Philosophiae Doctor - PhD / Semen analysis is the cornerstone for the investigation of male infertility. Semen quality can be influenced by geographical location, age, ejaculatory abstinence, and season. In 2010, the WHO published criteria for human semen characteristics that were markedly lower than those previously reported. Many reports have discussed the methodology used by the WHO to set the 2010 reference values. Some of the limitations of the WHO (2010) study included an undefined ejaculatory abstinence period, the limited representation of different age groups, and a limitation in geographical representation as the study did not include any data from Africa and Middle East. Therefore, the current cohort study was designed to provide retrospective data on semen quality (Africa and Middle East) and reproductive hormones (Middle East) in patients who underwent semen analysis and endocrine investigation at Andrology Laboratories in South Africa and Qatar.

Philosophiae Doctor - PhD / The survival of free-ranging (in situ) African elephant and Southern white rhinoceros populations are currently being challenged on a daily basis in Africa. Reproductive health is considered a vital component of species conservation. Conservation of the last mega land mammals may ultimately require intervention by breeding management or combined with assisted reproductive technologies (ART). There is a strong case for gathering baseline information, both physiological and biological, of any species, as opportunities arise. During this study a total number of 21 ejaculates collected over two seasons from 12 free-ranging African elephant bulls were characterised, as well as 10 ejaculates collected from 10 free-ranging Southern white rhinoceros bulls from two populations. Ejaculates were collected from adult bulls by means of electroejaculation under anaesthesia. Routine semen analysis was combined with Computer-aided sperm analysis (CASA), Computer-aided sperm morphology analysis (CASMA), Transmission electron microscopy (TEM) and Genomics as diagnostic tools. Additionally, sperm functionality within different media was investigated and sperm subpopulation classification according to the motion pattern displayed. The results presented is based on the evaluation and classification of ≈ 45 000 individual African elephant spermatozoa and ≈ 18 000 individual Southern white rhinoceros spermatozoa. The average elephant ejaculate contained a total number of 47 x 10⁹ spermatozoa (volume of 56 ± 38mL x concentration of 818 ± 750 x 10⁶/mL) that recorded a total motility of 81 ± 29% of which 62 ± 26% were progressively motile. CASA recorded velocities for curvilinear velocity (VCL 241 ± 58μm/s), straight-line velocity (VSL 173 ± 181μm/s) and average path velocity (VAP 201 ± 54μm/s), and kinematics at straightness of track (STR 86 ± 85%), linearity of track (LIN 67 ± 16%), amplitude of lateral head displacement (ALH 4 ± 0.75μm) and beat cross frequency (BCF 21 ± 3Hz). Structural analysis revealed 68 ± 11% of the spermatozoa were viable (intact plasma membrane) and 77 ± 11% maintained acrosome integrity. Ejaculates contained 55 ± 14% morphologically normal spermatozoa, CASMA measured sperm head lengths at 6.83 ± 0.26μm and width 3.32 ± 0.18μm (total head area of 20.17 ± 1.96μm²) of which 38.95 ± 0.92% is covered by an acrosomal cap. The average rhinoceros ejaculate contained a total number of 1.1 x 10⁹ spermatozoa (volume of 24 ± 24mL x concentration of 83 ± 96 x 10⁶/mL) that recorded a total motility at 82 ± 8% of which 28 ± 23% were progressively motile. CASA recorded velocities for VCL (85 ± 29μm/s), VSL (44 ± 25μm/s) and VAP (69 ± 30μm/s, and kinematics at STR (63 ± 14%), LIN (51 ± 16%), ALH (2 ± 0.16μm) and BCF (16 ± 6Hz). Structural analysis revealed 73 ± 10% of the spermatozoa were viable (intact plasma membrane) and 76 ± 4% maintained acrosome integrity. Ejaculates contained 62 ± 14% morphologically normal spermatozoa, CASMA measured sperm head lengths at 5.5 ± 0.17μm and width 2.9 ± 0.19μm (total head area of 14.8 ± 1.43μm²) of which 36.3 ± 0.59% is covered by an acrosomal cap. Based on a Boolean argument and CASA data exploration it was possible to derive elephant and rhinoceros CASA cut-off criteria to sort between activated and hyperactivated motile spermatozoa. For the genomic component of this study, the CatSper1 (Loxodonta africana) gene was identified,sequenced and verified in a free-ranging (natural) African elephant population. Multivariate analysis(MVA) was applied to examine the associations between the semen and sperm parameters and the traits they accounted for in this study. Our understanding of wildlife reproductive sciences can substantially progress as the analytical techniques applied and the combination thereof is expanded. This investigation presents a new set of comprehensive semen and sperm threshold values for future investigations.

Depuis quelques décennies une diminution de la qualité du sperme a été constatée dans la majorité des pays occidentaux. Si les raisons de cette diminution sont encore incomplètement comprises, des facteurs environnementaux pourraient y contribuer grandement. Parmi eux, l’exposition à certains matériaux, comme les nanomatériaux et l’aluminium, a grandement augmenté durant cette même période. Notre objectif a donc été d’évaluer l’impact de ces expositions sur la fonction de reproduction masculine. Pour ce faire, nous avons réalisé deux études de la biodistribution testiculaire des particules après injection par voie intramusculaire sur un modèle de souris, une étude clinique visant à évaluer la charge du plasma séminal en nanoparticules et à la corréler avec le nombre de spermatozoïdes dans l’éjaculat et une étude clinique visant à évaluer l’impact de la charge spermatique en aluminium sur les paramètres spermatiques. Les résultats de ces travaux couplés à l’analyse de la littérature nous rassurent sur la reprotoxicité masculine des nanoparticules mais montrent l’existence d’un impact potentiellement négatif de l’aluminium. Dans les deux cas, de nouvelles études devront poursuivre l’exploration de ces questions / In recent decades a decrease in sperm quality was observed in most Western countries. While the reasons for this decline are not yet fully understood, environmental factors could greatly contribute. Among them, exposure to materials such as nanomaterials and aluminium, has greatly increased during this period. Therefore, our goal was to assess these exposures impact on male reproductive function. To do so, we conducted two studies to determine testis biodistribution of particles after intramuscular injection in a mouse model. We also conducted two clinical studies, first one to assess the burden of nanoparticles in seminal plasma and to correlate it with sperm count and the second one to evaluate the impact of the aluminum load in semen on sperm parameters. The results of these works and the analysis of literature reassure us about male reproductive toxicity of nanoparticles but revealed a potentially negative impact of aluminum. In both cases, new studies will be needed to further explore these issues

Thesis (MScMedSc (Obstetrics and Gynaecology))--University of Stellenbosch, 2005. / Chapter 1 contains a review dealing with nuclear and morphological changes during spermatogenesis and spermatozoa transport with emphasis on the maturation of spermatozoa, capacitation, acrosome reaction and the interaction with the cumulus oophorus complex (COC). The oocyte and cumulus oophorus complex is also discussed particularly on the topic of maturity (oocyte and cumulus maturity). Also presented is a review of the fluorescent binding agents, namely Fluorescein Isothiocyanate labeled with Pisum sativum (FITC-PSA), Chlorotetracycline test (CTC) and Chromomycin A3 (CMA3). Chapter II describes all the materials and methods used during this study. Routine semen analysis is described with emphasis on normal spermatozoon morphology according to strict criteria. The evaluation of capacitation and acrosome reaction (AR) using the CTC and PSA-FITC staining methods as well as the evaluation of spermatozoon nuclear chromatin packaging using the CMA3 staining method is described. Chapter III represents the results recorded in this study. Compared with those spermatozoa cultured in medium alone, spermatozoa exposed to the cumulus mass were more likely to be capacitated and acrosome reacted, with a distinct increase in chromatin packaging quality. A general discussion of the results and future applications are discussed in Chapter IV. In short An in vitro model for spermatozoa penetration through the cumulus oophorus was established. The model can be applied to investigate the effect of the cumulus oophorus on sperm functions and to assist in the selection of functional sperm for intracytoplasmic sperm injection therapy. All relevant references are presented in Chapter V .

Introdução: O lúpus eritematoso sistêmico (LES) é uma doença crônica autoimune com predomínio no sexo feminino e com evidente impacto em sua fertilidade. Por sua vez, em homens com LES foi observado alterações nos parâmetros seminais e nos níveis de hormônios sexuais. A análise seminal somente apresenta baixa correlação com potencial de fertilidade dos pacientes. Recentemente, a análise da integridade do DNA do espermatozoide tem mostrado melhor capacidade prognóstica para predizer a fertilidade do que os parâmetros seminais convencionais. Objetivo: Avaliar a fragmentação do DNA espermático de homens com LES sem azoospermia. Métodos: Vinte e oito pacientes homens, consecutivos, com LES (pelos critérios da ACR) e 34 controles foram avaliados conforme dados demográficos e de exposição ambiental, avaliação urológica, perfil hormonal e avaliação seminal (incluindo a fragmentação do DNA espermático). Aspectos clínicos, escores de atividade e dano cumulativo da doença e aspectos do tratamento também foram analisados. Resultados: Mediana da idade [33(20-52) vs. 36.5(25-54) anos, p=0.329] e frequência de varicocele (25% vs. 32%, p=0.183) foram similares entre o grupo de pacientes e o grupo controle. Na análise da fragmentação do DNA do espermatozoide observou-se quantidades significativamente mais altas de células classe III [44(9-88) vs. 16.5(0-80)%,p=0.001] e células classe IV [10.5(3-86) vs. 7(0-36)%,p=0.039] no grupo com LES. O índice de fragmentação do DNA espermático também foi significativamente mais alto em pacientes com LES [62(31-97) vs. 25.5(0-100)%, p < 0.001]. Parâmetros seminais convencionais (incluindo contagem espermática, motilidade e morfologia) foram similares em ambos os grupos. Dentro de grupo de pacientes com LES não foi observada correlação entre o índice de fragmentação do DNA espermático com idade, duração da doença, SLEDAI-2K e SLICC/ACR-DI ou dose cumulativa de predinisona, hidroxicloroquina, metotrexato, azatioprina, micofenolato mofetil ou ciclofosfamida intravenosa (CIC) (p > 0.05). Análises adicionais evidenciaram que motilidade espermática total foi significativamente menor no grupo que fez uso de CIC [64%(15-83) vs. 72%(57-86)%, p=0.024]. O índice de fragmentação do DNA espermático foi semelhante nos dois grupos [52.5(31-95) vs. 67.5(34-97)%, p=0.185]. Conclusões: Homens com SLE sem azoospermia apresentam maior índice de fragmentação do DNA espermático sem alteração dos parâmetros seminais ou hormonal. CIC não parece ter papel significativo nesta alteração. Estudos prospectivos futuros são necessários para determinar o impacto desta alteração na fertilidade destes pacientes / Introduction: Systemic lupus erythematosus (SLE) is a chronic autoimmune disease with a female predominance and have clear impact on fertility. In male SLE has been shown to alter seminal parameters and sexual hormonal levels. Recently, sperm DNA integrity analysis has shown better prognostic performance to predicting male fertility than seminal parameters. Objective: To evaluate sperm DNA fragmentation in non-azoospermic male SLE patients. Methods: Twenty-eight consecutive male SLE patients (ACR criteria) and 34 healthy controls were evaluated for demographic/exposures data, urologic evaluation, hormone profile and seminal analysis (including sperm DNA fragmentation). Clinical features, disease activity/damage scores and treatment were also assessed. Results: Median age [33(20-52) vs. 36.5(25-54) years, p=0.329] and frequency of varicocele (25% vs. 32%, p=0.183) were similar in SLE patients and healthy controls. Sperm DNA fragmentation showed significantly higher levels of cells class III [44(9-88) vs. 16.5(0-80)%, p=0.001] and cell class IV [10.5(3-86) vs. 7(0-36)%,p=0.039] in SLE. Sperm DNA fragmentation Index was also significantly higher in SLE patients [62(31-97) vs. 25.5(0-100)%, p < 0.001]. Conventional sperm parameters (including sperm count, motility and morphology) were similar in both groups. In SLE patients no correlations were observed between sperm DNA fragmentation index with age, disease duration, SLEDAI-2K and SLICC/ACR-DI scores, and cumulative dose of prednisone, hydroxychloroquine, intravenous cyclophosphamide (IVCYC), methotrexate, azathioprine and mycophenolate mofetil (p > 0.05). Further analysis of SLE patients treated with and without IVCYC showed that total sperm motility was significantly lower in the former group [64%(15-83) vs. 72%(57-86)%, p=0.024]. Sperm DNA fragmentation index was alike in both groups [52.5(31-95) vs. 67.5(34-97)%, p=0.185]. Conclusions: To our knowledge, this is the first demonstration that male non-azoospermic SLE patients have increased sperm DNA fragmentation without evident gonadal dysfunction. IVCYC does not seem to be a major determinant for this abnormality. Future prospective study is necessary to determine the impact of this alteration in these patients\' fertility

Apesar da maioria dos estudos mostrarem que parâmetros seminais anormais, redução do volume testicular e elevação dos níveis de radicais livres de oxigênio (ROS) no sêmen estão associados à presença de varicocele em homens inférteis, a maioria dos homens com varicocele apresenta os parâmetros seminais normais e são capazes de estabelecer gravidez em suas esposas. Nesse sentido, a avaliação desses parâmetros ainda não foi adequadamente estudada em homens com fertilidade comprovada e que apresentam varicocele. Esse estudo teve como objetivo estudar o impacto da varicocele clínica em homens férteis avaliando o volume testicular, parâmetros tradicionais da análise seminal e níveis de radicais livres de oxigênio no sêmen. Esses parâmetros foram avaliados em 113 homens férteis sem varicocele, 43 homens férteis com varicocele e 38 pacientes inférteis com varicocele. A medida do volume testicular foi realizada utilizando-se orquidômetro de Prader. Os parâmetros seminais foram avaliados manualmente segundo os critérios da Organização Mundial da Saúde (OMS) e a morfologia avaliada conforme os critérios da OMS e estrito de Kruger. Os níveis de ROS foram mensurados em sêmen puro utilizando-se a quimioluminescência. Os pacientes inférteis com varicocele apresentaram diminuição dos parâmetros seminais, redução do volume testicular e elevação dos níveis de ROS no sêmen em comparação com os homens férteis, com ou sem varicocele. Os homens férteis com varicocele não apresentaram diferenças significativas em nenhum dos parâmetros avaliados quando comparados aos homens férteis sem varicocele. No entanto, os homens férteis com varicocele grau 3 apresentaram menor concentração espermática e níveis de ROS no sêmen mais altos quando comparados àqueles com grau 1 ou 2. Além disso, o grau de varicocele nos homens férteis apresentou correlação negativa com a concentração espermática e positiva com os níveis de ROS no sêmen. A dosagem dos ROS no sêmen poderá ajudar, principalmente, na avaliação dos homens com varicocele que apresentam parâmetros seminais normais, e quando o potencial de fertilidade ainda não foi testado. A dosagem de ROS no sêmen pode representar um marcador mais sensível e específico no diagnóstico precoce da disfunção testicular, principalmente, nos homens férteis com varicocele grau 3 / Although the majority of the studies showed that abnormal seminal parameters, reduced testicular volume and high levels of seminal reactive oxygen species (ROS) are frequently associated with varicocele in infertile population, the majority of the men with varicocele present normal seminal parameters and are able to establish a pregnancy. In view of that, the evaluation of these semen parameters in fertile men with varicocele has not yet been well documented. The objective of our study was to evaluate the impact of clinical varicocele in fertile men assessing testicular volume, routine seminal parameters and seminal ROS levels. These parameters were evaluated in 113 fertile men without varicocele, 43 fertile men with clinical varicocele and 38 infertile patients with varicocele. Testicular volume was measured with a Prader orchidometer. Seminal parameters were assessed according to World Healthy Organization (WHO) guidelines and morphology according to WHO as well as Krugers strict criteria. Measurement of seminal ROS in neat semen was performed using a chemiluminescence assay. Infertile men with varicocele presented lower semen parameters, reduced testicular volume and higher seminal ROS levels compared with fertile men with or without varicocele. Although fertile men with varicocele presented all parameters evaluated similar to fertile men without varicocele, fertile men with varicocele grade 3 presented lower sperm concentration and higher seminal ROS levels compared with fertile men with varicocele grade 1 or 2. In addition, varicocele grade infertile men presented a negative correlation with sperm concentration and a positive correlation with seminal ROS levels. Men with unknown fertility status presenting with palpable varicocele and normal seminal parameters, the presence of increased ROS levels may be indicative of an early recognition of those subjects who will experience a progressive decrease in fertility potential if left untreated, especially in those presenting with varicocele grade 3

Many Neotropical primate species are endangered of extinction. Reproductive biotechnologies can consistently contribute to species conservation, being among them the genetic resource banks. However, success in the use of such technologies relies on advances in the knowledge of basic reproductive biology of a given species. During these processes, assessment of sperm quality is of supreme importance. We focused our work in captive golden-headed lion tamarins (Leontopithecus chrysomelas), housed at São Paulo Zoological Park Foundation. Leontopithecus sp. is a very important target for the development of reproductive techniques aiming conservation, as all species from this genus are classified as endangered or critically endangered of extinction by IUCN. For the evaluation of sperm quality in L. chrysomelas, techniques like sperm-binding assay, and the non-fluorescent staining for evaluation of acrosome integrity fast-green/rose-bengal, plasma membrane integrity eosin-nigrosin and mitochondrial activity 3.3 diaminobenzidine were validated for fresh semen. We have also validated an acrosome integrity evaluation technique coomassie blue staining, for thawed samples due to the inadequacy of the method used for fresh semen. Some Neotropical primates species show variation in semen quality over the year. In order to assess semen quality for this species and to verify if golden-headed lion tamarin presents this variation, we collected semen in dry and rainy seasons, during different periods of breeding season in the interval of conceptive periods and in the end of the second conceptive period. Testicular measurements were taken prior each semen collection, and a significant difference was found in total testicular volume between seasons, with major volumes found during dry season (p = 0.0011). Samples collected during rainy season showed higher percentage of total motility (x = 93.85) and intact plasma membrane (x = 95.54) (p = 0.0149 and p = 0.0279, respectively), in comparison with dry season (total motility x = 90.96 and intact plasma membrane x = 92.46). Even with those differences, semen from both seasons presented good quality. The success rate for collection with penile vibrostimulation technique was of 100%, and all samples obtained were constituted by only a coagulated fraction. Despite not achieving coagulum dissolution, we were able to access a high number of spermatozoa by adding Biggers-Whitten-Whittingham (BWW) medium to the coagulum for 30 min at 37°C, and performed all proposed analyzes. The same samples used of fresh analyzes were subsequently submitted to cryopreservation using two different commercial extenders BotuBOV&reg; (BB) and Freezing Medium with Glycerol and Gentamicin Test Yolk Buffer&reg; (TYB). Thawed semen samples were analyzed with the same methods previously validated and were also evaluated for the susceptibility to oxidative stress through quantification of induced thiobartituric acid reactive substances (TBARS). For all seminal parameters assessed, no significant difference was observed between BB and TYB, except that BB presented a higher percentage of intact acrosome (p = 0.0101) and concentration of TBARS (p = 0.0005). Despite those differences, thawed semen from both extenders performed similarly at spermegg binding assay. From our work, we observed that the proposed methods were successfully validated for L. chrysomelas. Fresh semen quality was very high and both extenders were successful in cryopreserving semen. / Muitas espécies de primatas neotropicais são ameaçadas de extinção. As biotecnologias da reprodução podem contribuir consistentemente para a conservação das espécies, estando entre elas os bancos de recurso genético. Entretanto, o sucesso no uso de tais tecnologias depende de avanços no conhecimento da biologia reprodutiva básica de uma certa espécie. Durante esse processo, a avaliação da qualidade espermática é de suprema importância. Nós concentramos nosso trabalho em micos-leões-de-cara-dourada (Leontopithecus chrysomelas), mantidos na Fundação Parque Zoológico de São Paulo. Leontopithecus sp. é um alvo importante para o desenvolvimento de técnicas reprodutivas com o propósito da conservação, sendo que todas as espécies do gênero são classificadas como ameaçadas ou criticamente ameaçadas de extinção pela IUCN. Para a avaliação da qualidade espermática em L. chrysomelas, técnicas como o teste de ligação do espermatozoide, e as colorações não fluorescentes para a avaliação de integridade de acrossomo fast-green/rosa bengala, integridade de membrana plasmática eosina-nigrosina e atividade mitocondrial 3,3 diaminobenzidina foram validados para sêmen fresco. Nós também validamos uma técnica de avalição de integridade de acrossomo corante azul de coomassie, para amostras descongeladas devido a inadequação do método utilizado em sêmen fresco. Algumas espécies de primatas neotropicais apresentam variação na qualidade seminal ao longo do ano. Para avaliar a qualidade do sêmen para essa espécies e para verificar se mico-leão-de-cara-dourada possui essa variação, nós colhemos sêmen nas estações de seca e chuva, durante diferentes períodos da estação reprodutiva no intervalo dos períodos conceptivos e no final do segundo período conceptivo. As medições dos testículos foram realizadas previamente a cada colheita de sêmen, e uma diferença significativa foi encontrada no volume testicular total entre as estações, com maiores volumes encontrados durante a estação de seca (p = 0,0011). As amostras colhidas durante a estação de chuva mostraram uma maior porcentagem de motilidade total (x = 93,85) e integridade de membrana plasmática (x = 95,54) (p = 0,0149 e p = 0,0279, respectivamente), em comparação com a estação seca (motilidade total x = 90,96 e integridade de membrana plasmática x = 92,46). Mesmo com essas diferenças, sêmen de ambas as estações apresentaram boa qualidade. A taxa de sucesso das colheitas com a técnica de vibroestimulação peniana foi de 100%, e todas as amostras eram constituídas de somente uma fração coagulada. Apesar de não obtermos a dissolução completa do coágulo, nós pudemos acessar um grande número de espermatozóides pela adição do meio Biggers-Whitten-Whittingham (BWW) ao coágulo por 30 minutos a 37°C, e realizamos todas as análises propostas. As mesmas amostras usadas nas avaliações do sêmen fresco foram subsequentemente submetidas à criopreservação usando dois diluidores comerciais diferentes BotuBOV&reg; (BB) e Freezing Medium with Glycerol and Gentamicin Test Yolk Buffer&reg; (TYB). As amostras descongeladas foram analisadas com os mesmos métodos previamente validados e também foram avaliadas para a suscetibilidade ao estresse oxidativo pela quantificação de substâncias reativas ao ácido tiobarbitúrico (TBARS). Para todos os parâmetros seminais avaliados, não houveram diferenças significativas entre BB e TYB, exceto que o BB apresentou porcentagem de acrossomos íntegros (p = 0.0101) e concentração de TBARS (p = 0.0005) maiores. Apesar dessas diferenças, as amostras descongeladas de ambos diluidores mostraram resultados similares no ensaio de ligação do espermatozoide à membrana perivitelínica do ovo de galinha. Do nosso trabalho, podemos observar que os métodos propostos foram validados com sucesso para L. chrysomelas. A qualidade do sêmen fresco foi muito alta e ambos diluidores obtiveram sucesso na criopreservação do sêmen.

INTRODUÇÃO: A prevalência global da obesidade duplicou desde 1980, alcançando mais de 600 milhões de adultos obesos em 2014. Em paralelo a este aumento, alguns autores relataram uma tendência mundial de declínio da concentração de espermatozoides no ejaculado nas últimas décadas. Consequentemente, surge a hipótese da relação entre esses dois fatos temporais. Dentre todas as formas de tratamento da obesidade, a cirurgia bariátrica é a que apresenta melhores resultados em termos de perda de peso e melhor efetividade a longo-prazo. Há evidências na literatura que sugerem que a obesidade é capaz de alterar os níveis dos hormônios reprodutivos e função erétil. No entanto, seus efeitos sobre os parâmetros seminais clássicos e no índice de fragmentação do DNA espermático (IFD) são pouco estabelecidos. Além disso, o impacto da cirurgia bariátrica sobre estas alterações ainda não foi devidamente esclarecido. OBJETIVO: Identificar os efeitos da obesidade sobre a função erétil, hormônios reprodutivos, função testicular e IFD de homens obesos. Adicionalmente, avaliar se a cirurgia bariátrica é capaz de afetar estes mesmos parâmetros. MÉTODOS: Este estudo foi organizado em duas fases. Na primeira fase, indivíduos com índice de massa corpórea (IMC) menor que 30 kg/m2 e fertilidade comprovada foram comparados a pacientes obesos em programação para cirurgia bariátrica, com IMC maior que 35 kg/m2. Todos os pacientes foram submetidos a dosagem de hormônios reprodutivos, análise seminal e aferição do IFD pelo método do cometa alcalino. O questionário do Índice Internacional de Função Erétil (IIEF-5) foi aplicado. Na segunda fase, parte dos indivíduos obesos foi submetida à cirurgia bariátrica e todos foram convocados para reavaliação em 6 meses. RESULTADOS: Homens obesos apresentam maiores níveis séricos de estradiol, LH e FSH, e menores níveis séricos de testosterona total (TT) quando comparados com homens férteis eutróficos. Além disso, apresentam piores parâmetros seminais, com redução do volume ejaculado, concentração seminal, número total de espermatozoides no ejaculado, motilidade total, motilidade progressiva e percentual de espermatozoides com morfologia normal, e elevação do IFD. Na fase 2, homens obesos submetido a cirurgia bariátrica apresentaram elevação dos valores de FSH, SHBG, TT e testosterona livre (TL), e redução dos níveis de prolactina sérica. Além disso, apresentam queda da concentração seminal e do número total de espermatozoides no ejaculado, e queda na fragmentação do DNA espermático. Controles obesos, por outro lado, não apresentaram alterações nas variáveis estudadas. Além disso, maior intensidade da perda de peso esteve associada com alterações mais importantes do número total de espermatozoides no ejaculado, e dos níveis séricos de TT, TL, SHBG e FSH. CONCLUSÕES: A obesidade é capaz de induzir alterações importantes dos hormônios reprodutivos, parâmetros seminais e IFD. A cirurgia bariátrica, entretanto, pode reverter os efeitos deletérios sobre os hormônios reprodutivos e fragmentação do DNA, mas pode trazer piora de parâmetros seminais em 6 meses de seguimento / INTRODUCTION: Worldwide obesity prevalence has duplicated since 1980, reaching more than 600 million obese adults in 2014. Parallel to this increase, several authors have reported a global tendency of ejaculated sperm concentration decline in the last decades. Therefore, comes to light the hypothesis that these two temporal trends are related. Among all forms of obesity treatments, the one that reaches the best results and long-term effectiveness is the bariatric surgery. Growing evidence in the literature suggests that obesity is capable of altering reproductive hormones levels and erectile function. Effects on classic semen parameters and DNA fragmentation index (DFI), however, have not been properly established. OBJECTIVE: Determine the effects of obesity over erectile function, reproductive hormones, testicular function, and DFI in obese men. Additionally, evaluate if bariatric surgery can also impact those parameters. METHODS: This study was divided into two phases. On the first phase, individuals with body mass index (BMI) lower than 30 kg/m2 and proven fertility were compared to obese men, with BMI higher than 35 kg/m2, waiting to be submitted to bariatric surgery. Reproductive hormones evaluation, semen analysis and evaluation of DFI by the alkaline comet assay were performed in all patients. The International Index of Erectile Function (IIEF-5) was used to assess erectile function. On phase two, part of the obese patients was submitted to bariatric surgery, and all were invited to 6-month revaluation. RESULTS: Obese men have higher blood levels of estradiol, LH and FSH, and lower levels of total testosterone (TT) when compared to eutrophic fertile men. Additionally, they present worse semen parameters, with a reduction in ejaculated volume, sperm concentration, total number of sperm in the ejaculate, total motility, progressive motility, and percentage of normal sperm, and higher sperm DFI. On phase two, obese men submitted to bariatric surgery showed higher FSH, SHBG, TT, and free testosterone (FT) levels, and reduction of blood prolactin levels. Moreover, they exhibited a reduction in sperm concentration and total ejaculated sperm count, and a reduction in sperm DNA fragmentation. Obese controls, however, did not experience changes in the studied variables. Finally, the intensity of weight loss was associated with greater changes in total ejaculated sperm count and in TT, TL, SHBG and FSH blood levels. CONCLUSIONS: Obesity can induce important changes in reproductive hormones, semen parameters, and DFI. Bariatric surgery, however, can revert the deleterious effects in reproductive hormones and DFI, but can result in worsening of semen parameters on 6-month follow-up

<p>Numerous studies on mammalian spermatozoa have reported large variations in the dimensions of the main sperm structural components, namely the head, midpiece and flagellum. These variations in sperm architecture are believed to be adaptations for functioning of spermatozoa in complex environments outside the male reproductive system. The midpiece of the mammalian&nbsp / permatozoon contains a varied number of mitochondria, but the reason for the marked difference in the size and structure of this sperm component is not clear. This study&nbsp / confirmed the variations in the sperm morphometry of seven selected mammalian species and revealed unique features of the sperm midpiece and sperm mitochondria of these seven species. Evaluation of several sperm kinematic parameters revealed the unique swimming characteristics of the different spermatozoa. The importance of using standardized motility&nbsp / parameters was highlighted as well as the assessment of different subpopulations of spermatozoa in order to produce more reliable and comparable data. Investigating the role of sperm mitochondria in human sperm&nbsp / metabolism indicated that these organelles are related to sperm function in terms of sperm motility. Furthermore, it was suggested that glycolysis and mitochondrial respiration are linked processes and that both are important for the maintenance of human sperm motility. By optimizing and employing standardized experimental procedures and analysis techniques, this study was&nbsp / able to confirm the species specificity of almost all the sperm parameters evaluated, while also elucidating the phylogenetic relatedness of the non-human primate species. In conclusion, the present study has confirmed that the various midpiece morphometry parameters are related to the remaining sperm morphometry parameters as well as to the sperm kinematic parameters.&nbsp / These proposed associations between the various sperm parameters were used to explain the sperm velocity of two hypothetical and morphologically different sperm structures. Therefore, the results of the current study support the idea of co-evolution between sperm components in mammalian spermatozoa and propose that the midpiece morphometry parameters that are selected for in these spermatozoa are midpiece volume, total number of mitochondrial gyres, thickness of the mitochondrial sheath and mitochondrial height.</p>

<p>Numerous studies on mammalian spermatozoa have reported large variations in the dimensions of the main sperm structural components, namely the head, midpiece and flagellum. These variations in sperm architecture are believed to be adaptations for functioning of spermatozoa in complex environments outside the male reproductive system. The midpiece of the mammalian&nbsp / permatozoon contains a varied number of mitochondria, but the reason for the marked difference in the size and structure of this sperm component is not clear. This study&nbsp / confirmed the variations in the sperm morphometry of seven selected mammalian species and revealed unique features of the sperm midpiece and sperm mitochondria of these seven species. Evaluation of several sperm kinematic parameters revealed the unique swimming characteristics of the different spermatozoa. The importance of using standardized motility&nbsp / parameters was highlighted as well as the assessment of different subpopulations of spermatozoa in order to produce more reliable and comparable data. Investigating the role of sperm mitochondria in human sperm&nbsp / metabolism indicated that these organelles are related to sperm function in terms of sperm motility. Furthermore, it was suggested that glycolysis and mitochondrial respiration are linked processes and that both are important for the maintenance of human sperm motility. By optimizing and employing standardized experimental procedures and analysis techniques, this study was&nbsp / able to confirm the species specificity of almost all the sperm parameters evaluated, while also elucidating the phylogenetic relatedness of the non-human primate species. In conclusion, the present study has confirmed that the various midpiece morphometry parameters are related to the remaining sperm morphometry parameters as well as to the sperm kinematic parameters.&nbsp / These proposed associations between the various sperm parameters were used to explain the sperm velocity of two hypothetical and morphologically different sperm structures. Therefore, the results of the current study support the idea of co-evolution between sperm components in mammalian spermatozoa and propose that the midpiece morphometry parameters that are selected for in these spermatozoa are midpiece volume, total number of mitochondrial gyres, thickness of the mitochondrial sheath and mitochondrial height.</p>