Effect of Three Semen Extenders on Sperm Quality and In Vitro Fertilization Rates of Fresh and Cryopreserved Sperm Collected from Llama (Lama glama) Vas Deferens.

The advances in Assisted Reproductive Technologies (ARTs) applied in South American camelid species are still scarce. The aim of this study was to compare the effects of three semen extenders, before and after the cryopreservation of spermatozoa obtained from the vas deferens, on sperm quality parameters and in vitro fertilization rates of llama (Lama glama) oocytes. Mature fertile llama males (Lama glama; n = 6; age: 48-60 mo.; BCS: ~2.7) were included in the study. Sperm samples were collected from each male using the surgical technique of the vas deferens deviation. Then, the sperm samples were pooled and diluted with the Tris-EY, Andromed®, or BioxCell® extender in order to subsequently carry out the sperm cryopreservation process. The sperm quality assessment related to each extender was performed before and after cryopreservation with regard to sperm morphological abnormalities, acrosome integrity, sperm viability, membrane permeability, and sperm motility traits. Moreover, in vitro fertilization (IVF) procedures were carried out to evaluate the in vitro fertility of the cryopreserved sperm samples using each extender. Overall, significant differences were observed before and after cryopreservation regarding acrosome integrity, sperm viability, membrane permeability, and sperm motility traits among the extenders used, where Tris-EY and Andromed® were better than BioxCell® (p < 0.05); however, no differences were observed regarding the sperm morphological abnormalities among extenders (p > 0.05). Moreover, multiple differences were observed with regard to the velocity and linearity kinematic parameters obtained by computerized analysis before and after the cryopreservation process, irrespective of the extender used (p < 0.05). Finally, differences were observed regarding the in vitro fertilization rates among the different extender-derived samples (p < 0.05). In conclusion, the sperm quality using Tris-EY and Andromed® was better before and after cryopreservation compared to that using BioxCell®. Although the number of fertilized oocytes obtained after the IVF process between Tris-EY and Andromed® was similar, Andromed®-derived samples showed the best sperm quality results before and after cryopreservation. This indicates that the cryopreservation extender is a determining factor in significantly improving in vitro fertilization rates when using sperm samples obtained from vas deferens in llama (Lama glama) males.

Seasonal autoregressive integrated moving average (SARIMA) time-series model for milk production forecasting in pasture-based dairy cows in the Andean highlands.

Milk production in the Andean highlands is variable over space and time. This variability is related to fluctuating environmental factors such as rainfall season which directly influence the availability of livestock feeding resources. The main aim of this study was to develop a time-series model to forecast milk production in a mountainous geographical area by analysing the dynamics of milk records thorough the year. The study was carried out in the Andean highlands, using time-series models of monthly milk records collected routinely from dairy cows maintained in a controlled experimental farm over a 9-year period (2008-2016). Several statistical forecasting models were compared. The Mean Absolute Error (MAE), Root Mean Square Error (RMSE), and Mean Absolute Percent Error (MAPE) were used as selection criteria to compare models. A relation between monthly milk records and the season of the year was modelled using seasonal autoregressive integrated moving average (SARIMA) methods to explore temporal redundancy (trends and periodicity). According to white noise residual test (Q = 13.951 and p = 0.052), Akaike Information Criterion and MAE, MAPE, and RMSE values, the SARIMA (1, 0, 0) x (2, 0, 0)12 time-series model resulted slightly better forecasting model compared to others. In conclusion, time-series models were promising, simple and useful tools for producing reasonably reliable forecasts of milk production thorough the year in the Andean highlands. The forecasting potential of the different models were similar and they could be used indistinctly to forecast the milk production seasonal fluctuations. However, the SARIMA model performed the best good predictive capacity minimizing the prediction interval error. Thus, a useful effective strategy has been developed by using time-series models to monitor milk production and alleviate production drops due to seasonal factors in the Andean highlands.

Age-Related Differences in Testosterone Concentration and Its Relation to Testicular Biometrics, Hemodynamics, and Fertility in Alpacas (Vicugna pacos).

The goal of this study was to investigate the age-related differences in testosterone concentration and its relation to testicular biometrics, testicular blood flow, and fertility in alpacas (Vicugna pacos). Fifteen alpaca males with different ages (young (YM; ~12-14 mo.), n = 5; intermediate (IM; ~24 mo.), n = 5; and old (OM; ≥36 mo.), n = 5) were enrolled in the study. Blood samples were taken from each alpaca male and the circulating plasmatic testosterone concentration (TC; ng/mL) was determined using ELISA analysis. The testicular traits related to bio-morphometric parameters (the length (L), width (W), area (A), and volume (TV)) were assessed using B-mode ultrasonography. Pulse-wave/power Doppler ultrasonography was used to obtain the circulatory dynamic values (testicular hemodynamics) before the beginning of natural service mating. Significant differences were observed in TC among the age groups, increasing as the age of the males increased (2.47 ± 0.31, 8.45 ± 1.53, and 22.66 ± 2.15 for YM, IM, and OM, respectively; p < 0.05); however, no differences were observed regarding the testicular B-mode ultrasonographic parameters (L, W, and A) (p > 0.05). Positive correlations were observed between TV and testicular L, W, and A (r = 0.96, r = 0.95, and r = 0.96, respectively; p ≤ 0.001). Pulse-wave-Doppler-derived parameters such as the pulsatility index (PI) and the resistive index (RI), as well as the total vascularity area (TVA) assessed by power Doppler, were similar in all of the age groups studied (p > 0.05). General linear model (GLM) analysis showed a relationship between TC and TV (OR = 0.95; p = 0.04), as well as between TC and TVA (OR = 0.99; p= 0.02). Finally, no differences were observed regarding the pregnancy rate among the different age groups (p > 0.05). In conclusion, TC increased as the age of the alpaca males increased. Although TC was related to TV and TVA, the pregnancy rates obtained from individuals belonging to the different age groups were similar, indicating that TC, TV, and TVA were not determining factors in assessing the potential age-related fertility differences in alpaca males.

Phenotypic Characterization of Creole Cattle in the Andean Highlands Using Bio-Morphometric Measures and Zoometric Indices.

Several Creole cattle biotypes can be found in the Andean highlands, and most of them are considered as being in risk of extinction. The main aim of the present study was to perform a phenotypic characterization of the Creole cattle in the Andean highlands using bio-morphometric measures and zoometric indices. Individuals from three different biotypes (Black 'Negro' (n = 57), Colour-Sided 'Callejón' (n = 20), and Brindle 'Atigrado' (n = 18)) from an experimental research center located in the Peruvian highlands were enrolled in the study. In total, seventeen morphometric parameters were evaluated and ten zoometric indices were calculated in each biotype. To test the relationship between biometric traits, correlation analyses were carried out between morphometric parameters. Differences were observed regarding different morphometric variables such as head length (HL) and rump length (RL) among cattle biotypes (p ≤ 0.05). The coefficient of variation (CV; %) regarding different morphometric parameters ranged between 11.32 for neck length (NL) and 3.63 for height at withers (HaW), which indicated low-moderate variability among morphometric variables. Differences were observed in the longitudinal pelvic index (LPI) when different zoometric indices were compared among biotypes (p ≤ 0.05). The CV regarding different zoometric indices, which ranged between 10.78 for the cephalic index (CEI) and 5.05 for LPI, indicated low variability among indices. No differences were observed in any other morphometric parameter or zoometric index among cattle biotypes or genders (p > 0.05). Finally, multiple correlations were observed between morphometric variables (p ≤ 0.05). In conclusion, it was determined that Peruvian Andean Creole cattle can be considered as a dairy-related biotype with a slight tendency for beef production (dual-purpose). The great homogeneity regarding zoometric characteristics among biotypes and genders may indicate that the Andean Creole cattle have been maintained quite isolated, avoiding the genetic influence of other foreign breeds. Finally, the phenotypic characterization including bio-morphometric measurements and zoometric indices obtained from the different Creole bovine biotypes is crucial in order to begin different conservation programs to preserve cattle breeds in the Peruvian Andean highlands.

Ovarian Follicular Dynamics and Its Functional Significance in Relation with Follicle Deviation, Vaginal Cytology, and Hormone Profiles in Llamas (Lama glama).

The reproductive physiology in camelid species has its particularities. The present study aimed to characterize the ovarian follicular dynamics and its functional significance in relation to follicular deviation, vaginal cytological characteristics, and sexual hormone profiles in llamas as the first report in South American camelids. Non-pregnant, multiparous llamas (Lama glama; n = 10; age: 48−72 mo.; BCS: 2.5−3.0) were enrolled in the study. The ultrasonographic assessment was carried out transvaginally and follicular ablation was performed (day 0) when follicles were larger than 7 mm. The follicle number and diameter were scored daily throughout the process for a proper evaluation of the deviated follicles and to monitor the presence of new follicle pools (1.5 to 2.5 mm diameter). Vaginal cytological evaluation (parabasal, intermediate, and superficial cells) was performed every other day until day 6. Endocrine profiles (17ß estradiol, anti-Mullerian hormone, testosterone, and progesterone) during pre- and post-follicular deviation were determined by using the ELISA assay. Differential follicular dynamics both in the presence of a single dominant follicle (DF) and in codominance during the follicular deviation process were detected in llamas (p < 0.05). The percentage of superficial cells was the most related to the follicular wave phase. However, the percentage of parabasal, intermediate, and superficial cells was not related to the phases of follicular growth, dominance, and regression (p > 0.05). Differential patterns among the different hormone concentration levels regarding the 17ß estradiol, anti-Mullerian hormone, progesterone, and testosterone during follicular deviation were observed, with the latter being significantly different along the deviation process (p < 0.05). In conclusion, the use of vaginal cytology assessment would not be sufficient to determine the follicular phases in llamas. Therefore, complementary analyses, such as ultrasonography and endocrine assessment, are strongly recommended to determine follicular dynamics during the follicular deviation.

Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing.

The viscous seminal plasma (SP) is currently a major impediment to the handling of ejaculate and the development of some biotechnologies in South American camelids. The vas deferens-collected spermatozoa of alpacas is a useful technique to avoid this problem. On the other hand, SP contains a large protein component that has been implicated in the function of spermatozoa within the female reproductive tract. In this sense, the low fertility achieved using transcervical insemination with frozen-thawed spermatozoa in alpacas could be improved by adding SP. This study aimed to evaluate the effect of the whole SP on some in vitro parameters of alpaca spermatozoa after the freezing-thawing-process and the fertility after artificial insemination. It would contribute to a better understanding of the interaction between thawed sperm cells and SP. Spermatozoa were obtained by surgically diverted vas deferens. The samples were diluted with a Tris-based extender, packaged in straws, and frozen. At thawing, each straw was divided into two post-thawing conditions: with the addition of 10% of PBS (control) or with 10% SP (treatment). The sperm cells were evaluated using dynamic parameters, sperm cell morphology, and morphometry. Fertility was assessed by an artificial insemination trial. All in vitro parameters were analyzed by ANOVA. A heterogeneity test was scheduled for the fertility trial. After the freezing-thawing process, motility and plasma membrane functionality was improved when SP was added. No differences were found for post-thaw viability between the control and treatment samples. The percentage of normal cells was higher with SP at post-thawing, and a decrease of the presence of bent tailed spermatozoa with a droplet in the SP group was observed. The length of the head spermatozoa was 3.4% higher in the samples with PBS compared to those in which SP was added. Females pregnant at day 25 post-insemination were 0/12 (with SP inside the straw) and 1/10 (without SP inside the straw). In conclusion, the presence of 10% SP at post-thawing improves sperm cells' motility, functionality, and morphology, indicating that it would be beneficial to improve the frozen-thawed alpaca's physiology spermatozoa. More fertility trials must be developed to increase this knowledge.