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In vitro production of embryos has gained prominence as a tool for use in wildlife conservation programmes in situ and ex situ. However, the development of this technique depends on steps that include ovarian stimulation, collection and oocyte maturation. The purpose of this study was to assess the feasibility of an ovarian stimulation protocol for follicular aspiration, the efficiency of videolaparoscopy for follicular aspiration and test a medium for in vitro oocyte maturation for the species Mazama gouazoubira. Five females were submitted to repeated ovarian stimulation (hormone protocol using controlled internal drug release), and estradiol benzoate on D0 and eight injections of follicle-stimulating hormone, once every 12 h, from D4 onwards at 30-day intervals. Fourteen surgical procedures were performed in superstimulated females, resulting in the collection of 94 oocytes and an average of 17.1 ± 9.1 follicles observed, 13.5 ± 6.6 follicles aspirated and 7.2 ± 3.7 oocytes collected per surgery. After collection, the oocytes were submitted to in vitro maturation for 24 h and stained with Hoechst 33342 dye to evaluate their nuclear status; 64.5% of the oocytes reached MII and 16.1% were spontaneously activated by parthenogenesis. The nuclear status of oocytes that did not undergo in vitro maturation was evaluated; 80.9% were found to be immature.

Made available in DSpace on 2022-04-29T08:29:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Summary In vitro production of embryos has gained prominence as a tool for use in wildlife conservation programmes in situ and ex situ. However, the development of this technique depends on steps that include ovarian stimulation, collection and oocyte maturation. The purpose of this study was to assess the feasibility of an ovarian stimulation protocol for follicular aspiration, the efficiency of videolaparoscopy for follicular aspiration and test a medium for in vitro oocyte maturation for the species Mazama gouazoubira. Five females were submitted to repeated ovarian stimulation (hormone protocol using controlled internal drug release), and estradiol benzoate on D0 and eight injections of follicle-stimulating hormone, once every 12 h, from D4 onwards at 30-day intervals. Fourteen surgical procedures were performed in superstimulated females, resulting in the collection of 94 oocytes and an average of 17.1 ± 9.1 follicles observed, 13.5 ± 6.6 follicles aspirated and 7.2 ± 3.7 oocytes collected per surgery. After collection, the oocytes were submitted to in vitro maturation for 24 h and stained with Hoechst 33342 dye to evaluate their nuclear status; 64.5% of the oocytes reached MII and 16.1% were spontaneously activated by parthenogenesis. The nuclear status of oocytes that did not undergo in vitro maturation was evaluated; 80.9% were found to be immature. NUPECCe - Núcleo de Pesquisa e Conservação de Cervídeos Departamento de Zootecnia Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista Departamento de Zootecnia Centro de Ciências Agrárias Universidade Federal da Paraíba Departamento de Medicina Veterinária Faculdade de Zootecnia e Engenharia de Alimentos Universidade de São Paulo NUPECCe - Núcleo de Pesquisa e Conservação de Cervídeos Departamento de Zootecnia Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista

Made available in DSpace on 2018-11-26T15:44:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-06-01 The red brocket (Mazama americana) is a South American deer with a wide geographical distribution that presents different chromosomal variants depending on their location. At least six different cytotypes belonging to two distinct evolutionary lineages have been described. This study aimed to verify the existence of postzygotic reproductive isolation between cytotypes of M. americana by comparative evaluation of pure and hybrid males. Seven 18-month-old bucks were submitted to seminal collection and evaluation and testicle histological evaluations. The pure males showed normal parameters for sperm quality and testicular histology. Hybrids from the same evolutionary lineage (= 10 chromosomes different from progenitors) presented azoospermia and evidence of testicular degeneration. Despite the striking morphological similarities, we can conclude that populations with more distinct karyotypes possess an effective reproductive barrier; moreover, there is evidence that reproductive isolation mechanisms exist between some closer karyotypes, corroborating the hypothesis that M. americana is best characterized as a superspecies. Thus, the future description of several new species for this taxon is expected, since the tendency is to establish efficient mechanisms of postzygotic reproductive isolation, preventing the introgression and fusion of genomes from different populations through chromosome variation. Summary Sentence Differences equal to or greater than seven chromosomes produce an effective reproductive isolation between red brocket deer cytotypes and a depression in fertility occurs among karyotypes with at least two chromosome diferences. Univ Fed Piaui, Lab Histol & Embriol, Teresina, PI, Brazil Univ Estadual Paulista, Nucleo Pesquisa & Conservacao Cervideos, Jaboticabal, SP, Brazil Univ Fed Mato Grosso, Inst Ciencias Biol & Saude, Campus Univ Araguaia, Pontal Do Araguaia, MT, Brazil Univ Estadual Paulista, Nucleo Pesquisa & Conservacao Cervideos, Jaboticabal, SP, Brazil

In recent years, genome banks have grown as a way of maintaining the genetic variability of populations. However, the quality of gamete cryopreservation will determine their efficiency. This study aimed to evaluate prefreeze and postthaw sperm motility, vigor, membrane integrity, and morphology of semen of red brocket deer (Mazama americana) using three extenders: E1, Tris-Yolk; E2, Tes-Tris-Yolk; and E3, Tes-Tris-Yolk-Equex. Six bucks were used, and three collections per buck were performed at 90-day intervals. Before freezing, semen volume, ejaculate concentration, motility, vigor, membrane integrity, and sperm morphology were evaluated. To compare the effect exerted by the extenders after sample thawing, further analyses of sperm motility, vigor, membrane integrity, and morphology were performed. Mean ejaculate volume and sperm concentration were 365.33 +/- 120.5 microL and 2,675.73 +/- 810.4 sperm/mL, respectively. Prefreeze motility for the extenders showed no significant differences (approximately 60%). Postthaw motility (E1 = 16.33 +/- 5.5, E2 = 5.44 +/- 5.2, E3 = 24.66 +/- 10.0) was significantly different between E2 and E3, whereas postthaw vigor (E1 =2.66 +/- 0.8, E2= 1.89 +/- 1.2, E3 = 3.83 +/- 0.4) was greater for E3 (Por = 0.05). Analysis of postthaw membrane integrity revealed no significant differences between the extenders regarding counts of cells presenting intact membranes; however, E3 promoted the lowest number of cells with damaged membranes and higher cell counts for partially damaged membranes (Por = 0.05). Analysis of sperm morphology revealed an increase in severe abnormalities when using E2 and E3 (P0.05). However, observation verified that counts of altered cells were lower using E3 than E2, suggesting a protective effect of Equex. These findings indicate that E3 promoted better semen quality postthaw. However, the performance of this extender in protecting sperm cells of M. americana during freezing was lower than that verified for other species. Thus, further studies are needed for characterization of red brocket deer semen and the optimization of extender for sperm cryopreservation.

This study aimed to validate the enzyme immunoassay (EIA) for fecal progestin quantification of the species Mazama americana, define its excretion profile during periods of gestation and postpartum and determine the gestation period and resumption of postpartum ovarian activity in this species in captivity. Fecal samples were collected twice a week during gestation and every day in the postpartum period, and analyzed using EIA. The mean concentrations (±SEM) of fecal progestins during gestation were 2180.0±299.1ng/g in early pregnancy (week 1-11), 3271.4±406.9ng/g in middle pregnancy (week 12-22) and 5592.0±1125.8ng/g in late pregnancy (week 23-32). The gestation period determined for the species was 220.9±1.2 days. The concentration of progestins reached its peak prior to parturition and returned to baseline levels in 4±0.31 days after parturition. In the postpartum period, the mean concentrations of fecal progestins were 1564.2±182.6ng/g in the interval between parturition and resumption of ovarian activity, 469.8±24.5ng/g in the inter-luteal phase and 2401.7±318.5ng/g during the luteal phase, such that the postpartum period and the luteal phase differed from the inter-luteal phase. Fecal progestin profiling permitted the detection of ovulation 26.9±3.4 days after parturition in all the hinds studied and estimation of the mean duration of the estrous cycle, 21.3±1.1 days. Analysis established that concentrations of progestins above 3038.76ng/g diagnosed pregnancy, a value determined from the week 12 of gestation. Moreover, the quantification of fecal progestins by EIA proved to be an important tool for noninvasive endocrine monitoring and to obtain reproductive data on the species M. americana in captivity.

This study aims to evaluate the ovulation rate and the presence of functional corpora lutea after treatment by three different protocols designed to cause superovulation in brown brocket deer. Six female received an intravaginal device containing 0.33 g of progesterone (CIDR®) for 8 days, followed by 0.5 mg injection of estradiol benzoate at the time of insertion and 265 µg of cloprostenol at the time of removal. Afterwards, the hinds were divided into three groups (n = 2): Treatment A received injection of 600 IU eCG on Day 4 after CIDR® insertion; Treatment B received injection of 300 IU eCG at the same time; and Treatment C received injection of 250 IU FSH dissolved in PVP, also on Day 4 post-insertion. The treatments were crossed over with 44-48 day intervals after CIDR® removal, such that all the deer were submitted to all three treatments. The mean ovulation rate (Treatment A = 3.40 ± 0.68, Treatment B = 1.40 ± 0.24, Treatment C = 0.80 ± 0.49), total ovarian stimulation (Treatment A = 4.80 ± 1.02, Treatment B = 1.80 ± 0.37, Treatment C = 1.40 ± 0.60), and mean CL diameter (Treatment A = 7.33 ± 0.76 mm, Treatment B = 3.94 ± 0.19 mm, Treatment C = 2.18 ± 0.49 mm) in Treatment A were significantly higher than the mean ovulation rates, total ovarian stimulation, and mean CL diameter in Treatments B and C. The mean fecal progesterone metabolites at the luteal phase in Treatment A (6,277.94±2,232.47 ng/g feces) was significantly different from Treatment C (1,374.82±401.77 ng/g feces). Thus, although fertility was not evaluated directly, Treatment A proved capable of induce superovulation in the species Mazama gouazoubira, presenting the greatest mean ovulation rates, with the formation of functional corpora lutea.

This study aimed to establish a protocol for synchronization of estrus in brown brocket deer (Mazama gouazoubira). Two groups of hinds (n = 3) were submitted to two different protocols: Treatment 1 received an intravaginal progesterone (CIDR®) device for 8 days, followed by 265 μg injection of cloprostenol at the time of removal; and Treatment 2 received two injections of 265 μg of cloprostenol 11 days apart. After 30 days, each group of three hinds received the other treatment. Treatment efficacy was evaluated by reproductive behavior, fecal progestin and estrogen concentration and the observation of CL by laparoscopy 6 days after the end of estrus. All the hinds (100%) had estrous behavior upon the completion of treatment, but a significant difference occurred between the time of onset, 70.5 ± 5.0 h for Treatment 1 and 52.3 ± 5.6 h for Treatment 2. The mean estrus duration time (34.7 ± 4.50 and 37.0 ± 8.11 h), ovulation rates (5/6 and 4/6), mean CL size (4.85 ± 0.74 and 3.21 ± 0.19 mm) and mean fecal progestin concentration at 6 days after the end of estrus (865.53 ± 76.59 and 1073.35 ± 106.82 ng/g feces) were not significantly different between treatments. There was no difference in fecal estrogen concentrations throughout the treatment and the greatest values of the estrogen:progestin ratio coincided with estrous behavior. Although fertility was not evaluated directly, both treatments were effective in synchronizing estrus in the species M. gouazoubira, with the formation of functional corpora lutea.

Made available in DSpace on 2014-12-03T13:11:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-03-04Bitstream added on 2014-12-03T13:22:36Z : No. of bitstreams: 1 WOS000334458900002.pdf: 1785302 bytes, checksum: 67376de2e4b2cb31a6e4a38aec22f56a (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background: The red brocket deer, Mazama americana, has at least six distinct karyotypes in different regions of South America that suggest the existence of various species that are today all referred to as M. americana. From an evolutionary perspective, the red brockets are a relatively recent clade that has gone through intense diversification. This study sought to prove the existence of post-zygotic reproductive isolation in deer offspring between distinct chromosome lineages. To achieve this, inter-cytotype and intra-cytotype crosses were performed, which resulted in both F1 hybrid (n = 5) and pure offspring (n = 3) in captivity.Results: F1 females were analyzed in terms of their karyotypes, ovarian histology, estrous cycles and in vitro embryo production. Pure females presented parameters that were similar to those previously reported for M. Americana; however, the parameters for hybrid females were different. Two hybrids were determined to be sterile, while the remaining hybrids presented characteristics of subfertility.Conclusions: The results support the existence of well-established reproductive isolation among the most distant karyotype lineages and elucidates the need to define all karyotype variants and their geographical ranges in order to define the number of species of red brocket. Univ Estadual Paulista, UNESP, FCAV, Dept Zootecnia,NUPECCE Nucl Pesquisa & Conservaca, BR-14884900 Jaboticabal, SP, Brazil UNESP, FCAV, Programa Posgrad Med Vet Reproducao Anim, BR-14884900 Jaboticabal, SP, Brazil Fac Vet Med, Lab Embryol & Biotechn Reprod, BR-91501970 Porto Alegre, RS, Brazil Univ Estadual Paulista, UNESP, FCAV, Dept Zootecnia,NUPECCE Nucl Pesquisa & Conservaca, BR-14884900 Jaboticabal, SP, Brazil UNESP, FCAV, Programa Posgrad Med Vet Reproducao Anim, BR-14884900 Jaboticabal, SP, Brazil

Made available in DSpace on 2014-12-03T13:11:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-03-19Bitstream added on 2014-12-03T13:22:36Z : No. of bitstreams: 1 WOS000334703000001.pdf: 385890 bytes, checksum: 461e9e117c2e56211fe64f5764411b18 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Background: Stress is a limiting factor in assisted reproduction in wild animals maintained in captivity. However, the knowledge of assisted reproduction techniques for wild animals is useful for future in situ and ex situ conservation programs. Thus, this study evaluated the ovulation rate, presence of functional corpora lutea and fecal glucocorticoid levels following treatments promoting superovulation in captive brown brocket deer.Methods: The crossover design used six hinds, allocated to two groups (n = 6): eCG Treatment, CIDR for 8 days, followed by 0.25 mg of EB on day 0, 700 IU of eCG on day 4 following device insertion and 265 mug of PGF2alfa on day 8; and FSH Treatment, CIDR for 7.5 days, followed by 0.25 mg of EB on day 0, 130 mg of FSH in 8 equal doses and 265 mug of PGF2alfa on day 7.5. Induced adrenal activity and treatment efficacy were evaluated by corpora lutea (CL) counts and fecal glucocorticoid and progestin concentration (ng/g feces) analyses for five different phases: Pre, two days before treatment; Early, first four days of treatment; Late, last four days of treatment; Total, entire treatment period; and Post, five days posttreatment.Results: eCG Treatment resulted in the highest number of CL (P lower than 0.05). There was no significant difference for fecal glucocorticoid concentrations in five different time periods between the treatments; however Pre fecal glucocorticoid concentrations (90.06+/-19.64) were significantly different from Late (200.76+/-26.39) within FSH Treatment. The mean fecal progestin concentration and mean ovulation rate were higher in eCG Treatment (4293.69+/-769.47, 7.0+/-1.8) than in FSH Treatment (1571.26+/-240.28, 2.6+/-0.8) (P lower than or equal to 0.05).Conclusions: Although the eCG Treatment induced a good superovulatory response, with the formation of functional corpora lutea, we cannot yet affirm that we have established a suitable protocol for induction of SOV in the species M. gouazoubira because approximately 65% of the deer showed premature regression of the corpora lutea. Moreover, multiple FSH applications in FSH Treatment resulted in a low ovulation rate and induced an increase in fecal glucocorticoid levels. Univ Estadual Paulista, Fac Ciencias Agra & Vet, Deer Res & Conservat Ctr, Dept Zootecnia,NUPECCE,Nuleo Pesquisa & Conservac, BR-14884900 Jaboticabal, SP, Brazil Univ Estadual Paulista, Fac Ciencias Agra & Vet, Deer Res & Conservat Ctr, Dept Zootecnia,NUPECCE,Nuleo Pesquisa & Conservac, BR-14884900 Jaboticabal, SP, Brazil