Abstract The acquisition of meiotic and developmental competence in oocytes is a highly regulated process that includes post-transcriptional mechanisms mediated by microRNAs (miRNAs). To characterize these changes, we compared mRNA and miRNA expression profiles between non-matured, and oocytes submitted to IVM in canines using RNA sequencing. The oocytes were obtained from antral follicles dissected from canine ovaries. Total RNA was isolated from pooled oocytes from each condition and subjected to parallel mRNA and miRNA sequencing, followed by differential expression and pathway enrichment analyses. Our findings reveal transcriptional change after in vitro culture, showing significant differential expression (FDR < 0.05, |log2FC| > 1). Non-matured oocytes displayed an expression pattern associated with transcriptional preparation and structural organization, while IVM oocytes showed expression patterns consistent with cytoplasmic remodeling and metabolic activation. The miRNA profile changed with maturation, accompanied by a selective increase in miRNAs associated with post-transcriptional control of the cell cycle and structural pathways. Enrichment analysis revealed significant changes in signaling networks associated with cell adhesion, cytoskeletal organization, and communication. Components of the Hippo signaling pathway displayed altered transcript abundance following IVM, suggesting that structural and regulatory cues present within the oviduct may not be fully reproduced in culture. Our findings provide new insights into the molecular constraints of canine IVM that may serve as biomarkers for improving oocyte quality and reproductive technology outcomes in dogs.
The aim of this study was to compare the effectiveness of different analgesic protocols in reducing physiological stress markers in Santa Inês ewes undergoing non-surgical embryo recovery (NSER). In Experiment 1 (n = 32), ewes received a combination of butorphanol and meloxicam, whereas in Experiment 2 (n = 34), animals were treated with butorphanol combined with dipyrone and hyoscine. In Experiment 1, ewes received butorphanol (0.1 mg/kg IM) associated with meloxicam (1 mg/kg IV). In Experiment 2, animals were treated with butorphanol (0.1 mg/kg IM) combined with a dipyrone-hyoscine mixture (5 g dipyrone + 0.04 g hyoscine; 5 mL IM and 5 mL IV). Controls received equivalent volumes of saline. In Experiment 2, dipyrone-hyoscine was re-administered 24 h post-procedure. Ewes were synchronized, superovulated, and subjected to cervical dilation before NSER. Heart rate, respiratory rate, and blood biomarkers were recorded before sedation (BS), after sedation (AS), before cervical transposition (BCT), immediately after collection (IAC), and at 0.5, 1.5, 3, 6, 12, 24, and 48 h after collection (AC). In Experiment 1, respiratory rate was affected by treatment over time, it was greater (P=0.048) in control ewes than in ewes that received butorphanol associated with meloxicam at AS and 0.5hAC. No other variable differed significantly between groups. In Experiment 2, there was an interaction between treatment and time in heart rate. Heart rate was greater in ewes treated with butorphanol combined with a dipyrone-hyoscine mixture than in control ewes (P = 0.001), and there was also a significant interaction between treatment and time (P = 0.001). It was greater in treated ewes than in control ewes at AS, IAC, 0.5hAC, and 1.5hAC. No significant effects were detected for the remaining parameters. Under the tested conditions, neither analgesic protocol significantly reduced physiological indicators of stress.
This study investigated the patterns of sperm kinetics in two fish species, Piaractus mesopotamicus and Pseudoplatystoma reticulatum and their relationship with seminal quality. Semen samples were collected after hormonal induction, with the aim of identifying indicators of seminal quality and their relationship with reproductive success. The results revealed marked interspecific differences: P. reticulatum exhibited longer-lasting motility (83 ± 31 s) with stable linear movement patterns after activation, while P. mesopotamicus showed more intense but shorter-lasting motility (41 ± 8 s), exhibiting initial circular trajectories that transitioned to linear patterns. In P. reticulatum, positive correlations were found between total motility (MOT), progressivity (PROG), velocities: curvilinear (VCL), straight-line (VSL) and average (VAP), with hatching rate (HR). These are suggested to be reliable predictors of seminal quality. For P. mesopotamicus, multivariate analysis (PCA) indicated that sperm quality was associated with a specific combination of VCL, VAP, and amplitude and lateral displacement of the head (ALH), although no single parameter was directly correlated with HR. These differences reflect distinct evolutionary adaptations to their lotic reproductive environment, associated with effective oocyte fertilization capacity and larval hatching rate. P. mesopotamicus is adapted for rapid fertilization, with higher sperm speed, while P. reticulatum demonstrates more sustained sperm motility. These results highlight the need for species-specific approaches and demonstrate that the simple evaluation of sperm kinetic parameters in fish does not allow seminal qualification, requiring detailed knowledge of sperm movement by species, essential information to optimize the nutritional and reproductive management of breeders, and seminal cryopreservation protocols.
Our study explored the histological and histo-functional characteristics of the testis and seminal vesicles of Jaculus jaculus, collected from the Biskra (34° 51' 0" N 5° 43' 59.999" E) and M'sila regions in Algeria (35°42'20.99" N 4°32'30.98" E) from autumn to early spring, corresponding to activity and transitional periods of the reproductive cycle. An immunohistochemical analysis of matrix metalloproteinases (MMP-2 and MMP-9) was conducted on the seminal vesicles and testes during the active reproductive. The study utilized an indirect immunohistochemistry protocol with amplification through streptavidin-biotin-peroxidase. Histologically, seminiferous tubules in Jaculus jaculus, captured during the transitional period of the reproductive cycle, multinucleated giant cells of germinal origin were observed during the transitional phase of the reproductive cycle, reflecting a process of seasonal testicular remodelling. The histological structure of seminal vesicles appears highly characteristic of the studied species, with the vesicular epithelium organized into complex acinar glands surrounded by basal lamina and connective tissue. Gland cells secrete a heterogeneous product. In the testis, cells of the seminiferous epithelium express MMP-2 and MMP-9 in their cytoplasm, with a strong expression of MMP-2; however, both proteins are absent from the extracellular matrix. During the active period, in seminal vesicles, a significant immunohistochemical signal of MMP-2 and MMP-9 was observed in epithelial cells, smooth muscle cells, with no immunoexpression in the extracellular matrix. MMP-2 and MMP-9 are expressed in a pattern consistent with a potential role in the seasonal reproductive cycle.
Somatic cells serve as essential genetic resources for biobanking. Traditionally, jaguar fibroblast-like cells are frozen in cryovials using slow-freezing protocols. However, this approach necessitates access to ultra-low temperature freezers and requires substantial storage capacity. This study aimed to evaluate the efficacy of a rapid-freezing protocol as a feasible alternative for the freezing of jaguar fibroblasts. Following cultivation to the third passage, cells were frozen using either slow-freezing (SF) or rapid-freezing (RF). For SF, cells were frozen in cryovials (-1 ºC/min), whereas RF involved rapid-freezing (-60 ºC/min) using nitrogen vapor and storage in 0.25 mL straws. Post-thaw, cells were re-cultured to assess recovery. Cell viability was assessed via light microscopy (Trypan blue), fluorescence microscopy (acridine orange + propidium iodide), and flow cytometry (Hoechst 33342 + propidium iodide) immediately post-thaw and upon reaching confluency (> 80%). Fresh culture viability assessed by Trypan blue, acridine orange, and flow cytometry was 98.2 ± 0.3%, 88.7 ± 3.9%, and 94.2 ± 2.2%, respectively. No significant differences in viability were observed between SF and RF immediately post-thaw or after subsequent cultivation. All three assays revealed significant differences (p < 0.05) between cells assessed immediately after thawing and those assessed at confluence. Upon reaching confluence, there was no significant difference (p > 0.05) between fresh and frozen-thawed cells. These findings indicate that RF in straws is a viable alternative to SF for jaguar fibroblast cryopreservation, offering optimized storage density for genetic resource banks.
During equine semen cryopreservation, most of the seminal plasma is removed, making sperm more susceptible to oxidative stress. This study evaluated the effect of resveratrol supplementation on the quality of frozen-thawed equine sperm. Semen from ten stallions was frozen using a control INRA 96 extender and extenders supplemented with 5-, 10-, 100-, and 150- µM resveratrol. Post-thaw evaluations included motility, kinematic parameters, morphology, plasma and acrosomal membrane integrity, mitochondrial potential, lipid peroxidation, nitrite, hydrogen peroxide, malondialdehyde levels, total reactive oxygen species (ROS), chromatin protamine deficiency, chromatin condensation, sperm binding to bovine oviduct explants, and gene expression of apoptosis-related genes such as B cell lymphoma 2 (BCL2) and BCL2-associated X (BAX), mitochondrial ROS modulator 1 associated with mitochondrial ROS production (ROMO1), sperm acrosome-associated 3 related with sperm binding capacity to the zona pellucida (SPACA3), and DNA damage repair gene 8-oxoguanine DNA glycosylase 1 (OGG1). Data were analyzed by ANOVA and Tukey test (P < 0.05). The 10 µM resveratrol treatment significantly increased sperm motility, mitochondrial activity, SPACA3 expression, and the number of sperm bound to oviduct explants compared to the control. Additionally, 10 µM resveratrol reduced total ROS, ROMO1, and BAX gene expression, indicating reduced oxidative stress and apoptosis. In conclusion, 10 µM resveratrol supplementation improved sperm metabolic activity, enhanced oviduct binding capacity, and demonstrated antioxidant and anti-apoptotic effects. Thus, incorporating resveratrol into freezing extenders could be a promising strategy to improve the fertilizing capacity of equine sperm in artificial insemination programs.
This study aimed to investigate one- year- old male rat sensitivity to fetal programming effects of subnutrition during gestation and lactation. Thirty primiparous female 3 month old Wistar rats were all mated by the same male. Pregnant females were organised in two experimental groups: control (CG n= 15; ad libitum food access) and restricted group (RG n= 15; food intake 50% of the control group). Morphometric and immunohistochemical techniques were employed to determine the number of Sertoli cells and both the distribution and relative abundance of androgen receptor in Sertoli, Leydig, and myoid cells. Additionally, the proportions of proliferating and apoptotic cells were analyzed. Testosterone concentration was also measured in blood serum and determined by a direct solid-phase radioimmunoassay. Our findings indicate that nutritionally restricted animals at early developmental stages exhibited reduced body weight (CG 592,9±33,4*** vs RG 527,5±23,5), testicular weight (CG 1,82±0,05*** vs RG 1,69±0,06), and body length (CG 47,2±0,7*** vs RG 45,1±1,6). We also observed in the same experimental group a lower number of Sertoli cells both per cross section of the seminiferous tubule (CG 23,4±0,32*** vs RG 21,8±0,34) and total per testicle (CG 134,6±24,9 x 104* vs RG 111,4±14,4 x 104). We found no treatment effect on the volume density of the testicular interstitium. No differences were observed between the groups in the proportion of proliferating and apoptotic cells. Finally, undernourished animals exhibited higher concentrations of testosterone in blood serum (CG 2,90 ± 1,51* vs RG 5,25 ± 3,53), and their myoid cells demonstrated increased expression of androgen receptors. In conclusion, our findings suggest that undernutrition during early life stages fetally programs the rat, negatively affecting its body development, altering testosterone production and testicular histophysiology in adults older than one year of age.
This study aimed to evaluate the applicability of fetal heart rate variability (FHRvar%), umbilical artery resistivity index (RI-UmbArt), and maternal body temperature (Temp, °C) for predicting parturition in bitches. Fifteen bitches (1-6 years old) were included. Gestational age was estimated using fetal biometry (inner chorionic cavity or biparietal diameter), and during the last week of pregnancy the animals were evaluated twice daily until parturition. Data were retrospectively grouped according to the hours before parturition (HBP). The evaluated parameters were FHRvar% (measured over 5-10 minutes using pulsed Doppler abdominal ultrasonography), RI-UmbArt (assessed by triplex Doppler in three fetuses per session), and Temp (rectal thermometry). Data were statistically compared and correlated with HBP. FHRvar% showed a gradual increase (P = 0.010) beginning at 84 HBP, reaching a peak between 24 and 12 HBP, whereas Temp significantly decreased during the last 24 HBP (P < 0.001). Both FHRvar% and Temp were significantly correlated with HBP (P < 0.001); however, these correlations were weak (Pearson's r = -0.338 and -0.491, respectively). RI-UmbArt showed no significant variation across HBP (P = 0.711). An FHRvar% > 31.5% predicted parturition within 24 hours with a sensitivity of 65% and a specificity of 67%, whereas a Temp < 37.5°C predicted parturition with 69.6% sensitivity and 77.8% specificity. In conclusion, although FHRvar% and maternal body temperature are influenced by the proximity of parturition, they may not be reliable predictors of parturition timing, while RI-UmbArt appears to remain unchanged close to delivery.
Colostrogenesis and lactogenesis influenced by age require further understanding in Zebu herds, due to the longevity of breeding cows. Thus, we aimed to compare colostrum and milk physicochemical and proteinogram of Dairy Gir cows and calves' passive immunity transfer by grouping 28 cows and their calves by maternal age: Young (n=6; aged 24-36 months), Adult (n=15; 37-91 months) and Senior (n=7; 96-137 months) groups. Colostrum and milk were collected at first milking (immediately after calving), transition milk (3rd milking, after 24h of calving) and full milk (10th milking, after 5 days of calving) for analysis of density, total solids, pH, immunoglobulin (IgG) and proteinogram (lactoferrin, albumin, heavy and light chain IgG, β-lactoglobulin and α-lactalbumin). Calves' blood was collected after 3 days of birth to evaluate serum total solids, total protein, liver profile, light and heavy chain IgG. Data were statistically analyzed by LSD Test at P<0.05. Young Group had higher (P<0.05) colostrum total solid and density compared to Senior Group. Transition milk of young cows had lower (P<0.05) pH than seniors. Calves of senior cows presented higher (P<0.05) serum albumin than young and adult. Young cows had IgG decrease from colostrum to transition milk, whereas adult cows presented fall (P<0.05) of IgG throughout milking. In conclusion, age of Dairy Gir cows does not affect colostrum and milk immunological quality, nor calves' immunity transfer. However, it determines protein composition and dynamic changes, especially in young and senior cows.
The anti-Mullerian hormone (AMH) or Mullerian Inhibiting Substance (MIS), produced by the Sertoli cells induces the regression of Mullerian ducts in male embryos. Granulosa cells from ovarian follicle are homologous to Sertoli cells and produce AMH. The aim of this study was the immunolocalization of AMH in the testicles and ovaries of bovine fetuses at different ages. Gonads from bovine fetuses between 4 and 7 months (24-98 CRL) were collected in a local slaughterhouse, fixed in 10% formaldehyde, and processed for conventional histology and embedded in paraffin cut with a rotative microtome, deparaffinized, and subjected to immunohistochemistry using the anti- AMH 1:50 (SC 28912, Santa Cruz Biotechnology) according to the manufacturer's instructions. Immunostaining was observed in the cytoplasm of Sertoli cells, within the sexual cords (developing seminiferous tubules) at all ages and was not observed in gonocytes and interstitial tissue. In the ovary there was light staining in granulosa cells but not in the theca cells, however, intense staining was observed in the cytoplasm of the oocyte of primordial, primary, secondary and antral follicles, at all ages analyzed, suggesting that despite being produced by granulosa cells, AMH concentrates in the oocyte cytoplasm. The importance of AMH in sexual differentiation and action in Muller's ducts is well understood, however there are no studies on the role of this protein in gonadal development, this being the first report of AMH immunolocalization in fetal bovine testis and ovaries, requiring further studies on the action and importance of AMH in the gonadal development of bovine fetuses.
Postpartum uterine disease is a major cause of impaired bovine reproduction and is usually managed with intrauterine antibiotics. Alternative non‑antibiotic therapy is needed to support uterine health without contributing to antimicrobial resistance. The goal of this study was to characterize histomorphometric and ultrastructural changes of the bovine endometrium after intrauterine infusion of ozonated sunflower oil during early puerperium, and to assess its in vitro antimicrobial activity against uterine bacteria. Eighteen primiparous cows received a single intrauterine treatment with ozonated or non‑ozonated oil (O3 and non‑O3 groups; n=9/group) on day 10 postpartum. Endometrial biopsies were collected before and 15 days post-treatment (D0 and D15) for light microscopy and scanning electron microscopy. Uterine cytology was used to evaluate subclinical endometritis. Minimum inhibitory and bactericidal concentrations (MIC and MBC) of the treatments were determined against uterine isolates (Escherichia coli, Staphylococcus spp., Streptococcus spp. and Arcanobacterium pyogenes). At D15, the number of endometrial glands was higher in the O3 group than in the non‑O3 group (32.6±2.5 vs. 11.0±1.0; P<0.01), whereas glandular diameter, luminal area and cell number per gland were lower in O3 cows (P<0.01), indicating enhanced gland proliferation with reduced glandular dilatation. No degenerative histopathological changes, such as periglandular fibrosis or endometrial atrophy, were detected in either group, and scanning electron microscopy showed preserved apical ultrastructure without adverse effects of ozonated oil. Ozonated oil inactivated all uterine bacteria (MIC and MBC ≤0.18 µg mL‑1), while non‑ozonated oil showed no antimicrobial activity. Both groups showed a marked reduction in neutrophil percentage by D15, but histological evidence of persistent inflammation was more pronounced in non‑O3 cows. These findings indicate that intrauterine ozonated sunflower oil is a promising non‑antibiotic therapy to support postpartum uterine involution and endometrial recovery in dairy cows and may contribute to improved reproductive performance.
This study investigated the effects of lauric acid (LA) supplementation during in vitro maturation (IVM) of bovine oocytes on nuclear maturation and subsequent embryonic development. Cumulus-oocyte complexes (COCs) were matured in medium containing LA at concentrations of 50, 100, 150, 200, or 300 µM, or without LA (control). Embryo development following in vitro fertilization (IVF) and culture (IVC) was assessed. Based on optimal outcomes in the first experiment (A total of 1,558 oocytes were used across 6 - 8 replicates per treatment group),200 µM LA was selected to evaluate its protective role under cellular stress. Oocytes (n = 1,255; five replicates) were matured with or without 200 µM LA and exposed to heat stress (41 °C) or oxidative stress (1 mM H2O2). LA supplementation significantly improved blastocyst rates, with 100 - 200 µM groups showing higher formation rates (≥ 37.1%) compared to control (21.3%) (p < 0.05). Blastocyst quality, based on A-B grade embryos, was also enhanced in the 200 µM (51.1%) and 300 µM (56.5%) groups versus control (19.2%) and 50 µM (16.9%) (p < 0.05). Stress exposure reduced maturation and blastocyst rates. Oxidative stress significantly decreased maturation (OS: 44.9%) and blastocyst development (18.6%) compared to control (72.3% and 28.4%, respectively), while LA treatment improved these outcomes (OS-LA: 51.5% and 31.5%) (p < 0.05). Under heat stress, LA showed a non-significant trend toward higher blastocyst rates (24.8%) compared to stress alone (16.2%). Under normal conditions, 200 µM LA significantly enhanced blastocyst yield (44.2%) versus control (28.4%) (p < 0.05). Data were analyzed using ANOVA with post-hoc tests, and differences were considered significant at p < 0.05. These results indicate that LA supplementation during IVM enhances bovine embryo developmental competence and partially mitigates oxidative stress-induced impairment.
This study evaluated the toxicity of cryoprotectants and the performance of low-rate freezing protocols for oocytes and embryos of Piaractus brachypomus, a Neotropical fish of increasing relevant for Brazilian aquaculture and genetic conservation. Eight cryoprotectant solutions based on methanol (MeOH) or dimethyl sulfoxide (Me2SO), combined with 0.25 M sucrose in L-15 or HBSS media, were tested for oocytes toxicity at 28 °C and post-freezing viability. In spite of histological and scanning electron microscopy (SEM) analyses indicated preservation of gross morphological features, none of the cryopreserved oocytes supported embryonic development, indicating loss of functional viability following cryoprotectant exposure and freezing. For embryos, two low-rate freezing protocols were evaluated: Protocol 1 (P1E), based on gradual cooling to -13 °C, and Protocol 2 (P2E), involving linear cooling to -60 °C followed by storage in liquid nitrogen. In P1E, eight treatments using 3.1 M MeOH combined with different concentrations of polyvinylpyrrolidone (PVP) or sucrose were tested. The highest proportion of morphologically viable embryos (15.3%) was obtained with MeOT + 0.45 M sucrose (SC5), which different significantly from the other treatments. In contrast, no morphologically viable embryos were recovered after P2E, likely due to inadequate dehydration and intracellular ice formation. Overall, high concentrations of permeant cryoprotectants and prolonged equilibration times were detrimental to oocyte and embryo viability, while sucrose showed better cryoprotective performance than PVP. Even though protocols tested were insufficient to ensure consistent post-thaw viability, the partial success observed in P1E under MeOH and sucrose combinations provides a relevant experimental basis for future refinement of conservation strategies and contributes to development of ex situ germoplasm conservation approaches for P. brachypomus and other Neotropical species.
We aimed to evaluate puberty induction, follicular dynamics, and hormonal profiles of progesterone (P4) and estradiol (E2) in prepubertal Nelore and Murrah heifers subjected to synchronization for fixed-time artificial insemination (FTAI). Six Nelore heifers (14 to 17 months old; 320 to 350 kg) and six Murrah heifers (14 to 17 months old; 422 to 523 kg) confirmed as prepubertal by ultrasonography due to the absence of a corpus luteum were used. The protocol included an intravaginal P4 device (0.5 mg) on D0, along with the administration of estradiol benzoate (EB) and prostaglandin F2α. In Nelore, the device was removed on D7, followed by the administration of estradiol cypionate, prostaglandin F2α, and eCG, with insemination occurring 48-56 hours after intravaginal device removal (D9). In Murrah, the device was removed on D9, followed by prostaglandin F2α and Ecg administration, with insemination on D12 after GnRH administration (D11). Blood samples were collected (Nelore: D7 and D9; Murrah: D9 and D12) to measure hormone levels, and ultrasonography monitored follicular dynamics and ovulation. The ovulatory follicle diameter was 7.83 ± 2.25 mm (Nelore) and 7.7 ± 1.63 mm (Murrah), with no significant differences (P > 0.05). The dominant follicle diameter at P4 device removal was 6.5 ± 1.59 mm (Nelore) and 6.9 ± 4.97 mm (Murrah). The mean P4 levels were 0.76 ng/mL (Nelore) and 1.42 ng/mL (Murrah) at P4 device removal, and 1.00 ng/mL and 0.52 ng/mL at the moment of FTAI. The mean E2 levels were 9.28 pg/mL (Nelore) and 26.75 pg/mL (Murrah) at the P4 device removal, and 11.20 pg/mL and 11.59 pg/mL at the moment of FTAI. Ovulation rates were 100% in Nelore and 50% in Murrah. These results highlight the importance of tailoring FTAI protocols to the specific reproductive characteristics of each breed.
Capybaras (Hydrochoerus hydrochaeris) are increasingly synanthropic throughout South America, where their rapid population growth in human-altered landscapes heightens conflicts and, critically, elevates the public health risk of Brazilian Spotted Fever (BSF), a lethal Rickettsia rickettsii zoonosis transmitted by Amblyomma sculptum ticks. Since capybara density is tightly coupled with pathogen circulation, non-lethal, scalable management strategies are essential. We evaluated the long-term efficacy and safety of the single-dose GonaCon™ immunocontraceptive vaccine in free-living, sexually mature female capybaras (n=6 treated, n=3 control) ~ 33 months. All treated females demonstrated sustained reproductive suppression (absence of parturition or estrus behavior; p < 0.01). Efficacy was confirmed by multi-modal evidence: plasma progesterone (P4) and estradiol (E2) concentrations were significantly suppressed, showing reductions of 41% and 36% by Day 27 post-vaccination, respectively (p < 0.005; Cohen's d > 1.5). Histological analysis validated HPG axis disruption, revealing significantly lower ovarian weights (p=0.002) and suppressed folliculogenesis, with treated ovaries possessing drastically fewer antral follicles (mean 2.3 ± 0.5 vs. 12.7 ± 1.2 in controls; p<0.001). The treatment was safe, with the only adverse effect being a transient, localized swelling in 4/6 animals that resolved within two weeks. Importantly, treated females maintained critical social behaviors, including alloparental care. These findings provide strong evidence for the sustained, multi-year efficacy of GonaCon in a large wild rodent, supporting its use as a humane, non-surgical tool for density management and BSF risk mitigation under the One Conservation Paradigm.
Understanding diseases that affect the male reproductive system of dogs and cats is important for improving their health and quality of life. Imaging techniques, such as ultrasound and X-rays, play a fundamental role in the evaluation of this system. They help veterinarians detect problems in the testicles, prostate, and other reproductive organs of dogs and cats. In recent years, advanced ultrasound methods, such as Doppler, elastography, and contrast-enhanced ultrasound, have become more accessible and allow earlier and more accurate diagnosis. Although definitive diagnosis of neoplastic lesions depends on cytological or histopathological examinations, radiography and advanced ultrasonography contribute significantly to early detection and malignancy characterization. This literature review explores the main imaging techniques used in the investigation of malignant lesions in the male reproductive tract of dogs and cats, namely radiography, B-mode and Doppler ultrasonography, elastography, and contrast-enhanced ultrasonography (CEUS). The goal is to help veterinarians make better decisions and improve care for their patients. The integration of these modalities has become well established in human medicine and shows growing applicability in veterinary practice, allowing for real-time, non-invasive assessment of the testes, prostate, penis, and scrotal structures. The compiled data highlight the potential of combining different imaging modalities to improve diagnostic accuracy and provide clinical and surgical support in the management of malignant lesions affecting the male reproductive tract in dogs and cats.
Bees are essential pollinators with species differing morphologically and physiologically. Understanding the variations in reproductive parameters between phenotypes is crucial. This study compares the sperm characteristics of Italian (Apis mellifera ligustica) and Africanized drones (Apis mellifera L.) raised in the Caatinga biome. Nine sexually mature Italian drones and sixteen Africanized drones from different colonies were used. Semen was collected using the endophallus eversion technique and diluted in saline solution (1:20). The parameters analyzed included: motility (optical microscopy), sperm viability (Hoechst 33342; propidium iodide), functional integrity of the plasma membrane (hypo-osmotic test), morphology and morphometry (Rose Bengal), and scanning electron microscopy. The results were expressed as mean ± standard error. Statistical analyses included the Shapiro-Wilk test to the normality of residuals and the Bartlett test to verify homoscedasticity. Comparisons between groups were performed using the Mann-Whitney and Student's t-tests (P < 0.05). Both phenotypes presented 90% sperm motility with viability of 82.4 ± 2.5% for Italians and 81.1 ± 2.4% for Africanized ones; the functional integrity of the plasma membrane was 93.4 ± 1.8% and 91.6 ± 1.5%, respectively. Regarding morphology, the percentage of normal sperm was 10.89 ± 1.66% for Italian and 12.06 ± 1.01% for Africanized, with the curled tail being the most predominant feature of sperm morphology. No statistically significant differences (P > 0.05) were observed for the above-mentioned parameters. Sperm head morphometry was significantly larger (P < 0.05) in Italian (10.04 ± 0.03 µm) compared to Africanized (9.33 ± 0.04 µm). Scanning electron microscopy analysis revealed no ultrastructural differences between phenotypes. In conclusion, there is a high degree of similarity in sperm parameters of both phenotypes under the same environmental conditions, indicating the feasibility of applying similar reproductive strategies.
Approximately 85% of embryos produced in Brazil are in vitro (IVP). However, the success of IVP is still quite diverse, and several factors need to be coordinated to obtain a live calf from an oocyte. This study aims to evaluate the influence of the donor breed (Nellore vs. Senepol) and the seasons of year (Dry vs. Rainy) at the time of follicular aspiration (OPU) on the number of viable oocytes and embryos produced, and the recipients pregnancy rate per embryo transfer (P/ET). Data from 368 donors was evaluated, 198 Senepol and 170 Nellore, resulting in a total of 21,758 oocytes collected by OPU. During the period a total of 4,740 embryos were produced (2,135 Senepol and 2,605 Nellore) and 2,124 fresh embryo transfers were analyzed. A negative effect of dry season was observed on number of total and viable oocytes for Nellore. The embryo production rate was lower for Senepol in both periods. Nellore breed had lower embryo production rate in the rainy period. The P/TE was lower for embryos from Senepol donors during dry season, while the P/TE of embryos from Nellore donors was not affected by season. In conclusion, the dry season negatively influences total and viable oocyte collection in Nellore donors, while in Senepol donors remained constant throughout the year. The embryo production of Nellore donors is negatively influenced by rainy season, although it remains higher than the Senepol donors throughout the year, and P/TE of the embryos of Senepol donors is reduced during the dry season.
The reproduction of some fish species in captivity is only possible by imposing artificial reproductive procedures, and the manipulation of fish for these purposes is a stressor. Thus, anesthesia can reduce stress during handling. However, it is necessary to investigate the possible side effects on breeding and the general health of the animal. Therefore, we aimed to investigate the impact of tricaine methanesulfonate (MS-222) and eugenol (EUG) at concentrations of 300 mg L-1 and 40 mg L-1, respectively, on several stress- and reproductive-related parameters in Rhamdia quelen anesthetized before semen collection. After hormonal induction, 24 sexually mature males (534.4 ± 259 g) were randomly assigned to treatment groups and semen and blood samples were collected. Anesthesia recovery and induction time were extended in fish anesthetized with EUG; however, plasma cortisol concentrations did not differ among the treatment groups. EUG caused higher DNA fragmentation in blood cells than in the control group (without anesthesia). EUG increased the monocyte count compared to the other experimental groups. MS-222 showed a lower sulfhydryl group (SH) quantitation than the other experimental groups. The anesthetics used before semen collection in this study had no adverse effects on the fertilization or hatching capacity of anesthetized R. quelen. MS-222 (300 mg L-1) and EUG (40 mg L-1), despite being related to hematological and semen changes, did not negatively affect the reproductive capacity of R. quelen males. Thus, anesthesia is recommended for R. quelen during semen collection, considering reproductive parameters and animal welfare.
Dichlorvos (DDVP) is a widely used organophosphate insecticide. The reports of its toxicity and induction of oxidative stress have warranted the search for an antidote. Melatonin (MLT), a hormone found naturally in the body is known to mitigate oxidative stress. Therefore, the present study aimed to investigate the protective effects of melatonin on dichlorvos-induced toxicity in the testes and epididymis. Sixty (60) 10 weeks of male Wistar rats (160 ± 10g) randomly grouped into four (n=15, A-D) were used and treated as follows: A (Control): corn oil (1 mL/kg body weight), B: MLT (10 mg/kg), C: DDVP (1.6 mg/kg), and D: DDVP and MLT. Except for the MLT intra-peritoneal treatment, other test samples were administered orally for 45 days. Histomorphometry and histological examinations were conducted on the testes and epididymis 24h, 14 days, and 45 days after treatment. There was a significant decrease in the epithelial length of the epididymis of rats treated with DDVP at 24h and 45 days post-treatment compared to the control. MLT significantly increase the mean epididymal epithelial length at 24h and 45 days post-treatment in the co-treated group. The DDVP-induced significant reduction in the testes' luminal diameter was reversed by MLT in the co-treated group. MLT, DDVP (45th-day treatment), and DDVP+MLT (14th and 45th-day treatment) increased the epididymal luminal diameters significantly compared to the control. Meanwhile, DDVP altered both the testicular and epithelial architectures and caused atrophy of the seminiferous tubules and spermatogenic arrest with atrophy of epithelial tubules, especially following 14- and 45-day treatment. These lesions were reversed in the co-exposed rats by MLT. In conclusion, melatonin mitigates the derangements induced by dichlorvos in the testes and epididymis.
