Lívia Batista Campos

Cryopreservation of collared peccaries (Tayassu tajacu) semen using a powdered coconut water (ACP-116c) based extender plus various concentrations of egg yolk and glycerol.

The objective was to determine the effectiveness of a powdered coconut water-based extender (ACP-116c), plus various concentrations of egg-yolk and glycerol, as an alternative for cryopreservation of collared peccary semen. Twelve ejaculates were obtained from captive adult males by electroejaculation, and evaluated for sperm motility, kinetic rating, viability, morphology, and functional membrane integrity. The ejaculates were apportioned into aliquots that were diluted in Tris plus 10% egg yolk and 3% glycerol, or in ACP-116c plus 10 or 20% egg yolk and 1.5 or 3% glycerol. Samples were frozen in liquid nitrogen and, after 1 mo, thawed at 37 °C for 1 min. After thawing, samples were evaluated as reported for fresh semen, and also for sperm membrane integrity (fluorescent probes) and kinematic parameters (computerized analysis). Results were presented as means ± SEM. Freezing and thawing decreased sperm characteristics relative to fresh semen. Overall, ACP-116c plus 20% egg yolk and 3% glycerol provided better (P < 0.05) sperm motility and kinetic rating (48 ± 6.1% and 2.8 ± 0.2, respectively) after thawing than Tris extender (30.4 ± 5.7% and 2.4 ± 0.2). However, there were no differences (P > 0.05) among treatments with regard to the other sperm characteristics. Based on computerized motion analysis, total (26.5 ± 5.9%) and progressive (8.1 ± 2.2%) motility were best preserved (P < 0.05) with the above-mentioned treatment. In conclusion, a coconut water-based extender, ACP-116c, plus 20% egg yolk and 3% glycerol, was effective for cryopreservation of semen from collared peccaries.

Monitoramento de ciclo estral em catetos (Pecari tajacu) criados em cativeiro em condições semiáridas

Collared peccaries (Peccary tajacu) are among the most hunted species in Latin America due the appreciation of their pelt and meat. In order to optimize breeding management of captive born collared peccaries in semiarid conditions, the objective was to describe and correlate the changes in the ovarian ultrasonographic pattern, hormonal profile, vulvar appearance, and vaginal cytology during the estrus cycle in this species. During 45 days, females (n=4) were subjected each three days to blood collection destined to hormonal dosage by enzyme immunoassay (EIA). In the same occasions, evaluation of external genitalia, ovarian ultrasonography and vaginal cytology were conducted. Results are presented as means and standard deviations. According to hormonal dosage, six estrous cycles were identified as lasting 21.0 ± 5.7 days, being on average 6 days for the estrogenic phase and 15 days for the progesterone phase. Estrogen presented mean peak values of 55.6 ± 20.5 pg/mL. During the luteal phase, the high values for progesterone were 35.3 ± 4.4 ng/mL. The presence of vaginal mucus, a reddish vaginal mucosa and the separation of the vulvar lips were verified in all animals during the estrogenic peak. Through ultrasonography, ovarian follicles measuring 0.2±0.1 cm were visualized during the estrogen peak. Corpora lutea presented hyperechoic regions measuring 0.4±0.2 cm identified during luteal phase. No significant differences (P>0.05) between proportions of vaginal epithelial cells were identified when comparing estrogenic and progesterone phases. In conclusion, female collared peccaries, captive born in semiarid conditions, have an estral cycle that lasts 21.0±5.7 days, with estrous signs characterized by vulvar lips edema and hyperemic vaginal mucosa, coinciding with developed follicles and high estrogen levels.

Monitoramento do ciclo estral de cutias (Dasyprocta leporina Lichtenstein, 1823) através de citologia esfoliativa vaginal e ultrassonografia

ABSTRACT.- Campos L.B.,Peixoto G.C.X., Lima G.L., Castelo T.S., Souza A.L.P., Oliveira M.F. & Silva A.R. 2015. [ Monitoring of the estrous cycle of agoutis ( Dasyprocta leporina Li-chtenstein, 1823) by vaginal exfoliative cytology and ultrasonography. ] Monitoramen-to do ciclo estral de cutias ( Dasyprocta leporina Lichtenstein, 1823) atraves de citologia esfoliativa vaginal e ultrassonografia. Pesquisa Veterinaria Brasileira 35(2):188-192. Labora-torio de Conservacao de Germoplasma, Universidade Federal Rural do Semi-Arido, BR-110 Km 47, Costa e Silva, Mossoro, RN 59625-900, Brazil. E-mail: legio2000@yahoo.comThe objective of the study was to monitor the estrous cycle in agoutis ( Dasyprocta lepo-rina ) bred in captivity in Brazilian semiarid. During 70 days, five agoutis were daily subjec-ted to vaginal exfoliative cytology, and the ovarian ultrasound monitoring was conducted every three days. A total of 8 estrous cycles were completely monitored, lasting 28.2±0.7 days, ranging from 24 to 31 days. By vaginal exfoliative cytology, there was predominance of superficial cells at proestrus and estrus phases (P 0.05). Follicles during the estrogenic phases (proestrus and estrus) were identified, with an average diameter of 1±0.5mm. In only 12.5% of luteal phases, cor-pora lutea measuring 1.4±0.9mm were identified. We conclude that the association of vagi-nal cytology and ovarian ultrasonography is a useful alternative for monitoring the estrous cycle and identifying the estrogenic phases in

Use of Non-invasive Methods for Evaluating the Testicular Biometry in Collared Peccaries (Pecari tajacu Linnaeus, 1758)

The aim of this study was to compare the accuracy of two methods used to estimate testicular volume in the collared peccary. Calliper and ultrasonographic measurements of testicular dimensions (length, width and height) of both testes were taken on five adult collared peccaries. The testicular volume was calculated by Lamberts empiric formula: length (L) × width (W) × height (H) × 0.71, the formula of an ellipsoid L × W × H × 0.52, and Hansens formula: L × W2 × 0.52. The calculated volumes were then compared with the actual ones, which were estimated by water displacement. The mean of true testicular volume was 22.65 ± 1.52 ml. Lamberts formula estimated testicular volume more accurately when ultrasound measurements were taken. However, when the calliper was the methodology used, the results were closest to the true volume, especially when Ellipsoid formula and Hansens formula were applied, and underestimated the true volumes by 1.53 ± 1.75 ml and 1.53 ± 1.65 ml, respectively. This specific application of technologies in wild animals has the potential to revolutionize the selection process for the collared peccary entering artificial insemination or natural breeding programmes.

Comparison of different extenders on the recovery and longevity of epididymal sperm from Spix's yellow-toothed cavies (Galea spixii Wagler, 1831)

The aim of this study was to evaluate the performance of cavy (Galea spixii) epididymal sperm following addition to TES or TRIS extenders and using a thermal resistance test (TRT), as well as fluorescence analysis as a complementary method to predict the viability of these gametes. Nine testicle-epididymis complexes were used for sperm collection using a flotation method. Epididymis tails were sliced and one was immersed in 3 ml of TRIS buffer, and the other in 3 ml of TES, for 5 min. After sperm recovery, the samples were subjected to a TRT which involved incubation in a water bath at 37°C for 3 h. During incubation, sample parameters were assessed at 0, 15, 30, 60, 90, 120, 150 or 180 min intervals. Results indicated that the TRIS diluent was more efficient than TES (P < 0.05) for the maintenance of sperm parameters in Spixs yellow-toothed cavies over the whole TRT, maintaining sperm longevity for an extended time. In conclusion, we indicate the use of TRIS diluent for recovery and maintenance of longevity of epididymal sperm from cavies (G. spixii).

Estrous synchronization in captive collared peccaries (Pecari tajacu) using a prostaglandin F2α analog.

We verify the efficiency of a protocol for estrus synchronization in captive female collared peccaries (Pecaricari tajacu) using the prostaglandin analog D-cloprostenol. Five adult female collared peccaries received an intramuscular administration of 60 µg D-cloprostenol, which procedure was repeated after a 9-day interval. For 10 days after second the D-cloprostenol administration, females were monitored for changes in external genitalia, ovarian ultrasonography, vaginal cytology and reproductive hormonal dosage. As a result, four females synchronized their estrous at 9.5 ± 0.5 days after the second administration of the prostaglandin analog. Such females showed external signs of estrus, including vulvar opening, hyperemic vaginal mucosa, and vaginal mucus, concomitant with an increase in the proportion of superficial cells (52.2 ± 9.9%) verified through vaginal cytology. An estrogen peak of 22.7 ± 3.4 pg/ml was detected by hormonal dosage, and the presence of anechoic follicles measuring 0.29 ± 0.05 × 0.32 ± 0.07 mm were detected in the ovary by ultrasonography. Given these findings, we suggest that D-cloprostenol may be effective for use in estrus synchronization in collared peccaries.

Environmental Factors Related to a Semiarid Climate Influence the Freezability of Sperm from Collared Peccaries (Pecari tajacu Linnaeus, 1758)

The influence of environmental factors in a semiarid climate on characteristics of fresh and frozen/thawed sperm collected from collared peccaries (Pecari tajacu) was assessed. Semen from 11 male collared peccaries was collected by electroejaculation during the peaks of the dry and rainy periods while rainfall indices, air temperatures, relative humidity levels, and wind speeds were measured. The number, motility, morphology, osmotic response, and membrane integrity of sperm in the collected ejaculates were assessed. Samples were then frozen in liquid nitrogen, thawed, and reassessed. The rainfall index of the rainy period (73.2 mm) was significantly higher than that of the dry period (13.6 mm) and the relative humidity was significantly higher during the rainy period (74.6%) than it was during the dry period (66.8%). Air temperature and wind speed did not differ between the two periods. Characteristics of sperm in the fresh samples were not affected by environmental parameters. In contrast, computerized analysis revealed that sperm in samples frozen during the rainy period exhibited better post-thaw membrane integrity (28.6 ± 6%), motility (29.5 ± 7.7%), and rapid sperm population (13.7 ± 6.2%) than did sperm in samples frozen during the dry period (23.4 ± 3% membrane integrity, 14.6 ± 4.1% motility, and 4.1 ± 1.2% rapid sperm; p < 0.05). Other characteristics of the frozen/thawed sperm did not differ depending on the period in which they were collected. We demonstrated that environmental parameters did not affect the quality of fresh sperm, but could influence the freezability of sperm collected from collared peccaries raised under a semiarid climate.

Epididymal sperm from Spix’s yellow-toothed cavies sperm successfully cryopreserved in Tris extender with 6% glycerol and 20% egg yolk

As a non-threatened hystricognath rodent species, Spixs yellow-toothed cavies can be used as a model for the development of assisted reproductive techniques for the conservation of closely related species. The objective was to establish a functional protocol for cryopreservation of epididymal sperm from these cavies. Twelve sexually mature males, ∼2 y old and weighing ∼300 g, were euthanized. Sperm were recovered by retrograde flushing of the vas deferens and cauda epididymis with Tris extender. Thereafter, sperm were extended in Tris plus 20% egg yolk, with 3%, 6% or 9% glycerol or dimethyl sulfoxide (DMSO), placed in 0.25 mL straws and cryopreserved in liquid nitrogen. Sperm concentration, motility (using computer-assisted sperm analysis; CASA), plasma membrane integrity, osmotic response, morphology and sperm binding-ability were determined in fresh and frozen-thawed sperm. For most sperm endpoints, glycerol was a more desirable cryoprotectant than DMSO. Data (mean ± SEM) were similar with use of 3%, 6%, and 9% glycerol (P > 0.05) in osmotic response (40.66 ± 6.3%, 42.5 ± 7.1%, and 39.5 ± 5.0% respectably), and membrane integrity (55.17 ± 5.5%, 68.4 ± 4.1%, and 59.1 ± 4.9% respectably). Among concentrations assessed, the use of 6% glycerol resulted in the greatest (P < 0.05) post-thaw values for total motility (60.9 ± 4.4%), rapid subpopulation motility (27.7 ± 3.1%) and sperm-binding capability (227.0 ± 20.2). In conclusion, epididymal sperm from the Spixs yellow-toothed cavies (G. spixii) are optimally cryopreserved in Tris extender with 6% glycerol and 20% egg yolk.