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G Wirtu - One of the best experts on this subject based on the ideXlab platform.

  • Daily or thrice weekly handling of Eland antelope (Taurotragus oryx): effects on serum cortisol level.
    Research in veterinary science, 2013
    Co-Authors: P. M. Pennington, C E Pope, Robert A. Godke, J Galiguis, R.a. Maclean, L.r. Gentry, G Wirtu
    Abstract:

    We evaluated the cortisol response of adult female Eland (n=8) that were handled in hydraulic chute daily or 3×/week. Females were divided into two groups and each group (n=4) successively received two estrous cycle synchronization treatments: (1) two injections of prostaglandin (PG-PG) F2α at 11 day intervals and (2) oral administration of altrenogest for 7 days and an injection of PGF2α on day 7 (Alt-PG). Blood samples were collected 3×/week during the synchronization (Synch) and expected luteal phase (Nonintensive) periods, and daily during the expected time of induced (Intensive 1) or natural (Intensive 2) estrus. Overall, mean cortisol levels were highest during Intensive 1, followed by Intensive 2, Synch and Nonintensive periods. Individual Eland were the most significant source of variation for cortisol level. The frequency of handling and the synchronization treatment significantly affected cortisol levels in 3/8 and 4/8 females, respectively. In conclusion, in response to increased frequency of handling, Eland cortisol levels rose transiently and returned to baseline within few days after more intensive handling. Thus, the Eland females were tolerant to and recovered from the effects of repeated daily handling.

  • 201 endoscope guided transfer of sperm injected oocytes into the oviducts of Eland and bongo antelopes
    Reproduction Fertility and Development, 2010
    Co-Authors: G Wirtu, R A Godke, D L Paccamonti, B L Dresser, R Maclean, J Galiguis, B E Eilts, U Besenfelder, G T Gentry
    Abstract:

    We have previously established methods of gamete collection in Eland and Bongo antelopes; however, blastocyst development following IVF, intracytoplasmic sperm injection (ICSI), or somatic cell nuclear transfer (SCNT) has been sporadic, thus few developmentally competent embryos have been produced for uterine transfer. In the present study, we evaluated the possibility of oviductal transfer of presumptive zygotes. Estrous cycle synchronization and ovarian superstimulation were done as follows. Females were treated with altrenogest (0.11 g/os daily) for 7 days. The FSH Folltropin® was dissolved in 30% polyvinylpyrrolidone and administered, i.m. on Days 5 and 7 of altrenogest treatment at 266 mg and 134 mg, respectively. Prostaglandin F2α (Lutalyse®, 25 mg, i.m.) was administered on Day 7. Transvaginal ultrasound-guided oocyte retrieval was conducted on Day 9 as described previously (Wirtu G et al. 2009 Anim. Reprod Sci. 111, 160-172). Eleven oocytes were recovered from 2 Elands and 4 from 1 Bongo. One oocyte of each species was degenerate at the time of recovery. Oocytes were subjected to IVM for approximately 22 h, when Piezo drill-assisted ICSI was done using frozen-thawed Eland or Bongo spermatozoa. Oocytes were activated (5% ethanol, 5 min) and cultured overnight in CR1aa medium supplemented with BSA. Presumptive zygotes were subjected to endoscopic oviductal transfer at 21 to 24 h after ICSI. The oviductal transfer was adapted from the technique developed in domestic cattle (Wetscher F et al. 2005 Theriogenology 64, 30-40). Animals were sedated by i.m. administration of xylazine HCl and butorphanol tartrate and restrained in a hydraulic chute (Tamer®) that was used to squeeze and lift the females. An epidural block (5 mL of lidocaine) was induced after cleaning the rectum, perineal region, and the injection site. A uterine relaxant, isoxsuprine HCl (10 mg, i.v), was then administered. The Brem/Besenfelder set for laparoscopic bovine embryo transfer (Karl Storz Endoscope, Karl Storz GmbH & Co. KG, Tuttlingen, Germany) was used for the oviductal transfer. Briefly, a cannula (12.5 mm in diameter, 49.5 cm long) fitted with a blunt-tip obturator was introduced into the vagina and positioned, during transrectal manipulation, at the mid-dorsal aspect of the vaginal vault. The blunt-tip obturator was replaced with a sharp-tip obturator, which was used to puncture through the vaginal wall for entry into the pelvic cavity. The trocar was subsequently replaced with an inner sheath containing a telescope (5.5 mm in diameter, 54 cm long) and a pre-loaded transfer tubing. Visualization of the ovaries and oviducts required insufflation of the peritoneal cavity. Transfer was done into the oviduct ipsilateral to the recently ovulating ovary. Ten Eland oocytes had extruded the first polar body and 8 that survived ICSI were transferred into the right oviduct of an Eland female. Two Bongo oocytes had extruded the first polar body; both survived ICSI and were transferred to the right oviduct of a Bongo female. Pregnancy diagnosis is pending. This minimally invasive method of accessing the oviduct has the potential to advance the application of assisted reproductive technologies in large nondomestic ungulates.

  • ultrasonographic guided retrieval and in vitro maturation of Eland taurotragus oryx and bongo tragelaphus eurycerus isaaci antelope oocytes
    Animal Reproduction Science, 2009
    Co-Authors: G Wirtu, R A Godke, D L Paccamonti, Earle C Pope, B L Dresser
    Abstract:

    The limited availability of gametes is a major factor hindering the development and application of assisted reproductive technologies (ART) in large non-domestic ungulates. This is partly due to the small number of captive animals and handling difficulties associated with procedures for gamete recovery. In the present study, results are reported of multi-year studies on ovarian stimulation and oocyte retrieval by ultrasonographic-guided transvaginal follicular aspiration and subsequent in vitro maturation (IVM) in Eland and bongo antelopes. All procedures were conducted on sedated females handled in a hydraulic chute without inducing general anesthesia. Five estrous synchronization/ovarian stimulation protocols were evaluated and data are presented on 73 and 15 procedures in Eland and bongo, respectively. Repeating procedures (≤once/month) on the same female did not affect ovarian response or number oocytes recovered in either species. Eland females, but not the ovarian stimulation treatment, affected ovarian response. Ovarian stimulation treatment affected oocyte recovery rate in Eland, but not in bongo. In both species, ovarian hormone stimulation treatment affected the distribution of follicles by size and the status of expansion of the cumulus cell investment of oocytes, but not the frequency of metaphase II oocytes during IVM. The timing of extrusion of the first polar body during IVM was more synchronous in bongo than in Eland oocytes. It is concluded that Transvaginal oocyte retrieval (TVOR) can be safely and repeatedly applied in gonadotropin-treated Eland and bongo females to recover oocytes that can mature in vitro. The methods described for the present study can be adapted to improve the availability of non-domestic ungulate oocytes for basic and applied studies.

  • 169 BEHAVIORAL ASPECTS OF ESTRUS IN THE COMMON Eland ANTELOPE, TAUROTRAGUS ORYX
    Reproduction Fertility and Development, 2009
    Co-Authors: G Wirtu, C E Pope, Robert A. Godke, J Galiguis, P. M. Pennington, R.a. Maclean, J. Mercado, Dale L. Paccamonti, Betsy L. Dresser
    Abstract:

    Knowledge of the estrous cycle and behavior is important in managing captive breeding programs. However, such information is negligible in spiral-horned antelopes, including the Eland. In the present study, conducted between September 2007 and March 2008, we sought to characterize mounting activity and courtship behavior in a group of eight adult Eland females. Estrus was induced in groups of four females by treatment with 25 mg PGF2 ∝ (IM, Lutalyse®, Pharmacia and Upjohn Company, Kalamazoo, MI, USA) administered after 7 days of daily oral progestin (5 mL of 2.2% altrenogest, DPT Laboratories, San Antonio, TX, USA) or 11 days after an initial treatment of PGF2 ∝ (25 mg). The eight females received each treatment in a crossover design. Females were induced and monitored during the induced and subsequent natural estrus for 34 to 38 days. Observations for estrus behavior were initially assisted by using Estrotect™ heat detector patches (Rockyway, Inc., www.estrotect.com). Since January 2008, the HeatWatch® electronic heat detection system (CowChips, Denver, CO, USA) and an androgenized Eland female were used to assist with heat detection. For androgenization, 8 pellets of Synovex-H® (Fort Dodge Animal Health, Fort Dodge, IA) comprising a total dose of 1600 mg testosterone propionate and 160 mg estradiol benzoate were implanted (SC) on the convex side of the ear. To confirm mounting activity detected by Estrotect™ or HeatWatch®, Eland behavior was recorded continuously using an 8-channel real time DVR. Chi-square analysis was used to test the association between time of day (day or night) and the frequency of standing to be mounted. The cycle length (n = 6 females) was the time interval (d) between the first mounts at the induced and natural estrus. The time of mounts was determined for a total of 52 mounts. More than half of the mounts (n = 32 or 61.5%) occurred between 6 pm and 6 am (night); however, there was no association between the number of mounts and time of the day (p = 0.886). Analysis of mounts by quarters of the day showed that mounts were evenly distributed between early afternoon (noon to 6 pm: 29%), late afternoon (6 pm to midnight: 33%) and early morning (midnight to 6 am: 29%) but were less frequent during late morning, between 6 am and noon (10%) possibly due to interference by human activities. The number of mounts observed per female ranged from 1 to 20. Mounts lasted for 2 seconds or less. The duration of estrus ranged from 2.1 to 29.0 hours. Typical courtship behaviors included prolonged head butting, flehmen response and following the female in estrus before mounting. The average length of the estrous cycle, based on mounting activities, was 19.3 d (range: 17–24). Although further studies are required in herds with Eland males, the present results suggest that estrus detection for captive breeding or application of reproductive technologies in the Eland should be spread around different times of the day and night. We have also demonstrated that an electronic mount detection system can be applied to assist with determination of estrus in the Eland.

  • cloned embryos from semen part 2 intergeneric nuclear transfer of semen derived Eland taurotragus oryx epithelial cells into bovine oocytes
    Cloning and Stem Cells, 2008
    Co-Authors: Liesl Nelthemaat, G Wirtu, A Cole, B L Dresser, Earle C Pope, M. C. Gómez, Monica Lopez, Jill A Jenkins, K R Bondioli, R A Godke
    Abstract:

    The production of cloned offspring by nuclear transfer (NT) of semen-derived somatic cells holds considerable potential for the incorporation of novel genes into endangered species populations. Because oocytes from endangered species are scarce, domestic species oocytes are often used as cytoplasts for interspecies NT. In the present study, epithelial cells isolated from Eland semen were used for intergeneric transfer (IgNT) into enucleated bovine oocytes and compared with bovine NT embryos. Cleavage rates of bovine NT and Eland IgNT embryos were similar (80 vs. 83%, respectively; p > 0.05); however, development to the morula and blastocyst stage was higher for bovine NT embryos (38 and 21%, respectively; p or = 8 cells at 84 hpa, while 32% of the bovine NT embryos had > or = 8 cells at the same interval. However, 100 and 66% of bovine NT and Eland IgNT embryos, respectively, that had > or = 8 cells synthesized DNA. From these results we concluded that (1) semen-derived epithelial cell nuclei can interact and be transcriptionally controlled by bovine cytoplast, (2) the first cell-cycle occurred in IgNT embryos, (3) a high frequency of developmental arrest occurs before the eight-cell stage in IgNT embryos, and (4) IgNT embryos that progress through the early cleavage stage arrest can (a) synthesize DNA, (b) progress through subsequent cell cycles, and (c) may have the potential to develop further.

Radim Kotrba - One of the best experts on this subject based on the ideXlab platform.

  • Habituation of common Eland (Taurotragus oryx) to intensive routine handling, and the effect of immunocastration thereon
    Applied Animal Behaviour Science, 2021
    Co-Authors: Abubakar S. Musa, Radim Kotrba, Tersia Needham, Silvie Neradilova, Andre Ganswindt, Francisco Ceacero
    Abstract:

    Abstract The temperament of captive common Eland (Taurotragus oryx) during handling is crucial for their management, as they have been identified as an ideal antelope for game meat farming, and are well-represented in captive antelope populations world-wide. Generally speaking, common Eland are considered undomesticated, and thus maintain their natural prey instincts, making it necessary to habituate them to routine handling. Immunocastration can be implemented as part of the herd management strategy in mixed or single-sex Eland groups, and might reduce aggressive behaviour of males by blocking the production of testosterone, thereby improving docility for ease of handling. Within this study, data was collected to determine the influence of routine handling and immunocastration on the temperament of common Eland during handling. Twenty-nine common Eland were divided into two groups: sub-adults (n = 15; ca. 2 years old; 182.9 ± 59.37 kg) and juveniles (n = 14; ca. 6 months old; 94.18 ± 24.76 kg), and they were routinely handled every two weeks over a four-month period (10 handling events). Within each age group, females, males, and immunocastrated males were present. Immunocastration treatment was administered during the second and fourth handling events (2 mL Improvac®/ animal). During handling, animals were individually driven through the raceway and finally restrained in a squeeze chute, where routine biological samples and measurements were collected. Faecal androgen metabolite (fAM) concentrations were measured for each event, for males only. Each animal was focally sampled for specific behaviours during the handling. A temperament score was obtained by observing the general state of the animal from the point of entering the raceway, until its exit from the squeeze chute. Generalized linear mixed models tested the influence of immunocastration, week, group, fAM concentration, and body weight on the temperament score. Immunocastration did not affect the temperament score, the general state in the squeeze chute, the order of entering the handling system, and the chute exit. However, the temperament score and the state in the squeeze chute improved overtime (with animals being less nervous). Moreover, animals with low temperament scores (calmer) entered the system earlier, and exited the system calmly over the repeated handling period. The results show that routine handling through a raceway and squeeze chute system leads to habituation, and improves the ease of handling in common Eland. However, immunocastration did not affect the temperament of the male common Eland during handling.

  • Sensory characteristics of the longissimus thoracis et lumborum and biceps femoris muscles from male and female common Eland (Taurotragus oryx)
    Meat science, 2019
    Co-Authors: Tersia Needham, Radim Kotrba, Johannes G. Laubser, Daniel Bureš, Heather E. Smyth, Louwrens C. Hoffman
    Abstract:

    The sensory attributes of the longissimus thoracis et lumborum (LTL) and biceps femoris (BF) muscles were compared for male (n = 6) and female (n = 6) Eland. Descriptive sensory analysis showed that the meat from cows, and the BF muscle, had greater overall flavour scores, primarily characterized as beef-like flavour (r = 0.926). Unfavourable aroma and flavour attributes received low scores, indicating good potential for fresh Eland meat to be marketed commercially. The two muscles showed separation from one another regarding both sensory and physical attributes, which should be considered for their commercial sale. Thus, sex had minor influences on the sensory eating quality of Eland meat; however, the BF and LTL muscles were considered tough and further ageing thereof should be evaluated.

  • Seroprevalence of Neospora caninum and Toxoplasma gondii in exotic ruminants and camelids in the Czech Republic
    Parasitology Research, 2017
    Co-Authors: Eva Bártová, Radim Kotrba, Kateřina Kobédová, Jiří Lamka, Roman Vodička, Kamil Sedlák
    Abstract:

    Neospora caninum and Toxoplasma gondii are the protozoan parasites with definitive hosts from order Carnivora . Due to vertical transmission, both parasites can cause abortions and neonatal mortality that lead to significant productive and economic losses in the domestic ruminants. The aim of this study was to describe N. caninum and T. gondii seroprevalence in the group of frequently farmed captive exotic ruminants ( n  = 184) including Bovidae (barbary sheep, bezoar goat, common Eland, American bison, water buffalo, and yak) and Camelidae (bactrian camel, guanaco, llama, and alpaca). Antibodies were tested by indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). Higher prevalence of T. gondii antibodies (31% in IFAT and 52% in ELISA) was detected compared to N. caninum (24% in IFAT and 17% in cELISA). Mixed infection was found in 18 (10%) and 22 (12%) animals by IFAT and ELISA, respectively. Higher seroprevalence of both N. caninum and T. gondii was found in Camelidae compared to Bovidae. To author knowledge, this is the first detection of T. gondii and N. caninum antibodies in common Elands and bezoar goats.

  • Effect of species-specific differences in chromosome morphology on chromatin compaction and the frequency and distribution of RAD51 and MLH1 foci in two bovid species: cattle (Bos taurus) and the common Eland (Taurotragus oryx)
    Chromosoma, 2016
    Co-Authors: Hana Sebestova, Radim Kotrba, Miluse Vozdova, Svatava Kubickova, Halina Cernohorska, Jiri Rubes
    Abstract:

    Meiotic recombination between homologous chromosomes is crucial for their correct segregation into gametes and for generating diversity. We compared the frequency and distribution of MLH1 foci and RAD51 foci, synaptonemal complex (SC) length and DNA loop size in two related Bovidae species that share chromosome arm homology but show an extreme difference in their diploid chromosome number: cattle ( Bos taurus , 2n = 60) and the common Eland ( Taurotragus oryx , 2n_male = 31). Compared to cattle, significantly fewer MLH1 foci per cell were observed in the common Eland, which can be attributed to the lower number of initial double-strand breaks (DSBs) detected as RAD51 foci in leptonema. Despite the significantly shorter total autosomal SC length and longer DNA loop size of the common Eland bi-armed chromosomes compared to those of bovine acrocentrics, the overall crossover density in the common Eland was still lower than in cattle, probably due to the reduction in the number of MLH1 foci in the proximal regions of the bi-armed chromosomes. The formation of centric fusions during karyotype evolution of the common Eland accompanied by meiotic chromatin compaction has greater implications in the reduction in the number of DSBs in leptonema than in the decrease of MLH1 foci number in pachynema.

  • intramuscular fat characteristics of namibian common Eland tragelaphus oryx
    African Journal of Wildlife Research, 2015
    Co-Authors: L C Hoffman, Diana L Van Schalkwyk, K W Mcmillin, Radim Kotrba
    Abstract:

    The influence of sex on the intramuscular fat characteristics of Eland harvested from the highland savanna in Namibia was investigated. No significant differences were observed between sexes for moisture, protein and ash content in the longissimus dorsi et lumborum (LDL). Fat content in LDL differed (P = 0.022) for male (1.2%) and female (2.0%) Eland. Stearic acid was the most abundant fatty acid (26.1%) in males and differed from females (21.7%) whilst oleic acid was the most abundant fatty acid (39.1%) in females. Females contained more MUFA (41.6%) than males (26.4%). Eland had a PUFA to SFA ratio of 4–5. Males did not differ from females for the n-6:n-3 ratio (2.7–2.9). No significant differences for sex were observed in cholesterol content. The chemical composition of Eland can serve as a reference standard for future studies as well as be of value for food labelling and marketing strategies.

B L Dresser - One of the best experts on this subject based on the ideXlab platform.

  • 201 endoscope guided transfer of sperm injected oocytes into the oviducts of Eland and bongo antelopes
    Reproduction Fertility and Development, 2010
    Co-Authors: G Wirtu, R A Godke, D L Paccamonti, B L Dresser, R Maclean, J Galiguis, B E Eilts, U Besenfelder, G T Gentry
    Abstract:

    We have previously established methods of gamete collection in Eland and Bongo antelopes; however, blastocyst development following IVF, intracytoplasmic sperm injection (ICSI), or somatic cell nuclear transfer (SCNT) has been sporadic, thus few developmentally competent embryos have been produced for uterine transfer. In the present study, we evaluated the possibility of oviductal transfer of presumptive zygotes. Estrous cycle synchronization and ovarian superstimulation were done as follows. Females were treated with altrenogest (0.11 g/os daily) for 7 days. The FSH Folltropin® was dissolved in 30% polyvinylpyrrolidone and administered, i.m. on Days 5 and 7 of altrenogest treatment at 266 mg and 134 mg, respectively. Prostaglandin F2α (Lutalyse®, 25 mg, i.m.) was administered on Day 7. Transvaginal ultrasound-guided oocyte retrieval was conducted on Day 9 as described previously (Wirtu G et al. 2009 Anim. Reprod Sci. 111, 160-172). Eleven oocytes were recovered from 2 Elands and 4 from 1 Bongo. One oocyte of each species was degenerate at the time of recovery. Oocytes were subjected to IVM for approximately 22 h, when Piezo drill-assisted ICSI was done using frozen-thawed Eland or Bongo spermatozoa. Oocytes were activated (5% ethanol, 5 min) and cultured overnight in CR1aa medium supplemented with BSA. Presumptive zygotes were subjected to endoscopic oviductal transfer at 21 to 24 h after ICSI. The oviductal transfer was adapted from the technique developed in domestic cattle (Wetscher F et al. 2005 Theriogenology 64, 30-40). Animals were sedated by i.m. administration of xylazine HCl and butorphanol tartrate and restrained in a hydraulic chute (Tamer®) that was used to squeeze and lift the females. An epidural block (5 mL of lidocaine) was induced after cleaning the rectum, perineal region, and the injection site. A uterine relaxant, isoxsuprine HCl (10 mg, i.v), was then administered. The Brem/Besenfelder set for laparoscopic bovine embryo transfer (Karl Storz Endoscope, Karl Storz GmbH & Co. KG, Tuttlingen, Germany) was used for the oviductal transfer. Briefly, a cannula (12.5 mm in diameter, 49.5 cm long) fitted with a blunt-tip obturator was introduced into the vagina and positioned, during transrectal manipulation, at the mid-dorsal aspect of the vaginal vault. The blunt-tip obturator was replaced with a sharp-tip obturator, which was used to puncture through the vaginal wall for entry into the pelvic cavity. The trocar was subsequently replaced with an inner sheath containing a telescope (5.5 mm in diameter, 54 cm long) and a pre-loaded transfer tubing. Visualization of the ovaries and oviducts required insufflation of the peritoneal cavity. Transfer was done into the oviduct ipsilateral to the recently ovulating ovary. Ten Eland oocytes had extruded the first polar body and 8 that survived ICSI were transferred into the right oviduct of an Eland female. Two Bongo oocytes had extruded the first polar body; both survived ICSI and were transferred to the right oviduct of a Bongo female. Pregnancy diagnosis is pending. This minimally invasive method of accessing the oviduct has the potential to advance the application of assisted reproductive technologies in large nondomestic ungulates.

  • ultrasonographic guided retrieval and in vitro maturation of Eland taurotragus oryx and bongo tragelaphus eurycerus isaaci antelope oocytes
    Animal Reproduction Science, 2009
    Co-Authors: G Wirtu, R A Godke, D L Paccamonti, Earle C Pope, B L Dresser
    Abstract:

    The limited availability of gametes is a major factor hindering the development and application of assisted reproductive technologies (ART) in large non-domestic ungulates. This is partly due to the small number of captive animals and handling difficulties associated with procedures for gamete recovery. In the present study, results are reported of multi-year studies on ovarian stimulation and oocyte retrieval by ultrasonographic-guided transvaginal follicular aspiration and subsequent in vitro maturation (IVM) in Eland and bongo antelopes. All procedures were conducted on sedated females handled in a hydraulic chute without inducing general anesthesia. Five estrous synchronization/ovarian stimulation protocols were evaluated and data are presented on 73 and 15 procedures in Eland and bongo, respectively. Repeating procedures (≤once/month) on the same female did not affect ovarian response or number oocytes recovered in either species. Eland females, but not the ovarian stimulation treatment, affected ovarian response. Ovarian stimulation treatment affected oocyte recovery rate in Eland, but not in bongo. In both species, ovarian hormone stimulation treatment affected the distribution of follicles by size and the status of expansion of the cumulus cell investment of oocytes, but not the frequency of metaphase II oocytes during IVM. The timing of extrusion of the first polar body during IVM was more synchronous in bongo than in Eland oocytes. It is concluded that Transvaginal oocyte retrieval (TVOR) can be safely and repeatedly applied in gonadotropin-treated Eland and bongo females to recover oocytes that can mature in vitro. The methods described for the present study can be adapted to improve the availability of non-domestic ungulate oocytes for basic and applied studies.

  • cloned embryos from semen part 2 intergeneric nuclear transfer of semen derived Eland taurotragus oryx epithelial cells into bovine oocytes
    Cloning and Stem Cells, 2008
    Co-Authors: Liesl Nelthemaat, G Wirtu, A Cole, B L Dresser, Earle C Pope, M. C. Gómez, Monica Lopez, Jill A Jenkins, K R Bondioli, R A Godke
    Abstract:

    The production of cloned offspring by nuclear transfer (NT) of semen-derived somatic cells holds considerable potential for the incorporation of novel genes into endangered species populations. Because oocytes from endangered species are scarce, domestic species oocytes are often used as cytoplasts for interspecies NT. In the present study, epithelial cells isolated from Eland semen were used for intergeneric transfer (IgNT) into enucleated bovine oocytes and compared with bovine NT embryos. Cleavage rates of bovine NT and Eland IgNT embryos were similar (80 vs. 83%, respectively; p > 0.05); however, development to the morula and blastocyst stage was higher for bovine NT embryos (38 and 21%, respectively; p or = 8 cells at 84 hpa, while 32% of the bovine NT embryos had > or = 8 cells at the same interval. However, 100 and 66% of bovine NT and Eland IgNT embryos, respectively, that had > or = 8 cells synthesized DNA. From these results we concluded that (1) semen-derived epithelial cell nuclei can interact and be transcriptionally controlled by bovine cytoplast, (2) the first cell-cycle occurred in IgNT embryos, (3) a high frequency of developmental arrest occurs before the eight-cell stage in IgNT embryos, and (4) IgNT embryos that progress through the early cleavage stage arrest can (a) synthesize DNA, (b) progress through subsequent cell cycles, and (c) may have the potential to develop further.

  • isolation culture and characterisation of somatic cells derived from semen and milk of endangered sheep and Eland antelope
    Reproduction Fertility and Development, 2007
    Co-Authors: Liesl Nelthemaat, G Wirtu, B L Dresser, M. C. Gómez, P. Damiani, Kenneth R. Bondioli, Leslie A. Lyons
    Abstract:

    Semen and milk are potential sources of somatic cells for genome banks. In the present study, we cultured and characterised cells from: (1) cooled sheep milk; (2) fresh, cooled and frozen-thawed semen from Gulf Coast native (GCN) sheep (Ovis aries); and (3) fresh Eland (Taurotragus oryx) semen. Cells attached to the culture surface from fresh (29%), cooled (43%) and slow-frozen (1 degrees C/min; 14%) ram semen, whereas no attachment occurred in the fast-frozen (10 degrees C/min) group. Proliferation occurred in fresh (50%) and cooled (100%) groups, but no cells proliferated after passage 1 (P1). Eland semen yielded cell lines (100%) that were cryopreserved at P1. In samples from GCN and cross-bred milk, cell attachment (83% and 95%, respectively) and proliferation (60% and 37%, respectively) were observed. Immunocytochemical detection of cytokeratin indicated an epithelial origin of semen-derived cells, whereas milk yielded either fibroblasts, epithelial or a mixture of cell types. Deoxyribonucleic acid microsatellite analysis using cattle-derived markers confirmed that Eland cells were from the semen donor. Eland epithelial cells were transferred into Eland oocytes and 12 (71%), six (35%) and two (12%) embryos cleaved and developed to morulae or blastocyst stages, respectively. In conclusion, we have developed a technique for obtaining somatic cells from semen. We have also demonstrated that semen-derived cells can serve as karyoplast donors for nuclear transfer.

  • 249 intracytoplasmic sperm injection of Eland taurotragus oryx and bongo tragelaphus eurycerus antelope oocytes
    Reproduction Fertility and Development, 2007
    Co-Authors: G Wirtu, C E Pope, A Cole, D L Paccamonti, M C Gomez, B L Dresser
    Abstract:

    Intracytoplasmic sperm injection (ICSI) is an assisted reproductive technique applicable in cases of limited male gamete availability. Moreover, it bypasses barriers of the oocyte, thus avoiding poorly understood species-specific capacitation events affecting sperm–egg interaction. In the present study, we evaluated the application of conventional and piezo drill-assisted ICSI and whether subsequent chemical activation is required for initiating embryonic development in Eland (Taurotragus oryx) and bongo (Tragelaphus eurycerus) oocytes. Oocytes were collected using transvaginal ultrasound-guided follicular aspiration after gonadotropin-induced ovarian stimulation and incubated in modified TCM-199 medium (Gomez et al. 2000 Reprod. Fertil. Dev. 12, 423) containing 10% FBS. After 3 to 24 h, the cumulus cell layers were removed either by repeated mouth-pipetting and/or by using hyaluronidase. Oocytes with an extruded first polar body were used for ICSI and the other oocytes were returned to culture and evaluated every six hours Piezo drill-assisted (Kimura and Yanagimachi 1995 Biol. Reprod. 52, 709) or conventional (Gomez et al.) ICSI were done as described previously using glass pipettes with internal tip diameters of 9–10 µm. We used frozen–thawed or freshly collected spermatozoa that were kept in HEPES-buffered Tyrode's medium (Gomez et al.) for up to 24 h. Four to 6 h after ICSI, 3 activation treatments were examined: (1) none; (2) 7% ethanol, 5 min; or (3) calcium ionophore (5 µM, 5 min) followed by DMAP (2 mM, 4 h). Then we cultured oocytes in a humidified atmosphere of 5% O2, 5% CO2, and 90% N2 at 38.5°C in one of 3 media: SOF, α-MEM, or CR1aa containing essential and nonessential amino acids and FBS. Fifty-three of 70 (76%) Eland oocytes survived after piezo-ICSI, and 13 of 16 (81%) survived after conventional ICSI. For bongo oocytes, 27 of 30 (90%) survived piezo-ICSI and all (n = 8) survived after conventional ICSI. Table 1 outlines cleavage data on Day 2. Generally, embryonic development was arrested at about 10 cells. In summary, Eland and bongo oocytes can survive both conventional and piezo drill-assisted ICSI. Activation treatments do not appear to be a prerequisite for initiating cleavage after ICSI in Eland and bongo antelope oocytes. Table 1.Cleavage of Eland and bongo antelope oocytes after conventional or piezo-ICSI and three activation treatments

C E Pope - One of the best experts on this subject based on the ideXlab platform.

  • Daily or thrice weekly handling of Eland antelope (Taurotragus oryx): effects on serum cortisol level.
    Research in veterinary science, 2013
    Co-Authors: P. M. Pennington, C E Pope, Robert A. Godke, J Galiguis, R.a. Maclean, L.r. Gentry, G Wirtu
    Abstract:

    We evaluated the cortisol response of adult female Eland (n=8) that were handled in hydraulic chute daily or 3×/week. Females were divided into two groups and each group (n=4) successively received two estrous cycle synchronization treatments: (1) two injections of prostaglandin (PG-PG) F2α at 11 day intervals and (2) oral administration of altrenogest for 7 days and an injection of PGF2α on day 7 (Alt-PG). Blood samples were collected 3×/week during the synchronization (Synch) and expected luteal phase (Nonintensive) periods, and daily during the expected time of induced (Intensive 1) or natural (Intensive 2) estrus. Overall, mean cortisol levels were highest during Intensive 1, followed by Intensive 2, Synch and Nonintensive periods. Individual Eland were the most significant source of variation for cortisol level. The frequency of handling and the synchronization treatment significantly affected cortisol levels in 3/8 and 4/8 females, respectively. In conclusion, in response to increased frequency of handling, Eland cortisol levels rose transiently and returned to baseline within few days after more intensive handling. Thus, the Eland females were tolerant to and recovered from the effects of repeated daily handling.

  • 169 BEHAVIORAL ASPECTS OF ESTRUS IN THE COMMON Eland ANTELOPE, TAUROTRAGUS ORYX
    Reproduction Fertility and Development, 2009
    Co-Authors: G Wirtu, C E Pope, Robert A. Godke, J Galiguis, P. M. Pennington, R.a. Maclean, J. Mercado, Dale L. Paccamonti, Betsy L. Dresser
    Abstract:

    Knowledge of the estrous cycle and behavior is important in managing captive breeding programs. However, such information is negligible in spiral-horned antelopes, including the Eland. In the present study, conducted between September 2007 and March 2008, we sought to characterize mounting activity and courtship behavior in a group of eight adult Eland females. Estrus was induced in groups of four females by treatment with 25 mg PGF2 ∝ (IM, Lutalyse®, Pharmacia and Upjohn Company, Kalamazoo, MI, USA) administered after 7 days of daily oral progestin (5 mL of 2.2% altrenogest, DPT Laboratories, San Antonio, TX, USA) or 11 days after an initial treatment of PGF2 ∝ (25 mg). The eight females received each treatment in a crossover design. Females were induced and monitored during the induced and subsequent natural estrus for 34 to 38 days. Observations for estrus behavior were initially assisted by using Estrotect™ heat detector patches (Rockyway, Inc., www.estrotect.com). Since January 2008, the HeatWatch® electronic heat detection system (CowChips, Denver, CO, USA) and an androgenized Eland female were used to assist with heat detection. For androgenization, 8 pellets of Synovex-H® (Fort Dodge Animal Health, Fort Dodge, IA) comprising a total dose of 1600 mg testosterone propionate and 160 mg estradiol benzoate were implanted (SC) on the convex side of the ear. To confirm mounting activity detected by Estrotect™ or HeatWatch®, Eland behavior was recorded continuously using an 8-channel real time DVR. Chi-square analysis was used to test the association between time of day (day or night) and the frequency of standing to be mounted. The cycle length (n = 6 females) was the time interval (d) between the first mounts at the induced and natural estrus. The time of mounts was determined for a total of 52 mounts. More than half of the mounts (n = 32 or 61.5%) occurred between 6 pm and 6 am (night); however, there was no association between the number of mounts and time of the day (p = 0.886). Analysis of mounts by quarters of the day showed that mounts were evenly distributed between early afternoon (noon to 6 pm: 29%), late afternoon (6 pm to midnight: 33%) and early morning (midnight to 6 am: 29%) but were less frequent during late morning, between 6 am and noon (10%) possibly due to interference by human activities. The number of mounts observed per female ranged from 1 to 20. Mounts lasted for 2 seconds or less. The duration of estrus ranged from 2.1 to 29.0 hours. Typical courtship behaviors included prolonged head butting, flehmen response and following the female in estrus before mounting. The average length of the estrous cycle, based on mounting activities, was 19.3 d (range: 17–24). Although further studies are required in herds with Eland males, the present results suggest that estrus detection for captive breeding or application of reproductive technologies in the Eland should be spread around different times of the day and night. We have also demonstrated that an electronic mount detection system can be applied to assist with determination of estrus in the Eland.

  • 274 Lysolecithin treatment of Eland, bongo, and bovine spermatozoa and cleavage of bovine oocytes after interspecies intracytoplasmic sperm injection
    Reproduction Fertility and Development, 2008
    Co-Authors: G Wirtu, C E Pope, M. C. Gómez, R.a. Maclean, Dale L. Paccamonti, Betsy L. Dresser
    Abstract:

    Compared to success rates in human, intracytoplasmic sperm injection (ICSI) is inefficient in ungulate species. Although factors such as injection of membrane-intact sperm and toxic effects of acrosome contents are suspected causes, the reasons for the inefficiency are unclear. A recent report in mice demonstrated that ICSI using spermatozoa treated with a physiological detergent, lysolecithin, improved oocyte activation, cleavage, and offspring production after embryo transfer (Morozumi K et al. 2006 PNAS 109, 17 661–17 666). The objectives of the present study were to evaluate the effects of detergent treatment on motility and membrane integrity of frozen thawed Eland, bongo and bovine spermatozoa and to examine sperm decondensation/embryonic cleavage following ICSI of in vitro-matured bovine oocytes. In experiment 1, sperm motility was observed on a warm microscope stage during exposure to 3 lecithin concentrations, 0.04, 0.02, and 0.01%, and the time at which 100% of the spermatozoa lost motility was recorded. In experiment 2, spermatozoa were exposed to 0.02% lecithin for 22 s, and the membrane integrity and acrosome status of spermatozoa were determined using a combined trypan blue-Giemsa staining (Nagy et al. 1999 Theriogenology 52, 1153–1159). In experiment 3, bovine oocytes were injected, using the piezo drill, with lecithin-treated (0.02%, immobilized) or untreated (piezo pulse immobilized) Eland, bongo, or bovine spermatozoa and subsequently cultured for 2 days in CR1aa containing 3 mg mL–1 BSA. Each experiment was replicated at least 3 times. Lecithin induced time- and concentration-dependent loss of sperm motility. The average time to loss of motility in 100% of the spermatozoa at 0.04, 0.02, and 0.01% lecithin was 107, 222, and 344 s in bovine; 82, 135, and 179 s in Eland; and 65, 115, and 158 in bongo, respectively. Data on membrane integrity (intact or nonintact) and acrosome status (reacted or nonreacted) of detergent-treated or control spermatozoa are shown in Table 1. Sperm head decondensation and embryonic cleavage were observed following homologous and interspecies (antelope into bovine) ICSI of lecithin-treated or control spermatozoa. In conclusion, lecithin treatment induced concentration and time-dependent loss of motility and was effective in damaging the sperm membrane and acrosome in Eland, bongo, and domestic bulls. Eland and bongo spermatozoa underwent decondensation and activated bovine oocytes after interspecies ICSI. Table 1.

  • Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and Eland antelope.
    Reproduction fertility and development, 2007
    Co-Authors: Liesl Nel-themaat, G Wirtu, C E Pope, Betsy L. Dresser, M. C. Gómez, P. Damiani, Kenneth R. Bondioli, Leslie A. Lyons, Robert A. Godke
    Abstract:

    Semen and milk are potential sources of somatic cells for genome banks. In the present study, we cultured and characterised cells from: (1) cooled sheep milk; (2) fresh, cooled and frozen-thawed semen from Gulf Coast native (GCN) sheep (Ovis aries); and (3) fresh Eland (Taurotragus oryx) semen. Cells attached to the culture surface from fresh (29%), cooled (43%) and slow-frozen (1 ◦ C/min; 14%) ram semen, whereas no attachment occurred in the fast- frozen (10 ◦ C/min) group. Proliferation occurred in fresh (50%) and cooled (100%) groups, but no cells proliferated after passage 1 (P1). Eland semen yielded cell lines (100%) that were cryopreserved at P1. In samples from GCN and cross- bred milk, cell attachment (83% and 95%, respectively) and proliferation (60% and 37%, respectively) were observed. Immunocytochemical detection of cytokeratin indicated an epithelial origin of semen-derived cells, whereas milk yielded either fibroblasts, epithelial or a mixture of cell types. Deoxyribonucleic acid microsatellite analysis using cattle-derived markers confirmed that Eland cells were from the semen donor. Eland epithelial cells were transferred into Eland oocytes and 12 (71%), six (35%) and two (12%) embryos cleaved and developed to morulae or blastocyst stages, respectively. In conclusion, we have developed a technique for obtaining somatic cells from semen. We have also demonstrated that semen-derived cells can serve as karyoplast donors for nuclear transfer.

  • 249 intracytoplasmic sperm injection of Eland taurotragus oryx and bongo tragelaphus eurycerus antelope oocytes
    Reproduction Fertility and Development, 2007
    Co-Authors: G Wirtu, C E Pope, A Cole, D L Paccamonti, M C Gomez, B L Dresser
    Abstract:

    Intracytoplasmic sperm injection (ICSI) is an assisted reproductive technique applicable in cases of limited male gamete availability. Moreover, it bypasses barriers of the oocyte, thus avoiding poorly understood species-specific capacitation events affecting sperm–egg interaction. In the present study, we evaluated the application of conventional and piezo drill-assisted ICSI and whether subsequent chemical activation is required for initiating embryonic development in Eland (Taurotragus oryx) and bongo (Tragelaphus eurycerus) oocytes. Oocytes were collected using transvaginal ultrasound-guided follicular aspiration after gonadotropin-induced ovarian stimulation and incubated in modified TCM-199 medium (Gomez et al. 2000 Reprod. Fertil. Dev. 12, 423) containing 10% FBS. After 3 to 24 h, the cumulus cell layers were removed either by repeated mouth-pipetting and/or by using hyaluronidase. Oocytes with an extruded first polar body were used for ICSI and the other oocytes were returned to culture and evaluated every six hours Piezo drill-assisted (Kimura and Yanagimachi 1995 Biol. Reprod. 52, 709) or conventional (Gomez et al.) ICSI were done as described previously using glass pipettes with internal tip diameters of 9–10 µm. We used frozen–thawed or freshly collected spermatozoa that were kept in HEPES-buffered Tyrode's medium (Gomez et al.) for up to 24 h. Four to 6 h after ICSI, 3 activation treatments were examined: (1) none; (2) 7% ethanol, 5 min; or (3) calcium ionophore (5 µM, 5 min) followed by DMAP (2 mM, 4 h). Then we cultured oocytes in a humidified atmosphere of 5% O2, 5% CO2, and 90% N2 at 38.5°C in one of 3 media: SOF, α-MEM, or CR1aa containing essential and nonessential amino acids and FBS. Fifty-three of 70 (76%) Eland oocytes survived after piezo-ICSI, and 13 of 16 (81%) survived after conventional ICSI. For bongo oocytes, 27 of 30 (90%) survived piezo-ICSI and all (n = 8) survived after conventional ICSI. Table 1 outlines cleavage data on Day 2. Generally, embryonic development was arrested at about 10 cells. In summary, Eland and bongo oocytes can survive both conventional and piezo drill-assisted ICSI. Activation treatments do not appear to be a prerequisite for initiating cleavage after ICSI in Eland and bongo antelope oocytes. Table 1.Cleavage of Eland and bongo antelope oocytes after conventional or piezo-ICSI and three activation treatments

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