The meat market has enormous importance for the world economy, and the quality of the product offered to the consumer is fundamental for the success of the sector. In this study, we analyzed a database which contained information on 2470 animals from a commercial farm in the state of São Paulo, Brazil. Of this total, 2181 animals were genotyped, using 777,962 single-nucleotide polymorphisms (SNPs). After quality control analysis, 468,321 SNPs provided information on the number of genotyped animals. Genome-wide association analyses (GWAS) were performed for the characteristics of the rib eye area (REA), subcutaneous fat thickness (SFT), shear force at 7 days’ ageing (SF7), and intramuscular fat (IMF), with the aid of the single-step genomic best linear unbiased prediction (ssGBLUP) method, with the purpose of identifying possible genomic windows (~1 Mb) responsible for explaining at least 0.5% of the genetic variance of the traits under analysis (≥0.5%). These genomic regions were used in a gene search and enrichment analyses using MeSH terms. The distributed heritability coefficients were 0.14, 0.20, 0.18, and 0.21 for REA, SFT, SF7, and IMF, respectively. The GWAS results indicated significant genomic windows for the traits of interest in a total of 17 chromosomes. Enrichment analyses showed the following significant terms (FDR ≤ 0.05) associated with the characteristics under study: for the REA, heat stress disorders and life cycle stages; for SFT, insulin and nonesterified fatty acids; for SF7, apoptosis and heat shock proteins (HSP27); and for IMF, metalloproteinase 2. In addition, KEGG (Kyoto encyclopedia of genes and genomes) enrichment analysis allowed us to highlight important metabolic pathways related to the studied phenotypes, such as the growth hormone synthesis, insulin-signaling, fatty acid metabolism, and ABC transporter pathways. The results obtained provide a better understanding of the molecular processes involved in the expression of the studied characteristics and may contribute to the design of selection strategies and future studies aimed at improving the productivity of Nellore cattle.

Developmental competence is obtained by a series of morphological and molecular changes during mammalian oocyte growth within the ovulatory follicle. This entails the accumulation of cytoplasmic transcripts that will be used throughout the early stages of development prior to embryonic genome activation, a process known as ooplasm maturation. Furthermore, during follicular growth, epigenetic maturation occurs, which is essential for appropriate embryo development. We believe that transcripts and DNA methylation differ between blastocyst oocytes and those that cleaved but were arrested on day three. We devised a retrospective technique to identify transcripts in oocyte, cumulus, and granulosa cells, as well as DNA methylation connected with oocyte competence, in this work. We dissected and harvested ovarian follicles to achieve this purpose. We extracted and flash frozen the granulosa cells after rupturing them. The oocytes were put in maturation media droplets, and the cumulus cells and polar body were removed and kept the following day. To prevent spermatozoon interference, we chemically activated the oocytes and tracked their development (until they reached the blastocyst stage). We went back to their biopsies, cumulus cells, and polar bodies and did RNA-seq (biopsies and cumulus) and single polar body WGBS (polar bodies) when we collected the results 7 days later, i.e. 1-) embryos that cleaved but stopped development (termed CL) or 2-) embryos that cleaved and progressed until the blastocyst stage (termed BL). Additionally, after transcriptome results from oocyte-biopsy and cumulus cells, the granulosa cells from their individual oocytes were sequenced as a noninvasive strategy. This study is a follow-up to our previous work, “ Assessment of Total Oocyte Transcripts Representation in bovine Using Single Ooplasm Biopsy with High Reliability. ” Following sequencing, we discovered that the two groups, BL x CL, were transcriptionally different in granulosa and biopsy samples, although cumulus cells were a poor predictor of oocyte competence. By analyzing the differentially expressed genes, we discovered multiple genes and pathways related to oocyte competency, demonstrating the efficacy of our method. Despite no change in morphology, these alterations in pathways and genes show that the oocyte CL group was transcriptionally and epigenetically delayed, with ferroptosis and necroptosis processes activated. The oocyte BL group demonstrated numerous molecular signaling, oocyte meiosis, GnRH signaling, G-protein cascade, and RNA stability pathways. In network analysis, we discovered GNAS , an imprinted gene and one of the most important essential genes. The transcripts from granulosa cells confirm the oocyte results. Nonetheless, transcriptional variations in granulosa cells were far greater than those in oocytes (97% × 34% variance), implying a completely distinct transcriptome in the follicular niche. In terms of the WGBS, we discovered differentially methylated areas linked with oocyte competency, as well as transcriptome results confirming the structure’s ability to predict outcome. These findings might be beneficial in clinical settings for those undergoing infertility therapy. In the oocyte, we discovered a complex transcriptional and epigenetic regulation network. Furthermore, mature cumulus transcription produced information that differed from the true content of the MII oocyte and granulosa cells before maturity. Our findings underscore the significance of maternal transcripts and epigenetic maturation in early parthenogenesis, as well as the use of granulosa cells as early indicators of competence.

Background: Considering the difficulties in wound healing in horses, the possibility of using a biocompatible adhesive for wound suturing presents itself as an asset for equine surgery. The tissue adhesive, which is composed of ethyl-2-cyanoacrylate, offered advantages when used in human wounds, such as the absence of discomfort, reduction of surgical and healing time, a physical barrier against environmental infectious agents, resistance to exudates, and reduction of dehiscence. The objective of this study was to evaluate the efficacy and biocompatibility of ethyl-2-cyanoacrylate as a skin adhesive in equine surgical wounds and to compare it with the use of sutures in the healing process. Materials, Methods & Results: A total of 8 horses in 2 experimental stages, neck and croup, were used and divided into 2 groups at each stage. On both sides of the neck, 7-cm-long skin incisions were made, which were sutured with nylon thread (SG; suture group; left side of neck) or glued with ethyl-2-cyanoacrylate (AG; tissue adhesive group; right side of neck). The same procedure was performed on both sides of the croup of these animals, the left side being SG and the right side, AG. Macroscopic assessments of the surgical wounds of both groups were performed on days 1 (D1), 3 (D3), 7 (D7), 10 (D10), and 12 (D12) after surgery with standardized photographs regarding spacing and position. Animals with 70% to 100% dehiscence of the wound, i.e., with a length of more than 4.9 cm, were considered as total dehiscence and excluded from the evaluations. Aspects such as touch sensitivity, tissue consistency, volume increase and temperature, healing process, and signs of biomaterial rejection were observed. On the twelfth day after surgery (D12), a fragment of healed skin was harvested for microscopic evaluation. Tissue adhesive was considered a faster and easier method compared to SG, with advantages such as barrier function against microorganisms and the fact that it does not need to be removed. However, the major disadvantage was the high rate of dehiscence. Histopathological analysis showed a greater inflammatory infiltrate at SG, suggesting a reaction to the presence of the adhesive, in addition to being a more aggressive method of synthesis. Discussion: Ethyl 2-cyanoacrylate proved to be easy and quick to apply, with the time required for synthesis being significantly shorter compared with suturing, and caused no discomfort during the transoperative period. However, it was difficult to apply in cases of heavy bleeding because the product polymerized rapidly on contact with blood, which promoted good hemostasis but led to unsatisfactory synthesis of the wound, which may have contributed to its subsequent dehiscence. There was a significant difference between the studied groups in the presence of dehiscence, as the SG had no dehiscence compared with 50% of dehiscence in the AG as a whole (n = 16), with 37.5% considered total dehiscence and 12.5% partial wound dehiscence. One possible explanation is the high tensile force in the region, possibly due to excessive movement of the wound site, combined with the wound size of 7 cm, which is probably too large for the use of tissue adhesive alone, without deep relaxing sutures. In the present study, skin samples taken on day 12 from each group were examined. The results show that there was a greater amount of inflammatory cells in the SG than in the AG. The presence of the nylon thread as a foreign body and the greater aggressiveness in performing the suture may contribute to prolonging the inflammatory phases of the tissue repair process, as was found in this group. It was also possible to observe a greater amount of tissue reaction around the area of the AG scar, i.e. perivascular and periglandular (sebaceous and sweat glands), indicating contact dermatitis due to the presence of tissue glue. Keywords: healing, horses, wounds, surgical adhesive, cyanoacrylates.

Anais do Simpósio Internacional do Cavalo Atleta (SIMCAV) (978-65-5941-211-2) - Avaliação Da Utilização Do Etil-2-Cianoacrilato Na Síntese De Pele Em Equinos

Understanding the entire transcriptional and epigenetic landscape is facilitated by the application of omics in a number of ways. Today, omic instruments are more affordable and easier to implement. In human research, for instance, single-omics are a reality and are used extensively to generate vast quantities of data. This method permits the comprehensive reconstruction of transcriptome and epigenetic markers removing bias from pooled samples. In tandem with the evolution of machines and protocols, algorithms and genome annotation have undergone continuous improvement. The genome annotation of domestic animals is inferior to that of humans, rodents, and less complex organisms. In the case of heifers, the reference is incomplete, with significant gaps and only a portion of the noncoding transcripts. The purpose of this study is to validate our compartmentalized single oocyte biopsy by comparing a small fraction of bovine oocytes, 1%, to the entire oocyte at the Metaphase II stage. In addition, we examined the use of four database sources (NCBI, ENSEMBL, UCSC, and NONCODE) to produce a merged non-redundant gene alignment and counting in order to enhance gene detection and normalization, resulting in a more accurate method to comprehend the entire landscape. This study is a continuation of our research titled “ Retrospective model utilizing biopsies, granulosa cells, and polar body to predict oocyte competence in bovine ” in which this method was used to retrospectively compare biopsy oocytes collected during the MII phase. With the addition of NONCODE information, gene normalization was significantly enhanced. In addition, our analysis identified 4560 noncoding genes from NONCODE references. ENSEMBL and NCBI have nearly the same number of annotated genes (16,423 vs. 17,804), but using ENSEMBL as a reference, 2356 genes were able to be normalized and identified. Proceeding to biopsy x oocyte analysis, we were able to detect a greater number of genes in oocytes than in biopsy, where the preponderance was from NONCODE sources (68). Despite these minor differences, the high correlation of expression between them (89%) was consistent and proved to be a valuable instrument for studying the oocyte without destroying it.