High percentage of midpiece defects in Brangus bull sperm with no reduction in sperm kinematics.

The study investigated midpiece defects in sperm from a 5-year-old Brangus bull with a high rate of semen batch rejection, due to morphologically abnormal sperm, with no reduction in sperm kinematics. A comprehensive evaluation was conducted over a 16-month period, involving 28 ejaculates. Notably, despite the high proportion of midpiece defects (average 37.73%, from 3% to 58%), the study revealed stable sperm production, with no discernible differences in the kinematic data before and after cryopreservation. Electron microscopy identified discontinuities in the mitochondrial sheath, characteristic of midpiece aplasia (MPA). The anomalies were attributed to be of genetic origin, as other predisposing factors were absent. Additionally, the electron microscopy unveiled plasma membrane defects, vacuoles and chromatin decondensation, consistent with previous findings linking acrosome abnormalities with midpiece defects. The findings underscored the necessity of conducting thorough laboratory evaluations before releasing cryopreserved semen for commercialization. Despite substantial morphological alterations, the initial semen evaluation data indicated acceptable levels of sperm kinematics, emphasizing the resilience of sperm production to severe morphological changes. This case report serves as a contribution to the understanding of midpiece defects in bull sperm, emphasizing the need for meticulous evaluation and quality control in semen processing and commercialization.

Hipoplasia testicular em Mugil curema / Testicular hypoplasia in Mugil curema

Mugil curema valenciennes, 1836 (Osteichthyes: Mugilidae) é um peixe cosmopolita e é conhecido popularmente como parati no Sudeste e Sul do Brasil e como tainha no Nordeste, sendo encontrado em mar aberto e em estuários. Esta espécie não expressa dimorfismo sexual entre machos e fêmeas externamente. Machos e as fêmeas possuem um par de gônadas (testículos e ovários, respectivamente), as quais são alongadas, fusiformes e simétricas. Este trabalho descreve a ocorrência de hipoplasia testicular em um peixe da espécie Mugil curema, procedente da Baía da Babitonga, Santa Catarina, Brasil. Foi identificada hipoplasia testicular unilateral, a qual macroscopicamente, estava diminuído de volume e, microscopicamente, apresentava atividade espermatogênica ausente e redução do número de espermatozoides. Não foi possível determinar os fatores que possam ter contribuído para o desenvolvimento de tal alteração reprodutiva. Porém, como a espécie animal pode ter suas características de vida e reprodutiva influenciadas pela ação antrópica, é importante um estudo contínuo da vida dos peixes nesse ambiente. Existem poucos estudos direcionados à espécie, principalmente aos aspectos reprodutivos. Desta forma, este relato contribuirá para elevar o histórico de informações a respeito desta espécie na região.

Mugil curema valenciennes, 1836 (Osteichthyes: Mugilidae) is a cosmopolitan fish popularly known as parati in Southeast and South Brazil and as mullet in Northeast, being found in the open sea and estuaries. This species does not express sexual dimorphism between males and females externally. Males and females have a pair of gonads (testes and ovaries, respectively), which are elongated, fusiform, and symmetrical. This paper describes the occurrence of testicular hypoplasia in a fish of the species Mugil curema, from Babitonga Bay, Santa Catarina, Brazil. Unilateral testicular hypoplasia was identified, which macroscopically showed a decrease in testis volume and, microscopically, the absence of spermatogenic action and a reduction in the number of spermatozoa. It was not possible to determine the factors that may have contributed to the development of such reproductive alteration. However, as the living and reproductive traits of this animal species may be influenced by anthropic action, a continuous study of its life in this environment is important. There are few studies directed at this species, mainly on its reproductive aspects. Thus, this report will contribute to raising the information historic about this species in the region.

Scrotal thermoregulation and sequential sperm abnormalities in buffalo bulls (Bubalus bubalis) under short-term heat stress.

Heat stress reduces the reproductive capacity of bulls raised in tropical climate. However, the reestablishment of scrotal thermoregulation and the dynamics of sperm defects emergence after stress are not completely known in buffaloes. Thus, the study aimed to evaluate the effect of short-term heat stress over scrotal thermoregulation and sperm attributes, relating them to spermatogenesis stages. Five buffalo bulls went through scrotal insulation during 48 h (from day 0 to day 2). Semen samples were collected every 7 days (from day -7 to day 49) and analyzed about the progressive motility, viability, and sperm morphology. Heat stress significantly destabilized scrotal thermoregulation (P < 0.001). Scrotal temperature was from 4.2 to 6.3 °C lower than the core body temperature, except on insulation days (P < 0.001), and returned to the basal condition five days after the removal of the stressing stimulus. More significant deleterious effects were observed in sperm morphology than in cell concentration, motility, and viability. The chronology of morphologic defects expression demonstrated tail defects (days 7-14), cytoplasmic droplets (days 14-28), and head defects (day 28), returning to pre-insulation condition 35 days after the thermal challenge. Thus, hyperthermia harmed more intensely spermatozoa in epididymal transit, elongated spermatids, and secondary spermatocytes. It is concluded that water buffalo bulls present a peculiar manifestation of sperm morphology after short-term stress, indicating an important difference related to the bovine species. Therefore, during the andrological evaluation of buffalo bulls, it is necessary to avoid the allometric extrapolation between these species.

Recategorisation of body mass index to achieve andrological predictive power: a study in more than 20 000 patients.

The aim of this study was to recategorise body mass index (BMI) in order to classify patients according to their risk of semen abnormalities. Patients (n=20563) presenting at an andrology laboratory were classified into five groups according to BMI: underweight (BMI <20kg m-2), normal weight (BMI 20-24.9kg m-2), overweight (BMI 25-29.9kg m-2), obese (BMI 30-39.9kg m-2) and morbidly obese (BMI >40kg m-2). Semen quality was evaluated to determine: (1) differences between groups using analysis of variance (ANOVA); (2) the chances of semen abnormalities (using generalised linear models, Chi-squared tests and odds ratios); (3) reference BMI values with andrological predictive power (multivariate conglomerate analyses and multivariate analysis of variance (MANOVA)); and (4) expected values of abnormalities for each new group resulting from BMI recategorisation. Morbidly obese and underweight patients exhibited the highest decrease in semen quality and had higher chances of semen abnormalities. The smallest number of sperm abnormalities was found at a BMI of 27kg m-2. Four reference values were identified, recategorising BMI into four groups according to their risk of semen abnormalities (from lowest to highest risk): Group1,BMI between 20 and 32kg m-2; Group2, BMI <20 and BMI >32-37kg m-2; Group3, BMI >37-42kg m-2; and Group4, BMI >42kg m-2. A BMI <20 or >32kg m-2 is negatively associated with semen quality; these negative associations on semen quality increase from a BMI >37kg m-2 and increase even further for BMI >42kg m-2. The BMI recategorisation in this study has andrological predictive power.

Evaluation of reproductive parameters in male Neotropical bats during dry and rainy months in a specific area of the Cerrado biome.

The aim of this study was to analyse the reproductive aspects of male bats of three common species of the Phyllostomidae family: Artibeus lituratus, Platyrrhinus lineatus and Sturnira lilium, during dry and rainy months in a specific area of the Cerrado biome. Body weight was significantly higher during the dry months for S. lilium. The gonadosomatic index (GSI) and testicular weight were not significantly different between dry and rainy periods. The tubular parameters were significantly bigger in A. lituratus than in the other two species during both periods. No difference in the tubular/interstitial ratio was observed in any of the species during both periods. In both periods, all sperm cells and germ cell developmental stages were visible on seminiferous tubules whereas sperm cells were observed in epididymides of all sampled animals. The percentage of morphologically normal sperm was low (35%-60%), with no difference between periods. Spermatozoa from A. lituratus presented a leaf-shaped head, while the head was round-shaped in the other two species. In conclusion, our data suggest that males from the three studied species did not present reproductive latency during the most critical weather periods (dry and rainy months) in the metropolitan region of Brasilia, Brazil.

Disruption of bovine sperm functions in the presence of aplastic midpiece defect.

BACKGROUND: Bulls are of great importance in the productive chain and for this reason they should have a good semen quality. There is no doubt that sperm morphology is very important to bull fertility, although little is known about how exactly the abnormal morphologies may affect sperm functions. OBJECTIVES: To detail the morphological description of the aplastic midpiece defect (AMD), as well as to understand its consequences for male fertility based on membrane and acrosome status, mitochondrial membrane potential and DNA integrity parameters. MATERIALS AND METHODS: The bulls were divided into two groups: control, consisting of satisfactory potential breeders (n = 3); and AMD, consisting of unsatisfactory potential breeders with a high percentage of AMD (n = 3). Bulls were evaluated by the breeding soundness evaluation; five ejaculates were collected from each animal and analyzed by flow cytometry. RESULTS: Spermatozoa from AMD group exhibited lower sperm motility and vigor (p < 0.05). In addition, it also exhibited lower mitochondrial membrane potential (p < 0.05), a higher percentage of spermatozoa with DNA fragmentation (p < 0.05), lower acrosome and plasma membrane integrity (p < 0.05), and higher lipid bilayer sperm membrane disorganization (p < 0.05) in comparison with control bulls. DISCUSSION: These findings may be due to oxidative stress and a reduction of the energy production capacity in addition to an alteration in the structural composition of the sperm cell. Moreover, semen with a high percentage of AMD may also be undergoing apoptosis. CONCLUSION: Bulls with a high percentage of AMD in their semen are not suitable for reproduction. Furthermore, it suggests there is a putative genetic basis for this sperm defect.

Macrozoospermia associated with mutations of AURKC gene: First case report in Latin America and literature review.

A Chilean 35-year-old male patient with a history of primary infertility made an appointment at the Unit of Reproductive Medicine at Clínica Las Condes, Santiago, Chile. Multiple semen analyses revealed abnormal sperm morphology as the most prevalent finding. Multiflagellated and macrocephalic spermatozoa were observed and indicated a possible macrozoospermic phenotype. The constant presence of abnormal sperm morphology led the scope of the study to include Aurora Kinase C (AURKC) gene sequencing. The patient was diagnosed with a homozygous mutation of this gene. The mutation was detected in exon 6, type c.744C>G+/+ (P.Y248*) variant. As previously described in the Human Gene Mutation Database (HGMD), this pathogenic variant is associated with macrozoospermia. Although this mutation is not the most frequently observed, it is the first of its kind reported in Latin America.

Genetic parameters for fertility traits in Nellore bulls.

Nellore is the main cattle breed used in Brazil, being the largest commercial herd in the world. Beyond the importance of male reproductive efficiency for farm profit, the use of reproductive techniques, mainly artificial insemination, turns the evaluation of male reproductive traits even more important. Estimation of genetic parameters increases the knowledge on traits variances and allows envisaging the possibility of the inclusion of new traits as selection criterion. Genetic parameters for fifteen traits that can be classified as testicular biometry or physical and morphological semen traits were estimated for a Nellore bull population ranging from 18 to 36 months. Single-trait and bi-trait animal models were used for (co)variance components estimation. The contemporary group was considered as fixed effect and age at measurement as covariable. Scrotal circumference presented heritability of 0.47 ± 0.12. This value is similar to the heritabilities found for all testicular biometry traits (0.34-0.48). Sperm progressive motility, which has a direct effect on bull fertility, presented low heritability (0.07 ± 0.08). Major and total sperm defects presented moderate to high heritabilities (0.49 ± 0.18 and 0.39 ± 0.15, respectively), indicating that great genetic gain can be obtained through selection against sperm defects. High and positive genetic correlations were observed among testicular biometry traits, which also presented favourable genetic correlations with physical and morphological traits of the semen with magnitude ranging from high to low. Scrotal circumference presented moderate to high and favourable genetic correlations with sperm progressive motility, sperm turbulence, major sperm defects and total sperm defects. Thus, the selection for scrotal circumference results in favourable correlated genetic response for semen quality. The results show that the use of scrotal circumference as reference trait for bull fertility is appropriate, since it presents high heritability and favourable genetic correlation with semen quality.

Reference values (RV), MNS and MTS for morphological characteristics of normal and abnormal human spermatozoon describing Teratozoospermic Index (TZI) & Sperm Deformity Index (SDI) in Sudanese infertile couples using CASA System / Valores de referencia (RV), MNS y MTS para las características morfológicas de espermatozoides humanos normales y anormales que describen el Índice Teratozoospérmico (TZI) y el Índice de Deformidad del Esperma (SDI) en parejas infértiles de Sudán que utilizan el sistema CASA

A scientific determination of fertilization ability of the sperm can be made by motility, viability and morphological examinations that are called as potential fertility criteria. Some previous studies documented only the total morphological defect rates without including the morphologic subjects. As it is recorded in many studies, the success of IUI, ICSI and IVF depends mainly on the quality of the collected semen. 1) Determine and classify the common morphological defects and rates of sperms. 2) Analyze the effect of age on the sperm abnormalities and to assess the impact of environment on changing normal sperm characteristics. 3) Determine the common types of semen abnormalities (Asthenoterato-zoospermia, Asthenozoospermia, Azoopyospermia, Azoospermia, Hypospermia, Oligoasthenoteratozoospermia, Oligoasthenozoopyospermia, Oligoasthenozoospermia and Teratozoospermia). 4) Determine the range of teratozoospermic index (TZI) and sperm deformity index (SDI). A prospective case series study for 566 men of infertile couples who were requesting semen analysis was applied to those came to the ULTRALAB laboratory center main campus and ULTRALAB branch of the Sudan Assisted Reproduction Center (SARC), Khartoum, Sudan. Data collection was done for 20 months from April 1st, 2010 to December 31st, 2011. analysis of semen samples was done using Computer Assisted Semen Analysis (CASA). 1) The study demonstrates that there are differences in normality and abnormality of sperm morphology parameters of sperm between Sudanese population and other studies including WHO standards. 2) SDI and TZI were higher in overall semen as well as in abnormal semen samples. 3) Common semen abnormalities were oligoasthenoteratozoospermia, asthenoteratozoospermia and asthenozoospermia. There are differences in sperm morphology of both normal and abnormal parameters between Sudanese population and other studies including WHO standards.

Se realizó una determinación científica de la capacidad de fertilización de los espermatozoides mediante motilidad, viabilidad y exámenes morfológicos denominados criterios de fertilidad potencial. Algunos estudios previos documentan solamente el total de las tasas de defectos morfológicos sin incluir a los sujetos morfológicos. Como se registra en muchos estudios, el éxito de IUI, ICSI y FIV depende principalmente de la calidad del semen recolectado. Los objetivos de este trabajo fueron: 1) Determinar y clasificar los defectos morfológicos comunes y las tasas de espermatozoides; 2) Analizar el efecto de la edad en las anomalías de los espermatozoides y evaluar el impacto del medio ambiente en el cambio de las características normales de los espermatozoides; 3) Determinar los tipos comunes de anomalías en el semen (astenoterato-zoospermia, astenozoospermia, azoopiepermia, azoospermia, hipospermia, oligoastenoteratozoospermia, oligoastenozoopermia, oligoastenozoospermia y teratozoospermia); 4) Determinar el rango del índice teratozoospérmico (TZI) y el índice de deformidad del esperma (IDE). Se realizó un estudio prospectivo de series de casos en 566 hombres, de parejas infértiles, que solicitaban análisis de semen en laboratorios ULTRALAB y la sucursal de ULTRALAB del Centro de Reproducción Asistida de Sudán (SARC), Jartum, Sudán. La recolección de datos se realizó durante 20 meses, desde el 1 de abril de 2010 hasta el 31 de diciembre de 2011. El análisis de las muestras de semen se realizó mediante el análisis de semen asistido por computadora (CASA). Existen diferencias en la normalidad y anormalidad de los parámetros de la morfología de los espermatozoides entre la población sudanesa y otros estudios, incluidas las normas de la Organización Mundial de la Salud. Los parámetros SDI y TZI fueron más altos en el semen general, así como en muestras de semen anormales. Las anomalías comunes del semen fueron oligoastenatoatoospermia, astenoteratozoospermia y astenozoospermia. Existen diferencias en la morfología de los espermatozoides de los parámetros normales y anormales entre la población sudanesa y otros estudios que incluyen las normas de la Organización Mundial de la Salud.

Low concentrations of lead decrease the sperm fertilization ability by altering the acrosome reaction in mice.

Lead (Pb) exposure at high concentrations is associated with poor sperm quality, acrosome alterations, and low fertilization rate. Sperm capacitation and the acrosome reaction (AR) are required for successful fertilization. Actin polymerization is crucial for correct capacitation, and small GTPases, such as RhoA, Rac1, and Cdc42, are involved. This study aimed to evaluate the effects of Pb on sperm fertilization ability, capacitation, AR, and the mechanisms involved in mice exposed to low Pb concentrations. CD1 mice were exposed to 0.01% Pb2+ for 45 days through their drinking water and their spermatozoa were collected from the cauda epididymis-vas deferens to evaluate the following: AR (oAR: initial, sAR: spontaneous, and iAR: induced) using the PNA-FITC assay, sperm capacitation (P-Tyr levels), actin polymerization (phalloidin-TRITC), MDA production (stress oxidative marker), the RhoA, Rac1, and Cdc42 protein levels, and the in vitro fertilization (IVF). After the treatment, the blood Pb (PbB) concentration was 9.4 ±â€¯1.6 µg/dL. Abnormal sperm morphology and the oAR increased (8 and 19%, respectively), whereas the iAR decreased (15%) after a calcium ionophore challenge, and the actin polymerization decreased in the sperm heads (59%) and tails (42%). Rac1 was the only Rho protein to significantly decrease (33%). Spermatozoa from the Pb-treated mice showed a significant reduction in the fertilization rate (19%). Our data suggest that Pb exposure at environmental concentrations (PbB < 10 µg/dL) decreases the acrosome function and affects the sperm fertilization ability; this is probably a consequence of the low Rac1 levels, which did not allow adequate actin polymerization to occur.

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)

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.

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.

Validation of the CellRox Deep Red ® fluorescent probe to oxidative stress assessment in equine spermatozoa

Considering the importance of ROS influence on sperm functionality and some limitations in sperm oxidative stress assessment methods, a field to studies of new techniques are still open. In this sense, the aim of this study is to validate the ROS detection technique through the CellRox Deep Red Reagent®probe in stallion sperm. Four stallions were used and the analyses were conducted on four replicates of semen samples from each of stallion (n = 16). The results of the polynomial regression presented a quadratic effect, high determination coefficient value (R2 = 0.88) and high significant P value (P < 0.0001). The CellRox Deep Red® fluorescent probe is able to detect reactive oxygen species in equine sperm, indicating accurately the occurrence of oxidative stress in stallion semen.

Validation of the CellRox Deep Red ® fluorescent probe to oxidative stress assessment in equine spermatozoa

Considering the importance of ROS influence on sperm functionality and some limitations in sperm oxidative stress assessment methods, a field to studies of new techniques are still open. In this sense, the aim of this study is to validate the ROS detection technique through the CellRox Deep Red Reagent®probe in stallion sperm. Four stallions were used and the analyses were conducted on four replicates of semen samples from each of stallion (n = 16). The results of the polynomial regression presented a quadratic effect, high determination coefficient value (R2 = 0.88) and high significant P value (P < 0.0001). The CellRox Deep Red® fluorescent probe is able to detect reactive oxygen species in equine sperm, indicating accurately the occurrence of oxidative stress in stallion semen.(AU)

Morfología espermática: comparación de las observaciones realizadas por 10 expertos / Sperm morphology: comparison of observations made by 10 experts

Introducción: La morfología espermática es uno de los parámetros más importantes que se evalúan durante el análisis seminal, aunque es un parámetro que presentan alta variabilidad inter-laboratorios. Por lo tanto el objetivo de este trabajo fue determinar la variación interindividuo al analizar la morfología espermática observada por 10 expertos. Materiales y métodos: En este estudio descriptivo prospectivo se planteo una encuesta de 133 preguntas, cada una presentaba una fotografía de un espermatozoide con el fin de clasificarlo como normal o anormal, y de ser anormal se debía especificar su anormalidad (cabeza, pieza intermedia, cola y/o gota citoplasmática). Esta encuesta se distribuyo a los laboratorios y personal experto en el tema de evaluación seminal, se recolectaron 10 encuestas entre septiembre y agosto de 2016. Resultados: De las 132 fotografías, 23 (17%) representaban células normales, mientras que 109 (83%) representaban espermatozoides con algún tipo de anormalidad. El coeficiente de variación para el numero de aciertos totales fue de 22.4%, para aciertos normales fue de 66% y para los aciertos anormales del 4.5%. La variación para las células normales totales fue de 42.3%, mientras que la variación para los anormales totales fue solo del 6.5%. Conclusión: Aunque la variabilidad entre los observadores es similar a la reportado en otros estudios, es necesario crear redes de cooperación que permitan estandarizar los procedimientos de determinación de la morfología espermática entre los diferentes laboratorios del país.

Introduction: Sperm morphology is one of the most important parameters that are evaluated during the seminal analysis, although it is a parameter that presents high inter-laboratory variability. Therefore the objective of this study was to determine the interindividual variation when analyzing the sperm morphology observed by 10 experts. Material and Methods: In this prospective descriptive study a survey of 133 questions was presented, each photograph show of a spermatozoon in order to classify it as normal or abnormal, and if abnormal, its abnormality (head, intermediate piece, tail and/or cytoplasmic droplet). This survey was distributed to laboratories and experts in the subject of seminal evaluation, 10 surveys were collected between September and August 2016. Results: Of the 132 photographs, 23 (17%) represented normal sperm, while 109 (83%) represented spermatozoa with some type of abnormality. The coefficient of variation for the number of total hits was 22.4%, for normal hits was 66% and for abnormal hits of 4.5%. The variation for the total normal cells was 42.3%, while the variation for the total abnormal cells was only 6.5%. Conclusion: Although the variability among observers is similar to that reported in other studies, it is necessary to create cooperation networks that allow standardization of the procedures for determining the sperm morphology between the different laboratories in the country.

Could vitamin C and zinc chloride protect the germ cells against sodium arsenite?

Arsenic (As) is commonly associated with natural and human processes such as volcanic emissions, mining and herbicides production, being an important pollutant. Several studies have associated As intake with male fertility reduction, thus the aim of the present study was to evaluate whether vitamin C and/or zinc would counteract As side effects within the testicles. Adult male Wistar rats were divided into six experimental groups: control, sodium arsenite (5 mg/kg/day), vitamin C (100 mg/kg/day), zinc chloride (ZnCl2; 20 mg/kg/day), sodium arsenite + vitamin C and sodium arsenite + ZnCl2. Testicles and epididymis were harvested and either frozen or routinely processed to be embedded in glycol methacrylate resin. As reduced the seminiferous epithelium and tubules diameter due to germ cell loss. In addition, both the round spermatids population and the daily sperm production were reduced. However, ZnCl2 and vitamin C showed to be effective against such side effects, mainly regarding to sperm morphology. Long-term As intake increased the proportions of abnormal sperm, whereas the concomitant intake of As with zinc or vitamin C enhanced the proportions of normal sperm, showing that such compounds could be used to protect this cell type against morphological defects.

Head morphometry and chromatin instability in normal boar spermatozoa and in spermatozoa with cytoplasmic droplets

In boar studs, morphological analyses are used to evaluate sperm quality and there by categorize ejaculates as either approved or rejected. Normally, morphological characteristics correlate with chromatin disorders, but studies to date have only considered the average of abnormalities; cells were not segregated as normal or abnormal. The aim of this study was to assess whether the presence of cytoplasmic droplets was associated with morphometric characteristics and chromatin instability of spermatozoa heads. Morphological analyses were performed on semen from 11 boars using phase contrast microscopy (200 cells per sample). Normal cells were differentiated from those with cytoplasmic droplets and both types were evaluated separately. Photomicrographs were acquired ofnormal spermatozoa (Group NOR, N = 1,207) as well as spermatozoa with proximal and distal cytoplasmic droplets (Group DROP, N = 725). Sperm-head morphometry and chromatin structure were evaluated using the toluidine blue technique. Spermatozoa heads in the DROP group were longer (8.37 ± 0.60 × 8.31 ± 0.53; P = 0.025), narrower (4.16 ± 0.21 × 4.19 ± 0.19; P = 0.03), And more symmetric on the sides (0.973 ± 0.012 × 0.971 ± 0.011; P = 0.007) than were spermatozoa heads of the NOR group. The DROP group also had a greater average ellipticity (0.335 ± 0.034 × 0.329 ± 0.031; P= 0.0004),a greater percentage of decondensed chromatin (2.71 ± 3.87 × 2.28 ± 1.38; P < 0.0008), and a greater chromatin heterogeneity (4.66 ± 1.40 × 4.40 ± 1.42;P < 0.0001). A greater frequency of semen collection results in a shorter period of cell maturation and this probably affected the degree of chromatin condensation and the cytoplasmic droplet migration, with concomitant effect onthe head morphometry measurements.(AU)

Head morphometry and chromatin instability in normal boar spermatozoa and in spermatozoa with cytoplasmic droplets

In boar studs, morphological analyses are used to evaluate sperm quality and there by categorize ejaculates as either approved or rejected. Normally, morphological characteristics correlate with chromatin disorders, but studies to date have only considered the average of abnormalities; cells were not segregated as normal or abnormal. The aim of this study was to assess whether the presence of cytoplasmic droplets was associated with morphometric characteristics and chromatin instability of spermatozoa heads. Morphological analyses were performed on semen from 11 boars using phase contrast microscopy (200 cells per sample). Normal cells were differentiated from those with cytoplasmic droplets and both types were evaluated separately. Photomicrographs were acquired ofnormal spermatozoa (Group NOR, N = 1,207) as well as spermatozoa with proximal and distal cytoplasmic droplets (Group DROP, N = 725). Sperm-head morphometry and chromatin structure were evaluated using the toluidine blue technique. Spermatozoa heads in the DROP group were longer (8.37 ± 0.60 × 8.31 ± 0.53; P = 0.025), narrower (4.16 ± 0.21 × 4.19 ± 0.19; P = 0.03), And more symmetric on the sides (0.973 ± 0.012 × 0.971 ± 0.011; P = 0.007) than were spermatozoa heads of the NOR group. The DROP group also had a greater average ellipticity (0.335 ± 0.034 × 0.329 ± 0.031; P= 0.0004),a greater percentage of decondensed chromatin (2.71 ± 3.87 × 2.28 ± 1.38; P < 0.0008), and a greater chromatin heterogeneity (4.66 ± 1.40 × 4.40 ± 1.42;P < 0.0001). A greater frequency of semen collection results in a shorter period of cell maturation and this probably affected the degree of chromatin condensation and the cytoplasmic droplet migration, with concomitant effect onthe head morphometry measurements.

Leucocitos seminales y calidad espermática de hombres en estudio de infertilidad / Seminal leukocytes and sperm quality in men involved in infertility study

Introducción: los leucocitos en exceso en el semen humano pueden estar asociados a la disminución de la calidad espermática. La elevada incidencia de leucocitos en el semen de hombres que acuden a nuestro servicio, pudiera estar vinculada al deterioro de las variables de calidad espermática, y vincularse con problemas de infertilidad masculina. Objetivo: determinar la frecuencia de leucocitospermia en el semen de hombres que consultan por infertilidad, e identificar si existe asociación entre la presencia de leucocitospermia y alteraciones en las variables de calidad del semen. Métodos: se realizó un estudio descriptivo transversal, en el cual se incluyeron 136 hombres, con edades entre 20 y 45 años, que acudieron al examen de calidad del semen, procedentes de las consultas de atención a parejas en estudio de infertilidad, del Instituto Nacional de Endocrinología, en La Habana, y que no refirieron síntomas de infección genitourinaria clínicamente activa, ni causas demostrables que afectaran la calidad seminal. El análisis del semen, para determinar las variables seminales, se realizó siguiendo los lineamientos de la OMS, incluyendo la técnica de peroxidasa para determinar la concentración de leucocitos seminales. Se consideraron los aspectos éticos en la investigación y los resultados se analizaron mediante la aplicación de la prueba de chi cuadrado. Un valor de p< 0,05 se consideró estadísticamente significativo.Resultados: de las 136 muestras de semen evaluadas, se encontraron leucocitospermia en 31 (22,7 por ciento). Todos los indicadores de calidad seminal fueron menores en presencia de leucocitospermia, aunque solo se obtuvo una diferencia significativa en relación con la concentración espermática (p< 0,05). Conclusiones: la frecuencia de leucocitospermia en la muestra estudiada es prevalente y asociada con un deterioro estadísticamente significativo de la concentración espermática(AU)

Introduction: excessive amount of leukocytes in the human semen can be related to lower sperm quality. The high incidence of leukocytes in semen from males who went to our service could be linked to deterioration of the sperm quality variables and to male infertility problems.Objective: to determine the frequency of leukocytospermia in the semen of men who visited our infertility service and to identify the possible association between the leukocytospermia and altered quality variables of semen. Methods: a cross-sectional and descriptive study of 136 men aged 20 to 45 years, who were performed the semen quality exam and came from the service provided to couples under infertility study in the National Institute of Endocrinology in Havana. These men had not stated any symptom of clinically active urogenital infection and no proven causes that might affect the semen quality. The semen analysis to determine the seminal variables complied with the WHO guidelines including the peroxidase technique to estimate the seminal leukocyte concentrations. The ethical aspects of research were respected whereas the Chi-square test served to analyze the results; the p< 0.05 was considered as statistically significant. Results: of 136 evaluated semen samples, leukocytospermia was found in 31 (22.7 percent). All the seminal quality indicators were lower in leukocytospermia, although the significant difference was only found in the sperm concentration (p< 0.05). Conclusions: the frequency of leukocytospermia in the studied sample is prevalent and associated to statistically significant deterioration of the sperm concentration(AU)

Leucocitos seminales y calidad espermática de hombres en estudio de infertilidad / Seminal leukocytes and sperm quality in men involved in infertility study

Introducción: los leucocitos en exceso en el semen humano pueden estar asociados a la disminución de la calidad espermática. La elevada incidencia de leucocitos en el semen de hombres que acuden a nuestro servicio, pudiera estar vinculada al deterioro de las variables de calidad espermática, y vincularse con problemas de infertilidad masculina. Objetivo: determinar la frecuencia de leucocitospermia en el semen de hombres que consultan por infertilidad, e identificar si existe asociación entre la presencia de leucocitospermia y alteraciones en las variables de calidad del semen. Métodos: se realizó un estudio descriptivo transversal, en el cual se incluyeron 136 hombres, con edades entre 20 y 45 años, que acudieron al examen de calidad del semen, procedentes de las consultas de atención a parejas en estudio de infertilidad, del Instituto Nacional de Endocrinología, en La Habana, y que no refirieron síntomas de infección genitourinaria clínicamente activa, ni causas demostrables que afectaran la calidad seminal. El análisis del semen, para determinar las variables seminales, se realizó siguiendo los lineamientos de la OMS, incluyendo la técnica de peroxidasa para determinar la concentración de leucocitos seminales. Se consideraron los aspectos éticos en la investigación y los resultados se analizaron mediante la aplicación de la prueba de chi cuadrado. Un valor de p< 0,05 se consideró estadísticamente significativo. Resultados: de las 136 muestras de semen evaluadas, se encontraron leucocitospermia en 31 (22,7 por ciento). Todos los indicadores de calidad seminal fueron menores en presencia de leucocitospermia, aunque solo se obtuvo una diferencia significativa en relación con la concentración espermática (p< 0,05). Conclusiones: la frecuencia de leucocitospermia en la muestra estudiada es prevalente y asociada con un deterioro estadísticamente significativo de la concentración espermática(AU)

Introduction: excessive amount of leukocytes in the human semen can be related to lower sperm quality. The high incidence of leukocytes in semen from males who went to our service could be linked to deterioration of the sperm quality variables and to male infertility problems. Objective: to determine the frequency of leukocytospermia in the semen of men who visited our infertility service and to identify the possible association between the leukocytospermia and altered quality variables of semen. Methods: a cross-sectional and descriptive study of 136 men aged 20 to 45 years, who were performed the semen quality exam and came from the service provided to couples under infertility study in the National Institute of Endocrinology in Havana. These men had not stated any symptom of clinically active urogenital infection and no proven causes that might affect the semen quality. The semen analysis to determine the seminal variables complied with the WHO guidelines including the peroxidase technique to estimate the seminal leukocyte concentrations. The ethical aspects of research were respected whereas the Chi-square test served to analyze the results; the p< 0.05 was considered as statistically significant. Results: of 136 evaluated semen samples, leukocytospermia was found in 31 (22.7 percent). All the seminal quality indicators were lower in leukocytospermia, although the significant difference was only found in the sperm concentration (p< 0.05). Conclusions: the frequency of leukocytospermia in the studied sample is prevalent and associated to statistically significant deterioration of the sperm concentration(AU)