Investigating the need for antibiotic supplementation to the extender used for semen cryopreservation in collared peccaries.

The objective was to investigate the effects of semen freezing extender supplementation with antibiotics on bacterial load of semen samples, sperm functional and morphological metrics in the collared peccary. Fresh ejaculates from 10 males were extended in Tris-egg yolk-glycerol supplemented or not (control) with gentamicin (70 µg/mL) streptomycin-penicillin (SP; 1 mg/mL-1000 IU/mL) or and cryopreserved in liquid nitrogen. Bacterial load, sperm motility patterns, morphology, membrane functionality and integrity, mitochondrial activity, chromatin integrity and sperm-binding ability were evaluated in fresh and frozen-thawed samples. Regardless of the use of antibiotics, the sole cryopreservation provoked a significant decrease (P < 0.05) in bacterial load compared to fresh samples (from average values > 1 x 106 CFU/mL to <0.4 × 106 CFU/mL). Post-thawing sperm kinetic parameters were not affected by the absence or presence of different antibiotics, except for beat cross frequency that was significantly (P < 0.05) impaired by SP supplementation compared to the group without antibiotics. After thawing, sperm morphology, membrane functionality and integrity, and mitochondrial activity were also not affected by the presence or absence of antibiotics; however, a significant decrease was observed in the group without antibiotics (P < 0.05) in comparison to fresh samples. Regarding sperm-binding ability, there were no differences among the different groups. While collared peccary semen could be efficiently cryopreserved in the absence of antibiotics in the extender, the use of both gentamicin or the streptomycin-penicillin combination is recommended as effective antibiotic supplementation for a further control of bacterial loads without affecting sperm parameters.

Single injection of eCG/hCG leads to successful estrous synchronization in the collared peccary (Pecari tajacu Linnaeus, 1758).

The establishment of protocols for the control of the ovarian function of collared peccaries is recommended for the development of assisted reproductive techniques. The goals were to (1) compare a gonadotropin combination with prostaglandin analogue to synchronize timing of onset of estrus among animals, and (2) elucidate the effects of the most desirable protocol for performing an artificial insemination study and macroscopic evaluation of the ovaries. Three of five females treated with a double administration of 120 µg prostaglandin (cloprostenol) at a 9-day interval expressed symptoms of estrus 9 days after the second injection. One female presented estrus after 6 days, whereas other did not respond to the treatment. All females (5/5) treated with a single dose containing 400 IU eCG and 200 IU hCG manifested estrus 6 days after the hormone injection. In a second experiment, ten females that were estrous synchronized using eCG/hCG, were artificially inseminated with fresh semen and monitored for pregnancy every 30 days. Although there was no detection of fetuses by ultrasonic examination, seven females (7/10) had greater than basal progesterone values for 60 days after the treatments were imposed. Ovaries from two females treated with eCG/hCG were collected 6 days post-injection. There was confirmation of an ovarian stimulation as a result of the presence of 88 and 25 antral follicles, as well as three and eight hemorrhagic structures in ovaries of each female, respectively. It, therefore, is proposed that eCG/hCG can be used as an effective treatment for estrous synchronization in collared peccaries.

Cryopreservation of collared peccary (Pecari tajacu L., 1758) epididymal sperm using extenders based on Tris and powdered coconut water (ACP®-116c).

SummaryThe aim of this study was to establish a functional freezing-thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed. In addition, to evaluate the survival of frozen-thawed sperm, a thermal resistance test (TRT) was executed. Samples preserved using Tris were in better condition compared with those preserved using ACP®, showing higher values for most assessments performed, including CASA and the TRT (P<0.05). After determining Tris to be the better of the two extenders, additional samples were thawed using different thawing rates (37°C/60 s, 55°C/7 s, 70°C/8 s). Sperm thawed at 37°C/60 s had the greatest preservation (P<0.05) of viability (54.1 ± 5.9%) and functional membrane integrity (43.2 ± 5.4%), and had higher values for various CASA parameters. In conclusion, we suggest the use of a Tris-based extender added to egg yolk and glycerol for the cryopreservation of epididymal sperm obtained from collared peccaries. In order to achieve better post-thawing sperm quality, we suggest that samples should be thawed at 37°C/60 s.

Monitoramento do ciclo estral de cutias (dasyprocta leporina lichtenstein, 1823) através de citologia esfoliativa vaginal e ultrassonografia / Monitoring of the estrous cycle of agoutis (dasyprocta leporina lichtenstein, 1823) by vaginal exfoliative cytology and ultrasonography

O objetivo deste estudo foi monitorar o ciclo estral em cutias (Dasyprocta leporina) criadas em cativeiro no semiárido brasileiro. Durante 70 dias, cinco cutias foram diariamente submetidas a citologia esfoliativa vaginal, e o monitoramento ultrassonográfico ovariano foi realizado a cada três dias. Um total de 8 ciclos estrais foi completamente monitorado, com duração de 28,2±0,7 dias, variando de 24 a 31 dias. Pela citologia esfoliativa vaginal, houve uma predominância de células superficiais nas fases de proestro e estro (P<0,05), seguida da predominância de células intermediárias no metaestro (P<0,05) e de células parabasais no diestro (P<0,05). Por ultrassonografia, não houve diferenças na morfologia ovariana durante as diferentes fases do ciclo estral (P>0,05). Os folículos foram identificados durante as fases estrogênicas (proestro e estro), com diâmetro médio de 1±0,5mm. Em apenas 12,5% das fases luteais, corpos lúteos medindo 1,4±0,9mm foram identificados. Conclui-se que a associação da citologia vaginal e da ultrassonografia ovariana constitui uma alternativa viável para o monitoramento de ciclos estrais e identificação das fases estrogênicas em cutias da espécie Dasyprocta leporina.

The objective of the study was to monitor the estrous cycle in agoutis (Dasyprocta leporina) bred in captivity in Brazilian semiarid. During 70 days, five agoutis were daily subjected 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), followed by the predominance of intermediate cells in the metestrus (P<0.05) and parabasal cells in diestrus (P<0.05). By ultrasound, there were no differences in ovarian morphology during the different phases of the estrous cycle (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, corpora lutea measuring 1.4±0.9mm were identified. We conclude that the association of vaginal cytology and ovarian ultrasonography is a useful alternative for monitoring the estrous cycle and identifying the estrogenic phases in Dasyprocta leporina.

Objective assessment of the cryoprotective effects of dimethylformamide for freezing goat semen.

The aim of this work was to assess the cryoprotective effects of dimethylformamide (DMF) for freezing goat semen, using an objective analysis by computer-assisted sperm analysis (CASA). Twenty-one ejaculates (seven per animal) were collected from three stud bucks with the aid of an artificial vagina and immediately evaluated for gross and microscopic characteristics. The semen was diluted in two steps with a Tris-egg yolk extender containing 6% glycerol or 6% DMF, frozen in 0.50-mL straws, and stored in liquid nitrogen. Samples were accessed for sperm morphology, sperm membrane structural and functional integrity, and by CASA, immediately after thawing. There were differences (P<0.05) between glycerol and DMF with regard to subjective progressive motility (23.9±2.2% vs. 16.6±2.0%), objective progressive motility (3.5±0.4% vs. 1.8±0.3%), linearity (53.9±1.6% vs. 48.1±1.4%) and amplitude of lateral head (2.3±0.1 vs. 2.9±0.1 mm), which confirmed the efficiency of glycerol. In conclusion, dimethylformamide could be used as an alternative cryoprotectant for goat semen freezing. However it was showed that no benefits were derived by using dimethylformamide to replace glycerol at an equal 6% concentration.

Determination of semen characteristics and sperm cell ultrastructure of captive coatis (Nasua nasua) collected by electroejaculation.

Despite the wide geographical distribution of coati (Nasua nasua) from the south of Canada to the north of Argentina, studies regarding the reproductive characteristics of this species are extremely limited. The objective of this study was to describe the various characteristics of coati semen by morphometric and ultrastructural analysis. Five mature males were anesthetized and electroejaculated for the collection of semen. Semen was immediately evaluated for color, volume, pH, sperm motility, vigor, morphology, acrosomal integrity, percentage of live cells and hypo-osmotic response by light microscopy. Sperm cell morphometry and ultrastructural analyses were also performed. Observations of seminal characteristics determined by electroejaculation in captive coatis represent a valuable baseline dataset for establishing fertility standards and provide background information that may be useful for assisted breeding programmes in members of the Procyonidae family.