRH induced a marked rise in serum LH and testosterone within 15 and 30 min respectively, indicating that these two hormones were coupled. Both LH and testosterone profiles in Group II males mimicked those in Group I; concentrations of cortisol in Group II males immobilized on one occasion were similar to those of Group I animals sampled from January-May but appeared to be less than values measured from June-December. These data demonstrate that the clouded leopard, compared to other felids, produces markedly elevated concentrations of cortisol, which are likely related to an aggressive behavioral temperament.(ABSTRACT TRUNCATED AT 400 WORDS)

Acclimatization↗

PubMed1986

Influence of cryoprotective diluent on post-thaw viability and acrosomal integrity of spermatozoa of the African elephant (Loxodonta africana).

Electroejaculates from free-ranging, African elephants were frozen to test various seminal diluents, freezing methods and thawing media on post-thaw sperm viability and structural integrity. In Study I, each ejaculate was tested with each of 7 cryoprotective diluents. After cooling to 5 degrees C and equilibration on ice (4 degrees C) for 120 min, each aliquant was pellet frozen on solid CO2, stored in liquid nitrogen and thawed (37 degrees C) in saline or tissue culture solution. Amongst all diluents, post-thaw sperm motility, motility duration in vitro (37 degrees C) and acrosomal integrity were greatest (P less than 0.05) when diluent BF5F was used. Thawing medium had no effect on results. In Study II, the optimal diluent from Study I (BF5F) was compared with the diluent SGI. Results were not affected by a 90- or a 150-min cooling-equilibration interval in an electronic cooler (5 degrees C); however, post-thaw sperm motility rating and duration of motility in vitro were greater (P less than 0.01) with the pellet than the straw container freezing method. When the pelleting method was used, diluents BF5F and SGI provided comparable cryoprotection. Duration of post-thaw motility was enhanced 2-fold and up to 12 h by maintaining thawed semen at 21 rather than 37 degrees C (P less than 0.05). All diluents provided some protection on acrosomal integrity, but the overall proportion of intact acrosomes after thawing was markedly less in Study II, apparently as a result of the slower initial cooling rate (approximately 1.5 degrees C/min) compared to that of Study I (approximately 6.5 degrees C/min). This study demonstrates the feasibility of cryopreserving semen from free-ranging African elephants and indicates that spermatozoa must effectively survive freezing when the BF5F or SGI diluent is used in conjunction with the pelleting method.

Acrosome↗