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In vivo, the temperature inside preovulatory follicles of cows is approximately 1 °C lower than rectal temperature. However, standard bovine oocyte in vitro maturation (IVM) protocols use 38.5 °C based on rectal temperature. This study evaluated the effect of reducing IVM temperature to 37.5 °C on the proteomic profile of oocytes compared to the routine 38.5 °C. Nuclear maturation rate and cumulus cell (CC) expansion (30 COCs per group, 21 replicates) were assessed by observing the first polar body and using a subjective scoring method (0-4). Total nitrite concentrations in the culture medium were measured using the Griess method. Differential proteomics was performed using LC-MS/MS on pooled oocyte samples (500 matured oocytes per group, three replicates), followed by gene ontology enrichment, protein-protein interaction, and putative miRNA target analyses. No significant differences were observed between the groups in nuclear maturation, CC expansion, or nitrite concentration (P > 0.05). A total of 806 proteins were identified, with 7 up-regulated and 12 down-regulated in the treatment group compared to the control. Additionally, 12 proteins were unique to the control group, and 8 were unique to the treatment group. IVM at 37.5 °C resulted in the upregulation of proteins involved in protein folding and GTP binding, and the downregulation of enzymes with oxidoreductase activity and proteins involved in cytoskeletal fiber formation. Furthermore, 43 bovine miRNAs potentially regulating these genes (DES, HMOX2, KRT75, FARSA, IDH2, CARHSP1) were identified. We conclude that IVM of bovine oocytes at 37.5 °C induces significant proteomic changes without impacting nuclear maturation, cumulus cell expansion, or nitrite concentration in the IVM medium.

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Canine mammary tumours (CMT) have histological, clinicopathological and molecular resemblances to human breast cancer (HBC), positioning them as viable models for studying the human disease. CMT initiation and progression occur spontaneously in immune-competent animals, which challenge the suggested limitations of genetically modified mice, also enabling the evaluation of immunotherapies in canine patients. Dogs have shorter life expectancy compared to humans, and cancer advances more rapidly in this species. This makes it possible to perform studies about the clinical efficacy of new therapeutic modalities in a much shorter time than in human patients. The identification of biomarkers for tumour subtypes, progression and treatment response paves the way for the development of novel therapeutic and diagnostic approaches. This review addresses the similarities between CMT and HBC and the molecular signatures identified in CMT samples that have been explored to date. We proposed a detailed molecular exploration of the CMT stroma using state-of-the-art methods in transcriptomics and proteomics. Using CMT as an analog for HBC not only helps to understand the complexities of the disease, but also to advance comparative oncology to the next level to prove the claim of dogs as a valid translational model.

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Abstract Objective: Seminal cryopreservation causes significant damage to the sperm; therefore, different methods of cryopreservation have been studied. The aim of the study was to compare the effects of density gradient processing and washing/centrifugation with seminal plasma removal for cryopreservation in semen parameters. Methods: Seminal samples of 26 normozoospermic patients were divided into 3 parts: with seminal plasma; after washing/centrifugation; and after selection through density gradient. The samples were cryopreserved for at least two weeks. Motility, sperm count, morphology and viability were evaluated before cryopreservation and after thawing. Results: Density gradient processing selected motile and viable sperm with normal morphology in fresh samples (p<0.05). Cryopreservation negatively affected all sperm parameters regardless of the processing performed, and even if the sperm recovery was lower in the density gradient after the thawing, progressive motility, total motility, viability and morphology remained higher (p<0.05). Conclusion: Cryopreservation significantly compromises sperm parameters (motility, morphology, viability). In normozoospermic patients, the density gradients select better quality spermatozoa compared to other processing methods; this benefit was kept after thawing.

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O controle da qualidade seminal produzido por centrais e unidades de disseminação genética permite a maximização da produção de doses inseminantes por ejaculado, bem como organiza o acompanhamento da produtividade de cada macho. Este acompanhamento é crítico tanto quando o reprodutor jovem é introduzido na rotina de coleta de sêmen quanto na determinação do momento do descarte e reposição. Essencialmente, a avaliação do sêmen em suínos é a mesma há décadas, porém novas metodologias de análise individual da célula como a citometria de fluxo e de constituintes de tecidos e fluídos como a proteômica e metabolômica já estão trazendo avanços na andrologia animal. Na presente revisão, são abordados os aspectos técnicos, vantagens e limitações destas três análises avançadas aplicadas ao sêmen suíno, discutindo a sua implementação e impactos no manejo reprodutivo da espécie.(AU)

The quality control of the semen doses produced by artificial insemination centers and allows the maximization of the production of inseminating doses per ejaculate, as well as organizes the monitoring of the productivity of each male. This follow-up is critical when the young male is initiated in routine semen collection and in determining the moment of male replacement. Essentially, semen evaluation in pigs has been the same for decades, but new methodologies for individual cell analysis such as flow cytometry and tissue and fluid constituents such as proteomics and metabolomics are already bringing advances in animal andrology. In this review, the technical aspects, advantages, and limitations of these three advanced analyzes applied to swine semen are addressed, discussing their implementation and impacts on the reproductive management of the species.(AU)

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Sperm cells rely on different substrates to fulfil thei energy demand for different functions and diverse moments of their life. Species specific mechanism involve both energy substrate transport and their utilization: hexose transporters, a protein family of facilitative passive transporters of glucose and other hexose, have been identified in spermatozoa of different species and, within the species, their localization has been identified and, in some cases, linked to specific glycilitic enzyme presence. The catabolism of hexose sources for energy purposes has been studied in various species, and recent advances has been made in the knowledge of metabolic strategies of sperm cells. In particular, the importance of aerobic metabolism has been defined and described in horse, boar and even mouse spermatozoa; bull sperm cells demonstrate to have a good adaptability and capacity to switch between glycolysis and oxidative phosphorylation; finally, dog sperm cells have been demonstrated to have a great plasticity in energy metabolism management, being also able to activate the anabolic pathway of glycogen syntesis. In conclusion, the study of energy management and mitochondrial function in spermatozoa of different specie furnishes important base knowledge to define new media for preservation as well as newbases for reproductive biotechnologies.

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Sperm cells rely on different substrates to fulfil thei energy demand for different functions and diverse moments of their life. Species specific mechanism involve both energy substrate transport and their utilization: hexose transporters, a protein family of facilitative passive transporters of glucose and other hexose, have been identified in spermatozoa of different species and, within the species, their localization has been identified and, in some cases, linked to specific glycilitic enzyme presence. The catabolism of hexose sources for energy purposes has been studied in various species, and recent advances has been made in the knowledge of metabolic strategies of sperm cells. In particular, the importance of aerobic metabolism has been defined and described in horse, boar and even mouse spermatozoa; bull sperm cells demonstrate to have a good adaptability and capacity to switch between glycolysis and oxidative phosphorylation; finally, dog sperm cells have been demonstrated to have a great plasticity in energy metabolism management, being also able to activate the anabolic pathway of glycogen syntesis. In conclusion, the study of energy management and mitochondrial function in spermatozoa of different specie furnishes important base knowledge to define new media for preservation as well as newbases for reproductive biotechnologies.(AU)

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Background: Superficial necrolytic dermatitis (SND), hepatocutaneous syndrome (HCS), metabolic epidermal necrosis (MEN), and necrolytic migratory erythema (NME) are useful terms to describe a disease that likely has a multifactorial etiopathogenesis. SND is a rare and fatal disease characterized by skin lesions and liver disease. Common skin lesions include hyperkeratosis, fissures, erosion, ulceration, crusting, exudation from the paws, face, perianal regions, and pressure points. This case report aimed to report the case of a bitch that developed the rare Superficial Necrolytic Dermatitis disease, emphasizing the clinical signs of the disease, and the importance of complementary exams such as abdominal ultrasound and skin biopsy for the definitive diagnosis. Case: A 9-year-old, mixed-breed, neutered female was referred for clinical examination with 5 months history of hyperkeratosis and ulceration of the paw pads, presenting pain, lameness and weight loss. Abdominal ultrasound revealed a liver with heterogeneous echotexture, mixed echogenicity, irregular and poorly delimited margins with hypoechoic nodules throughout like honeycombs. The gallbladder was visualized with a moderately thick layer. Histological analysis confirmed the diagnosis of SND. Skin biopsies showed an increase in thickness of the epidermis due to irregular hyperplasia and proliferation of keratinocytes in the basal layer of the epidermis, pallor of the spinous layer of the epidermis and important parakeratosis. Due to the progression of the disease, significant worsening of the patient's clinical condition and pain, associated with the impossibility of cure, the animal was submitted to euthanasia. A necropsy was performed to allow assessment of the liver and pancreas. The biopsies showed a severe proliferative chronic hepatitis, steatosis and cholestasis associated with pancreatitis and necrotic multifocal proliferative fibrinopurulent areas in the pancreas. Discussion: Clinical signs such as lethargy, inappetence, weight loss, as well as the dermatological signs presented by this bitch are nonspecific clinical signs and require a deeper clinical, pathological and histopathological diagnostic investigation to reach the diagnosis of this disease. The definitive diagnosis is made on the basis of a characteristic honeycomb pattern in the liver or associated with a neoplastic finding in the pancreas on ultrasound examination and confirmed by histopathological evaluation of skin biopsies. Palliative treatment with corticosteroid anti-inflammatories, improvement in feed quality, with higher nutritional and protein intake and intravenous amino acid supplementation are suggested by some authors as treatment alternatives. However, not all owners can afford a costly lifetime treatment. With the progressive worsening of the condition, many owners opt for euthanasia as a way to shorten the suffering of the animal. This decision is not an easy one to make. Despite the poor prognosis of the disease, treatment options should be tried by veterinarians and owners prior to the option of euthanasia. However, new affordable nutritional and pharmacological strategies to treat or control the disease are needed in order to improve quality of life of SND patients.

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O estresse oxidativo é caracterizado pelo desequilíbrio entre a produção de espécies reativas de oxigênio e a atuação de sistemas antioxidantes da própria célula e tecido. O espermatozoide de mamíferos apresenta metabolismo oxidativo e por isso é um potencial produtor destas espécies reativas que podem causar danos a estrutura lipídicas, proteínas e mesmo ao DNA espermático. Na espécie suína, o plasma seminal confere importante proteção antioxidante ao espermatozoide ejaculado no útero da fêmea, onde os desafios à sua sobrevivência são inúmeros. Contudo, esta defesa natural do plasma seminal não é suficiente para garantir o máximo de resultado de fertilidade com o sêmen submetido a processos de refrigeração ou congelação e utilizado na inseminação artificial. Na presente revisão, serão abordados os temas estresse oxidativo e metabolismo espermático, bem como apresentados alguns dados sobre a aplicação de antioxidantes na andrologia suína.

Oxidative stress is characterized by an imbalance between the production of reactive oxygen species and the action of antioxidant systems present in the cell and tissue. Mammalian sperm have oxidative metabolism and are therefore a potential producers of these reactive species which can lead to damage to lipids, proteins and even sperm DNA. In the swine species, seminal plasma provides an important antioxidant protection to spermatozoa that are ejaculated and deposited in the uterus, where the challenges to their survival are numerous. However, this natural seminal plasma defense is not sufficient to guarantee maximum fertility when semen is processed for artificial insemination, and later used after preservation by refrigeration or freezing. In this review, the topics related to oxidative stress and sperm metabolism will be addressed, and results about the application of antioxidants in swine andrology will be presented as well as.

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Os primeiros estudos de espermatozoides com citometria de fluxo com espermatozoides iniciaram no final da década de 1970. Com os avanços tecnológicos, hoje contamos com equipamentos com alta sensibilidade e eficiência que, em conjunto com amplo catálogo de sondas fluorescentes, podemos mensurar com alta precisão características celulares. Como exemplo, destacam-se dano em membrana plasmática, atividade mitocondrial, produção de espécies reativas de oxigênio, dano ao DNA espermático e muito mais. Na presente revisão, as potencialidades e limitação para a implementação da citometria de fluxo na análise seminal de espécies domésticas são exploradas e comentadas.

The first flow cytometry studies with spermatozoa were published in the late 1970s. With the technological advances in the following years, today we have equipments with high sensitivity and efficiency that, together with a large number of commercially available fluorescent probes, allow us to measure different cell characteristics with high accuracy. Some of the most evaluated characteristics are plasma membrane damage, mitochondrial activity, production of reactive oxygen species, sperm DNA damage, and much more. In this review, the potentials and limitations for the implementation of flow cytometry in the seminal analysis of domestic species are explored and commented.

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The protein disulphide isomerase A1 (PDIA1) is an important chaperone involved in protein quality control and redox regulation. Also, the ability of PDIA1 to bind to oestrogens suggests that it may play a role in epididymal maturation and male fertility. The goals of this study were to (a) verify the possible interaction between 17ß-estradiol and equine PDIA1 using bioinformatics; (b) identify and quantify PDIA1 protein in equine cauda epididymis throughout peripuberty; and (c) determine whether the amounts of PDIA1 in equine seminal plasma and spermatozoa are associated with fertility. Using in silico analysis, we were able to predict the tertiary structure of equine PDIA1 and to demonstrate the interaction between 17ß-estradiol and the putative binding site in domains b and b'. Colts under 24 months of age had lower relative amounts of PDIA1 in cauda epididymal fluid in comparison with older males (p < .01). No difference was observed in seminal plasma PDIA1 between fertile and subfertile stallions. Our study demonstrates that PDIA1 expression in the epididymis increases during peripuberty. However, in the adult stallion, its quantity in seminal plasma is not associated with fertility.

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Seminal plasma (SP) contributes to sperm physiology and metabolism, prevents premature capacitation, and protects sperm against oxidative stress. In order to evaluate the impact of heat stress in the semen of tropically adapted Brangus breed and in their seminal plasma proteome, we studied the effects of scrotal insulation for 72 h. Semen samples from six bulls, between 7 and 8 years of age, were collected prior to scrotal insulation (pre-insulation), and at 4 and 11 wk after insulation. Seminal plasma samples were analyzed by 2D SDS-PAGE and liquid chromatography coupled with mass spectrometry (LC-MS/MS). Insulation caused decrease in vigour, gross and total motility after 4 wk of scrotal insult (P < 0.001). Total defects in sperm were higher after 4 wk compared to pre-insulation and 11 wk after scrotal insulation (P < 0.001). The analysis of the 2D protein profile of the SP resulted in the identification 183 unique protein spots in all gels evaluated. There was no difference in mean number of protein spots amongst time points. Eight protein spots were more abundant in SP after scrotal insulation, returning to the same expression level at 11 wk post-insulation. One spot had higher abundance at 11 wk post-insulation, and one spot had decreased abundance 4 wk after insulation. The ten protein spots with differential abundance amongst time points were identified as Seminal plasma protein PDC-109, Seminal plasma protein A3, Seminal plasma protein BSP-30 kDa, Spermadhesin-1 and Metalloproteinase inhibitor 2. The validation of these five proteins as biomarkers for thermal testicular stress in Brangus breed would allow the development of new biotechnologies that could improve bovine semen analysis in breeding systems in tropical and subtropical conditions. A close association between the identified BSP and Spermadhesin-1 was evidenced in protein-protein interaction analysis. Based on gene ontology analysis, variation in sperm function after insulation could be explained by variation in the expressed proteins in the SP. Further studies are required to verify if these proteins could be used as biomarkers for the identification of bulls with increased seminal resistance to heat stress in Brangus breed.

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The modern pig industry relies on extensive use of artificial insemination with cooled semen. It is important that semen doses maintain their quality during processing, transport and storage before insemination to guarantee maximum fertility rates. However, ejaculates may respond differently to liquid preservation at 17 °C, despite the optimal quality assessed before cooling. Thus, the aim of this study was to identify differences in seminal plasma proteome of ejaculates with a higher or lower seminal resistance to storage at 17 °C. A total of 148 ejaculates from 65 sexually mature healthy boars were classified as: High Resistance to cooling (HR, total motility > 60% at 144h) and Low resistance to cooling (LR, total motility <60 at 72h). To identify differentially expressed seminal plasma proteins between HR and LR ejaculates, ten ejaculates of each group were analyzed by 2D SDS-PAGE and ESI-Q-TOF mass spectrometry. The proteins associated with HR ejaculates were cathepsin B (spot 2803 and 6601, p < 0.01); spermadhesin PSP-I (spots 3101 and 3103, p < 0.05); epididymal secretory protein E1 precursor (spot 2101, p < 0.05) and IgGFc binding protein (spot 1603, p < 0.01). The protein associated with LR group was the Major seminal plasma PSPI (spot 9103, p < 0.01). To our knowledge, this is the first report of the association of boar seminal plasma proteins to semen resistance to cold storage at 17 °C. These results suggest the use of these proteins as biomarkers for semen resistance to preservation at 17 °C.

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In the present study, we describe the proteome of porcine cauda epididymis fluid and spermatozoa by means of Multidimensional Protein Identification Technology (MudPIT). Ten sexually mature healthy boars were surgically castrated and epididymides were dissected to obtain the cauda epididymal content. Polled protein extracts of cauda epididymal fluid (CEF) and spermatozoa (CESperm) were loaded in an Agilent 1100 quaternary HPLC and peptides eluted from the microcapillary column were electro-sprayed directly into a LTQ Orbitrap XL mass spectrometer. Using bioinformatics, identified proteins were classified by their molecular functions, involvement in biological processes and participation in relevant metabolic pathways associated with spermatozoa physiology, fertility potential and protection. A total of 645 proteins were identified in the CEF, with epididymal-specific lipocalin-5, beta-hexosaminidase subunit beta precursor and phosphatidylethanolamine-binding protein 4 being the most abundant proteins found. A total of 2886 proteins were identified in the CESperm proteome with 81 proteins being considered more abundant (spectral counts > 100). CEF and CESperm data were compared and 345 proteins were present in both proteomes. Phosphatidylethanolamine-binding protein 4 precursor was the only protein found most abundant in both CEF and CESperm proteomes. Based on Gene Ontology analysis, we identified CEF and CESperm proteins associated with sperm protection against ROS and immune mediated response, glycosaminoglycan degradation, ubiquitin-proteasome system, metabolic process and maturation, modulation of acrosome reaction and ZP binding and oocyte penetration. These results provide a better comprehension about the molecular process and biological pathways involved in sperm epididymis maturation and establishment of the cauda epididymis sperm reservoir.

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BACKGROUND: Hypothermic storage at 5°C has been investigated as an alternative to promote the prudent use of antibiotics for boar artificial insemination doses. However, this temperature is challenging for some ejaculates or boars. OBJECTIVE: The present study aimed to identify putative biomarkers for semen resistance to hypothermic storage at 5°C by comparing the seminal plasma proteomes of boars with high and low seminal resistance to preservation at 5°C. MATERIALS AND METHODS: From an initial group of 34 boars, 15 were selected based on the following criteria: ejaculate with ≤20% abnormal spermatozoa and at least 70% progressive motility at 120 hours of storage at 17°C. Then, based on the response to semen hypothermic storage at 5°C, boars were classified into two categories: high resistance-progressive motility of >75% in the three collections (n = 3); and low resistance-progressive motility of <75% in the three collections (n = 3). Seminal plasma proteins were analyzed in pools, and differential proteomics was performed using Multidimensional Protein Identification Technology. RESULTS: Progressive motility was lower at 120 hours of storage in low resistance, compared to high resistance boars (P < .05). Acrosome and plasma membrane integrity were not affected by the boar category, storage time, or their interaction (P ≥ .104). Sixty-five proteins were considered for differential proteomics. Among the differentially expressed and exclusive proteins, the identification of proteins such cathepsin B, legumain, and cystatin B suggests significant changes in key enzymes (eg, metalloproteinases) involved in spermatogenesis, sperm integrity, and fertilizing potential. DISCUSSION AND CONCLUSION: Differences in the seminal plasma suggest that proteins involved in the proteolytic activation of metalloproteinases and proteins related to immune response modulation could disrupt key cellular pathways during spermatogenesis and epididymal maturation, resulting in altered resistance to chilling injury. Further in vivo studies focusing on the immunological crosstalk between epithelial cells and gametes might explain how the immune regulators influence sperm resistance to hipothermic storage.

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A adequada analise do sêmen na espécie suína é fator chave para a obtenção da máxima eficiência reprodutiva. Com o avanço tecnológico na avaliação andrológica, a implementação sistemas computadorizados de analise seminal (motilidade espermática, concentração, morfologia e vitalidade celular) já é realidade em muitas centrais de produção de doses inseminantes. Tal evolução permitiu uma melhor categorização dos machos com relação a qualidade seminal e a resistência das doses a preservação por refrigeração. Com isto, abriu-se uma janela de oportunidade para investigar de forma mais apurada os componentes seminais que podem contribuir para a diferenças reprodutivas entre machos. É neste cenário que os estudos em proteômica do sêmen suíno vêm contribuindo para um melhor entendimento dos papeis das proteínas do plasma seminal e dos espermatozoides na fertilidade de cachaços. Nesta revisão, serão discutidos os diferentes métodos de aplicação da proteômica na andrologia, bem como as principais proteínas já associadas às características seminais e à fertilidade.

The proper semen analysis in the boar is a key factor for maximum reproductive efficiency. With the technological advances in andrology, the implementation of computer assisted sperm analyzers (sperm motility, concentration, morphology and cell viability) is a reality in most of the artificial insemination centers. Such evolution allowed a better classification of the sires regarding their seminal quality and resistance to cooling. As a consequence, a window of opportunity opened for a detailed assessment of the seminal components which could influence the reproductive differences among sires. In this scenario, the proteomic studies of the boar semen have been contributing to a better understanding of the roles of seminal plasma and sperm proteins in boar fertility. In this review, the different approaches of proteomics in andrology will be discussed, as well main proteins already associated with seminal characteristics and fertility.

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A adequada analise do sêmen na espécie suína é fator chave para a obtenção da máxima eficiência reprodutiva. Com o avanço tecnológico na avaliação andrológica, a implementação sistemas computadorizados de analise seminal (motilidade espermática, concentração, morfologia e vitalidade celular) já é realidade em muitas centrais de produção de doses inseminantes. Tal evolução permitiu uma melhor categorização dos machos com relação a qualidade seminal e a resistência das doses a preservação por refrigeração. Com isto, abriu-se uma janela de oportunidade para investigar de forma mais apurada os componentes seminais que podem contribuir para a diferenças reprodutivas entre machos. É neste cenário que os estudos em proteômica do sêmen suíno vêm contribuindo para um melhor entendimento dos papeis das proteínas do plasma seminal e dos espermatozoides na fertilidade de cachaços. Nesta revisão, serão discutidos os diferentes métodos de aplicação da proteômica na andrologia, bem como as principais proteínas já associadas às características seminais e à fertilidade.(AU)

The proper semen analysis in the boar is a key factor for maximum reproductive efficiency. With the technological advances in andrology, the implementation of computer assisted sperm analyzers (sperm motility, concentration, morphology and cell viability) is a reality in most of the artificial insemination centers. Such evolution allowed a better classification of the sires regarding their seminal quality and resistance to cooling. As a consequence, a window of opportunity opened for a detailed assessment of the seminal components which could influence the reproductive differences among sires. In this scenario, the proteomic studies of the boar semen have been contributing to a better understanding of the roles of seminal plasma and sperm proteins in boar fertility. In this review, the different approaches of proteomics in andrology will be discussed, as well main proteins already associated with seminal characteristics and fertility.(AU)

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Despite the development of efficient boar semen extenders, there is still room for improvement of new formulas using new molecules that could increase fertilisation outcomes and substitute cryoprotectants and antibiotics. The goal of this work was to evaluate if the essential oils from the leaves of Myrrhinium atropurpureum and Cymbopogon citratus are suitable as additives in boar semen extender. The major compounds found in the essential oils from M. atropurpureum were 1,8-cineole (37.37%) and terpinolene (19.18%); and geranial (49.8%) and neral (33.24%) in essential oil of C. citratus. The addition of 1% and 0.1% of both essential oils to extended semen had immediate spermicidal effects (p < 0.05). Lower concentrations were tested and no cytotoxic effect was observed when M. atropurpureum essential oil was added at 0.001%. Differently, essential oil from C. citratus reduced sperm motility, membrane functionality and integrity and mitochondrial membrane potential even in concentrations as low as 0.001%. Also, addition of essential oils in low concentrations had no inhibitory effect against Escherichia coli and Pseudomonas aeruginosa growth. We conclude that the essential oils from C. citratus and M. atropurpureum, rich in monoterpenes, are cytotoxic to swine spermatozoa, therefore unsuitable as semen extender additives.

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The epididymis is a complex organ where spermatozoa acquire motility and ability to fertilize theegg. Epididymal maturation lasts 1 or 2 weeks and exposes the immature spermatozoa to a sequentially modified milieu, promoting intense interactions with secretions by the epididymal epithelium. Sperm surface modifications in response to interactions with epididymal secretions are key steps to achieve fertility ability. However, the precise molecular mechanisms that convert an immotile and infertile gamete into a highly motile cellcapable of fusion with an oocyte are still unknown. Recent data on proteomics and transcriptomics of epididymal fluid and epididymosomes brought new ideas of the physiological roles of proteins and miRNAs in epididymal maturation in spermatozoa. This review focuses on the recent discoveries on epididymal fluid composition and its role on sperm maturation and preservation, linking to their survival and fertility potential.(AU)

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The epididymis is a complex organ where spermatozoa acquire motility and ability to fertilize theegg. Epididymal maturation lasts 1 or 2 weeks and exposes the immature spermatozoa to a sequentially modified milieu, promoting intense interactions with secretions by the epididymal epithelium. Sperm surface modifications in response to interactions with epididymal secretions are key steps to achieve fertility ability. However, the precise molecular mechanisms that convert an immotile and infertile gamete into a highly motile cellcapable of fusion with an oocyte are still unknown. Recent data on proteomics and transcriptomics of epididymal fluid and epididymosomes brought new ideas of the physiological roles of proteins and miRNAs in epididymal maturation in spermatozoa. This review focuses on the recent discoveries on epididymal fluid composition and its role on sperm maturation and preservation, linking to their survival and fertility potential.

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The use of butafosfan in combination with cobalamin modulates many cellular metabolic functions in several species. Its use enhances productive and reproductive performance and reduces stress responses in animals. Despite all these attributes, so far there have been no controlled studies to evaluate the effects of butafosfan and cobalamin on the quality of stallion semen. The purpose of this study was to evaluate the action of butafosfan in combination with cobalamin on the quality of fresh and cooled stallion semen. Four healthy stallions were kept in the same place and under the same management conditions during the entire experiment. Stallions were randomly assigned to two treatment groups in a 2x2 crossover design. Group A stallions were treated with an intramuscular injection of butafosfan twice a week for 80 days, while group B did not receive any treatment. After that, both groups were not treated for another 80 days allowing a washout period for the treated group. Then, the groups were reversed, and group B was treated with butafosfan and group A acted as the control for another 80 days. Semen was collected twice a week, diluted in skim milk and evaluated for total sperm count, total and progressive sperm motility, membrane integrity (CFDA/PI staining) and membrane functionality (HOS test) at 0 and 24 hours after preservation at 5ºC. Data were analyzed by comparing the...(AU)

A utilização de Butafosfan em combinação com cobalamina modula uma série de funções metabólicas celulares em várias espécies. Seu uso melhora o desempenho produtivo e reprodutivo e reduz as respostas ao estresse em animais. Apesar de todos esses atributos, até agora não houve estudos controlados para avaliar os efeitos da substância Butafosfan e cobalamina sobre a qualidade do sêmen de garanhões. O objetivo deste estudo foi avaliar a ação do Butafosfan em combinação com cobalamina sobre a qualidade do sêmen fresco e resfriado de garanhões. Quatro garanhões saudáveis foram mantidos no mesmo lugar e sob as mesmas condições de manejo durante todo o experimento. Garanhões foram divididos aleatoriamente em dois grupos de tratamento em um desenho cruzado 2x2. Garanhões do grupo A foram tratados com uma injeção intramuscular de Butafosfan duas vezes por semana durante 80 dias, enquanto o grupo B não recebeu qualquer tratamento. Depois disso, ambos os grupos não foram tratados durante mais 80 dias, permitindo um período de washout para o grupo tratado. Em seguida, os grupos foram invertidos, o grupo B foi tratado com Butafosfan enquanto o grupo A agiu como controle por mais 80 dias. O sêmen foi coletado duas vezes por semana, diluído em leite desnatado e avaliado para a contagem total de espermatozoides, motilidade progressiva e total, integridade da membrana (coloração CFDA/PI) e...(AU)

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