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The ancestry of domestic species from their closest wild relatives is one of the most debated and intriguing topics in evolutionary genetics. This review synthesizes current scientific understanding of the phylogenetic relationships between wild mouflon populations and domestic sheep (Ovis aries). It delves into the complex ancestry, tracing the primary role of the Asiatic mouflon (Ovis gmelini) as the progenitor, while also addressing the debated contributions of other wild Ovis species. The report explores the insights gained from diverse genetic markers, including mitochondrial DNA haplogroups and comprehensive whole-genome sequencing, highlighting their strengths, limitations, and the resolution of phylogenetic discrepancies. The multi-faceted taming process is examined, discussing proposed evolutionary mechanisms such as the domestication syndrome and thyroid hormone hypotheses, alongside human-mediated selection for key phenotypic traits like horn morphology, coat type, and tail characteristics. Furthermore, the pervasive role of hybridization and introgression between wild and domestic populations is analyzed, detailing its impact on genetic distinctiveness, adaptive potential, and the critical implications for conservation strategies. Finally, the review addresses ongoing scientific debates, particularly concerning the taxonomic classification of European mouflon, and identifies crucial avenues for future research to further unravel the intricate evolutionary tapestry of Ovis species. To ensure taxonomic consistency and promote conservation, nomenclature should be updated across all public repositories. Following the widely accepted classification that recognizes its lineage from the Asian mouflon, the Corsican and Sardinian mouflon should be designated as Ovis gmelini musimon.

Background Cerium oxide nanoparticles (CeO 2 NPs) are able to store and release oxygen, conferring them scavenger activity against oxidative stress. However, their effects in reproductive systems are not yet well understood. The aim of the study was to investigate the effects of exposure of refrigerated ram semen to CeO 2 NPs for 96 h on the main structural and kinematic parameters of spermatozoa. Methods The ejaculates of 5 Sarda rams were collected, pooled and diluted in a soybean lecithin extender. Samples were exposed to increasing doses of CeO 2 NPs (0, 44 and 220 μg/mL) and stored at 4 °C for 96 h. Analyses of kinematic parameters (computer assisted sperm analysis, CASA), integrity of membranes (PI/PSA staining), ROS production (H 2 DCFDA staining) and DNA damage (sperm chromatin structure assay with acridine orange, SCSA) were performed every 24 h (0, 24, 48, 72 and 96 h of incubation). The experiment was carried out in 6 replicates. Data were analysed by repeated measures ANOVA with Bonferroni’s as post hoc test. When the assumption of normality was not met (ROS), non-parametric Kruskal-Wallis rank test was carried out. Results Exposure of ram spermatozoa to increasing doses of CeO 2 NPs had a beneficial effect on the main motility parameters from 48 h of incubation onward. Velocity of sperm cells was enhanced in the groups exposed to CeO 2 NPs compared to the control. Incubation with NPs had beneficial effects on the integrity of plasma membranes of spermatozoa, with higher percentage of damaged cells in the control group compared to the exposed ones. Production of ROS was not affected by exposure to NPs and its levels rose at 96 h of incubation. The integrity of DNA remained stable throughout the 96 h of storage regardless of co-incubation with NPs. Conclusions We reported beneficial effects of CeO 2 NPs on kinematic and morphologic parameters of ram semen, such as motility and membrane integrity following 96 h of exposure. Furthermore, we also proved no genotoxic effects of CeO 2 NPs. These effects could not be related to an antioxidant activity of CeO 2 NPs, since ROS levels in exposed cells were similar to those of unexposed ones.

This study investigated the genetic diversity of Griffon Vultures ( Gyps fulvus ) from two South - East European, and seven Mediterranean countries and islands, including Spain, which hosts the largest European population. Our sample, comprising 249 newly generated plus 92 public D-loop sequences, is the largest dataset so far analyzed for this species. We identified 14 novel haplotypes, with Hpt-A being predominant across populations. Exclusive haplotypes have been identified in the populations from Spain, Serbia, Israel, Croatia, and Sardinia, suggesting them as worthy of targeted conservation programs to preserve such haplotypes. Principal coordinate analysis identified a big cluster composed of populations from continental Europe and the Aegean islands that were clearly differentiated from Western Mediterranean and Middle East populations. This structure would suggest the Serbian population is potentially a good source for restocking efforts in the Eastern Mediterranean. In addition, results indicated that historical restocking actions with Spanish individuals have significantly influenced genetic composition in the insular population of Sardinia, and provided evidence consistent with possible hybridization between Griffon and Rüppell’s Vultures ( Gyps rueppellii ) in Spain. This raises concerns about the use of non-genotyped Spanish individuals for restocking in populations like the Eastern Mediterranean ones, where contact zones between the two species have never been reported.

Background Heavy metal cadmium (Cd) is a widespread environmental contaminant with a potential toxicity that might negatively affect female reproduction and fertility. It has been reported that Cd exposure impaired the quality of oocytes and led to a defective maturation and fertilization, through oxidative stress induction. Resveratrol (Res) is a natural polyphenol with strong antioxidant properties that exhibited protective role in preventing oocyte redox homeostasis disruption and quality decline. Here, we explored whether the addition of Res to in vitro maturation (IVM) medium might act as a protection against Cd-induced toxicity on ovine oocyte maturation and fertilization. Firstly, we evaluated the effect of supplementing IVM medium with two different Res concentrations (1 and 2 μmol/L) on nuclear maturation and fertilization of oocytes matured under CdCl 2 (2 μmol/L) exposure. Therefore, the concentration of 1 μmol/L Res was selected to analyse the effects of this compound on intracellular ROS levels, mitochondrial (mt) distribution and activity, chromatin configuration, cytoskeleton morphology, cortical granules (CGs) distribution and mRNA expression of genes associated with cellular response to oxidative stress (i.e. SIRT1, SOD 1, GPX1, GSR, CAT ) in Cd-exposed in vitro matured oocytes. Results We found that 1 μmol/L Res restored the reduced oocyte meiotic competence induced by Cd exposure as well as, Res sustained oocyte ability to be normally fertilized and decreased polyspermic fertilization at both tested concentrations. Moreover, we demonstrated that 1 μmol/L Res mitigated Cd-induced alterations of oocyte cytoplasmic maturation by reducing reactive oxygen species (ROS) accumulation, preventing mt dysfunction, maintaining the correct meiotic spindle and cortical F-actin assembly and the normal cortical granule distribution as well as up-regulating SIRT1 , SOD1 and GPX1 genes. Conclusions Taken together, our findings highlighted the beneficial influence exerted by Res in preventing Cd-induced disturbance of nuclear and cytoplasmic maturation and subsequent fertilization in ovine oocytes. Res treatment may help to establish defence strategies counteracting Cd-induced toxicity on the female gamete.

Suboptimal culture conditions during in vitro maturation (IVM) affect oocyte developmental competence and the viability of the resulting embryo. Three-dimensional (3D) culture systems provide a more biologically appropriate environment compared to traditional two-dimensional (2D) cultures. The aim of this study was to evaluate the effect of liquid marble (LM) microbioreactors as a 3D culture system on IVM and the subsequent embryo development of prepubertal goat oocytes. The cumulus–oocyte complexes (COCs) recovered from prepubertal goat ovaries underwent IVM in drops under oil (the 2D system and the control group) and in the 3D LM system (the LM group). After IVM, oocytes were parthenogenetically activated and cultured until the blastocyst stage. The control and LM groups showed similar rates of nuclear maturation (52.17% and 44.12%) and blastocyst formation (10.64% and 10.10%). Reactive oxygen species and glutathione levels and the density of transzonal projections (TZPs) in oocytes did not differ between groups. The LM system increased mitochondrial activity and modified the organization of these organelles in the oocyte cytoplasm compared to the control group. The LM microbioreactor demonstrated the ability to improve the mitochondrial status of the oocytes and was not harmful for oocyte IVM and subsequent embryo development. Therefore, LM could be used as a 3D cost-effective culture system for the IVM of prepubertal goat oocytes.

In recent years, the widespread environmental presence of microplastics (MPs) has raised major concerns regarding animal and human health, including potential risks to reproductive function and offspring. This study aimed to evaluate the effects of increasing concentrations of polystyrene MPs (PS-MPs; 0, 5, 50, or 100 μg/mL) on ovine cumulus-oocyte complexes (COCs) during maturation (IVM). Fluorescent microspheres were used for uptake assessment into COCs and cumulus cells (CCs) monolayers, whereas non-fluorescent PS-MPs were employed to evaluate potential toxic effects induced on CCs and oocytes. As regards CCs, increased PS-MPs uptake was highlighted at the highest exposure concentration (100 μg/mL), whereas no significant differences were observed in oocyte intracellular fluorescence intensity, compared to the control. The bioaccumulation increment in CCs monolayers was already visible after 6 h, both at 5 and 100 μg/mL, and confirmed at 24 h. The real-time PCR analysis in CCs revealed significant reductions in the expression levels of genes involved in antioxidant defense and alterations in those implicated in apoptosis. Finally, the TUNEL assay revealed a dose-dependent increase in CCs apoptotic index. Consequently, PS-MPs exposure impaired oocyte meiosis resumption by significantly reducing the maturation rates, particularly at 50 and 100 μg/mL, whereas no effects were observed at 5 μg/mL. Oocyte intracellular reactive oxygen species levels were significantly increased at all concentrations, whereas no differences in mitochondrial membrane potential were detected. The percentages of oocytes with abnormal configurations of meiotic spindle and cortical F-actin were found to be significantly increased, regardless of concentration. Finally, the cleavage rate was significantly reduced in oocytes exposed to 50 μg/mL, whereas no differences were found in the blastocyst rate at both 50 and 5 μg/mL. In conclusion, exposure of sheep COCs to PS-MPs during IVM reduced oocyte quality and developmental potential through alterations induced in the CCs, which turned out to be the main target of these environmental contaminants.

There is strong scientific evidence that exposure to environmental contaminants, such as heavy metal(loid)s (HMs), can impair female reproductive function. Pets, such as cats and dogs, who share the same habitat as humans, may be particularly useful sentinel models for detecting HMs in the ovary. In the present study, we compared the concentration of essential (Ems; Cu, Fe, Mn, Se, and Zn) and non-essential metal(loid)s (NEMs; Al, As, Cd, and Pb) in the ovarian tissues of free-ranging queens and bitches of different ages living in industrialized/highly polluted (south group) and non-polluted (north group) urban areas of the island of Sardinia, Italy. The results showed that both EMs and NEMs were present at detectable concentrations in feline and canine ovaries and their levels varied according to geographical areas and animal age. Among the EMs, Cu was found elevated in older queens and bitches inhabiting the southern area. Cadmium and lead were higher in feline and canine ovaries of older animals from the south compared to those living in the north. In addition, Cd and Pb concentrations increased in individuals of both species living in the south. These findings showed new perspectives for the use of pets as early warning sentinels of environmental pollution by HMs and for the risk of human exposure within a “One Health” approach. Pets may help to study the link between exposure to metals and female reproductive disturbances in mammals.

Background Storage conditions during transportation of explanted ovaries are a critical step in setting up fertility preservation protocols in both animal and human fields. Here, we evaluated the effects of ovary storage at 4 °C on the preservation of preantral follicles and oocytes retrieved from antral follicles using the domestic cat as model. Methods Ovaries were harvested from fifty-five healthy domestic queens during ovariectomy and stored at 4 °C for 0 (control), 24, 48, 72 and 96 h. In Experiment 1, the effects of the storage period at 4 °C on the morphology, cytoskeleton (α/β tubulin) and DNA integrity (phosphorylation of histone H2AX) of preantral follicles were investigated. In Experiment 2, oocytes recovered from antral follicles were matured and fertilized in vitro to evaluate their meiotic and developmental competence . Reactive oxygen species (ROS), glutathione (GSH) and lipid peroxidation were measured in matured oocytes. Results The results showed that: a) storage up to 24 h did not affect the morphology and the DNA integrity of preantral follicles; b) extended storage times caused progressive morphological abnormalities, disassembling of microtubules and DNA damage; c) storage up to 48 h did not influence in vitro meiotic maturation of oocytes nor cleavage after in vitro fertilization. However, only oocytes stored within the ovary for 24 h produced blastocysts in a percentage similar to control oocytes; d) GSH levels of in vitro matured oocytes did not change at any time during ovary storage; a progressive increase in ROS levels was detected from 48 h associated with elevated lipid peroxidation at 72 and 96 h of storage. Conclusions Storage of cat ovaries for up to 24 h caused minimal alteration of preantral follicles and oocytes. The extension of the storage period beyond 24 h progressively impaired the structure of follicles, and modified the oxidative status of in vitro matured oocytes and their developmental competence after in vitro fertilization. This information may help when setting up programs for fertility conservation, especially for wild feline species which die in geographic areas located far away from ARTs centers.

Cadmium (Cd), a highly toxic pollutant, impairs oocyte fertilization, through oxidative damage on cumulus cells (CCs). This study analysed the transcriptomic profile of CCs of cumulus-oocyte complexes (COCs) from adult and prepubertal sheep, exposed to Cd nanomolar concentration during in vitro maturation. In both age-groups, CCs of matured oocytes underwent RNA-seq, data analysis and validation. Differentially expressed genes (DEGs) were identified in adult (n = 99 DEGs) and prepubertal (n = 18 DEGs) CCs upon Cd exposure. Transcriptomes of adult CCs clustered separately between Cd-exposed and control samples, whereas prepubertal ones did not as observed by Principal Component Analysis. The transcriptomic signature of Cd-induced CC toxicity was identified by gene annotation and literature search. Genes associated with previous studies on ovarian functions and/or Cd effects were confirmed and new genes were identified, thus implementing the knowledge on their involvement in such processes. Enrichment and validation analysis showed that, in adult CCs, Cd acted as endocrine disruptor on DEGs involved in hormone biosynthesis, cumulus expansion, regulation of cell signalling, growth and differentiation and oocyte maturation, whereas in prepubertal CCs, Cd affected DEGs involved in CC development and viability and CC-oocyte communications. In conclusion, these DEGs could be used as valuable non-invasive biomarkers for oocyte competence.

Purpose Detection of chemical modifications induced by aging-related oxidative damage in mouse metaphase II (MII) oocytes by Raman microspectroscopy. Methods CD-1 mice at the age of 4–8 weeks (young mice) and 48–52 weeks (old mice), were superovulated and oocytes at metaphase II stage were recovered from oviducts. MII oocytes from young animals were divided into three groups: A) young oocytes, processed immediately after collection; B) in vitro aged oocytes, cultured in vitro for 10 h before processing; C) oxidative-stressed oocytes, exposed to 10 mM hydrogen peroxide for 2 min before processing. Oocytes from reproductively old mice were referred to as old oocytes (D). All the oocytes were analyzed by confocal Raman microspectroscopy. The spectra were statistically analyzed using Principal Component Analysis (PCA). Results PCA evidenced that spectra from young oocytes (A) were clearly distinguishable from those obtained from in vitro-aged, oxidative-damaged and old oocytes (B, C, D) and presented significant differences in the bands attributable to lipid components (C = C stretching, 1,659 cm −1 ; CH 2 bending, 1,450 cm −1 ; CH 3 deformation,1,345 cm −1 ; OH bending, C-N stretching, 1,211 cm -1 ) and protein components (amide I band,1,659 cm −1 ; CH 2 bending modes and CH 3 deformation, 1,450 cm −1 ; C-N and C-C stretching vibrations, 1,132 cm −1 ; phenylalanine’s vibration, 1,035 cm −1 ) Conclusions Raman spectroscopy is a valuable non-invasive tool for the identification of biochemical markers of oxidative damage and could represent a highly informative method of investigation to evaluate the oocyte quality.