Equine uterine sanitizer: in vitro inhibition of endometritis-causing microorganisms and its stability to dilute antibiotics.

Endometritis is the leading cause of mare subfertility. Most mares respond to standard therapy, but alternative therapies have been developed for mares failing to respond. This study aimed to investigate a commercially available, yet unassessed, product labeled as a uterine sanitizer to determine the in vitro antimicrobial activity against microorganisms associated with endometritis and its in vitro stability to dilute antibiotics. In experiment 1, the microdilution broth technique and antimicrobial effects were assessed against Escherichia sp, Staphylococcus sp., Klebsiella sp., Pseudomonas sp., and Candida sp. Percentage inhibition was calculated by comparing the optical density. The minimum inhibitory concentration (MIC) 100% was determined using the resazurin dye technique. MIC 50% and 90% were determined using a dose-response non-linear regression. In experiment 2, the uterine sanitizer was used to dilute commonly used antibiotics achieving a final volume of 90 mL at 5°C, 21°C, and 37°C. The pH was measured at 0, 1, 3, 6, and 24 h after dilution. The uterine sanitizer had inhibitory properties against all microorganisms; Escherichia sp. being the most susceptible, and Pseudomonas sp. the most resistant. The uterine sanitizer had an acidic pH=4; however, when combined with the antibiotics, the pH of the antibiotic remained unchanged with the different temperatures and did not precipitate. In conclusion, the uterine sanitizer showed antimicrobial effects against endometritis-causing microorganisms. The dilution of antibiotics in the uterine sanitizer was stable and this association could potentiate the antimicrobial effects. Uterine sanitizer's safety and clinical efficacy in vivo remain to be tested.

Exposure to acetaminophen impairs gametogenesis and fertility in zebrafish (Danio rerio).

Acetaminophen (ACE; paracetamol) is one of the most widely used nonsteroidal anti-inflammatory drugs worldwide and is often found in aquatic systems, where it can act on nontarget species and impair fish reproduction. This study aimed to investigate the effects of chronic exposure to environmentally relevant ACE concentrations (0.5, 5 and 50 µg/L) on multiple reproductive parameters in zebrafish (Danio rerio). Gametogenesis was analyzed using histology, morphometry, cell proliferation, and apoptosis. This study also evaluated sex steroids, and prostaglandin E2 (PGE2) levels, gene expression for sex steroids and PGE2 receptors, fertilization rate, and semen quality. In females, exposure to 5 and 50 µg/L ACE induced larger and more abundant vitellogenic follicles and increased follicular atresia. In these treatments, males showed a lower proportion and proliferation of undifferentiated spermatogonia and a higher proportion of TUNEL-positive differentiated spermatogonia, spermatids, and spermatozoa, resulting in lower sperm production. ACE increased 17ß-estradiol (E2) and reduced 11-ketotestosterone levels in the testis, whereas only E2 increased in the ovaries. In both sexes, gonadal PGE2 levels were reduced. ACE at 50 µg/L induced an increase in the gene expression of androgen, estrogen, and PGE2 receptors in the ovaries, and reduced expression in the testes. Results also showed lower egg production and fertilization rate from 28 days of exposure with reduced sperm quality. These results demonstrated that ACE impairs the reproductive performance of zebrafish, affecting multiple reproductive parameters, which may be caused by the synergistic action of the imbalance of sex steroids, with a reduction of PGE2 and its receptors.

Ozonioterapia, PRP, fitoterapia, misoprostol e imunomodulação: Novas abordagens terapêuticas para a solução de velhos problemas / Ozone therapy, PRP, phytotherapy, misoprostol and immunomodulation: New therapeutic approaches to solving old problems

A resistência aos antimicrobianos representa uma ameaça sanitária global crescente, que deve ser abordada com urgência na saúde pública, na produção animal, agrícola e no meio ambiente. Essa circunstância põe em risco a eficiência do tratamento e diminui a eficácia contra infecções causadas pelos mais diversos agentes etiológicos, resultando em enfermidades mais prolongadas afetando diretamente os índices de fertilidade. Dessa forma, o objetivo dessa revisão foi abordar diferentes formas terapêuticas para sub/infertilidade em equinos, com o intuito de minimizar os problemas acarretados pelo uso indiscriminado de fármacos, relatando novas terapias alternativas, tais como ozonioterapia, plasma rico em plaquetas, uso de prostaglandina E1 e imunomoduladores na tentativa de solucionar velhos problemas.(AU)

Antimicrobial resistance represents a growing global health threat that must be urgently addressed in public health, animal production, agriculture and the environment. This circumstance jeopardizes the efficiency of the treatment and decreases the effectiveness against infections caused by the most diverse etiological agents, resulting in more prolonged illnesses, directly affecting fertility rates. Thus, the objective of this review was to address different therapeutic forms for sub/infertility in horses, with the aim of minimizing the problems caused by the indiscriminate use of drugs, reporting new alternative therapies, such as ozone therapy, platelet-rich plasma, use of prostaglandin E1 and immunomodulators in an attempt to solve old problems.(AU)

Idiopathic subfertility in stallions. Clinical approaches to a complex problem

At least 30-40% of stallions in commercial breeding programs are moderately fertile and 8-12% are subfertile (0.5-3% with severe subfertility). From the total reported cases of the subfertility, in 2-20% of the stallions the cause is unknown or was not established. The objective of this work is to present the concept of subfertile stallion based on the current state of knowledge and advanced molecular diagnostic technologies. Low pregnancy rates have been reported in stallions with normal semen quality after conventional evaluation. Acrosome reaction (AR) is necessary for natural fertilization and impaired acrosome reaction (IAR) leads to subfertility or infertility in horses, however, AR test is not included in routine semen analysis. Genome-wide association study identified FKBP6 as a strong candidate gene responsible for this failure. The gene encodes for FK506 binding protein 6 (FKBP6) which is involved in sperm development and functions. We could conclude that the evaluation of the acrosomal status is essential in cases of stallions with good motility, concentration, morphology and viability but unexplained (idiopathic) subfertility or infertility. It is important to highlight the recent increase in reports of fertility problems in stallions related to disorders of genetic origin.(AU)

Equine genital leptospirosis: Evidence of an important silent chronic reproductive syndrome.

Leptospirosis may be associated with important syndromes in equines, including reproductive ones. Recently, our group demonstrated a reproductive syndrome of leptospirosis in ruminants (Bovine Genital Leptospirosis - BGL), that presents its own agent, particular diagnostic methods, and specific treatment and prevention, requiring special attention. We realized that the genital syndrome of leptospirosis may not be exclusive to ruminants and may also affect horses. The aim of the present study is to gather the available data on leptospirosis in equine reproduction and propose a new genital leptospirosis syndrome in horses, which we have named Equine Genital Leptospirosis (EGL). The main agent is most probably serovar Bratislava (serogroup Australis), whose role has been increasingly recognized in the etiology of infection, presenting a worldwide distribution. The most probable relevant transmission route in the EGL context is the sexual transmission. Subfertility, estrus repetition, and, less commonly, late-term abortions are the most common effects. Serology may not be adequate to diagnose this chronic silent reproductive leptospiral infection. As most studies regarding equine leptospirosis investigate urinary samples, EGL is vastly underdiagnosed, and the disease is little studied and underestimated. Therefore, PCR, particularly from cervicovaginal mucus (CVM) and uterine fragments, turns out to be a rapid and definitive diagnosis choice. In order to mitigate the effects of the disease in equines, triad antibiotic therapy, vaccination, and environmental management are essential. Adequate recognition of the particularities of this syndrome may help to highlight its occurrence and contribute to a broader understanding of its clinical manifestations, use of a correct diagnosis, and development of novel approaches for its control.

Preserving effect of Loboob (a traditional multi-herbal formulation) on sperm parameters of male rats with busulfan-induced subfertility.

OBJECTIVE: Male infertility secondary to exposure to gonadotoxic agents during reproductive age is a concerning issue. The aim of this experimental study was to determine the effect of Loboob on sperm parameters. METHODS: 55 healthy rats were selected, weighted and divided into five groups consisting of 11 rats each. The control group received no medication. Rats in Treatment Group 1 received 10mg/kg Busulfan and rats in Treatment Groups 2, 3, and 4 received 35,70 and 140 mg/kg Loboob respectively in addition to 10mg/kg Busulfan. Finally, the sperm parameters and weights of the rats were compared using the Kolmogorov-Smirnov, non-parametric Kruskal-Wallis, and Dunn-Bonferroni tests. RESULTS: All sperm parameters and weights were significantly decreased among rats receiving Busulfan. All dosages of Loboob were effective to enhance the motility of slow spermatozoa, while only in the rats given 70 and 140 mg/kg of Loboob saw improvements in progressively motile sperm percentages (0.024 and 0.01, respectively). Loboob at a dosage of 140mg/kg improved sperm viability. It did not improve normal morphology sperm or decrease immotile sperm counts. Loboob did not affect mean rat weight. CONCLUSIONS: Loboob offered a dose-dependent protective effect on several sperm parameters in rats with busulfan-induced subfertility.

In vitro and in vivo parameters for identification of landrace pigs with low reproductive performance / Parâmetros in vitro e in vivo para detecção de machos suínos Landrace com baixo desempenho reprodutivo

In pig farming, measurements of production parameters play a fundamental role in the success of the activity. Minimal differences in fertility between breeders can lead to less reproductive efficiency and, less productivity. However, assessing the fertility of each male and the early identification of subfertile males is a difficult task to be performed. Thus, the aim of this study was to evaluate the use of in vitro and in vivo parameters in the identification of subfertile males of the Landrace breed, aiming to collaborate with genetic improvement programs, routine optimization in the Genetic Diffusion Units (GDUs) and the results of performance. In experiment 1, an approach to identify males with subfertility was evaluated based on retrospective data. For this, the results (averages of birth rates, number of total births and average percentages of female and male piglets per litter) were evaluated for a total of 996 matings and 847 parturitions. The inseminations came from ejaculates of 32 males, who had at least 19 females inseminated with homospermic doses in the concentration of 2.5 x 109 total sperm from the same male. As for the birth rate (BR), an average of 85.47% ± 6.05 was observed with a group of median males, seven males that stood out and one individual (M32) with a performance of 58.06% ± 9.0. For the total number of piglets born (PB) the average was 13.41 ± 0.56, with three males with better performance and one (M32) with very poor performance (8.62 ± 0.59). In experiment 2, it was verified whether evaluations of inseminating doses (ID) of semen in vitro (motility and sperm morphology) after 96 hours of storage had correlations with fertility in vivo, which can be used to identify subfertile males. The evaluations were performed on 30 ejaculates regarding the means of BR and PB, considering only those who had at least 7 females inseminated. There were no correlations between the motility assessments and semen morphological changes and the reproductive parameters evaluated. The results obtained in vivo, referring to BR and PB, demonstrated that it was possible to identify differences between males, the individual (M32) had the worst results for the percentages of BR and PB. It is concluded that there are males of high and low fertility and that only the in vitro analyzes carried out in this study are not enough to categorize them, however, the evaluation of retrospective data was efficient for this purpose.(AU)

Na suinocultura moderna, as mensurações de parâmetros de produção têm papel fundamental para o sucesso da atividade. No entanto, a avaliação da fertilidade de cada macho e a identificação precoce de machos subférteis é uma tarefa difícil de ser realizada. O objetivo deste estudo foi avaliar a utilização de parâmetros in vivo e in vitro na identificação de machos subférteis da raça Landrace, visando colaborar com os programas de melhoramento genético, otimização da rotina nas Unidade de Difusão Genética (UDGs) e dos resultados a campo. No experimento 1, foi proposta uma abordagem de identificação dos machos subférteis tendo como base dados retrospectivos. Para isso, foram avaliados os resultados (médias das taxas de parto, número de nascidos totais e média das porcentagens de leitões fêmeas e machos por leitegada) de um total de 996 coberturas e 847 partos. As inseminações foram oriundas de ejaculados de 32 machos, que tiveram ao menos 19 fêmeas cobertas com doses homospérmicas na concentração de 2,5 x 109 de espermatozoides totais e obrigatoriamente do mesmo macho. Quanto a taxa de parto (TP) obtivemos uma média de 85,47% ± 6,05 e observou-se um grupo de machos medianos, sete machos que se destacaram positivamente e um indivíduo (M32) com um desempenho 58,06 ± 9,0. Para número de leitões nascidos totais (NT) obtivemos uma média de 13,41 ± 0,56 e notou-se três machos com melhor desempenho e um (M32) com péssimo desempenho (8,62 ± 0,59). No experimento 2, foi verificado se as avaliações das doses inseminantes (DI) de sêmen in vitro (motilidade e morfologia espermática) após 96 horas de armazenamento apresentaram correlação com a fertilidade in vivo. As avaliações foram realizadas em 30 ejaculados quanto às médias de TP e NT, considerando apenas ejaculados que tiveram ao menos 7 fêmeas inseminadas. Não foram verificadas correlações entre as avaliações de motilidade e alterações morfológicas do sêmen com os parâmetros produtivos avaliados. Os resultados obtidos in vivo, referentes a TP e NT, mostrou que foi possível identificar diferença entre os machos, onde o indivíduo (M32) apresentou os piores resultados para as porcentagens de TP e NT. Desta forma, pode-se concluir que existem machos de alta e baixa fertilidade e que somente as análises in vitro realizadas neste estudo não são suficientes para categorizá-los, no entanto, a avaliação de dados retrospectivos foi eficiente para esta finalidade.(AU)

Condição microbiológica após antissepsia da genitália externa em éguas / Microbiological condition after antisepsis of reproductive tract in mares

A endometrite é a principal responsável pela diminuição nas taxas de fertilidade e recuperação embrionária em éguas e em parte, pode ser associada a contaminações por falhas na antissepsia das biotécnicas reprodutivas aplicadas à espécie. O objetivo deste trabalho foi avaliar os níveis de contaminação decorrentes de diferentes protocolos de antissepsia da genitália externa de éguas doadoras de embrião. Éguas sadias tiveram a vulva higienizada conforme um dos três grupos experimentais: (G1) sabão de côco; (G2) detergente neutro e (G3) clorexidina degermante 2%. Foi realizada a coleta de material da região da vulva, vestíbulo e útero com auxílio de swab estéril. As amostras foram processadas com objetivo de se determinar os principais microgranismos presentes, além da avaliação de Unidades Formadoras de Colônia (UFC). O G1 mostrou-se mais efetivo quanto à redução nas contagens de UFC em todas amostras. Por outro lado, G2 apresentou eficiência de 62%, sendo o tratamento menos eficiente na redução da contaminação das diferentes regiões avaliadas. A escolha adequada do agente sanitizante utilizado na higienização de períneo previamente a intervenções ginecológicas são de suma importância afim de se evitar contaminações ascendentes que levem a endometrites.(AU)

Endometritis is mainly responsible for the decrease in fertility and embryonic recovery rates in mares and, in part, it can be associated with contaminations due to failures in the antisepsis of reproductive biotechniques applied to the specie. The objective of this study was to evaluate the levels of contamination resulting from different protocols for antisepsis of the external genitalia of embryo donor mares. Healthy mares had their vulva sanitized according to one of the three experimental groups: (G1) coconut soap; (G2) neutral detergent and (G3) 2% chlorhexidine degerming. It was made the collection of material from the vulva, vestibule and uterus region with the aid of a sterile swab. The samples were processed in order to determine the main microorganisms present, in addition to the evaluation of Colony Forming Units (CFU). G1 was more effective in reducing CFU counts in all samples. On the other hand, G2 showed an efficiency of 62%, being the treatment less efficient in reducing contamination in the different regions evaluated. The proper choice of the sanitizing agent used to clean the perineum prior to gynecological interventions is of paramount importance in order to avoid ascending contamination that lead to endometritis.(AU)

Condição microbiológica após antissepsia da genitália externa em éguas / Microbiological condition after antisepsis of reproductive tract in mares

A endometrite é a principal responsável pela diminuição nas taxas de fertilidade e recuperação embrionária em éguas e em parte, pode ser associada a contaminações por falhas na antissepsia das biotécnicas reprodutivas aplicadas à espécie. O objetivo deste trabalho foi avaliar os níveis de contaminação decorrentes de diferentes protocolos de antissepsia da genitália externa de éguas doadoras de embrião. Éguas sadias tiveram a vulva higienizada conforme um dos três grupos experimentais: (G1) sabão de côco; (G2) detergente neutro e (G3) clorexidina degermante 2%. Foi realizada a coleta de material da região da vulva, vestíbulo e útero com auxílio de swab estéril. As amostras foram processadas com objetivo de se determinar os principais microgranismos presentes, além da avaliação de Unidades Formadoras de Colônia (UFC). O G1 mostrou-se mais efetivo quanto à redução nas contagens de UFC em todas amostras. Por outro lado, G2 apresentou eficiência de 62%, sendo o tratamento menos eficiente na redução da contaminação das diferentes regiões avaliadas. A escolha adequada do agente sanitizante utilizado na higienização de períneo previamente a intervenções ginecológicas são de suma importância afim de se evitar contaminações ascendentes que levem a endometrites.

Endometritis is mainly responsible for the decrease in fertility and embryonic recovery rates in mares and, in part, it can be associated with contaminations due to failures in the antisepsis of reproductive biotechniques applied to the specie. The objective of this study was to evaluate the levels of contamination resulting from different protocols for antisepsis of the external genitalia of embryo donor mares. Healthy mares had their vulva sanitized according to one of the three experimental groups: (G1) coconut soap; (G2) neutral detergent and (G3) 2% chlorhexidine degerming. It was made the collection of material from the vulva, vestibule and uterus region with the aid of a sterile swab. The samples were processed in order to determine the main microorganisms present, in addition to the evaluation of Colony Forming Units (CFU). G1 was more effective in reducing CFU counts in all samples. On the other hand, G2 showed an efficiency of 62%, being the treatment less efficient in reducing contamination in the different regions evaluated. The proper choice of the sanitizing agent used to clean the perineum prior to gynecological interventions is of paramount importance in order to avoid ascending contamination that lead to endometritis.

Desafios e estratégias para identificação de machos subférteis em rebanhos suínos / Challenges and strategies for identification of subfertile boars

A subfertilidade de reprodutores suínos pode alcançar até 15% dos machos em um plantel, gerando perdas econômicas pela menor taxa de parto e menor número de leitões nascidos totais. O uso de doses ou inseminações heterospérmicas e o elevado número de espermatozoides para compensar características espermáticas do ejaculado, dificultam o diagnóstico desses machos na rotina produtiva. Além disso, há uma baixa associação dos exames espermáticos realizados nas centrais de produção de sêmen com os resultados in vivo. Ao longo dos anos, avaliações com a implementação dos sistemas computadorizados de análise de sêmen, proteômica do plasma seminal ou célula espermática, inseminação artificial em tempo fixo e a classificação dos machos pela capacidade de preservação das doses de sêmen durante o armazenamento foram metodologias utilizadas para classificar e identificar machos subférteis. O conhecimento de estratégias para classificação dos machos quanto a fertilidade pode auxiliar na tomada de decisão dos reprodutores a serem descartados e otimização dos melhores machos, proporcionando melhorias no desempenho reprodutivo.

Boar subfertility reach up to 15% of the males in a herd, which represent economic losses due to the lower farrowing rate, and fewer total piglets born. The use of heterospermic semen doses, heterospermic inseminations and the high number of sperm cells is used to compensate sperm characteristics of the ejaculates making difficult to identify these males in the productive routine. Furthermore, there is a low association of sperm analysis performed in the boar studs with in vivo results. Over the years, with the implementation of computerized systems for semen analysis, proteomics of seminal plasma or sperm cell, fixed-time artificial insemination, and classification of boars based on capacity for semen preservation during storage, were methodologies used to classify and identify subfertile boars. The knowledge of strategies for classification of boars regarding fertility aid the decision for culling or for optimizing the best boars allow an improvement on the reproductive performance. can help in the decision-making of breeders to be discarded and those where the user should be optimized, thus providing improvements in reproductive performance.

Clinical application of Doppler ultrasound in the diagnosis of fertility problems in equines / Aplicações da ultrassonografia Doppler no diagnóstico de problemas de fertilidade em equinos

The use of different ultrasound modalities (color, power and pulsed Doppler) in clinical research has enhanced knowledge of reproductive pathophysiology in equine, in addition to improving the clinical diagnosis of reproductive disorders in both the stallion and the mare. In the stallion, color Doppler ultrasound has improved the diagnostic potential of conventional ultrasound, improving the differential diagnosis of pathologies such as testicular torsion (decrease or absence of blood flow in the cord) and orchitis (increased blood flow in the cord). The function of the testicle is highly dependent on the blood flow it receives; therefore, any vascular alterations can affect both the quality and production of sperm. In recent years there have been a greater number of studies in which Doppler ultrasound is used as a tool in the diagnosis of testicular dysfunction and in the monitoring of medical and surgical treatments. In the mare, ultrasound technology has allowed numerous advances in basic research regarding the vascular hemodynamics of the ovary and uterus. Moreover, it has become an indispensable tool in reproductive practice. Among the clinical applications of this technique in mares are the evaluation of CL (corpus luteum) functionality, being a useful tool in the selection of recipients in embryo transfer programs or the use of power Doppler in the diagnosis of pregnancy as early as 7 days postovulation prior to flushing. Finally, the incorporation of color and pulsed Doppler ultrasound into the examination improves the diagnosis of mares with endometritis due to a pathological increase in uterine blood flow.

Ultrassonografia Doppler aplicada ao diagnóstico de distúrbios testiculares em garanhões / Doppler ultrasonography applied to the diagnosis of testicular disorders in stallions / Ultrasonografía Doppler aplicada al diagnóstico de disturbios testiculares en garañones

As injúrias vasculares afetam diretamente a produção e a qualidade das células espermáticas, portanto, o diagnóstico rápido dessas alterações é de extrema importância para evitar danos irreversíveis à reprodução. Desse modo, a ultrassonografia Doppler têm se mostrado um método eficaz no diagnóstico precoce de afecções reprodutivas relacionadas com distúrbios na perfusão sanguínea testicular. Além disso, possibilita o acompanhamento de tratamentos em curso, a fim de melhorar resultados terapêuticos e proporcionar melhor previsão de fertilidade aos garanhões. Em homens, já é um método empregado para diagnosticar distúrbios de fertilidade, entretanto, na veterinária os relatos ainda são escassos e o uso por andrologistas de equinos para este fim, é esporádico. O objetivo dessa revisão é apresentar os princípios da ultrassonografia Doppler e os benefícios para a andrologia de equinos, afim de ampliar o conhecimento a respeito da técnica e facilitar o diagnóstico de afecções durante exames reprodutivos de garanhões.(AU)

Vascular disturbance directly affects sperm production and quality, so the rapid diagnosis of these changes is extremely important to avoid irreversible damage to reproduction activity. Thereby, Doppler ultrasonography has been shown to be an effective method in the early diagnosis of reproductive disorders related to testicular blood perfusion. In addition, it's possible to monitor treatments in progress for the purpose of to improve therapeutic results and provide better prediction of fertility for stallions. In men, it is already a method used to diagnose fertility disorders, however, in the veterinarian routine the reports are still scarce and the use by equine andrologists for this purpose is sporadic. Thus, the aim of this review is to present the principles of Doppler ultrasonography and its benefits for the equine andrology, in order to expand knowledge about the technique and facilitate the diagnosis of diseases during reproductive examinations of stallions.(AU)

Las afecciones vasculares afectan directamente la producción y la calidad de las células espermáticas, por lo tanto, el diagnóstico temprano de estas alteraciones es de extrema importancia para evitar daños irreversibles en la reproducción. En este sentido, la ultrasonografía Doppler ha sido empleada como un método eficaz en el diagnóstico precoz de afecciones reproductivas relacionadas con disturbios en la perfusión sanguínea testícular. Además, este método posibilita el acompañamiento de tratamientos en curso, con el fin de mejorar resultados terapéuticos y obtener una predicción de la fertilidad de los garañones. En hombres, ya es um método empleado para el diagnóstico de disturbiosde la fertilidad, mientras, en veterinaria los relatos aún son escasos y el uso por andrólogos en el campo equino para este fin, es esporádico. El objetivo de esta revisión es presentar los principios de la ultrasonografía Doppler y las ventajas para la andrología equina, con la finalidad de ampliar los conocimientos sobre esta técnica y facilitar el diagnóstico de afecciones durante los examenes reproductivos de los garañones.(AU)

Endometrial expression of key genes related to fertility in repeat breeder and non-repeat breeder cows.

The aim of the present study was to compare the endometrial gene expression of epidermal growth factor receptor (EGFR), nodal growth differentiation factor (NODAL), prostaglandin-endoperoxide synthase 2 (PTGS2), oestrogen receptor 1 (ESR1) and progesterone receptor (PGR) in repeat breeder cows (RBC) and non-RBC during diestrus. Endometrial samples were collected by cytobrush technique and stored in RNA stabilizing solution at -20°C until RT-qPCR analysis. Differences in endometrial mRNA expression of selected genes were assessed by ANOVA and simple (r) and the partial correlations (rp) among selected genes were performed. Results demonstrated that mRNA expression of EGFR and NODAL were higher in RBC than in non-RBC (3 and 25-fold change, p < .01 and p < .01, respectively), while the mRNA expression of PTGS2 was lower (1.56-fold change, p < .01). Although there were no differences detected in the mRNA expression of ESR1 and PGR, there was a positive correlation between the expression of ESR1 and EGFR (0.84, p < .05) and a negative correlation between PGR and PTGS2 (-0.49, p < .05). In conclusion, the difference on the endometrial mRNA expression of the genes included in the study between RBC and non-RBC indicates a deregulation of important mechanisms that are vital to establish a successful pregnancy. Thus, the present study provides useful insight as a base for future studies to elucidate the causes of RBC.

Feline testicular ultrasonogram differentiates pre vs. postpubertal and normal vs. disrupted spermatogenesis.

The aims of this study were: to ultrasonograhically describe and compare testicular parenchyma echogenicity and heterogeneity using digital image analysis in: I) prepubertal (PREP), peripubertal (PERI) and mature (MAT) cats; II) Normal and abnormal mature felids. Secondary, the relationships between histomorphological and ultrasonographic attributes of the testes were also determined. I) Fourteen, PREP, PERI and MAT male cats were ultrasonographically examined and then castrated. II) Seven adult cats were ultrasonographically examined before and after a GnRH antagonist administration and then castrated. All the testes were grossly and histomorphometrically assessed. In the frozen digital images of the longitudinal ultrasound sections, 3 regions of interest (ROI, 1 mm2) were selected. Within each ROI the echogenicity and the heterogeneity of the testicular parenchyma were digitally analyzed. In experiment I, testicular volume (0.15 ± 0.0 vs. 0.49 ± 0.1 vs. 1.65 ± 0.1; P < 0.01) and gonadosomatic index (0.04 ± 0.0 vs. 0.05 ± 0.0 vs. 0.08 ± 0.0; P < 0.01), echogenicity (56.54 ± 0.75 vs. 81.87 ± 5.88 vs.94.67 ± 3.62; P < 0.01) and heterogeneity (10.2420 ± 1.3740 vs.13.65 ± 0.65 vs. 14.67 ± 1.49; P < 0.01) augmented throughout PRE, PERI, and MAT. In experiment II, testicular volume (1.00 ± 0.09 vs. 0.85 ± 0.09; P < 0.05), echogenicity (87.74 ± 1.53 vs. 83.32 ± 1.54; P 0.01) but not heterogeneity (14.09 ± 0.26 vs. 14.19 ± 0.29; P > 0.05) decreased in the post GnRH antagonist abnormal testes. For both experiments, testicular volume, seminiferous tubular diameter, percentage of spermatids as the most mature cell type, and luminal/intertubular ratio were highly correlated (P < 0.01) with their echotextural attributes. Computer-assisted image analysis of B mode ultrasonogram appears as a good indicator of pubertal development and mild alterations of spermatogenesis in felids.

Ultrassonografia Doppler aplicada ao diagnóstico de distúrbios testiculares em garanhões / Doppler ultrasonography applied to the diagnosis of testicular disorders in stallions / Ultrasonografía Doppler aplicada al diagnóstico de disturbios testiculares en garañones

As injúrias vasculares afetam diretamente a produção e a qualidade das células espermáticas, portanto, o diagnóstico rápido dessas alterações é de extrema importância para evitar danos irreversíveis à reprodução. Desse modo, a ultrassonografia Doppler têm se mostrado um método eficaz no diagnóstico precoce de afecções reprodutivas relacionadas com distúrbios na perfusão sanguínea testicular. Além disso, possibilita o acompanhamento de tratamentos em curso, a fim de melhorar resultados terapêuticos e proporcionar melhor previsão de fertilidade aos garanhões. Em homens, já é um método empregado para diagnosticar distúrbios de fertilidade, entretanto, na veterinária os relatos ainda são escassos e o uso por andrologistas de equinos para este fim, é esporádico. O objetivo dessa revisão é apresentar os princípios da ultrassonografia Doppler e os benefícios para a andrologia de equinos, afim de ampliar o conhecimento a respeito da técnica e facilitar o diagnóstico de afecções durante exames reprodutivos de garanhões.

Vascular disturbance directly affects sperm production and quality, so the rapid diagnosis of these changes is extremely important to avoid irreversible damage to reproduction activity. Thereby, Doppler ultrasonography has been shown to be an effective method in the early diagnosis of reproductive disorders related to testicular blood perfusion. In addition, it's possible to monitor treatments in progress for the purpose of to improve therapeutic results and provide better prediction of fertility for stallions. In men, it is already a method used to diagnose fertility disorders, however, in the veterinarian routine the reports are still scarce and the use by equine andrologists for this purpose is sporadic. Thus, the aim of this review is to present the principles of Doppler ultrasonography and its benefits for the equine andrology, in order to expand knowledge about the technique and facilitate the diagnosis of diseases during reproductive examinations of stallions.

Las afecciones vasculares afectan directamente la producción y la calidad de las células espermáticas, por lo tanto, el diagnóstico temprano de estas alteraciones es de extrema importancia para evitar daños irreversibles en la reproducción. En este sentido, la ultrasonografía Doppler ha sido empleada como un método eficaz en el diagnóstico precoz de afecciones reproductivas relacionadas con disturbios en la perfusión sanguínea testícular. Además, este método posibilita el acompañamiento de tratamientos en curso, con el fin de mejorar resultados terapéuticos y obtener una predicción de la fertilidad de los garañones. En hombres, ya es um método empleado para el diagnóstico de disturbiosde la fertilidad, mientras, en veterinaria los relatos aún son escasos y el uso por andrólogos en el campo equino para este fin, es esporádico. El objetivo de esta revisión es presentar los principios de la ultrasonografía Doppler y las ventajas para la andrología equina, con la finalidad de ampliar los conocimientos sobre esta técnica y facilitar el diagnóstico de afecciones durante los examenes reproductivos de los garañones.

Genital bovine leptospirosis: A new look for an old disease.

Bovine leptospirosis is often associated with host-adapted leptospires infections, such as strains belonging to the Sejroe serogroup. Although bovine leptospirosis by adapted strains may result in abortions, fetal death, premature births and the birth of weak and/or low-weight calves, this infection is more closely associated with subtler syndromes, such as subfertility and early embryonic death. In this way, this silent disease can go unnoticed and undiagnosed, compromising reproductive efficiency with a consequent decrease in the productivity of herds over long periods. For many years, genital tract infection has been considered a secondary effect of renal infection with the assumption being that leptospires are found in the genital tract due to bacteremia caused by renal colonization. In contrast to this hypothesis, there is some evidence in the literature that suggests that genital leptospirosis should be considered a specific syndrome dissociated from renal/systemic disease. Therefore, this paper aims to gather and critically analyze information about genital leptospirosis in cattle, considering the disease a distinct syndrome, herein denominated bovine genital leptospirosis (BGL), that requires a unique approach to diagnosis and treatment.

Efeito da adição de plasma seminal oriundo de animais de alta e baixa fertilidade na criopreservação de espermatozoides da cauda do epidídimo e do ejaculado de garanhões subférteis / Effect of seminal plasma addition from high and low fertility animals on the cryopreservation of epididymal tail and ejaculated sperm from subfertile stallions

O objetivo do presente estudo foi avaliar o efeito da adição de plasma seminal de garanhões de alta e baixa fertilidade sobre a congelabilidade e a viabilidade de espermatozoides do ejaculado (EJ) e do epidídimo (EP) de garanhões subférteis. Foram utilizados seis garanhões com histórico de subfertilidade. Após coleta, espermatozoides do ejaculado foram divididos em três alíquotas: BotuSêmen® (EJ-CT); plasma seminal de alta qualidade espermática (EJ-PS1); e plasma seminal de baixa qualidade espermática (EJ-PS2). O mesmo protocolo foi realizado com espermatozoides da cauda do epidídimo após orquiectomia (EP-CT; EP-PS1; EP-PS2). Foram realizadas avaliações da cinética espermática pelo CASA e análises de integridade de membrana, acrossoma, fragmentação de DNA, capacitação espermática e peroxidação espermática por citometria de fluxo. Não foram observadas diferenças na cinética espermática entre EJ e EP, logo após a descongelação. Porém, foi observada maior (P<0,05) porcentagem de células com membranas plasmática e acrossomal íntegras nos grupos EP (EP-CT:31,7±7,5b; EP-PS1:35,2±7,0b; EP-PS2:33,9±7,2b) em comparação aos grupos EJ (EJ-CT:15,1±4,9a; EJ-PS1:11,7±4,5a; EJ-PS2:13,1±5,2a). Adicionalmente, foram observadas diferenças no índice de fragmentação de DNA (EJ-CT:2,6±0,6a; EJ-PS1:2,4±0,8a; EJ-PS2:3,0±0,8a; EP-CT:1,4±0,4b; EP-PS1:1,2±0,3b; EP-PS2:1,3±0,2b). Concluiu-se que a adição de 20% de plasma seminal, oriundo de animais férteis ou subférteis, previamente à congelação de espermatozoides epidídimários de animais subférteis não interfere na qualidade espermática.(AU)

The aim of this study was to compare the effect of the addition of seminal plasma from high and low fertility stallions on sperm viability of frozen-thawed sperm cells from ejaculate and from epididymal tail of subfertile stallions. Six stallions with a history of subfertility were used. After collection, ejaculate spermatozoa were divided into three aliquots: Botu-Semen® (EJ-CT); High-quality seminal plasma (EJ-PS1); Low-quality seminal plasma (EJ-PS2). The same was done with sperm cells from epididymis tail after orchiectomy (EP-CT; EP-PS1; EP-PS2). Evaluations of sperm kinetics were assessed by CASA and membrane and acrosome integrity, DNA fragmentation, sperm capacitation and sperm peroxidation were assessed by flow cytometry. After thawing, no differences were observed between ejaculated sperm (EJ) and epididymal sperm (EP) in any CASA evaluations. However, higher (P< 0.05) percentage of cells with intact plasma and acrossomal membranes was observed in EP groups (EP-CT:31.7±7.5b; EP-PS1:35.2±7.0b; EP-PS2:33.9±7.2b) compared to EJ groups (EJ-CT:15.1±4.9a, EJ-PS1:11.7±4.5a, EJ-PS2:13.1±5,2a). In addition, differences in DNA fragmentation index were observed (EJ-CT:2.6±0.6a; EJ-PS1:2.4±0.8a; EJ-PS2:3.0±0.8a; CT:1.4±0.4b; EP-PS1:1.2±0.3b; EP-PS2:1.3±0.2b). It was concluded that the addition of 20% seminal plasma from fertile or subfertile animals prior to the freezing of epididymal spermatozoa from subfertile animals does not interfere in sperm quality.(AU)

Efeito da adição de plasma seminal oriundo de animais de alta e baixa fertilidade na criopreservação de espermatozoides da cauda do epidídimo e do ejaculado de garanhões subférteis / Effect of seminal plasma addition from high and low fertility animals on the cryopreservation of epididymal tail and ejaculated sperm from subfertile stallions

O objetivo do presente estudo foi avaliar o efeito da adição de plasma seminal de garanhões de alta e baixa fertilidade sobre a congelabilidade e a viabilidade de espermatozoides do ejaculado (EJ) e do epidídimo (EP) de garanhões subférteis. Foram utilizados seis garanhões com histórico de subfertilidade. Após coleta, espermatozoides do ejaculado foram divididos em três alíquotas: BotuSêmen® (EJ-CT); plasma seminal de alta qualidade espermática (EJ-PS1); e plasma seminal de baixa qualidade espermática (EJ-PS2). O mesmo protocolo foi realizado com espermatozoides da cauda do epidídimo após orquiectomia (EP-CT; EP-PS1; EP-PS2). Foram realizadas avaliações da cinética espermática pelo CASA e análises de integridade de membrana, acrossoma, fragmentação de DNA, capacitação espermática e peroxidação espermática por citometria de fluxo. Não foram observadas diferenças na cinética espermática entre EJ e EP, logo após a descongelação. Porém, foi observada maior (P<0,05) porcentagem de células com membranas plasmática e acrossomal íntegras nos grupos EP (EP-CT:31,7±7,5b; EP-PS1:35,2±7,0b; EP-PS2:33,9±7,2b) em comparação aos grupos EJ (EJ-CT:15,1±4,9a; EJ-PS1:11,7±4,5a; EJ-PS2:13,1±5,2a). Adicionalmente, foram observadas diferenças no índice de fragmentação de DNA (EJ-CT:2,6±0,6a; EJ-PS1:2,4±0,8a; EJ-PS2:3,0±0,8a; EP-CT:1,4±0,4b; EP-PS1:1,2±0,3b; EP-PS2:1,3±0,2b). Concluiu-se que a adição de 20% de plasma seminal, oriundo de animais férteis ou subférteis, previamente à congelação de espermatozoides epidídimários de animais subférteis não interfere na qualidade espermática.(AU)

The aim of this study was to compare the effect of the addition of seminal plasma from high and low fertility stallions on sperm viability of frozen-thawed sperm cells from ejaculate and from epididymal tail of subfertile stallions. Six stallions with a history of subfertility were used. After collection, ejaculate spermatozoa were divided into three aliquots: Botu-Semen® (EJ-CT); High-quality seminal plasma (EJ-PS1); Low-quality seminal plasma (EJ-PS2). The same was done with sperm cells from epididymis tail after orchiectomy (EP-CT; EP-PS1; EP-PS2). Evaluations of sperm kinetics were assessed by CASA and membrane and acrosome integrity, DNA fragmentation, sperm capacitation and sperm peroxidation were assessed by flow cytometry. After thawing, no differences were observed between ejaculated sperm (EJ) and epididymal sperm (EP) in any CASA evaluations. However, higher (P< 0.05) percentage of cells with intact plasma and acrossomal membranes was observed in EP groups (EP-CT:31.7±7.5b; EP-PS1:35.2±7.0b; EP-PS2:33.9±7.2b) compared to EJ groups (EJ-CT:15.1±4.9a, EJ-PS1:11.7±4.5a, EJ-PS2:13.1±5,2a). In addition, differences in DNA fragmentation index were observed (EJ-CT:2.6±0.6a; EJ-PS1:2.4±0.8a; EJ-PS2:3.0±0.8a; CT:1.4±0.4b; EP-PS1:1.2±0.3b; EP-PS2:1.3±0.2b). It was concluded that the addition of 20% seminal plasma from fertile or subfertile animals prior to the freezing of epididymal spermatozoa from subfertile animals does not interfere in sperm quality.(AU)

Chronic epididymitis and Grade III varicocele and their associations with semen characteristics in men consulting for couple infertility.

Chronic epididymitis and varicocele are frequently observed genital disorders in men consulting for couple infertility, but their impact on semen characteristics at the time of infertility consultation is still a matter of debate. We investigated 652 male partners of couples who had their first infertility consultation between 1999 and 2015 in Argentina. Men with chronic epididymitis (n = 253), Grade III varicocele (n = 106), and both conditions (n = 125) were compared with a control group (n = 168) composed of men without these disorders or any other recognized causes of male infertility. We showed that men who presented isolated chronic epididymitis were more likely to have high percentages of low sperm motility and abnormal sperm morphology as well as a high number of white blood cells. Men with isolated Grade III varicocele had low sperm production and motility and an increased percentage of abnormal sperm morphology. Finally, men who simultaneously presented chronic epididymitis with Grade III varicocele had a low sperm motility and increased percentage of abnormal sperm morphology as well as a high number of white blood cells. Physical examination of the genital organs may identify common disorders, potentially involved as causal factors of patient's infertility. These disorders are associated with specific seminal profiles that should help in identifying the best treatment from the available therapeutic options, effectiveness, safety, and allowing as much as possible natural conception.

Peça intermediária de espermatozoides bovinos: funções e anormalidades / Midpiece of bovine spermatozoa: functions and abnormalities

O reprodutor tem grande importância na cadeia produtiva por possuir maior potencial de difusão do seu material genético dentro do rebanho do que as fêmeas que produzem apenas uma cria por ano, em manejo de monta natural. Entretanto, este potencial somente se torna real quando os machos apresentam boa qualidade seminal, resultando em menores taxas de retorno ao estro. No entanto, apesar das características reprodutivas apresentarem baixa herdabilidade, é consensual que algumas patologias espermáticas possam apresentar características hereditárias e neste caso podem permanecer disseminadas nos rebanhos. Entre as anomalias espermáticas identificadas nos ejaculados de bovinos, as patologias de peça intermediária são classificadas como defeitos espermáticos maiores, comprometendo a fertilidade dos reprodutores, porém estas têm recebido pouca atenção. O objetivo deste trabalho de revisão é apresentar os principais estudos sobre a formação da peça intermediária, sua importância e como esta pode sofrer anormalidades, visto que ejaculados com presença destas anomalias apresentam baixa motilidade espermática e fertilidade.(AU)

Bulls are of immense importance in the productive chain because they have a greater potential of diffusion of genetic material within the herd than females that produce only one calf per year in natural mating management. However, this potential is only achieved when males present good seminal quality, resulting in lower rates of return to estrus. Although the reproductive traits have low heritability, it is consensual that some sperm pathologies may present hereditary characteristics and in this case they can remain disseminated in the herds. Among the sperm abnormalities identified in bovine ejaculates, the midpiece pathologies are classified as a major sperm defect, compromising bull fertility, but these pathologies have received little attention. The objective of this review is to present the main studies on the midpiece formation, its importance and how it can undergo abnormalities, since ejaculates with the presence of this pathology present low sperm motility and fertility.(AU)