The objective was to evaluate the effect of two methods of freezing on seminal quality and effect of seminal plasma as an inducer of ovulation in pregnancy percentages in inseminated alpacas. The semen collection was made post copula. After the collection of the ejaculates, motility and volume were assessed. The ejaculates with volume (≥1 ml) and motility (≥ 60%) were mixed (pool), 80 % was used (3 samples/pool) and 20 % (2 samples/pool) during the experiment. Then and diluted in Tris-base with 20% egg yolk. The samples were cooled 1.5 hours at 5 °C. At that temperature it was combined with the basic dilutor plus glycerol, obtaining a final concentration of 5% glycerol, then they were packed in 0.5 mL (13 x 106 spem/straw) straws to be frozen by horizontal and vertical methods. The seminal (semen + vaginal fluid) quality analysis was performed fresh and after cooling and thawing. Insemination was performed in two groups with thawed semen from two straws of the vertical method, the first group 20 females induced to ovulate with GnRH analog, the second group 20 females induced to ovulate with seminal plasma. When comparing, the results obtained of motility, viability, HOST and acrosomal integrity of fresh sperm, after the freezing process, decreased (p<0.05) compared to fresh and refrigerated samples. On the other hand, when comparing the freezing methods, the sperm values frozen by the vertical method were higher than those obtained by the horizontal method, with (p<0.05) in motility and HOST without (p>0.05) in acrosomal integrity and viability. The vertical semen freezing method can replace the horizontal method to obtain pregnancy.
The objective of this study was to evaluate the cryopreservation of alpaca spermatozoa obtained via post copula in a Tris extender with egg yolk from three avian species. Forty samples of eight alpacas were collected by the post-copula method. After the collection, we proceeded to evaluate sperm volume, color, motility and concentration. The 25 samples with volume 1 ml and total motility 60% were mixed to form pool (5 samples/pool), divided into three aliquots and diluted in Tris-base with 20% egg yolk from three avian species (hen, quail, paw). These diluted samples were refrigerated for 1,5 h at 5 °C. Once this temperature was reached, the 5% glycerol basic dilutor was added, balanced for 20 min, packed in 0,5 mL straws and frozen in liquid nitrogen vapours for 20 min. The thawed samples were evaluated at different incubation times at 37 °C: 0; 1,5 and 3 h. All parameters of fresh and thawed sperm quality were analyzed using the GLM procedure (ANOVA). The samples collected (fresh) showed a motility of 69,1%, viability of 82,8%, membrane functionality of 77,9% and acrosomal integrity of 85,8%. After the cooling process, no differences were observed between the different egg yolk when comparing the sperm characteristics evaluated (p>0,05). At thawing, motility and acrosomal integrity were superior (p<0,05) when hen and quail were used compared to paw egg yolk. At 1,5 and 3 h of incubation, motility and acrosomal integrity were superior (p<0,05) in the samples with hen and quail with respect to paw. In conclusion, the use of hen and quail provided better cryoprotective action than paw egg yolk in cryopreserved alpaca sperm and incubated at 37°C for 3 h
The objective of this study was to determine the effect of glycerol (GL), dimethyl sulfoxide (DMSO) and dimethylformamide (DMF) at temperatures of 5 °C and 16 °C of equilibrium on sperm quality, after cryopreservation. 6 males were used in the semen collection, motility parameters were evaluated with the CASA system, viability, acrosomal integrity and mitochondrial activity by flow cytometry in fresh and thawed semen. We collected 30 ejaculates of which 10 were discarded for not fulfilling (volume ≥ 1 ml and total motility ≥ 60 %), the 20 ejaculates were mesclo in a tube to form pool carrying out the assessment of the initial quality of fresh semen, obtaining a total motility (MT) 61,6 ± 3,4%, progressive motility (MP) 51,7 ± 3,7%, viability 78,1 ± 9.1%, mitochondrial activity (MA) 77,4 ± 7,7 and acrosomal integrity 88,9 ± 11,5%. After cryopreservation no equilibrium temperature effect was observed, independently of cryoprotectant in the parameters of seminal quality. Cryoprotective effect (P≤ 0.05) was observed, presenting superior values in the sperm conserved in GL with respect to the rest of cryoprotectants, with significant differences (P ≤ 0,05) in MT, MP, viability and AM and without significant differences in acrosomal integrity. The equilibrium temperature of 16 °C was effective for cryopreservation of alpaca sperm. In addition, the 5% GL preserved better the functional characteristics of sperm compared to 7% (DMF and DMSO).
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