The cryopreservation of jaguar semen must be improved to produce high-quality biobanking doses. Until now, the rare studies of semen freezing in the species have only evaluated glycerol, always with a significant reduction in sperm quality in thawed semen. The purpose of this study was to assess the efficacy of three cryoprotectants, dimethylsulfoxide (DMSO), glycerol (GLY), and methanol (MET), in the cryopreservation of jaguar semen in an LDL-based extender, as well as the effect of thawing temperature on dosage quality. Five mature males with a history of reproduction were used. On the males, an infrared thermal image (IRT) was captured, the spicules and testes were analyzed, and the CASA system was used to evaluate the quality of fresh and thawed sperm. The superficial IRT was 4.6 ± 1.2 °C cooler than the anal sphincter, and the semen measured between 27.3 and 28.7 °C shortly after exiting the urethra. The total motility of fresh sperm was 55.3 ± 22.6%, and progressive motility was 36.3 ± 18%. The total motility of thawed sperm was 5.28 ± 2.51%, 4.49 ± %2.49, and 0.51 ± 0.62% for DMSO, GLY, and MET, respectively. DMSO and GLY performed better than MET, and there was no difference in thawing temperature (37°C 30 s vs. 50°C 12 s). All animals exhibit a considerable level of morphological changes in sperm. Low amounts of total and progressive motility were found in the thawed sperm. Males with a high level of sperm morphological changes were found to be fertile, but the lone male with normospermia was infertile. Thus, we contest the applicability of the commonly used morphological classification for bovines to felid species.
The objective of this study was to reduce the effects of cryoinjury caused in bovine semen by cryopreservation. Ejaculates were collected from Nellore bulls and subjected to freezing in C (control), ozone (15, 30, and 60 µg mL-1 of ozone), quercetin (25, 50, and 100 µg mL-1 of quercetin), and carnosine groups (100, 200, and 300 ng mL-1 of carnosine). Samples were evaluated post-thaw (M0) and post-rapid thermoresistance test (M30) for sperm kinetics (total motility, progressive motility, curvilinear speed, linearity and amplitude of lateral head displacement) and cell structure viability (plasma membrane integrity, acrosomal integrity, mitochondrial potential, membrane fluidity, and lipid peroxidation). There were no differences (P > 0.05) between the control, quercetin, and carnosine-treated groups for the parameters evaluated at M0 and M30. In turn, supplementation with ozone resulted in lower values for sperm kinetics (P < 0.05) and lower mitochondrial potential at M30 (P < 0.05). Quercetin and carnosine at the concentrations used did not promote significant gains in frozen semen, nor did they demonstrate cytotoxicity. We expected to obtain positive results with the use of ozone. Nonetheless, the addition was harmful to the parameters of sperm kinetics, and its effect was not observed as a possible pro-antioxidant. We believe that the fact that the gas did not harm the sperm structure opens avenues for tests with lower dosages, since, by reducing its concentration, we could minimize the damage to sperm kinetics.
The cryopreservation reduces ram sperm quality, decreasing the pregnancy rate of ewes inseminated with thawed sperm. Hence, we aimed to improve the post-thaw quality of ram sperm replacing egg yolk on Tris-Glucose extender with different concentrations of LDL (2 or 8%), associated with the addition of 10 mM non-enzymatic antioxidants (ascorbic acid, hydroxytoluene butylate, ascorbyl palmitate, and trehalose). Semen samples were collected from six rams, split into different treatments, and frozen. After thawing, kinematic (CASA), structural (propidium iodide and carboxyfluorescein diacetate) and functional (hypoosmotic test) sperm membrane integrity was assessed. Total motility, VCL, and LIN were also assessed in thawed samples during 3 h of incubation (38 °C). The results showed that hydroxytoluene butylate at 10 mM in Tris-Glucose extender with 8% LDL improved velocity parameters immediately post-thaw compared with Tris-Glucose egg yolk extender, as well as prevented the reduction of total motility and VCL after incubation. There was no benefit of adding ascorbic acid and trehalose. Moreover, for the first time, it was shown the motility impairment promoted by ascorbyl palmitate to ram sperm.
In this study, it was aimed to determine the effect of sulforaphane (SFN) on rabbit semen cryopreservation. Semen collected from animals was divided into 5 equal volumes as Control, SFN 5 µM, SFN 10 µM, SFN 25 µM and SFN 50 µM groups. Afterwards, semen analyzes were performed. According to our results, there was no statistical difference between the groups at 4°C. However after freezing thawing, the highest total motility, progressive motility and rapid spermatozoa rate was seen in the 10 µM SFN group, while the lowest was observed in the 50 µM SFN group (P<0.05). Static sperm ratio was highest in the 50 µM group, while the lowest was observed in the 10 µM SFN group. When flow cytometry results examined the rate of acrosomal damaged and dead sperm was the lowest in the 10 µM SFN group, a statistical difference was observed between the control group (P<0.05). The highest rate of sperm with high mitochondrial membrane potential was seen in the 5 µM SFN and 10 µM SFN groups. Apoptosis and ROS rates were found to be lower in the experimental groups compared to the control groups (P<0.05). As a result, SFN supplementation at a dose of 10 µM increased the quality of sperm in the freezing and thawing processes of rabbit semen. In conclusion, 10 µM SFN improved the quality of cryopreservation of rabbit semen.
The aim of this study was to investigate uterine involution using ultrasonography techniques during postpartum. Postpartum ultrasonography evaluation (B-mode, color Doppler and Acoustic Radiation Force Impulse elastography) of the uterus was performed by transabdominal approach at immediate after birth and sequentially every 48 hours, during 30 days. The uterine echotexture did not present significant variations (P >0.05) being homogeneous in most evaluations; echogenicity of the uterus increased along the evaluation period (P =0.0452). Progressive and remarkable decrease of the total uterine diameter (UD) were observed (P <0.0001), especially during the first days postpartum. The thickness of uterine wall gradually decreased, as well the endometrial, myometrium and lumen diameters (P <0.0001). Uterine blood flow was assessed by Doppler and decreased during postpartum period, being significantly lower (P=0.0225) on the 30th day of postpartum. Uterine parenchyma presented as homogeneous dark areas (not deformable) on qualitative ultrasound elastography and the means shear velocity values of the uterine wall on quantitative elastography did not differ. This is the first study that evaluate the stiffness of uterine wall in healthy ewes, providing baseline data about quantitative and qualitative stiffness of the normal uterus, and it may be a useful tool for early diagnosis of uterine alterations during the postpartum period, using the reference parameter established for the assessment of uterine integrity during postpartum period.
Some sectors of animal production and reproduction have shown great technological advances due to the development of research areas such as Precision Livestock Farming (PLF). PLF is an innovative approach that allows animals to be monitored, through the adoption of cutting-edge technologies that continuously collect real-time data by combining the use of sensors with advanced algorithms to provide decision tools for farmers. Artificial Intelligence (AI) is a field that merges computer science and large datasets to create expert systems that are able to generate predictions and classifications similarly to human intelligence. In a simplified manner, Machine Learning (ML) is a branch of AI, and can be considered as a broader field that encompasses Deep Learning (DL, a Neural Network formed by at least three layers), generating a hierarchy of subsets formed by AI, ML and DL, respectively. Both ML and DL provide innovative methods for analyzing data, especially beneficial for large datasets commonly found in livestock-related activities. These approaches enable the extraction of valuable insights to address issues related to behavior, health, reproduction, production, and the environment, facilitating informed decision-making. In order to create the referred technologies, studies generally go through five steps involving data processing: acquisition, transferring, storage, analysis and delivery of results. Although the data collection and analysis steps are usually thoroughly reported by the scientific community, a good execution of each step is essential to achieve good and credible results, which impacts the degree of acceptance of the proposed technologies in real life practical circumstances. In this context, the present work aims to describe an overview of the current implementations of ML/DL in livestock reproduction and production, as well to identify potential challenges and critical points in each of the five steps mentioned, which can affect results and application of AI techniques by farmers in practical situations.
The aim of this study was to evaluate and correlate modifications of vaginal mucous impedance, vulvar temperature and ultrasonographic measurements (echobiometric parameters) to parturition in pregnant Saanen does. 30 does were selected for the study and submitted to an estrus synchronization protocol and natural mating. The females were evaluated daily from Day 143 of pregnancy to parturition. For the sonographic evaluations, the following structures were measured: biparietal diameter, thoracic diameter, abdominal diameter, ocular orbit, kidney length, kidney height, cardiac area, placentome length, cervical measurement and fetal heart rate; by means of two different approaches: transrectal and transabdominal, using a 7.5 MHz linear transducer. The vaginal mucous impedance was assessed using an electric estrous detector and vulvar temperature was measured using a non-contact infrared thermometer. Statistical analysis was performed using the R-project software and the significance level was set at 5% for all tests. 25 Saanen does became pregnant, resulting in 80.33% pregnancy rate. Fetal heart rate was negatively correlated to the hours to parturition (p<0,001; r-Pearson= -0,451), as well as vaginal temperature (p= 0,001; r-Pearson= -0,275), while cervical thickness was positively correlated to hours to parturition (p<0,001; r-Pearson= 0,490). The echobiometric parameters (biparietal diameter, thoracic diameter, abdominal diameter, ocular orbit, kidney length and height, cardiac area, placentome length), as well as vaginal mucous impedance did not vary throughout the timepoints of evaluation and did not correlate to the moment of parturition. It was concluded that the parameters of fetal heartbeat, vaginal temperature and cervical effacement in the last week of pregnancy provide valuable information regarding the proximity of parturition.