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Over the past decade long distance marathon running has become an important recreational activity. There is evidence that males with high levels of physical activity have some impairment of fertility. In order to investigate this further, 24 male marathon runners were studied over a period of a year. Each runner was assessed at regular intervals using hormonal profiles, anthropomorphic indices and semen evaluation. The training time and distance run increased progressively over the first five months of the study as the runners prepared for the Two Oceans marathon. Analysis of the serum hormonal profiles in this longitudinal study showed that the prolactin level increased when comparing the initial study month with the rest of the year and the progesterone level decreased. However the luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone and estradiol (E2) levels remained unchanged. When the runners were divided into a high and low training group according to the distance run in the preceding week, the only significant difference was the lower mean serum FSH level in the high training group. A decrease in semen volume was demonstrated as the training time increased. This trend was reversed as the runners' training decreased after the Two Oceans marathon. The percentage of morphologically normal spermatozoa showed an initial significant decrease in the first month of training. However, no significant difference was observed throughout the rest of year. An overall downward trend in semen motility in the first 5 months of the study was shown but this was only significant if the first and fifth study months were compared. The decrease in semen motility coincided with the period of maximum training. Since patients with an adequate sperm count but decreased motility have impaired fertility this finding is of considerable importance. In addition to the decrease in motility, there was a decrease in the percentage of morphologically normal spermatozoa when the initial month of low physical activity (December) was compared to all of the subsequent months analysed. This, too, is an important finding as the percentage of morphologically normal spermatozoa correlates directly with fertilisation and pregnancy rates. When the results were analysed in the high and low training months there was a significant difference in mean semen count and semen morphology. The mean count was higher in the high training group and this group also had a significantly higher normal morphology. However, there was no significant difference in semen volume and motility in the high and low training groups.

A nivel mundial se está dando un fenómeno que cada vez es más común, la infertilidad. En la actualidad la edad es considerada un factor determinante en la calidad seminal, existe una relación directa entre la edad y el aumento del daño en el ADN espermático. El objetivo de este estudio fue evaluar las características seminales en jóvenes universitarios mediante espermatogramas utilizando el sistema computarizado de análisis seminal C.A.S.A (Computer Assisted Sperm Analyzer, ISAS v1.2) para la evaluación morfológica. Se calcularon los estadísticos descriptivos, frecuencias y coeficientes de variación para todos los parámetros seminales procedentes de 30 jóvenes universitarios voluntarios de 18 a 30 años de edad. Con el fin de determinar si los hábitos de los jóvenes consideradas en la investigación tuvieron efecto en alguno de los parámetros seminales se realizó la prueba exacta de Fisher, en el caso de variables nominales, y una prueba de T de student o de U de Mann Whitney, previa verificación de la normalidad con la prueba de Shapiro-Wilk, en el caso de las variables cuantitativas. Todos los análisis se realizaron con un nivel de confianza de 95% en el software SPSS v.21. En conclusión no se encontraron asociaciones significativamente estadísticas entre los hábitos y los distintos parámetros seminales y se determinó que solo los criterios de pH, volumen, vitalidad, motilidad, concentración y recuento total, cumplen con los valores establecido por la Organización Mundial de Salud y la Sociedad Europea de Reproducción Humana y Embriología (ESHRE), a diferencia del 76.7% las muestras seminales que no cumple con los criterios de morfología. Se observó que existían alteraciones morfológicas en la cabeza y la pieza intermedia de los espermatozoides, comparándolo con los valores considerados normales por la OMS se obtuvo que las principales áreas afectadas fueron la longitud, el ancho, el área, la elipticidad y la elongación de la cabeza al igual que el ancho de la pieza intermedia, y al compararlo con los valores de la ESHRE se obtuvo que las principales áreas afectadas fueron el ancho, el área y la elipticidad de la cabeza. Worldwide is taking a phenomenon that is becoming more common, infertility. Today's age is considered a important factor in semen quality, there is a direct relationship between age and the increased damagein DNA sperm. The aim of this study wasto evaluate the seminal characteristics in young students by Spermograms and use a computerized semen analysis C.A.S.A (Computer Assisted Sperm Analyzer, ISAS v1.2) for morphological evaluation Descriptive statistics and frequencies for all semen parameters considered in this study were calculated. The coefficient of variation was calculated by dividing the standard deviation to the average and expressed in percentage. In order to determine whether the habits of young people considered in the investigation had any effec on semen parameters Fisher's exact test was performed, in the case of nominal variables, and student T test or Mann Whitney, after verification of normality with the Shapiro-Wilk test, in the case of quantitative variables. All analyzes were performed with 95% confidence in the SPSS v.21 software In conclusion not significantly statistical associations between the habits and different sperm parameters were found and determined that only the criteria of pH, volume, vitality, motility, concentration and total count, meet values set by the World Health Organization and the European Society of Human Reproduction and Embryology, unlike 76.7% of young people who do not meet the requirements criteria of morphology. It was found that there were obvious morphological changes in the anatomical parts of the head and the intermediate piece, comparing the valuesconsidered normal by WHO, was obtained that the main affected areas were the length, width, area, ellipticity and elongation of the head as the width of the intermediate piece, and when compared with the valuesthat were obtained ESHRE relevant affected areas were the wide area and the ellipticity of the head.

This study occurred in the Rural Academic Unit-Tiahuanaco installations, of the Bolivian Catholic University-La Paz, Bolviia, with the objective of evaluating macro- and microscopic characteristics of sperm ejaculation from bulbourethrectomized llamas. Six q'ara-variety male llamas of 3, 4, and 5 years of age were used over 8 weeks during which they were fed with natural and cultivated pastures. Ejaculate was collected with an artificial vagina with stimulation (libido) of male llamas by female llamas, for macro- and microscopic evaluation (volume, pH, color, appearance, motility, concentration, and sperm vitality). The results obtained were: average volume of 0.55[+or-]0.36 ml, with a CV of 20.2%; average pH of 7.53[+or-]0.42 with a CV of 4.9%; the ejaculate's color varied between crystal white, opaque white, and milky white at proportions of 50%, 25%, and 25%, respectively; average motility was 25.9[+or-]21.8% with a CV of 27.6%; average sperm concentration was 28.7x106[+or-]20.11x106 sperm/ml with a CV of 13%; average live sperm count was 31.8[+or-]24.4% with a CV of 25.3%; and the ejaculate an appearance of nonviscous fluid. The 4-year-old animals had excellent sperm ejaculations (macro- and microscopic characteristics) without differences between collection weeks.

Thesis (MScMedSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: The basic semen analysis plays a pivotal role in the diagnosis of male infertility and makes a significant contribution to the diagnostic process in andrology, gynecology and clinical urology. In 1902, the man considered to be ―the founding father of modern andrology‖ Edward Martin, proposed that an analysis of a semen sample should be incorporated into all infertility assessments. Following this suggestion in 1956, the scientist John MacLeod advanced the basic semen analysis from beyond a mere observation and introduced the importance of certain semen parameters such as morphology, motility and viscosity. The present day examination includes the analysis of certain established semen parameters, which can provide key information about the quality of a patient‘s semen and the functional competence of the spermatozoa. A semen analysis is also a valuable diagnostic tool in assessing possible disorders of the male genital tract and the secretory pattern of the male accessory sex glands. This information can help to determine the reproductive capacity of the male and can be used in conjunction with the partner to indicate the impact of male genital pathophysiology in the assessment of a couple‘s prospect for fertility. Patients attending the andrology laboratory at Tygerberg Academic Hospital for a semen analysis are referred based on primary, secondary or idiopathic infertility. Amongst these patients, an increase in semen viscosity has been observed over a period of time and created the need to assess the possible causes behind this trend. Despite viscosity being included in a routine spermiogram, it raises a considerable amount of concern as it is assessed semi-quantitatively. In the first part of this study, the possible correlation between seminal hyperviscosity and leukocytospermia was assessed. To achieve the most comprehensive assessment of viscosity, a new approach was used, which is a highly quantitative method to record viscosity in the international unit, centipoise (cP). The analysis of semen samples for possible leukocytospermia was approached by three methods the first of which was cytological. During this method granulocyte grading was performed on stained semen smears during the normal determination of morphology. The same approach was taken for the second method, whereby white blood cell concentrations were quantified with a leukocyte peroxidase test in the total sample group (n=200). Viscosity was compared between the samples classified as leukocytospermic positive or negative, according to the set reference values of the World Health Organisation (WHO). Correlation analysis between the two variables was also performed. In the biochemical approach of detecting leukocytospermia, an enzyme-linked immunoabsorbant assay (ELISA) was used to quantify the concentration of the extracellular polymorphonuclear (PMN) enzyme released from leukocytes. This test was performed on 124 randomly selected samples. All samples were fractionated before storage in liquid nitrogen, to allow for multiple assessments to be performed on each sample. The PMN elastase concentration was assessed against viscosity to investigate a possible correlation and relationship with the presence of leukocytospermia. All three methods of detecting possible infection showed a significantly positive relationship with increased viscosity in semen samples. The second approach in the study was to assess increased viscosity and leukocytospermia against parameters included in the spermiogram. An evaluation of hyperviscosity and its correlations to the various other semen parameters can allow for a detailed study into the effects that this anomaly may elicit. With the assessment of each of the sperm parameters against the leukocyte count and viscosity (cP), volume, concentration and morphology showed significance. To further the study, the third angle was to investigate a possible correlation between viscosity and the functional status of the male accessory sex glands. The biochemical approach of assessing the secretory patterns of the prostate and seminal vesicles against markers of infection can possibly further the understanding behind hyperviscous semen and leukocytospermia. Citric acid and fructose, secretory products of the prostate and seminal vesicles respectively, showed no significance when assessed against the leukocyte count and viscosity. However, this project was a pilot study and this approach offers an exciting avenue for further research. These research findings may provide a more comprehensive assessment of a man‘s fertility status. Seen in the context of patients attending the andrology laboratory of Tygerberg Academic Hospital, this is greatly needed as the majority of these patients cannot afford advanced assisted reproductive therapies. The introduction of a more accurate method of quantifying viscosity may possibly help to identify, diagnose and treat patients suffering from leukocytospermia in order to ultimately enhance their fertility potential. / AFRIKAANSE OPSOMMING: Die basiese semenanalise speel 'n belangrike rol in die diagnose van manlike infertiliteit en maak dus 'n betekenisvolle bydrae tot die diagnostiese proses in andrologie, ginekologie en kliniese urologie. In 1902 het Edward Martin, wat deur sommige navorsers as die vader van moderne andrologie beskou word, voorgestel dat 'n semenanalise deel moet vorm van alle infertiliteitsondersoeke. In 1956 het die wetenskaplike John MacLeod aanvoorwerk gedoen om die grondslag van 'n basiese semenanalise daar te stel, wat beteken het dat, in plaas van net 'n observasie studie te doen, 'n semenmonster kwantitatief analiseer moes word en dat parameters soos spermmorfologie, motiliteit en viskositeit as deel van die volledige analise gedoen moet word. Die hedendaagse analise sluit, behalwe die basiese semenparameters, ook inligting in oor die funksionele aspekte van spermatozoa. Die semenanalise is dus ook ‗n belangrike diagnostiese hulpmiddel om inligting rakende moontlike abnormaliteite in die manlike genitale traktus en die sekretoriese funksies van die manlike bykomstige geslagskliere te verskaf. Hierdie inligting kan help om 'n moontlike diagnose van die man se fertiliteitspotensiaal te maak. Terselftertyd kan dit ook tesame met die metgesel se reproduktiewe inligting meer lig werp op die impak van die man se genitale patofisiologie op die paartjie se fertilitetspotensiaal. Pasiënte wat die andrologielaboratorium van die Tygerberg Akademiese Hospitaal besoek word verwys op grond van primêre, sekondêre of idopatiese infertiliteit. Gedurende die laaste aantal jare is daar ‗n toename in voorkoms van verhoogde semenviskositeit onder hierdie groep pasiënte waargeneem. Dit het die behoefte laat ontstaan om die moontlike redes hiervoor te ondersoek. Ten spyte van die feit dat viskositeit deel vorm van die roetine semenanalise is dit tog kommerwekkend aangesien dit op 'n semi-kwantitatiewe manier bepaal word. In die eerste deel van hierdie studie is 'n moontlik korrelasie tussen seminale hiperviskositeit en leukositospermie ondersoek. Om die beste moontlike verwantskap te kon bepaal is 'n nuwe en hoogs kwantitatiewe metode gebruik om viskositeit in numeriese waardes volgens internasionale standaarde in centipoise (cP) te meet. Daar is van drie metodes gebruik gemaak om die teenwoordigheid van leukositospermie in 'n semenmonster te ondersoek. Die eerste metode was die sitologiese metode waar die teenwoordigheid van granulosiet op die gekleurde semensmeer tydens die standaard morfologie beoordeling bepaal word. Die tweede was deur middel van 'n leukosietperoksidase toets waarmee daar 'n kwantitatiewe telling gedoen kan word, soos teenwoordig in 'n voorbereide semenmonster. Hierdie twee bepalings is op die totale studiepopulasie van 200 pasiënte gedoen. Die viskositeit van monsters met of sonder die teenwoordigheid van leukositospermie, soos bepaal met die voorafgaande metodes en gebaseer op die WGO riglyne, is met mekaar vergelyk. Korrelasies is ook tussen hierdie twee veranderlikes en verskeie semenparameters van hierdie twee groepe gedoen. Die derde metode was 'n biochemiese ontleding met behulp van 'n ensiemgekoppeldeimmuunsorberende essai (ELISA) vir die bepaling van die ekstrasellulêre konsentrasie van polimorfonukleêre (PMN) elastase ensiem in die seminale plasma. Hierdie toets is op 124 lukraak gekose semenmonsters uitgevoer. Alle monsters is gefraksioneer voor berging in vloeibare stikstof om meervoudige analises van elke monster moontlik te maak. Die PMN elastase konsentrasies is vergelyk met die viskositeit van die semenmonsters vir 'n moontlike korrelasie en verwantskap met die teenwoordigheid van leukositospermie. Die resultate van al drie hierdie metodes, vir die moontlike bepaling van infeksie, het 'n betekenisvolle positiewe verwantskap met die toename in graad van viskositeit in semenmonsters aangetoon. Die tweede benadering van hierdie studie was om die viskositeitsgradering en die kwantitatiewe leukositopermie waardes te vergelyk met die semenparameters wat bepaal is tydens die semenanalise. Die doel van hierdie benadering was om enige verwantskap of effek van viskositeit asook die teenwoordigheid van witbloedselle op die semenparameters te ondersoek. Daar is betekenisvolle verwantskappe gevind tussen die viskositeitstatus van 'n semenmonster, die teenwoordigheid van witbloedselle en die semenparameters, soos motiliteit, morfologie en spermatosoa konsentrasie. Die derde benadering was om 'n ondersoek te doen na die moontlike verwantskap tussen viskositeit en die sekretoriese funksies van die manlike bykomstige geslagskliere, te wete die prostaat en seminale vesikula. Die biochemiese ondersoek na die sekresies van hierdie twee organe, naamlik fruktose en sitroensuur, is gedoen om te bepaal of die teenwoordigheid van infeksies van die manlike traktus, en waargeneem as leukositospermia, ook in verband gebring kan word met die viskositeitstatus van 'n semenmonster. Daar is geen verband gevind tussen die sekresies van hierdie twee kliere en die viskositeit van die semenmonsters nie. Aangesien hierdie deel van die studie net as 'n loodsprojek beskou is, is die biochemiese bepalings slegs op 'n beperkte aantal semenmonsters uitgevoer en kan hierdie tipe ondersoek as 'n moontlike verdere studie onderneem word. Hierdie navorsingsresultate kan lei tot ‗n meer omvattende assessering van mans se fertiliteitstatus. Dit is uiters noodsaaklik in die konteks van omstandighede van die pasiënte wat die andrologielaboratorium van die Tygerberg Akademiese Hospitaal besoek aangesien die meerderheid nie gevorderde in vitro behandeling kan bekostig nie. Die akkurate bepaling van 'n semenmonster se viskositeit kan dus moontlik waarde toevoeg tot die identifisering, diagnose en behandeling van pasiënte met leukositospermie om sodoende hulle fertiliteitspotensiaal te verbeter.

As espécies reativas de oxigênio (ERO) em baixos níveis são necessárias para as funções normais do espermatozoide, as quais estão envolvidas com a capacidade de fertilização. Entretanto, várias evidências demonstram que a produção excessiva de ERO leva ao estresse oxidativo, que por sua vez está relacionado à infertilidade masculina. Uma correlação positiva entre os níveis excessivos de ERO e concentrações anormais de espermatozoides, motilidade e morfologia tem sido descrita. Além disso, a capacidade antioxidante total do sêmen tem sido significativamente menor em casos de infertilidade comparado com casos férteis. O estresse oxidativo também tem sido relacionado ao envelhecimento e há evidências de que o acúmulo de radicais livres danifica o DNA do espermatozoide e prejudica a fertilização. A Organização Mundial da Saúde tem enfatizado a importância da avaliação do estresse oxidativo seminal e recomendado que cada laboratório estabeleça os valores de referência para os parâmetros do sêmen em sua população. O objetivo deste estudo foi avaliar o estado redox no sêmen humano e sua correlação com os parâmetros do espermograma. Para tanto, avaliou-se a medida de ERO (quimioluminescência, QL), a atividade antioxidante (QL e catalase), a peroxidação lipídica (malondialdeído, MDA), a medida dos produtos avançados de oxidação proteica (AOPP) e apoptose/necrose (anexina/iodeto de propídeo, citometria de fluxo) no sêmen humano de homens saudáveis e férteis (Controles; n=7). As metodologias foram padronizadas para aplicá-las à análise do sêmen de homens que estavam em investigação de infertilidade (Pacientes; n=23), e correlacionar os resultados com os parâmetros do espermograma. Os principais resultados mostraram 1) a medida de ERO no sêmen de Pacientes sem alterações no espermograma (Pacientesa; n=9) foi maior do aquela para sêmen dos Pacientes com alterações no espermograma (Pacientesb; n=14) e dos Controles, sugerindo comprometimento da qualidade do sêmen pelo aumento de ERO, mesmo com espermograma normal; 2) não houve diferenças entre os grupos quanto à peroxidação lipídica (MDA), aos AOPP, catalase e apoptose/necrose; 3) quanto às correlações entre os parâmetros analisados, observou-se: correlação positiva entre a medida de ERO no sêmen in natura e no sêmen lavado do grupo Pacientes, validando a utilização do sêmen in natura para esta metodologia; correlação positiva entre a medida de ERO in natura e a apoptose, número de espermatozoides e número de leucócitos somente para o grupo Pacientesb; correlação entre a medida de ERO in natura e a motilidade progressiva somente para o grupo Controle; correlação entre apoptose e número de espermatozoides nos grupos Pacientes e Pacientesa; estes resultados mostram correlações particulares em cada grupo e correlações compartilhadas, que caracterizam Controles e Pacientes. O perfil da presença e da ausência destas correlações no grupo Controle pode estabelecer um padrão de referência para as análises do estresse oxidativo no sêmen. Os resultados poderão contribuir para a aplicação da medida das ERO e das suas correlações com parâmetros do espermograma na análise de rotina do sêmen humano, para a investigação da infertilidade e de patologias do sistema reprodutor masculino. / Reactive oxygen species (ROS) at low levels are required for normal function of sperm, which are involved in fertilization capacity. However, evidences show that the excessive production of ROS leads to oxidative stress, which in turn is related to male infertility. A positive correlation between excessive levels of ROS and abnormal sperm concentration, motility and morphology has been described. Moreover, the total antioxidant capacity of the semen has been significantly lower in cases of infertility compared with fertile cases. Oxidative stress has also been associated with aging and there is evidence that the excess of free radicals damages the DNA of sperm and impairs fertilization. The World Health Organization has emphasized the importance of the evaluation of seminal oxidative stress and recommended that each laboratory establish reference values for the parameters of semen in their population. The aim of this study was to evaluate the redox state in human semen and its correlation with semen parameters. For this purpose, we evaluated the measurement of ROS (chemiluminescence, CL), the antioxidant activity (CL and catalase), lipid peroxidation (malondialdehyde, MDA), the advanced oxidation protein products (AOPP) and apoptosis / necrosis (annexin / propidium iodide flow cytometry) in semen of healthy, fertile men (Controls, n = 7). The methods were standardized to apply them to the analysis of semen of men who were in infertility investigation (Patients, n = 23), and to correlate the results with the parameters of sperm. The main results showed: 1) the measurement of ROS in the semen of patients with normal semen (Patientsa, n = 9) was higher than that for semen of patients with abnormal semen analysis (Patientsb, n = 14) and Controls, suggesting impairment of the quality semen by increasing ROS, despite normal semen; 2) no differences were found among the groups regarding to lipid peroxidation (MDA), the AOPP, catalase and apoptosis / necrosis; 3) concerning the correlations among the parameters analyzed, we observed: positive correlation between the measurement of ROS in semen in natura and washed semen of Patients group, validating the use of semen in natura for this method; positive correlation between the measurement of ROS in semen in natura and apoptosis, number of spermatozoa and leukocytes only for the Patientsb group; correlation between the measurement of ROS in semen in natura and progressive motility only for the Control group; correlation between apoptosis and number of spermatozoa in the groups Patients and Patientsa; these results show particular and shared correlations in each group, which characterize Controls and Patients. The profile of the presence and absence of these correlations in the Control group may establish a reference standard for the analysis of oxidative stress in the semen. These results may contribute to the use of the measurement of ROS and their correlations with semen parameters in routine analysis of human semen, for the investigation of infertility and disorders of the male reproductive system.

Introdução: Em torno de 15% dos casais apresentam o diagnóstico de infertilidade, sendo que 50% se devem a fatores masculinos. Diversos testes de função espermática são propostos para a avaliação da fertilidade, mas o espermograma é o principal exame para o diagnóstico de infertilidade masculina. O valor prognóstico das características seminais como concentração, morfologia e motilidade como marcadores de infertilidade masculina é confundido. A avaliação desses e a classificação quanto à normalidade dos mesmos permanece sendo tema de discussão frequente. Objetivos: Comparar as diferentes classificações de parâmetros seminais (OMS 1999, OMS 2010 e TMSC) e avaliar as características de pacientes em terapia de reprodução humana assistida que se relacionam com esses parâmetros. Métodos: Foi realizado estudo longitudinal, retrospectivo, baseado em revisão de banco de dados para avaliar as características e parâmetros seminais e comparar as mesmas conforme as classificações da OMS de 1999, OMS de 2010 e TMSC de 477 homens submetidos a investigação de infertilidade ou tratamentos de reprodução assistida na clínica Generar – Reprodução Humana, entre 2011 e 2015. Resultados: 401 pacientes tiveram alguma alteração pela OMS 1999, 223 pela OMS 2010 e 200 pela TMSC. O critério que mais alterou a condição de uma amostra seminal de anormal em 1999 para normal em 2010 foi a morfologia. Conclusão: Os parâmetros se tornaram menos rígidos de 1999 para 2010 alterando significativamente a proporção de indivíduos que deixaram de ser classificados como inférteis. A classificação baseada no TMSC pode ser útil na indicação de alguns tratamentos, mas não pode definir um indivíduo como fértil ou infértil em função de não levar em consideração a morfologia espermática. / Background: Approximately 15% of couples presenting the diagnosis of infertility, and 50% of cases are due to male factors. Several sperm function testing is proposed for the evaluation of male fertility, but sperm is the first test for the diagnosis of male infertility causes. The prognostic value of the seminal characteristics as concentration, morphology and motility as male infertility markers is often confused. Evaluation of semen parameters and classification for normality remains frequent topic of discussion. Objectives: Compare the different classifications of seminal parameters (OMS 1999, WHO 2010 and TMSC) and evaluate the characteristics of patients treated in assisted reproduction therapy relating to these parameters. Methods: Retrospective study based on chart review evaluated 477 semen samples from men undergoing investigation or infertility treatments in assisted reproduction between 2011 and 2015. Results: 401 patients were considered abnormal by the WHO 1999, 223 by WHO 2010 and 200 for TMSC. The criteria that most changed the classification was sperm morphology. Conclusion: The parameters have become less rigid 1999 to 2010 significantly changing the proportion of individuals who are no longer classified as infertile. The classification based on TMSC can’t define an individual as fertile or infertile regardless due to not take into account the sperm morphology, but may be helpful when it comes to the indication of the intrauterine insemination.

Introdução: Em torno de 15% dos casais apresentam o diagnóstico de infertilidade, sendo que 50% se devem a fatores masculinos. Diversos testes de função espermática são propostos para a avaliação da fertilidade, mas o espermograma é o principal exame para o diagnóstico de infertilidade masculina. O valor prognóstico das características seminais como concentração, morfologia e motilidade como marcadores de infertilidade masculina é confundido. A avaliação desses e a classificação quanto à normalidade dos mesmos permanece sendo tema de discussão frequente. Objetivos: Comparar as diferentes classificações de parâmetros seminais (OMS 1999, OMS 2010 e TMSC) e avaliar as características de pacientes em terapia de reprodução humana assistida que se relacionam com esses parâmetros. Métodos: Foi realizado estudo longitudinal, retrospectivo, baseado em revisão de banco de dados para avaliar as características e parâmetros seminais e comparar as mesmas conforme as classificações da OMS de 1999, OMS de 2010 e TMSC de 477 homens submetidos a investigação de infertilidade ou tratamentos de reprodução assistida na clínica Generar – Reprodução Humana, entre 2011 e 2015. Resultados: 401 pacientes tiveram alguma alteração pela OMS 1999, 223 pela OMS 2010 e 200 pela TMSC. O critério que mais alterou a condição de uma amostra seminal de anormal em 1999 para normal em 2010 foi a morfologia. Conclusão: Os parâmetros se tornaram menos rígidos de 1999 para 2010 alterando significativamente a proporção de indivíduos que deixaram de ser classificados como inférteis. A classificação baseada no TMSC pode ser útil na indicação de alguns tratamentos, mas não pode definir um indivíduo como fértil ou infértil em função de não levar em consideração a morfologia espermática. / Background: Approximately 15% of couples presenting the diagnosis of infertility, and 50% of cases are due to male factors. Several sperm function testing is proposed for the evaluation of male fertility, but sperm is the first test for the diagnosis of male infertility causes. The prognostic value of the seminal characteristics as concentration, morphology and motility as male infertility markers is often confused. Evaluation of semen parameters and classification for normality remains frequent topic of discussion. Objectives: Compare the different classifications of seminal parameters (OMS 1999, WHO 2010 and TMSC) and evaluate the characteristics of patients treated in assisted reproduction therapy relating to these parameters. Methods: Retrospective study based on chart review evaluated 477 semen samples from men undergoing investigation or infertility treatments in assisted reproduction between 2011 and 2015. Results: 401 patients were considered abnormal by the WHO 1999, 223 by WHO 2010 and 200 for TMSC. The criteria that most changed the classification was sperm morphology. Conclusion: The parameters have become less rigid 1999 to 2010 significantly changing the proportion of individuals who are no longer classified as infertile. The classification based on TMSC can’t define an individual as fertile or infertile regardless due to not take into account the sperm morphology, but may be helpful when it comes to the indication of the intrauterine insemination.

Introdução: Em torno de 15% dos casais apresentam o diagnóstico de infertilidade, sendo que 50% se devem a fatores masculinos. Diversos testes de função espermática são propostos para a avaliação da fertilidade, mas o espermograma é o principal exame para o diagnóstico de infertilidade masculina. O valor prognóstico das características seminais como concentração, morfologia e motilidade como marcadores de infertilidade masculina é confundido. A avaliação desses e a classificação quanto à normalidade dos mesmos permanece sendo tema de discussão frequente. Objetivos: Comparar as diferentes classificações de parâmetros seminais (OMS 1999, OMS 2010 e TMSC) e avaliar as características de pacientes em terapia de reprodução humana assistida que se relacionam com esses parâmetros. Métodos: Foi realizado estudo longitudinal, retrospectivo, baseado em revisão de banco de dados para avaliar as características e parâmetros seminais e comparar as mesmas conforme as classificações da OMS de 1999, OMS de 2010 e TMSC de 477 homens submetidos a investigação de infertilidade ou tratamentos de reprodução assistida na clínica Generar – Reprodução Humana, entre 2011 e 2015. Resultados: 401 pacientes tiveram alguma alteração pela OMS 1999, 223 pela OMS 2010 e 200 pela TMSC. O critério que mais alterou a condição de uma amostra seminal de anormal em 1999 para normal em 2010 foi a morfologia. Conclusão: Os parâmetros se tornaram menos rígidos de 1999 para 2010 alterando significativamente a proporção de indivíduos que deixaram de ser classificados como inférteis. A classificação baseada no TMSC pode ser útil na indicação de alguns tratamentos, mas não pode definir um indivíduo como fértil ou infértil em função de não levar em consideração a morfologia espermática. / Background: Approximately 15% of couples presenting the diagnosis of infertility, and 50% of cases are due to male factors. Several sperm function testing is proposed for the evaluation of male fertility, but sperm is the first test for the diagnosis of male infertility causes. The prognostic value of the seminal characteristics as concentration, morphology and motility as male infertility markers is often confused. Evaluation of semen parameters and classification for normality remains frequent topic of discussion. Objectives: Compare the different classifications of seminal parameters (OMS 1999, WHO 2010 and TMSC) and evaluate the characteristics of patients treated in assisted reproduction therapy relating to these parameters. Methods: Retrospective study based on chart review evaluated 477 semen samples from men undergoing investigation or infertility treatments in assisted reproduction between 2011 and 2015. Results: 401 patients were considered abnormal by the WHO 1999, 223 by WHO 2010 and 200 for TMSC. The criteria that most changed the classification was sperm morphology. Conclusion: The parameters have become less rigid 1999 to 2010 significantly changing the proportion of individuals who are no longer classified as infertile. The classification based on TMSC can’t define an individual as fertile or infertile regardless due to not take into account the sperm morphology, but may be helpful when it comes to the indication of the intrauterine insemination.

Submitted in partial fulfillment of the requirements for the degree of Master of Health Sciences in Clinical Technology, Durban University of Technology, Durban, South Africa, 2017. / Introduction The prevalence of infertility is increasing at an alarming rate globally. Many couples are afflicted with infertility due to an array of diseases, trauma and psychological stresses. Renal disease is one such pathophysiological condition which is increasing amongst the younger age group. Often the progression of chronic renal disease leads to end stage renal failure that requires a renal transplantation. Post renal transplant, immunosuppressive agents are routinely prescribed to prevent allograft rejection. Immunosuppressive agents are potent drugs that can have deleterious side effects on semen parameters. However, the effects of the immunosuppressive agents on semen parameters in the literature are unclear and require further investigation. It is, therefore, important to assess the effects of immunosuppressive agents on semen, especially the three vital aspects of sperm concentration, motility and morphology which form the basis of male reproduction. Aims and Objectives of study This was a prospective observational study evaluating the effects of different immunosuppressive regimens on sperm parameters in post renal transplant male patients. The main aspects of semen parameters such as sperm concentration, motility and morphology that determine reproductive potential were assessed in the study patients and compared to the gold standard of semen analysis according to the World Health Organisation (WHO) reference values. Methodology Thirty-four renal transplant patients were recruited from the databases of both private nephrologists in the greater Durban area and the academic renal unit at Inkosi Albert Luthuli Central Hospital. Following bioethical approval and informed consent, patients were required to produce a semen sample by masturbation. A questionnaire documenting the patient’s lifestyle, aetiology of renal disease, transplant date and immunosuppressive duration and regimen were recorded. The semen samples were analysed comprehensively according to the protocol on semen analysis recommended by the WHO. This included the macroscopic investigation (volume, appearance, colour, viscosity, liquefaction time and pH) and microscopic evaluation (sperm concentration, total motility, morphology, IgG/IgA and vitality). Sperm concentration, total motility, morphology and vitality were examined and recorded in duplicate to strengthen the validity of the results. A biostatistician analysed the data and determined the statistical analysis. Descriptive statistics determined values of semen parameters in renal transplanted males and in each race demographic. The one sample t-test analysed the statistical significance between the mean study values and the WHO reference values. The effect of the immunosuppressive agent on semen parameters was determined using multiple linear regressions whilst ROC analysis determined the sensitivity and specificity of sperm concentration, total motility and morphology in predicting pregnancy from the patients that fathered children post renal transplant. Results The mean sperm concentration and morphology in the study patients were 14.0 mill/ml (95% Confidence Interval (CI) 10.2 – 17.7) and 3.3% (95% CI 2.7 – 3.9), respectively. Although values obtained were minimally lower than the WHO reference values, these results were within the 95% CI of the WHO guidelines. Motility evaluation revealed higher values of 43.2% (95% CI 36.6 – 49.7). In contrast, sperm vitality was considerably decreased, 47.5% (95% CI 40.6 – 54.4). All semen parameters exhibited no statistical significance (one sample t-test) when analysed against the WHO reference values except for sperm morphology, (p = 0.025; p< 0.05) which showed decreased morphology irrespective of immunosuppressive regimen. Semen volume 1.7 ml (95% CI 1.3 – 2.0) and pH 7.7 (95% CI 7.6 – 7.9) were both within the WHO guidelines. Descriptive statistics according to racial demographics showed no differences in semen values. An almost perfect linear relationship existed between total sperm motility and vitality (r = 0.967). Multiple linear regressions of duration and dosages of immunosuppressive drugs tacrolimus and mycophenolate mofetil, could not predict the effect of the immunosuppressive agents on sperm concentration, total motility and morphology. There was a significant difference in morphology between those with and without children post renal transplant. Those with children post renal transplant exhibited a higher morphology value, (p = 0.001; p< 0.05). Sensitivity and specificity analysis of the patients with children post renal transplant concluded that morphology is the most optimal indicator and predictor of pregnancy (AUC = 0.854). Tacrolimus was the common immunosuppressive agent used in the four patients that fathered children. This was more evident in patients that underwent therapy with Sirolimus followed by Cyclosporin A (CsA) and changed to Tacrolimus as the last immunosuppressive agent used for maintenance therapy. Conclusion The ability to procreate in renal transplanted males has become increasingly difficult and emotionally challenging. In this study sperm concentration and morphology of renal transplanted males exhibited parameters similar to the general fertile population. Total motility possessed a higher range of values in contrast to sperm vitality which showed a significant decrease from the WHO reference values. The effect of immunosuppressive treatment on semen parameters could not be clearly defined due to the number of immunosuppressive regimens that patients were subjected to intermittently resulting in small sample sizes within each immunosuppressive regimen grouping. The majority of patients underwent a triple maintenance therapy of tacrolimus, MMF and prednisone. The dosage and duration of these tacrolimus and MMF was inconclusive in determining a beneficial or detrimental relationship on semen parameters. Morphology was shown to be the most significant indicator in predicting pregnancy in patients that fathered children. Tacrolimus was a common immunosuppressive agent used in the majority of patients that fathered children. It may have protective effects on sperm parameters as shown in patients that fathered children. This was a study with a small sample size and further investigations are required in a larger cohort of patients to assess individualized effects of the different immunosuppressive agents on sperm parameters. / M