Eugenol influences the expression of messenger RNAs for superoxide dismutase and glutathione peroxidase 1 in bovine secondary follicles cultured in vitro.

This study aimed to investigate the effects of eugenol on growth, viability, antrum formation and mRNA expression of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase 1 (GPX1) and peroxiredoxin 6 (PRDX6) in bovine secondary follicles cultured in vitro. To this end, bovine ovaries were collected from a local slaughterhouse and in the laboratory the follicles were isolated from the ovarian cortex. The follicles were then cultured in TCM-199+ alone or supplemented with different concentrations of eugenol (0.5, 5.0 and 50.0 µM). Follicular diameters and antrum formation were evaluated on days 0, 6, 12 and 18. Viability analysis was performed using calcein and ethidium homodimer. Real-time PCR was used to quantify mRNA levels for SOD, CAT, GPX1 and PRDX6 in cultured follicles. Follicular diameters and mRNA levels in follicles cultured in vitro were compared using analysis of variance and Kruskal-Wallis tests, while follicular survival and antrum formation were compared using the chi-squared test (P < 0.05). The results showed that secondary follicles cultured with eugenol maintained similar morphology and viability to follicles cultured in the control group. A progressive increase in follicular diameter was observed between days 0 and 12 for all treatments, except for follicles cultured with 50 µM eugenol. Eugenol (5.0 and 50.0 µM) increased mRNA levels for GPX1 in cultured follicles, but 0.5 µM eugenol reduced mRNA levels for SOD. The addition of eugenol did not influence mRNA expression for CAT and PRDX6. In conclusion, eugenol supplementation reduces mRNA levels for SOD and increases mRNA levels of GPX1 in bovine secondary follicles cultured in vitro.

Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro.

This study aimed to evaluate the effects of dexamethasone on development, viability, antrum formation and ultrastructural integrity of bovine secondary follicles cultured in vitro for 18 days. Bovine ovaries were obtained from slaughterhouses and secondary follicles of ~150-200 µm diameter were isolated and cultured in the laboratory in TCM-199+ alone or supplemented with different concentrations of dexamethasone (1, 10, 100 and 1000 ng/ml). Follicle viability was evaluated after the culture period, using calcein-AM (viable) and ethidium homodimer (nonviable). Follicle diameters and antrum formation were evaluated at days 0, 6, 12 and 18. Before or after in vitro culture, follicles were fixed for histological and ultrastructural analysis. Follicle diameters were evaluated using analysis of variance and Kruskal-Wallis test, while chi-squared test was used to evaluate the percentage of viable follicles and antrum formation (P < 0.05). Follicles cultured for 6 days with all treatments increased their diameters significantly, but there was no significant difference between treatments at the end of the culture period. In vitro cultured follicles showed antral cavity formation at the end of the culture period, but no influence of dexamethasone was seen. Ultrastructural analysis showed that follicles cultured with dexamethasone (1, 10, 100 and 1000 ng/ml) had well preserved granulosa cells. However, oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone showed signs of degeneration. It can be concluded that follicles cultured in vitro in the presence of dexamethasone demonstrated continuous in vitro growth, but oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone had poor ultrastructure.

Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during in vitro culture.

The present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles cultured in vitro for 12 days. To this end, secondary follicles were isolated (185-202 µm) and cultured in vitro in TCM-199+ medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2 in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal-Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P < 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles cultured in vitro.

Interactions Among Different Devices and Electrical Stimulus on the Electroejaculation of Captive Agoutis (Dasyprocta leporina).

The interactions among different electroejaculation devices associated with serial or continuous stimuli were investigated to improve the efficiency of the electroejaculation for semen collection in agoutis. Ten sexually matured male Dasyprocta leporina were restrained by the intramuscular administration of xylazine-ketamine association. Each individual was randomly subjected to four electroejaculation protocols, by combining two devices (one presenting longitudinal electrodes emitting square waves and other presenting ring electrodes emitting sine waves) and two electrical stimuli protocols (serial or continuous). A total of 40 attempts for electroejaculation were conducted in agoutis, being 10 per treatment. The most efficient treatment in providing ejaculates containing sperm (p < 0.05) was that using and electroejaculator connected to a probe with ring electrodes and associated with serial stimuli (4/7; 57%). In spite of semen parameters obtained by sine waves were adequate for using the samples for assisted reproduction, higher values for sperm motility and functional membrane integrity were obtained in the use of the square wave, independently of the electric stimulation protocol used (p < 0.05). In conclusion, we verified that the use of a device presenting a probe with ring electrodes and emitting sine waves, associated with a serial stimuli protocol, improves the efficiency for semen obtaining by electroejaculation in adults D. leporina.

Morphometry, Morphology and Ultrastructure of Ring-tailed Coati Sperm (Nasua nasua Linnaeus, 1766).

The ring-tailed coati (Nasua nasua) is a procyonid whose population is in sharp decline. Therefore, studies are needed to better understand the reproduction of this animal. For this reason, this study aimed to evaluate the morphology, morphometry and sperm ultrastructure of ring-tailed coati sperm. Four captive adult males were used for this study. Slides stained with Bengal Rose were used for the morphometric and morphologic analyses. The length and width of the head were measured, as well as the length of the midpiece and tail and the total length of the sperm. Scanning electron microscopy and transmission electron microscopy were used for the ultrastructural analyses. The most obvious morphological abnormalities observed were coiled tails (6.1 ± 8.7%) and the lack of acrosomes (5.4 ± 4.4%). Regarding the morphometry, the measurements of the head (length × width), midpiece (length) and tail (length) were (mean ± SD) 6.2 ± 0.4 × 8.1 ± 0.6 µm, 14.1 ± 0.5 and 63.9 ± 4.1 µm, respectively, and the total length of the sperm was 86.1 ± 4.3 µm. Through electron microscopy, the presence of electron-lucent points in the nucleus and the presence of approximately 55 mitochondrial spirals in the midpiece were identified. The data obtained in this study provide detailed information on the sperm characteristics of coatis and may inform future research on germplasm conservation, both for this species and other threatened procyonids.

Comparison of different glycerol and egg yolk concentrations added to tris-based extender for the collared peccaries (Tayassu tajacu) semen freezing.

This study aimed to evaluate various concentrations of egg yolk (5, 10, or 20%) in combination with different concentrations of glycerol (3% or 6%) added to a Tris-based extender on the post-thaw characteristics of sperm obtained from Tayassu tajacu. For this purpose, semen from 10 sexually male mature collared peccaries was collected by electroejaculation and evaluated for sperm motility, vigour, viability, morphology and functional membrane integrity. The ejaculates were initially extended in Tris-fructose plus egg yolk (5%, 10% or 20%). After cooling, the semen was added to Tris-egg yolk plus glycerol (6% or 12%), resulting in a final concentration of 3% or 6% glycerol of the extender. Straws were frozen using liquid nitrogen and thawed in a water bath at 37°C for 30 s. The frozen-thawed semen was evaluated as reported for fresh semen. After thawing, a significant decrease was verified for sperm motility and vigour, for all the samples in comparison with fresh semen. However, no differences were evidenced among treatments for any sperm characteristics evaluated (p > 0.05), except for the combination between 10% egg yolk and 6% glycerol, which provided the worst preservation of functional membrane integrity (p < 0.05). The interactions between higher concentrations of egg yolk (20%) and glycerol (6%) and also between lower concentrations of the same substances (5% egg yolk and 3% glycerol) added to the Tris-based extender negatively affected the preservation of the normal sperm morphology after thawing (p < 0.05). In conclusion, the use of Tris-based extender added to 10% or 20% egg yolk plus 3% glycerol is recommended for effective sperm cryopreservation in collared peccaries.

Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone.

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Immunolocalization of melatonin receptors in bovine ovarian follicles and in vitro effects of melatonin on growth, viability and gene expression in secondary follicles.

This study aims to investigate the (1) expression of melatonin receptors types 1A/B (MTNR1A/B) in bovine ovaries and (2) the in vitro effects of melatonin on secondary follicle development, antrum formation, viability, and expression of messenger ribonucleic acid (mRNA) for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase-1 (GPX1) and peroxiredoxin 6 (PRDX6). The expression of MTNR1A/B in bovine ovarian follicles was demonstrated by immunohistochemistry. To choose the most effective concentration of melatonin on follicular growth and viability, isolated secondary follicles were cultured individually at 38.5°C, with 5% CO2 in air, for 18 d in TCM-199+ alone or supplemented with 10-11, 10-9, 10-7 or 10-5 M melatonin. Then, melatonin receptor antagonist, luzindole, was tested to further evaluate the mechanisms of actions of melatonin, that is, the follicles were cultured in control medium alone or supplemented with 10-7 M melatonin, 10 µM luzindole and both 10-7 M melatonin and 10 µM luzindole. Follicular growth, morphology and antrum formation were evaluated at days 6, 12 and 18. At the end of culture, viability of secondary follicles was analyzed by calcein-AM and ethidium homodimer-1, and the relative levels of mRNA for SOD, CAT, GPX1 and PRDX6 were evaluated by real time polymerase chain reaction. Immunohistochemistry results showed expression of MTNR1A/B in oocyte and granulosa cells of primordial, primary, secondary and antral follicles. Secondary follicles cultured in medium supplemented with melatonin at different concentrations had well preserved follicles after 18 d of culture. Furthermore, follicles cultured in presence of 10-7 M melatonin presented significantly higher diameters than those cultured in other treatments. The presence of melatonin receptor antagonist, luzindole, blocked the effects of melatonin on follicular growth and viability. In addition, follicles cultured in medium containing only melatonin had significantly higher rates of antrum formation. Follicles cultured in medium containing only melatonin had higher relative levels of mRNA for CAT, SOD and PRDX-6 than those cultured with both melatonin and luzindole. Follicles cultured with luzindole only or both melatonin and luzindole had lower relative levels of mRNA for PRDX6 and GPX1 than those cultured control medium. In conclusion, melatonin promotes growth of bovine secondary follicles through its membrane-coupled receptors, while luzindole blocks the effects of melatonin on follicle growth and reduces the expression of antioxidant enzymes in cultured follicles.

Effects of epidermal growth factor and progesterone on development, ultrastructure and gene expression of bovine secondary follicles cultured in vitro.

The aims of this study were to investigate the effects of epidermal growth factor (EGF) and progesterone on the development, viability and the gene expression of bovine secondary follicle culture in vitro for 18 days. Secondary follicles (∼0.2 mm) were isolated from ovarian cortex and individually cultured at 38.5 °C, with 5% CO2 in air, for 18 days, in TCM-199+ (n = 63) alone (control medium) or supplemented with 10 ng/mL progesterone (n = 64), 10 ng/mL EGF (n = 61) or both EGF and progesterone (n = 66). The effects of these treatments on growth, antrum formation, viability, ultrastructure and mRNA levels for GDF-9, c-MOS, H1foo and cyclin B1 were evaluated, significantly different (p < 0.05). The results showed that there was a progressive increase in follicular diameter in all treatments, but only follicles cultured in medium supplemented with EGF had increased significantly in diameter when compared to follicles cultured in the control medium at the end of the culture period, significantly different (p < 0.05). A positive interaction between EGF and progesterone was not observed. In addition, the presence of EGF, progesterone or both in culture medium did not influence the rate of follicle survival and antrum formation. However, the presence of only progesterone in cultured medium increased the expression of mRNAs for GDF9 and cyclin B1 in oocytes. EGF also significantly increased the levels of mRNAs for cMOS and GDF9 when compared to follicles cultured in control medium. Ultrastructural analyzes showed that cultured follicles in all treatments maintained the integrity of granulosa cells. In conclusion, the EGF promotes the development of secondary follicles cultured in vitro for 18 days and increases the expression of cMOS and GDF9, while progesterone alone or in association with EGF have not a positive effect on follicular growth. However, progesterone increases the expression of GDF9 and cyclin B1 in oocytes.

Environmental effects on collared peccaries (Pecari tajacu) serum testosterone, testicular morphology, and semen quality in the Caatinga biome.

The objective of the study was to understand the influence of climatic variations in a semiarid environment on serum testosterone, testicular morphology and semen quality in collared peccaries (Pecari tajacu). Reproductive metrics (semen quality, testicular morphometry and testosterone serum profiles) of 10 mature males were measured monthly for 18 months. Meteorological data (rainfall, air temperature, relative humidity, wind speed and radiant heat load) also were recorded during the same period. Rainfall regimes were classified in different classes (Class 1: months with no rain; Class 2: months with up to 50 mm of rain; and Class 3: months with >50 mm of rain). Among rainfall classes, average air temperature (°C) and relative humidity (%) were different. Climatic changes between rainfall classes did not lead to overall variations of testicular size, testosterone production, and semen metrics. However, relative humidity recorded before semen collection (one day, one week, or over 51-55 days) was positively correlated (P < 0.05) with semen motility metrics (total motility, beat cross frequency and straightness) and sperm subpopulations (medium and static sperm), as well as with volume. Negative correlations (P < 0.05) were revealed between air temperature and the same semen motility patterns and volume. Additionally, radiant head load measured on the day of semen collection negatively influenced (P < 0.05) sperm straightness. This study demonstrates for the first time that no seasonal changes could be detected overt the 18-month period on the serum testosterone, testicular morphology and semen quality of collared peccaries raised in the Caatinga biome; however, it is expected that long term environmental changes will influence the reproductive physiology of species leaving in that habitat.

Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Complete Genome Sequence of the BeAn 58058 Virus Isolated from Oryzomys sp. Rodents in the Amazon Region of Brazil.

Here, we report the complete genome sequence of the BeAn 58058 virus (prototype) strain, isolated from a wild rodent Oryzomys sp. in the Utinga forest, Belém, state of Pará, Brazil in 1963. The genome of this virus showed similarity to the Poxviridae family, suggesting its inclusion in a possible new genus.

Use of Aloe vera-based extender for chilling and freezing collared peccary (Pecari tajacu) semen.

As an alternative for the conservation of collared peccary semen, this research aims at evaluating the use of Aloe vera (AV) extract as a cryoprotectant for semen chilling and freezing. Five ejaculates were divided in two aliquots that were diluted in Tris plus egg yolk (EY; 20%) or AV extract (20%) and chilled at 5 °C. In both treatments, an adequate semen conservation was achieved and values closer to 40% motile sperm with viability and osmotic response ranging from 20% to 40%, and normal morphology of 80% were found after 36 hours of storage. Moreover, 12 other ejaculates were diluted in Tris plus EY (20%) or AV extract (5, 10, or 20%) and glycerol (3%). Samples were frozen in liquid nitrogen and thawed after 1 week. After thawing, all the treatments containing EY or AV provided similar values for sperm morphology, viability, osmotic response, membrane integrity, sperm motility, amplitude of lateral head, beat cross frequency, and rapid, low, and static subpopulations, but the highest values for straightness and the lowest values for curvilinear velocity were found using 20% AV (P < 0.05). In conclusion, we found that AV extract at a 20% concentration could be used as an alternative substitute to EY in the formulation of Tris extenders for collared peccaries' semen chilling or freezing.

Individual variation related to testicular biometry and semen characteristics in collared peccaries (Tayassu Tajacu Linnaeus, 1758).

The aim of this research was to study the individual variation with regard to the morphometry of the testes evaluated by ultrasonography and semen characteristics and to verify the existence of relationship between these variables in collared peccaries. In addition, the testes of the animals were evaluated by histology in order to determine the proportion occupied by the seminiferous tubules. A total of 52 ejaculates were obtained from ten adult specimens that had been restrained by anesthesia. The testicular measurements (length, height, and width) were performed by ultrasonography, and the testicular volume was calculated according to Lambert's formula. The scrotal circumference was measured by encircling the thickest portion of the testicle with a graduated nylon tape. The semen was collected by electroejaculation. Testicular fragments were analyzed through classic histology for the determination of the area occupied by the seminiferous tubules. The results show a great amount of individual variation with regard to testicular morphometry and semen characteristics. No significant correlations were obtained between testicular measurements and semen characteristics. The histometric analysis revealed that 67.8% of the testes are occupied by seminiferous tubules. Results show that the measurement of testicular dimensions does not serve as an indicator of the quality of semen obtained by electroejaculation in collared peccaries, as there is no correlation between testicular morphometry and semen characteristics in this species that presents large variations among individuals.

Effects of chronic exposure of diets with reduced concentrations of aflatoxin and deoxynivalenol on growth and immune status of pigs.

This study investigated the growth and immune responses of pigs fed diets containing reduced concentrations of aflatoxin (AF) and deoxynivalenol (DON) from naturally contaminated corn. Sixty gilts (13.9 ± 0.2 kg of BW) were randomly assigned to 4 treatments (5 replicate pens per treatment and 3 pigs per pen): A (a control diet without detectable AF and DON); B (a diet with 60 µg of AF/kg and 300 µg of DON/kg); C (a diet with 120 µg of AF/kg and 600 µg of DON/kg); and D (a diet with 180 µg of AF/kg and 900 µg of DON/kg). Pigs were allowed ad libitum access to feed and water for 33 d. Feed intake and BW were measured weekly and pigs were bled (8 mL) on d 33 to measure the numbers of blood cells, to conduct liver function tests, and to measure immunological variables including IgG, IgM, interferon γ, IL4, IL6, and tumor necrosis factor α. One pig representing the average BW of each pen was killed to obtain the liver, kidneys, and spleen for weight, tissue color measurement, and histological evaluation of tissue damage. When compared with A, pigs in C and D tended to have reduced ADG (0.52 vs. 0.43 and 0.41 kg/d, respectively; P = 0.058) and ADFI (1.04 vs. 0.92 and 0.88 kg/d, respectively; P = 0.061). White blood cell count of pigs in D (23.4 × 10(3) cells/µL) was greater (P < 0.05) than those in A, B, and C (18.4, 18.5, and 16.8 × 10(3) cells/µL, respectively. Serum tumor necrosis factor α concentration of pigs in D (335 pg/mL) differed (P < 0.05) from those in A and C (299 and 290 pg/mL, respectively). Pigs in B and D had greater (P < 0.05) fibrosis in liver tissues than those in A. Collectively, this study shows that diets containing both AF and DON greater than 60 and 300 µg/kg, respectively, may reduce growth and decrease feed intake, whereas diets containing 120 µg of AF/kg and 600 µg of DON/kg may result in altered immune health, systemic inflammation, and partial liver damage, causing further reduction in growth of pigs.

Influence of the thawing rate on the cryopreservation of semen from collared peccaries (Tayassu tajacu) using Tris-based extenders.

The objective was to evaluate the influence of the thawing rate on the quality of frozen-thawed (cryopreserved in Tris-based extenders) semen obtained from collared peccaries (Tayassu tajacu). Semen from 13 sexually mature collared peccaries males were collected by electroejaculation, and evaluated for motility, vigor, sperm viability, membrane integrity, and sperm morphology. Semen was divided in two equal portions: the first was diluted in Tris-fructose and the other in Tris-glucose, with egg yolk (20%) and glycerol (3%) added to each portion. Extended semen was frozen in liquid nitrogen and thawed using two thawing protocols (37 degrees C for 1 min or 55 degrees C for 7 s, followed by an additional 30 s at 37 degrees C). There were no significant differences between the two extenders after extension, chilling, or glycerol addition. After thawing at 37 degrees C, there were 37.9 +/- 4.2% and 28.5 +/- 5.1% motile spermatozoa for samples extended in Tris-fructose and Tris-glucose, respectively, with 33.8 +/- 3.7% and 28.2 +/- 3.5% motile spermatozoa after thawing at 55 degrees C (no significant differences). Furthermore, there were no significant interactions between extenders and thawing protocols for any semen end point. In conclusion, semen from collared peccaries was successfully cryopreserved in Tris-based extenders and thawed with two protocols (37 degrees C for 1 min or 55 degrees C for 7 s).

Dimethylformamide is no better than glycerol for cryopreservation of canine semen.

The objective of this study was to compare the effects of dimethylformamide (DMF) and glycerol in canine (Canis lupus familiaris) semen cryopreservation based on postthaw motility and velocity evaluated by computer-assisted sperm analysis (CASA) and the effects on subjective progressive motility, percentage of live sperm, and plasma membrane functional integrity. The semen was diluted in two steps with an egg-yolk Tris extender containing 6% glycerol or DMF, frozen in 0.25-mL straws, and stored in liquid nitrogen. Immediately after thawing, samples were accessed for subjective sperm motility, sperm membrane functional integrity, percentage of live sperm, and evaluation by CASA. There were differences (P<0.05) between glycerol and DMF with regard to subjective progressive motility (43.1% vs. 21.5%), objective progressive motility (11.8% vs. 6.2%), velocity average pathway (31.1 vs. 23.1 microm/sec), and amplitude of lateral head (3.3 vs. 3.9 microm), which confirmed the efficiency of glycerol. In conclusion, objective analysis performed by CASA confirmed that no benefits were derived by using DMF to replace glycerol for cryopreservation of canine semen.

Evaluation of anesthetic protocol for the collection of semen from captive collared peccaries (Tayassu tajacu) by electroejaculation.

The objective of this study is to verify and compare the effects of acepromazine-tiletamine-zolazepam and propofol used in anesthetic protocols for semen collection by electroejaculation from captive collared peccaries. Ten sexually mature animals were physically restrained and anesthetized by either intravenous administration of tiletamine-zolazepam (2mg/kg) after acepromazine premedication, or a propofol dose of 5mg/kg. The onset of anesthetic recovery was determined by the animals regaining consciousness and attempting to stand. Semen was collected by electroejaculation and evaluated for volume, pH, sperm concentration, progressive motility, morphology, percentage of live cells and functional membrane integrity. Six anesthetized animals with the acepromazine-tiletamine-zolazepam protocol showed erection, but semen could be collected in only four (40%) attempts. Of the animals anesthetized using propofol, nine showed erection, and the ejaculates were collected in eight (80%) attempts. Furthermore, propofol afforded rapid recovery of animals, and ejaculates with enhanced sperm motility and functional membrane integrity as compared with those collected by the other protocol (P<0.05). In conclusion, use of propofol for anesthetic restraint of collared peccaries enhanced collection of semen by electroejaculation.