Artificial insemination is an important assisted reproductive technology that has been applied in several mammalian species. However, successful cryopreservation of semen of South American camelids has been limited, hindering the commercial application of artificial insemination in alpaca species. In this scenario, the addition of antioxidants to semen extenders provides a strategy to improve the freezability of mammalian sperm. Bioactive metabolites from natural extracts of black maca have shown valuable antioxidant properties. Thus, the objective of this study was to evaluate the effect of the addition of atomized black maca in the freezing medium of epididymal spermatozoa of alpacas. Fifteen pairs of epididymis were collected from a local slaughterhouse. Each sample was divided into six groups: (1) fresh, (2) yolk medium (YM), (3) 10 mg/mL maca, (4) 20 mg/mL maca, (5) 30 mg/mL maca, and (6) resveratrol (as an antioxidant control). Sperm cryopreservation was performed through the slow freezing method. Markers associated with functionality, such as motility, viability, and plasma membrane integrity, as well as markers associated with oxidative damage, such as DNA integrity, total ROS production, and mitochondrial function, were analyzed. The results show that the supplementation with black maca (20 mg/mL) improved the sperm motility, viability, plasma membrane integrity, and mitochondrial function evaluated according to an index of formazan deposits. Similarly, the ROS production decreased with maca at 20 mg/mL, although the DNA integrity did not show any differences among the groups. These results suggest that maca at 20 mg/mL has cytoprotective effects during freezing/thawing of epididymal sperm of alpaca species. Further research will be focused on assessing the effects of maca supplementation on semen extenders by using biomolecular markers (proAKAP4) associated with fertility.

The objective of this study was to evaluate the effect of the addition of atomized black Maca in the cryopreservation medium of epididymal spermatozoa of alpaca species. Fifteen pares of testes and epididymis from alpacas of reproductive age were collected from a local (Huancavelica) slaughterhouse and transported in NaCl 0,9% at 10°C to the laboratory. Each sample of epididymal sperm was divided into six groups: 1) Fresh, 2) Yolk medium (YM), 3) YM+10mg/ml Maca, 4) YM+20mg/ml Maca, 5) YM+30mg/ml Maca, and 6) YM+ Resveratrol (antioxidant positive control). Sperm cryopreservation was performed through the slow-freezing method. Markers associated with functionality such as motility, vitality and plasma membrane integrity. Cellular markers associated with oxidative damage, such as DNA fragmentation, total ROS production and lipid peroxidation, were analyzed before and after freezing. The results showed that the supplementation with black Maca at 20mg/ml improved the sperm motility, viability and integrity of the membrane. Similarly, total ROS production decreased with Maca at 10 mg/ml (25 ± 2.23%) and 20mg/ml (20 ± 2.52%) (P<0.05). However, DNA fragmentation and lipid peroxidation were similar in all groups. These results suggest that Maca at 20mg/ml has a protective effect during the freezing/thawing process of epididymal sperm of alpaca species.

Among the in vitro embryo production (IVP) procedures, the in vitro maturation (IVM) is an essential step for the oocyte to achieve nuclear and cytoplasmic maturity (competence). In this line, supplementation of the IVM medium with resveratrol have been associated with higher oocyte competence in other species. On the other hand, a precise analysis of transcript levels in mammalian gametes and embryos requires the use of reliable reference genes (RGs) for data normalization. However, RGs specific to IVM of alpaca oocytes have yet to be identified, creating a barrier to functional gene studies in this species. Therefore, this study aimed to identify suitable reference genes to proper analyze gene expression patterns during IVM of alpaca oocytes. Four candidate transcripts (Gapdh, Actb, Rplp0 and Ppia) were selected and their stability during IVM with and without exposure to resveratrol was analyzed using RefFinder software (https://github.com/fulxie/RefFinder). The transcript levels of two target genes (Bmp15 and Sirt1) were normalized by the selected reference genes. Our findings indicate that, among the four candidate reference genes, Rplp0 and Gapdh were the most stable transcripts. Additionally, we observed that transcript levels of Bmp15 were differentially expressed after IVM in the presence of resveratrol (p<0,05). We also demonstrated that supplementation with resveratrol modulates the expression patterns of transcripts associated with chromatin remodeling and folliculogenesis. Thus, this is the first report evaluating suitable reference genes for alpaca oocytes, providing valuable tools to better understand oocyte physiology at the gene expression level of South American camelids.

Atlases of gene expression are an essential resource to connect transcript abundance and structure with tissue biology. The goal of this study was to build a comprehensive atlas of gene expression in adult female domestic goats (Capra hircus). To this end, we have extracted and sequenced total RNA from 33 tissues retrieved from six Murciano-Granadina goats. On average, more than 54 million paired-end raw reads (2 ´ 150 bp) per sample were obtained, and 92.5% of them were successfully aligned to the goat reference ARS1 genome. Downstream analyses were performed to ensure the quality of the atlas of gene expression generated with the procedures mentioned before. Sample clustering revealed 7 outliers, which were eliminated from the data set. Moreover, a second PCA demonstrated that functionally related tissues tended to group together, forming well defined clusters, while strong transcriptional signatures were observed for encephalic, muscle and hypophyseal tissues. These findings are highly consistent with previous reports and provide evidence about the high reliability of the transcriptomic resource generated in the current work.