Animal Reproduction最新文献

Germ cell transplantation in fish is a promising technique for surrogate broodstock parents with broader application in aquaculture and conserving endangered and valuable genetic resources. Herein, we describe the establishment of an intrapapillary xenogeneic transplant of germ cells from sexually mature goldfish (C. auratus) males into common carp (C. carpio) males cytoablated with a thermochemical treatment (two doses of busulfan at 40 mg/kg at 35°C). To analyze the presence and development of donor germ cells in recipient testes, donor germ cells were labeled with PKH26, a fluorescent cell membrane dye, before transplantation. Our results demonstrated that thermochemical treatment caused effective spermatogenesis suppression and pronounced germ cell loss. Moreover, transplanted spermatogonial cells were able to colonize the recipients' testes, resume spermatogenesis, and generate spermatozoa within eight weeks after germ cell transplantation. These findings suggested that recipient testes provided suitable conditions for the survival, colonization, proliferation, and differentiation of donor spermatogonia from a related species. This study indicated that recipients' testes exhibited a high degree of plasticity to accept and support xenogeneic donor germ cells, which were able to form sperm in a short time frame. This approach has significant implications for assisted animal reproduction, biotechnology, conservation, and the production of valuable genetic resources and endangered fish species.

Cryoprotectants are required to reduce damage caused to the cells due to low temperatures during the cryopreservation. Antifreeze proteins (AFP) have a well-known role in cell membrane protection, while resveratrol is a potent antioxidant. This study assessed the effect of the association of resveratrol concentrations and AFP I in a ram semen extender. Pooled semen of four rams was allocated into six treatments in a factorial arrangement: (CONT, only the semen extender); only AFP I (ANT: 0.1 µg/mL of AFP I), only resveratrol, one treatment with two levels (10 µM/mL or 50 µM/mL of resveratrol); and two treatments with the interactions, with one AFP I and one of the two levels of resveratrol (0.1 µg/mL of AFP I with 10 µM/mL resveratrol; 0.1 µg/mL of AFP I with 50 µM/mL resveratrol). No interaction between factors was observed on sperm kinetics, plasma membrane integrity, hypo-osmotic test, and mitochondrial activity parameters. There was a high probability (P = 0.06) of reducing sperm cells with functional membrane percentage in the hypo-osmotic test and increasing the percentage of sperm with high mitochondrial activity (P = 0.07) was observed in AFP presence. An interaction of AFP and resveratrol was observed in non-capacitated sperm (P = 0.009), acrosomal reaction (P = 0.034), and sperm binding (P = 0.04). In conclusion, the association of resveratrol and AFP did not improve the quality of frozen-thawed semen and even promoted deleterious effects compared to their single addition in the semen extender. The supplementation of 50 µM/mL of resveratrol improved the outcomes of frozen-thawed ram sperm, being a potential cryoprotectant.

The number of antral follicles is considered an important fertility trait because animals with a high follicle count (HFC) produce more oocytes and embryos per cycle. Identification of these animals by genetic markers such as single nucleotide polymorphisms (SNPs) can accelerate selection of future generations. The aim of this study was to perform a genome wide association study (GWAS) on Nelore and Angus heifers with HFC and low (LFC) antral follicle counts. The groups HFC and LFC for genotyping were formed based on the average of total follicles (≥ 3 mm) counted in each breed consistently ± standard deviation. A total of 72 Nelore heifers (32 HFC and 40 LFC) and 48 Angus heifers (21 HFC and 27 LFC) were selected and the DNA was extracted from blood and hair bulb. Genotyping was done using the Illumina Bovine HD 770K BeadChip. The GWAS analysis showed 181 and 201 SNPs with genotype/phenotype association (P ≤ 0.01) in Nelore and Angus heifers, respectively. Functional enrichment analysis was performed on candidate genes that were associated with SNPs. A total of 97 genes were associated to the 181 SNPs in the Nelore heifers and the functional analysis identified genes (ROBO1 and SLIT3) in the ROBO-SLIT pathway that can be involved in the control of germ cell migration in the ovary as it is involved in lutheal cell migration and fetal ovary development. In the Angus heifers, 57 genes were associated with the 201 SNPs, highlighting Fribilin 1 (FBN1) gene, involved in regulation of growth factors directly involved in follicle activation and development. In summary, GWAS for Nelore and Angus heifers showed SNPs associated with higher follicle count phenotype. Furthermore, these findings offer valuable insights for the further investigation of potential mechanism involved in follicle formation and development, important for breeding programs for both breeds.

Semen collection methods vary greatly and rely on the practitioner's expertise, available materials, and the specific behavioral traits of the male animals involved. When it comes to domestic cats, wild felids, and canids, semen collection is particularly challenging. Thus, given the difficulty of semen collection in these species, pharmacological semen collection (PSC) stands out since it is a quick and straightforward method that does not require specific equipment. The PSC consists of administering α2-adrenergic receptor agonist drugs, mainly medetomidine, and dexmedetomidine, aiming semen release into the urethra with posterior urethral catheterization and sperm recovery. This technique was primarily described in domestic cats and wild felids, and despite the decreased seminal volume, the retrieved semen is highly concentrated and presents good quality. However, further studies are required to optimize semen collection in domestic dogs and wild canids. Therefore, the purpose of this review is to provide a comprehensive overview of the research developed on pharmacological semen collection (PSC) in the past few decades. The objective is to equip professionals with the essential knowledge required for the efficient application of this technique in both domestic and wild canids and felids and to make a valuable contribution to conservation efforts and the preservation of biodiversity, aligning with the principles of One Conservation.

Sperm motility and kinematics analysis are important to predict bull fertility. However, there are other molecules in the sperm with the ability to improve the pregnancy rate. For example, PLCZ1 is a sperm protein that plays a unique role in the activation of the zygote and is important for the survival of the embryo. The objective of this work was to compare the expression of PLCZ1 mRNA in sperm cells of Chihuahuan Criollo and European bulls in the winter and summer seasons, under a low-input system. Six (3.33 ± 0.43 years old) bulls (three Criollo, three European) were used. Gross and individual motility were measured in semen obtained by electrostimulation. The cell pack was pelletized by centrifugation and stored in liquid nitrogen. The sperm cells were purified and total RNA was extracted. cDNA was synthesized to perform qPCR and measure the relative level of PLCZ1 transcripts in each bull. There were no differences in individual motility, however, gross motility was lower (P < 0.05) in Criollo bulls, both in the winter (71.1 ± 2.8 vs. 76.6 ± 2.8%) and in the summer season (58.9 ± 2.8 vs. 77.7 ± 2.8%). PLCZ1 expression was 5.3 times higher (P < 0.05) in winter than in summer (5.09 ± 1.09 vs 0.959 ± 1.09). No difference (P>0.05) was found in the expression levels of PLCZ1 between both breeds (4.36 ± 1.09 vs 1.69 ± 1.09), for Criollo and European, respectively. Although the animals presented seminal motility within the recommended limits for insemination, the expression levels of PLCZ1 vary depending on the time of the year and this might impact the rate of successful pregnancies. Therefore, it is important to complement conventional analysis of seminal quality with molecular characteristics.

The objectives of the study were to (1) describe the kinematic parameters of spermatozoa (2) compare methods of evaluating sperm viability (3) validate assays of functionality and integrity of the sperm membrane and (4) evaluate possible changes between spermatozoa from the epididymis and the vas deferens of the greater rhea. Semen samples were recovered from 7 adult individuals. Sperm motility was characterized by adjusting the set-up for Computer-assisted semen analysis (CASA) to that new species. For sperm viability evaluation, smears of bromophenol blue and eosin-nigrosine dyes were used. Five solutions of different osmolarities were then tested for the hypoosmotic swelling test (HOST). The combination of fluorescent probes (propidium iodide - IP and Hoechst 33342) was also used to assess plasma membrane integrity. Data were presented as mean ± SEM. Rhea spermatozoa from the vas deferens had an overall motility of 14.6 ± 2.5%. The bromophenol blue staining technique revealed that 64.6 ± 5.2% sperm were viable, while that proportion was 72.1 ± 2.5% using eosin-nigrosine. An average of 77.6 ± 4.8% of spermatozoa reacted to the HOST with distilled water at 0 mOsm/l. Fluorescent probes indicated that 65.3 ± 2.6% of spermatozoa had intact membranes. Interestingly, no statistical differences were observed between the parameters analyzed in the epididymal spermatozoa and the vas deferens. These new assays set reference values that can now be used to further exploration of sperm handling conditions and freezing protocols in rheas.

The risk of pregnancy loss in mares leads to the use of exogenous hormones to help pregnancy maintenance. The objective was to evaluate the proportion of thyroid hormones and steroids in neonates, in the following postpartum period, born to mares fed with synthetic progesterone and to verify the existence of a correlation between the level of progesterone between mother and neonate. Twenty-seven mares and their foals were used. The animals were divided into 5 experimental groups: group 1 (control, without hormonal supplementation), group 2 (random samples fed to 120 days of pregnancy with long-term progesterone), group 3 (mares fed with short-term progesterone as of 280.º day of pregnancy), group 4 (mares fed with long-term progesterone as of 280.º day of pregnancy) and group 5 (mares fed with synthetic hormone [altrenogest] as of 280.º day of pregnancy). The animal's blood collection took place immediately after parturition for the hormonal measurement. The hormones measured in neonates were total T3, free T4, TSH, progesterone and cortisone. In mares, only levels of progesterone. The groups of neonates showed no difference on levels of total T3, free T4, TSH and progesterone. There was no difference on levels of progesterone in mares among the groups. Neonates from groups 4 and 5 had higher and lower cortisone levels, respectively. No neonate showed clinical change. There was also no correlation between levels of progesterone in mares and foals. Thus, hormonal supplementation with long-term progesterone as of 280 days of pregnancy leds to an increase in the neonate's cortisone levels, in the meantime, supplementation with altrenogest as of 280 days of pregnancy caused a decrease on cortisone levels in foals, despite clinical signs have not been observed on these animals.

This study aimed to investigate the effect of a short glutamate supply on the ovarian response in goats with low body condition scores. Twenty-one goats had their estrus and follicular waves synchronized using three injections of prostaglandin analog at seven-day intervals. Goats were allocated to groups receiving 10 mg/kg LW (iv) of glutamate administered in a single dose (group LBCG1, n = 7) or in two doses five days apart (group LBCG2, n = 7). The control group (LBC; n = 7) received saline solution. Glutamate treatment did not affect glucose, cholesterol, or glutathione peroxidase levels, body weight, or adipose deposits. During the experimental period, the LBCG2 group showed a higher (P < 0.05) number of follicles (> 3 mm) and an increase in follicle diameter (P < 0.05). Glutamate supply improved (P < 0.05) the intraovarian Doppler blood area size in the LBCG groups, and the second dose in LBCG2 also induced a higher (P < 0.05) systolic and diastolic peak of the ovary artery. After ovulation induction, LBCG2 exhibited a high (P < 0.05) volume of the corpus luteum and vascularized area. We concluded that the supply of two doses of glutamate five days apart was efficient in ovarian stimulation in goats with a low body condition.

The Brazilian Buriti oil presents low extraction costs and relevant antioxidant properties. Thus, this work aimed to analyze Buriti oil biomaterial (BB), within its physicochemical properties, biocompatibility and cellular integration, with the purpose to the use as a growth matrix for Goat Wharton's jelly mesenchymal stem cells. Biomaterials were produced from Buriti oil polymer (Mauritia flexuosa), for it's characterization were performed Infrared Region Spectroscopy (FTIR) and Thermogravimetric Analysis (TG and DTG). The biointegration was analyzed by Scanning Electron Microscopy (SEM) and histological techniques. In order to investigate biocompatibility, MTT (3-(4,5-dimetil-2-tiazolil)-2,5-difenil-2H-tetrazólio) test and hemolytic activity tests were performed. The activation capacity of immune system cellswas measured by phagocytic capacity assay and nitric oxide synthesis . The BB presented an amorphous composition, with high thermal stability and high water expansion capacity, a surface with micro and macropores, and good adhesion of Wharton's jelly mesenchymal stem cells (MSCWJ). We verified the absence of cytotoxicity and hemolytic activity, in addition, BB did not stimulate the activation of macrophages. Proving to be a safe material for direct cultivation and also for manufacturing of compounds used for in vivo applications.