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1.
Asian Journal of Andrology ; (6): 479-483, 2021.
Article in English | WPRIM | ID: wpr-888459

ABSTRACT

The novel coronavirus disease (COVID-19) pandemic is emerging as a global health threat and shows a higher risk for men than women. Thus far, the studies on andrological consequences of COVID-19 are limited. To ascertain the consequences of COVID-19 on sperm parameters after recovery, we recruited 41 reproductive-aged male patients who had recovered from COVID-19, and analyzed their semen parameters and serum sex hormones at a median time of 56 days after hospital discharge. For longitudinal analysis, a second sampling was obtained from 22 of the 41 patients after a median time interval of 29 days from first sampling. Compared with controls who had not suffered from COVID-19, the total sperm count, sperm concentration, and percentages of motile and progressively motile spermatozoa in the patients were significantly lower at first sampling, while sperm vitality and morphology were not affected. The total sperm count, sperm concentration, and number of motile spermatozoa per ejaculate were significantly increased and the percentage of morphologically abnormal sperm was reduced at the second sampling compared with those at first in the 22 patients examined. Though there were higher prolactin and lower progesterone levels in patients at first sampling than those in controls, no significant alterations were detected for any sex hormones examined over time following COVID-19 recovery in the 22 patients. Although it should be interpreted carefully, these findings indicate an adverse but potentially reversible consequence of COVID-19 on sperm quality.


Subject(s)
Adult , Asthenozoospermia/virology , COVID-19/physiopathology , China , Gonadal Steroid Hormones/blood , Humans , Male , Progesterone/blood , Prolactin/blood , SARS-CoV-2 , Semen/physiology , Semen Analysis , Sperm Count , Sperm Motility , Spermatozoa/physiology , Time Factors
2.
Pesqui. vet. bras ; 40(12): 1048-1053, Dec. 2020. tab
Article in English | LILACS, VETINDEX | ID: biblio-1155032

ABSTRACT

Awareness of the physiological changes that occur when animals are subjected to climatic changes that are considered stressful is essential to maintain animal welfare and to be able to exploit their reproductive potential efficiently and rationally. The present study was carried out to evaluate climatic variables' influence on physiological parameters, and Murrah buffalo ejaculates reared in a humid tropical climate in the Amazon. The immediate analyzes pertinent to the physical and morphological characteristics of the ejaculates were carried out and corresponded in the rainy season (RS) volume of 3.4±2.0mL; the mass activity of 4.4±0.5; motility of 80.4±5.6%; vigor of 4.4±0.4; concentration of 657,300±237,865.1 x 106sptz/mL; major defects of 9.0±2.6%; minor defects of 11.2±3.9%; total defects 20.2±5.3% and sperm plasma membrane integrity (SPMI) 84.8±5.6%, whereas in the non-rainy season (nRS), the results were 4.0±2.1mL; the mass activity of 3.0±1.0; motility of 56.2±13.4%; vigor of 3.0±1.0; concentration of 586,000±291,925.9 x 106sptz/mL; major defects of 20.8±9.9%; minor defects of 27.5±6.3%; total defects 48.3±9.3% and SPMI of 57.9±12.4%. Furthermore, a statistical difference (P<0.05) was observed for the parameters mass activity, motility, vigor, major defects, minor defects, total defects, and sperm plasma membrane integrity between both periods. The data on heart frequency, superficial temperature (head, back, groin, and scrotal pouch) showed a statistical difference between both periods (P<0.05). To conclude is necessary specific management in the non-rainy season that thermal stress is not a determining factor in reducing the reproductive quality of buffaloes; it is necessary to use means to improve animal welfare; one alternative is to use baths regularly for these animals or provide constant access to areas of rivers or lakes, as well as shading, preventing the buffaloes from being directly exposed to the unfavorable thermal environment.(AU)


O conhecimento das alterações fisiológicas que ocorrem quando os animais são submetidos a alterações climatológicas consideradas estressantes é fundamental para manter o bem-estar animal, e poder explorar o seu potencial reprodutivo de forma eficiente e racional. O presente estudo foi realizado com o objetivo de avaliar a influência de variáveis climáticas sobre parâmetros fisiológicos e de ejaculados de búfalos, da raça Murrah, criados em clima tropical úmido da Amazônia. As análises imediatas pertinentes às características físicas e morfológicas dos ejaculados foram realizadas e corresponderam no período chuvoso (PCh) o volume de 3,4±2,0mL, turbilhonamento de 4,4±0,5; motilidade de 80,4±5,6%; vigor de 4,4±0,4; concentração de 657.300±237.865,1 x 106sptz/mL; defeitos maiores de 9,0±2,6%; defeitos menores de 11,2±3,9%; defeitos totais de 20,2±5,3% e integridade da membrana plasmática (IMP) de 84,8±5,6%, enquanto que no período não chuvoso (PnCh), os resultados foram de 4,0±2,1mL; turbilhonamento de 3,0±1,0; motilidade de 56,2±13,4%; vigor de 3,0±1,0; concentração de 586.000±291.925,9 x 106sptz/mL; defeitos maiores de 20,8±9,9%; defeitos menores de 27,5±6,3%; defeitos totais de 48,3±9,3% e IMP de 57,9±12,4%. Observou-se diferença estatística (P<0,05) para os parâmetros movimento de massa, motilidade, vigor, defeitos maiores, defeitos menores, defeitos totais e integridade da membrana plasmática entre os dois períodos. Dados de frequência cardíaca, temperatura superficial (cabeça, dorso, virilha e bolsa escrotal) diferiram estatisticamente entre os períodos (P<0,05). Conclui-se que se faz necessário usar de um manejo específico no período não chuvoso para que o estresse térmico não seja um fator determinante na redução da qualidade reprodutiva dos búfalos, para isto se faz necessário utilizar de meios para melhorar o bem-estar animal, sendo uma das alternativas fazer uso de banhos regularmente para estes animais, ou disponibilizar acesso constante destes a áreas de rios ou lagos, assim como sombreamentos, evitando que os búfalos fiquem expostos diretamente ao ambiente térmico desfavorável.(AU)


Subject(s)
Spermatozoa/physiology , Buffaloes/physiology , Semen Analysis/veterinary , Animal Welfare , Reference Parameters
3.
Arq. bras. med. vet. zootec. (Online) ; 72(5): 1727-1730, Sept.-Oct. 2020. tab
Article in English | LILACS, VETINDEX | ID: biblio-1131576

ABSTRACT

There is a need for various anesthetic agents to obtain sperm in the field of human and veterinary medicine. Propofol and midazolam are among the most preferred among these agents. The aim of this study was to determine how sperm paramaters are affected according to the anesthetic agent used. Propofol (2mg/kg) and midazolam (3,5-7,5mg/kg) were administered twice a day (morning-evening) for one week. As a result of this study, there was no statistical difference in sperm density and abnormal sperm rates (respectively P=0,673, P=0,479). Sperm motility rates are similar in the control and propofol groups, while the motility rate in the midazolam group is statistically lower. (Control group %85 - Midazolam group %68.75 - Propofol group %83.75), (P<0.05). As a result of this study, the confidence interval of propofol was higher than the other anesthetic agents used for sperm retrieval.(AU)


São necessários vários agentes anestésicos para obter espermatozoides no campo da medicina humana e veterinária. Propofol e midazolam estão entre os agentes preferidos. O objetivo deste estudo foi determinar como os parâmetros de esperma são afetados de acordo com o agente anestésico utilizado. Propofol (2mg / kg) e midazolam (3,5-7,5mg / kg) foram administrados duas vezes ao dia (manhã e noite) durante uma semana. Neste estudo, não houve diferença estatística na densidade espermática e nas taxas anormais de espermatozoides (respectivamente P = 0,673, P = 0,479). As taxas de motilidade espermática são semelhantes nos grupos controle e propofol, enquanto a taxa de motilidade no grupo midazolam é estatisticamente menor. (Grupo controle % 85 - grupo midazolam % 68,75 - grupo propofol % 83,75), (P <0,05). Neste estudo, o intervalo de confiança do propofol foi maior do que os outros agentes anestésicos utilizados na recuperação espermática.(AU)


Subject(s)
Animals , Male , Rats , Semen/chemistry , Spermatozoa/physiology , Midazolam/administration & dosage , Propofol/administration & dosage
4.
Rev. chil. obstet. ginecol. (En línea) ; 85(4): 312-323, ago. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1138627

ABSTRACT

OBJETIVO: El objetivo de este trabajo fue establecer el efecto de la borra de café sobre la movilidad y los parámetros funcionales de los espermatozoides humanos in vitro. MATERIALES Y MÉTODOS: La borra de café, un subproducto obtenido en establecimientos especializados en la preparación de café soluble a base de grano, se diluyo en tampón fosfato salino y se mezcló en proporciones iguales con las muestras de semen de 16 voluntarios aparentemente sanos. A cada muestra se le determinó el efecto sobre la movilidad espermática en función del tiempo (30, 60, 90 y 120 minutos, n=16) y sobre los parámetros funcionales (n=6) por medio de citometría de flujo: potencial de membrana mitocondrial, producción de especies reactivas de oxígeno y lipoperoxidación de la membrana espermática. RESULTADOS: La incubación de los espermatozoides con la borra de café evidencio un cambio positivo en la movilidad espermática. Adicionalmente, la incubación con la borra de café incremento significativamente el potencial de membrana mitocondrial en los espermatozoides. CONCLUSIÓN: La borra de café, seguramente debido a los compuestos antioxidantes, afecta positivamente la movilidad espermática aumentando el potencial de membrana mitocondrial. Por lo tanto, esto es un paso inicial en la búsqueda de un suplemento de origen natural que aumente la calidad seminal.


OBJECTIVE: The objective of this work is to establish the effect of spent coffee grounds on the motility and functional parameters of human spermatozoa, in vitro. MATERIALS AND METHODS: Spent coffee grounds, a by-product obtained in specialized establishments in the preparation of soluble coffee based on grain, was diluted in saline phosphate buffer and mixed in equal proportions with semen samples from 16 apparently healthy volunteers. Each sample was determined the effect on sperm motility as a function of time (30, 60, 90 and 120 minutes, n=16) and on functional parameters (n=6) by means of flow cytometry: mitochondrial membrane potential, reactive oxygen species production and membrane lipoperoxidation. RESULTS: The incubation of the spermatozoa with the spent coffee grounds showed a positive change in sperm motility. Additionally, incubation with spent coffee grounds significantly increased the mitochondrial membrane potential in human sperm cells. CONCLUSION: Spent coffee grounds, probably due to antioxidant compounds, positively affects sperm motility by increasing mitochondrial membrane potential. Therefore, this is an initial step in the search for a supplement of natural origin that increases seminal quality.


Subject(s)
Humans , Male , Adult , Young Adult , Semen/drug effects , Sperm Motility/drug effects , Spermatozoa/drug effects , Plant Extracts/pharmacology , Coffee/chemistry , Semen/physiology , Sperm Motility/physiology , Spermatozoa/physiology , Time Factors , In Vitro Techniques
5.
Arq. bras. med. vet. zootec. (Online) ; 72(4): 1163-1171, July-Aug. 2020. tab
Article in Portuguese | LILACS, VETINDEX | ID: biblio-1131502

ABSTRACT

Objetivou-se, no primeiro experimento, avaliar o efeito da velocidade de captura de imagens de 25Hz, 30Hz e 50Hz na cinética dos espermatozoides equinos criopreservados. Todas as velocidades mostraram-se adequadas para capturar o movimento espermático (P>0,05). No segundo experimento, objetivou-se avaliar o efeito da deposição de sêmen em lâmina sob lamínula, Leja®10 e 20, na cinética espermática. O uso de lâmina e lamínula foi superior às lejas para manter a LIN e o WOB (P<0,05). No terceiro experimento, objetivou-se avaliar o efeito das concentrações de 25, 50 e 100x106 na cinética espermática. As concentrações de 25 e 50 x106 foram superiores a 100x106 para preservar a LIN, a STR e a BCF e não afetar negativamente a motilidade (P<0,05). No quarto experimento, objetivou-se avaliar o efeito dos diluidores BotuCrio®, BotuSêmen®, TALP sperm e da solução fisiológica na cinética espermática. O BotuCrio® foi superior a todos os diluidores em preservar a BCF e os hiperativos (P<0,05). Conclui-se que o emprego da velocidade de captura entre 25 e 50Hz, a deposição do sêmen entre lâmina e lamínula e a rediluição em diluidor de congelação para atingir 25 a 50x106 de espermatozoides/mL são ideais para o SCA® avaliar, de forma fidedigna, o sêmen equino criopreservado.(AU)


The objective of the first experiment was to evaluate the effect of 25, 30 and 50Hz frame acquisition rate on equine cryopreserved sperm. All frame acquisition rates tested were adequate to capture the sperm movement (P>0.05). The aim of the second experiment was to evaluate the effect of chambers, slide-coverslip, Leja®10 and 20 on sperm movement. The use of slide-coverslip was superior to maintain LIN and WOB (P<0.05). The aim of the third experiment was to evaluate the effect of 25, 50 and 100x106 sperm/mL concentration on sperm movement. Concentrations of 25 and 50x106 sperm/mL were greater than 100x106 to preserve LIN, STR and BCF and did not adversely affect motility (P<0.05). The aim of the fourth experiment was to evaluate the effect of BotuCrio®, BotuSêmen®, TALP sperm and physiological solution on sperm movement. BotuCrio® was superior among other extenders in preserving BCF and hyperactive (P<0.05). It is concluded that the use of the frame acquisition rate between 25 and 50 Hz; the deposition of semen between slide and coverslip and new dilution in the freezing extender to 25-50x106 of sperm/mL is ideal to reliably evaluate cryopreserved equine semen by SCA®.(AU)


Subject(s)
Animals , Male , Sperm Motility , Spermatozoa/physiology , Image Processing, Computer-Assisted/methods , Semen Analysis/veterinary , Horses/physiology , Cryopreservation/veterinary
6.
Int. braz. j. urol ; 46(1): 116-123, Jan.-Feb. 2020. graf
Article in English | LILACS | ID: biblio-1056368

ABSTRACT

CASE STUDY 40-year-old male patient and 32-year-old female partner, with a history of primary infertility of two years duration. The workup revealed idiopathic mild oligoasthenotheratozoospermia, and no apparent female infertility factors. The couple has failed three intrauterine insemination (IUI) cycles, planning more IUI cycles but also considering in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI).


Subject(s)
Humans , Male , Spermatozoa/pathology , Oxidative Stress , Sperm Injections, Intracytoplasmic/methods , Oligospermia/pathology , Spermatozoa/physiology , Reproducibility of Results , Semen Analysis/methods , Fertilization/physiology
7.
Article in English | WPRIM | ID: wpr-878294

ABSTRACT

Objective@#This research was performed to evaluate the effect of tebuconazole (TBZ) on reproductive organs of male rats and to assess the protective role of combined essential trace elements in alleviating the detrimental effect of TBZ on male reproductive function.@*Methods@#For this purpose, 48 rats were exposed to 100 mg/kg TBZ, TBZ supplemented with zinc (Zn), selenium (Se), copper (Cu), and iron (Fe), TBZ + (Se + Zn); TBZ + Cu; or TBZ + Fe. The experiment was conducted for 30 consecutive days.@*Results@#TBZ caused a significant perturbation in mineral levels and reduction in reproductive organs weights, plasma testosterone level, and testicular antioxidant enzyme activities. The TBZ-treated group also showed a significant increase in sperm abnormalities (count, motility, and viability percent), plasma follicle-stimulating hormone and luteinizing hormone concentrations, lipid peroxidation, protein oxidation, and severe DNA degradation in comparison with the controls. Histopathologically, TBZ caused testis impairments. Conversely, treatment with trace elements, in combination or alone, improved the reproductive organ weights, sperm characteristics, TBZ-induced toxicity, and histopathological modifications in testis.@*Conclusion@#TBZ exerts significant harmful effects on male reproductive system. The concurrent administration of trace elements reduces testis dysfunction, fertility, and toxicity induced by TBZ.


Subject(s)
Animal Feed/analysis , Animals , Antioxidants/metabolism , Diet , Dietary Supplements/analysis , Fungicides, Industrial/adverse effects , Male , Minerals/metabolism , Mutagenicity Tests , Rats , Rats, Wistar , Spermatozoa/physiology , Testis/physiology , Trace Elements/metabolism , Triazoles/adverse effects
8.
Braz. j. biol ; 79(3): 438-445, July-Sept. 2019. tab, graf
Article in English | LILACS | ID: biblio-1001465

ABSTRACT

Abstract In this work, the seminal parameters of P. mesopotamicus were evaluated fresh and after cryopreservation, focusing on the sperm variables that affect the rates of fertilization, hatching and post-hatching parameters such as larval survival and morphology. The semen and oocytes from the animals were collected after extrusion, and seminal quality and oocyte fertilization were analyzed. Subsequently, a portion of each semen sample was cryopreserved and, after two days, the oocytes from three new females were fertilized with cryopreserved semen from the males. The analyzes showed that progressive motility, spermatic vigor, motility duration, number of normal sperm and secondary abnormalities were higher in fresh semen than in semen after thawing (P <0.0001). Similarly, fertilization and hatching rates and the percentage of normal and abnormal larvae in fertilized oocytes were higher when fresh semen was used (P <0.0001). The cryopreservation process affected the qualitative parameters of the semen of Piaractus mesopotamicus. The primary abnormality of spermatozoa was the main variable that influenced both fertilization and hatching rates, both in fresh and thawed semen. The second most important variable that influenced, particularly, thawed semen, was the spermatic vigor.


Resumo Neste trabalho, os parâmetros seminais de P. mesopotamicus foram avaliados fresco e após criopreservação, com foco nas variáveis espermáticas ​​que afetam as taxas de fertilização, eclosão e os parâmetros pós-eclosão como a sobrevivência e a morfologia das larvas. Os espermatozoides e os ovócitos dos animais foram coletados após a extrusão, e a qualidade seminal e a fertilização dos ovócitos foram analisados. Posteriormente, uma porção de cada amostra de semen foi criopreservada e, após dois dias, os ovócitos de três novas fêmeas foram fertilizados com semen criopreservado dos machos. As análises mostraram que a motilidade progressiva, o vigor espermático, a duração da motilidade, o número de espermatozoides normais e anormalidades secundárias foram maiores no semen fresco do que no semen após descongelamento (P <0,0001). Da mesma forma, as taxas de fertilização e eclosão e a porcentagem de larvas normais e anormais em ovócitos fertilizados foram maiores quando o semen fresco foi utilizado (P <0,0001). O processo de criopreservação afetou os parâmetros qualitativos do sêmen de Piaractus mesopotamicus . A anormalidade primária dos espermatozoides foi a principal variável que influenciou tanto a taxa de fertilização como a de eclosão, tanto no semen fresco como no semen descongelado. A segunda variável mais importante que influenciou, particularmente, o semen descongelado, foi o vigor espermático.


Subject(s)
Animals , Male , Female , Reproduction , Spermatozoa/physiology , Characiformes/physiology , Cryopreservation/veterinary , Fertilization
9.
Int. braz. j. urol ; 45(4): 815-824, July-Aug. 2019. tab, graf
Article in English | LILACS | ID: biblio-1019894

ABSTRACT

ABSTRACT Introduction Chronic hyperglycemia is caused by diabetes mellitus-committed genital morphophysiology, and oxidative stress is one of the main factors involved in this process. Alpha lipoic acid (ALA) can prevent metabolic and morphological changes in diabetic individuals. Objectives In present study, we evaluated the effects of regular ALA consumption on the spermatogenesis and histoarchitecture in the male genital system of diabetic rats. Materials and Methods Thirty-two Wistar rats were divided into groups: Control (CG); Diabetic Control (DCG), receiving commercial diet: ALA Group (ALAG) and Diabetic ALA Group (DALAG), fed diets with added ALA (300 mg/Kg bw). The diabetic groups received a single injection of streptozotocin (60 mg/kg). After sixty days of the diet, the animals were euthanized, and semen, testis and epididymis samples were collected. A histomorphometric analysis was performed to determine the epithelial height, tubular and luminal diameter, tubular and luminal area of seminiferous tubules and each epididymal region. Sertoli cells were evidenced using the antivimenti antibody and were quantified. The results were statistically analyzed by the ANOVA test. Results At the end of the experiment, the DALAG glycemia was significantly lower than DCG. The histomorphometric parameters of the seminiferous and epididymal tubules did not show improvement in the DALAG. However, there was an improvement in the DALAG in terms of the concentration, motility and percentage of spermatic pathologies, as well as in the number of Sertoli cells (p<0.001). Conclusions The results demonstrated that supplementation with the ALA antioxidant retards testicular lesions and preserve the process of spermatogenesis in diabetes.


Subject(s)
Animals , Male , Spermatozoa/drug effects , Testis/drug effects , Thioctic Acid/pharmacology , Diabetes Mellitus, Experimental/pathology , Epididymis/drug effects , Antioxidants/pharmacology , Sertoli Cells , Sperm Count , Sperm Motility/drug effects , Spermatogenesis/drug effects , Spermatogenesis/physiology , Spermatozoa/physiology , Testis/physiopathology , Testis/pathology , Immunohistochemistry , Random Allocation , Reproducibility of Results , Rats, Wistar , Diabetes Mellitus, Experimental/physiopathology , Epididymis/pathology
10.
Braz. j. biol ; 79(1): 144-148, Jan.-Mar 2019. graf
Article in English | LILACS | ID: biblio-984000

ABSTRACT

Abstract Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) is a potential vector of Chagas's disease and the comprehension of its reproductive biology is an important tool to control this insect. In the reproductive tract of female insects, the spermatheca plays a crucial role storing male spermatozoa after mating. Whithin insects the spermatheca shows a wide morphological diversity and the analysis of this characteristic can contribute to understand the reproductive biology of the species. This study describes the histology and histochemistry of the spermatheca of T. lecticularia. Females have a pair of elongated spermathecal reservoirs without associated accessory gland. The reservoir opens into the common oviduct via a narrow muscular duct. The reservoir epithelium has single layer of columnar secretory cells. The control of the release of spermatozoa from the spermatheca occurs via the muscular duct. The anatomical features of the spermatheca of T. lecticularia resemble those described of other Reduviidae. However, the histological and histochemical features of spermatheca observed in T. lecticularia were important to explain the maintenance of the viability of the spermatozoa stored.


Resumo Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) é um potencial vetor da doença de Chagas e a compreensão de sua biologia reprodutiva é um importante fator para seu controle populacional. No aparelho reprodutor feminino dos insetos, a espermateca desempenha a importante funcão de armazenar os espermatozoides após cópula. Nos insetos, a espermateca apresenta uma ampla diversidade morfológica e a análise destas características pode contribuir com o entendimento da biologia reprodutiva das espécies. Este estudo descreve histológica e histoquimicamente a espermateca de T. lecticularia. As fêmeas tem um par de espermatecas alongadas sem glândulas acessórias associadas. O reservatório conecta-se ao oviduto comum através de um ducto muscular curto que controla a liberação dos espermatozoides. O epitélio do reservatório possui uma camada de células secretoras colunares. As características anatômicas da espermateca de T. lecticularia são semelhantes às encontradas em outros Reduviidae. Entretanto, as características histológicas e histoquímicas observadas na espermateca são importantes para explicar a manutenção da viabilidade dos espermatozoides armazenados.


Subject(s)
Animals , Male , Female , Triatoma/physiology , Reproduction , Spermatozoa/cytology , Spermatozoa/physiology , Triatoma/cytology
11.
Int. braz. j. urol ; 44(1): 172-179, Jan.-Feb. 2018. tab, graf
Article in English | LILACS | ID: biblio-892958

ABSTRACT

ABSTRACT Purpose To assess the impact of sperm retrieval on the gonadal function of rats with impaired spermatogenesis by comparing testicular sperm extraction (TESE) to aspiration (TESA). The efficacy of these procedures to sperm obtainment was also compared. Materials and Methods A pilot study showed impaired spermatogenesis, but normal testosterone (T) production after a bilateral orchidopexy applied to 26 rats, which were randomly assigned into four groups: TESE (n=7), TESA (n=7), SHAM (n=6) and Control (n=6). The T levels were measured through comparative analysis after the orchidopexy. Results There was no statistical difference in the animal's baseline T levels after orchidopexy in comparison to the controls: the TESE and TESA groups, 6.66±4.67ng/mL; the SHAM group (orchidopexy only), 4.99±1.96ng/mL; and the Control, 4.75±1.45ng/mL, p=0.27. Accordingly, no difference was found in the postoperative T levels: TESE, 5.35±4.65ng/mL; TESA, 3.96±0.80ng/mL; SHAM, 3.70±1.27ng/mL; p=0.4. The number of sperm cells found through TESE (41.0±7.0) was significantly larger than that found through TESA (21.3±8.1, p=0.001). Moreover, higher tissue weight was found through TESE (0.09±0.02g versus 0.04±0.04g, p=0.04). Conclusions The testicular sperm capture performed in rats through extraction or aspiration, after orchidopexy, did not significantly decrease the T levels. The amount of sperm found through testicular sperm extraction was higher than that through testicular sperm aspiration.


Subject(s)
Animals , Male , Rats , Sperm Motility/physiology , Spermatogenesis/physiology , Spermatozoa/physiology , Testis/physiology , Sperm Retrieval/adverse effects , Testis/surgery , Testosterone/biosynthesis , Random Allocation , Pilot Projects , Rats, Wistar , Models, Animal , Orchiopexy/methods
12.
Int. j. morphol ; 35(4): 1337-1341, Dec. 2017. tab
Article in Spanish | LILACS | ID: biblio-893138

ABSTRACT

RESUMEN: El objetivo fue comparar la tasa de división (TD) y desarrollo embrionario (DE) con semen sexado X (SX) en FIV capacitado con Percoll vs. BO, y evaluar el efecto de dos concentraciones (5x106 vs. 10x106 espermatozoides/ml), capacitado con BO, comparado con el semen no-sexado (NS). Un avance importante es predeterminar el sexo en bovinos, es posible obtener mayor proporción de terneras a partir del citómetro de flujo con la capacidad de seleccionar los espermatozoides X por la diferencia del ADN (4 %, bovinos), con confiabilidad del 90 %. El SX aumenta la eficiencia reproductiva, permite la selección de hembras e incrementa la ganancia genética. Se obtuvieron complejos ovocito cúmulus (COC), de ovarios de matadero, se cultivaron para maduración en gotas de 100 µl de TCM-199 + 5 % de suero fetal bovino + 0.005 U/ml de (FSH-p) + 10 IU hCG/ml + 1µg Estradiol (E2)/ml, 15 COC/gota, cubiertas con aceite mineral, en incubadora (38,5 ºC, 5 % de CO2 y 95 % de humedad), 22 h. Posmaduración se dividieron en 4 grupos (G1, G2, G3, G4) y se realizaron dos experimentos simultáneamente: I) G1: NS a 5x106, G2: SX a 5x106, G3: SX a 10x106 espermatozoides/ml capacitados con BO. II) G2: SX a 4,5x106 espermatozoides/ml capacitados con BO y G4: SX capacitado con Percoll a 5x106 espermatozoides/ ml. Se colocaron 15 COC/gota de semen cubiertas con aceite mineral, en incubadora, 18 h. Para el desarrollo se colocaron en gotas de CR1aa, en incubadora. Se aplicó el test de χ2. La TD a las 48 h entre G1 y G3 no presentó diferencias significativas (p<0,05), sin embargo, en ambos grupos fue significativamente mayor al G2. En el DE al día 7 hubo diferencias significativas (p<0,05) a favor del G4. Se obtuvo mayor DE con el SX capacitado con Percoll respecto al BO y no hubo diferencias entre ambas concentraciones de semen.


SUMMARY: The objective of this study was to evaluate the in vitro fertility of bovine sexed semen (SX) capacitated with Percoll vs. BO. The division rate (DR), embryo development (ED) were evaluated in two concentrations 5x106 vs. 10x106 sperm/ml, capacitated with BO and compared with non-sexed semen (NS). Offspring sexing represents an important advance for livestock production. Flow cytometry separates X and Y spermatozoa by difference in DNA content (4 % greater in X) with 90 % effectiveness. The SX increases the reproductive efficiency, allows the selection of females and increases the genetic gain. Cumulusoocytes complexes (COC) were obtained from slaughterhouse ovaries. They were then cultured 22 hours for maturation in TCM199 + 5 % fetal calf serum + 0.005 IU/ml (FSH-p) + 10 IU hCG/ml + 1 mg Estradiol (E2)/ml, in 100 µl drops with mineral oil, in incubator (38.5 ºC, 5 % CO and 95 % humidity). Post maturation, 4 groups were randomly assigned (G1, G2, G3, G4) and were performed two experiments simultaneously: I) G1 was inseminated with NS at 5x106 sperm/ml, G2: SX at 5x106, G3: SX at 10x106 sperm/ml capacitated with BO. II) G2: SX at 5x106 sperm/ml capacitated with BO and G4: SX capacitated with Percoll at 5x106 sperm/ml. 15 COC/drop of capacitated semen covered with mineral oil and placed in an incubator for 18 hours. For development, they were placed in drops of CR1aa, in an incubator. Results were analyzed with the c square test. At 48 hours, there were no significant differences (p<0.05) in DR between G1 and G3, however, in both groups it was significantly greater than G2. At day 7 there were significant differences (p <0.05) in ED, greater in G4. At 48 hours, there were no significant differences (p<0.05) in DR between G1 and G3, however, in both groups it was significantly greater than G2. At day 7 there were significant differences (p <0.05) in ED, greater in G4. A higher ED was obtained with the SX capacitated with Percoll, with respect to BO and there was no difference between the two semen concentrations.


Subject(s)
Animals , Cattle , Embryonic Development , Fertilization in Vitro/veterinary , Sex Preselection/methods , Spermatozoa/physiology , Flow Cytometry , Sex Preselection/veterinary , Sperm Capacitation
13.
Acta cir. bras ; 32(9): 755-766, Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-886239

ABSTRACT

Abstract Purpose: To determine the effect of folic acid (FA) on experimental testicular ischemia/reperfusion (I/R) in rats. Methods: Sixty male Wistar rats were randomly divided into 6 groups. The control group received physiologic saline orally. The sham-operated group received physiologic saline orally then exposed to midline laparotomy without clamping the IR. The I/R rats received oral gavage of the saline then subjected to 1h ischemia /24h reperfusion, period. In folic acid (2mg/kg+IR) rats received oral gavage of the FA (2mg/kg) then subjected to 1h I/24h R. groups 5-6 received FA (5 and 10 mg/kg), then subjected to 1 h I/24 h, respectively. At the end of the study, semen samples were collected for spermatozoa characteristics. The left testis was removed for histological analysis and superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GPx) measurement. Results: Spermatozoa mobility, mortality (%) significantly decreased in I/R group (P<0.05). Dose dependent increase observed on spermatozoa mobility, mortality (%) using different levels of the FA (2, 5 and 10 mg/kg) treated rat (P<0.05). Tissue MDA levels significantly increased in I/R rat (P<0.05) while FA (2, 5 and 10mg/kg) in a dose dependent manner decreased I/R-induced MDA (P<0.05). Experimental I/R significantly decreased SOD and GPx activity (P<0.05). Administration of the FA (2, 5 and 10mg/kg) significantly increased tissue SOD and GPx activity in I/R rat (P<0.05). Seminiferous tubules degenerated and loss of spermatogenesis with few spermatocytes was observed in degenerated testis tubules in I/R rat. Orally administration of the FA (5 and 10 mg/kg) improved testis characteristics with few normal seminiferous tubules and spermatocyte in seminiferous tubules in experimental I/R-induced rat. Conclusion: The treatment of folic acid had a benefit effect against ischemia-reperfusion.


Subject(s)
Animals , Male , Rats , Testis/blood supply , Reperfusion Injury/drug therapy , Folic Acid/therapeutic use , Spermatozoa/drug effects , Spermatozoa/physiology , Random Allocation , Rats, Wistar , Disease Models, Animal
14.
Braz. j. biol ; 77(3): 553-557, July-Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-888767

ABSTRACT

Abstract Erythrolamprus poecilogyrus sublineatus (Cope, 1860), is a species widely distributed in the Pampa Domain, occurring in Rio Grande do Sul, Argentina and Uruguay, mainlyin the pampa region. In the coastal region of southern Brazil this is serpent is considered one of the most abundant. The purpose of the present study is to describe the techniques of sperm evaluation in vitro for E. poecilogyrus sublineatus in the coastal plain of Rio Grande do Sul, Brazil. After laparatomy the efferent vases were collected and the semen was diluted in 1ml Beltsville Thawing Solution. The characteristics of motility, membrane integrity, mitochondria, acrosome, DNA, cell viability and cellular functionality were evaluated. Fluorescent probes were used for the evaluation of sperm structure in epifluorescence microscope. With the techniques described, it was possible to identify intact and injured cells, enabling the determination of cell characteristics for the spring season (October and November). It was observed in the analyses that 80% of sperm cells were mobile and that 84.1 ± 8.0% of sperm membranes were intact. The standards found were of 48 ± 13.8% of intact acrosome, 73.6 ± 6.0 of perfect DNA and of 91.8 ± 4.0 of functional mitochondria. Thus, these values from the sperm analysis can be used as standards for the species Erythrolamprus poecilogyrus sublineatus.


Resumo Erythrolamprus poecilogyrus sublineatus (Cope, 1860), é uma espécie amplamente distribuída no Domínio Pampa, ocorrendo no Rio Grande do Sul, Argentina e Uruguai, principalmente na região dos pampas. Na região costeira do extremo sul do Brasil essa é uma das serpentes consideradas mais abundantes. O objetivo deste estudo é descrever as técnicas de avaliação espermática in vitro para E. poecilogyrus sublineatus da planície costeira do Rio Grande do Sul, Brasil. Após laparatomia os vasos eferentes foram coletados e o sêmen diluído em 1ml Beltsville Thawing Solution. Foram avaliadas as características de motilidade, integridade de membrana, mitocôndria, acrossoma, DNA, viabilidade celular e funcionalidade celular. Foram utilizadas sondas fluorescentes para avaliação das estruturas espermática em microscópio de epifluoescência. Com as técnicas descritas foram possível identificar células integras e lesadas, podendo determinar as características celulares para o período de primavera (outubro e novembro). Nas análises foi observado que 80% das células espermáticas estavam móveis e que 84,1 ± 8,0% das membranas espermáticas estavam íntegras. Os padrões encontrados para foram de 48 ± 13,8% de acrossoma íntegro, 73,6 ± 6,0 de DNA íntegro e de 91,8 ± 4,0 de mitocôndria funcional. Desta forma, esses valores das análises espermáticas podem ser utilizados como padrão para a espécie Erythrolamprus poecilogyrus sublineatus.


Subject(s)
Animals , Male , Snakes/physiology , Sperm Motility , Spermatozoa/physiology , Ejaculation , Brazil , Semen Analysis
15.
Int. j. med. surg. sci. (Print) ; 3(3): 893-902, sept. 2016.
Article in Spanish | LILACS | ID: biblio-1087451

ABSTRACT

Actualmente, cientos de crías se han gestado a través de inseminación artificial conespermatozoides sexados en producción animal. Desde 1992 se utiliza la citometría de flujo, técnica que permite diferenciar espermatozoides X e Y según su contenido de ADN. No existe, hasta el momento, ninguna otra técnica práctica para sexar espermatozoides manteniendo la capacidad fecundante. Los objetivos de esta revisión son explicar: (1) por qué los espermatozoides que contienen el cromosoma X o Y son similaresfenotípicamente, pero a la vez mantienen diferencias entre ellos, (2) los principios y procedimientos utiliza-dos para sexar espermatozoides mediante citometría de flujo y sorting (3) la precisión, velocidad y la eficiencia de los procedimientos actuales de sexaje espermático, (4) el daño espermático ocurrido durante el sexaje espermático y consecuentemente los efectos sobre la fecundidad.


Flow cytometry is a useful technology in the sexed sperm, which measures and analyzes simultaneously, multiple physical characteristics of the cell, as they flow in a stream flow, through a lightbeam. The measured properties are the size of a particle, relative internal granularity, relative complexity and relative fluorescence intensity. Currently, hundreds of calves have been gestated through artificial insemination with sexed sperm in animal production. Since 1992, flow cytometry has been used, a technique that allows spermatozoa X and Y differentiation by DNA content. There is no other practical technique forsperm sexing to keep sperm functionality. The objectives of this review are to explain: (1) why the sperm containing the X or Y chromosome are phenotypically similar, but differ among themselves, (2) the principlesand procedures used for sexing sperm by flow cytometry and sorting ( 3) accuracy, speed and efficiency ofcurrent procedure sperm sexing, (4) sperm damage occurred during sperm sexing and consequently the effects on fertility.


Subject(s)
Animals , Sex Preselection/methods , Sex Preselection/veterinary , Spermatozoa/physiology , Flow Cytometry/methods , Flow Cytometry/veterinary , Sex Chromosomes
16.
Pesqui. vet. bras ; 36(7): 657-664, jul. 2016. tab
Article in Portuguese | LILACS, VETINDEX | ID: lil-794777

ABSTRACT

Objetivou-se avaliar o efeito da adição de diferentes concentrações de melatonina no sêmen diluído de carneiros após criopreservação. Foram coletados 10 ejaculados de três carneiros adultos (n=30), por meio de vagina artificial para ovinos. Os ejaculados coletados foram diluídos em Tris-Gema de ovo, para a concentração final de 200x106 sptz/mL, mantidos em banho maria a 32°C, e a melatonina adicionada conforme os tratamentos: Controle; 100pM; 100nM; 100µM e 1mM de melatonina. Então, as amostras foram resfriadas em câmara fria a 5°C por duas horas, envasadas em palhetas de 0,5 mL e lacradas. Logo após, foram acondicionadas sob vapores do nitrogênio liquido, por 15 minutos, a 8cm da lâmina líquida e congeladas com nitrogênio líquido. As amostras foram analisadas quanto à motilidade espermática, integridade da membrana plasmática, membrana acrossomal, atividade mitocondrial, quantificação do estresse oxidativo e a capacidade de ligação. As variáveis foram submetidas à análise de variância e as médias foram comparadas pelo teste de Tukey a 5% de probabilidade. A motilidade total e progressiva dos espermatozoides descongelados foi maior nas amostras tratadas com 100pM de melatonina (62,99 e 45,07% respectivamente; P<0,05) quando comparado aos demais tratamentos. A adição das diferentes concentrações de melatonina no sêmen diluído, com exceção da concentração de 1 mM, apresentou maior percentual de células com membrana plasmática íntegra, quando comparadas com o controle (P<0,05). O percentual de espermatozoides com integridade da membrana do acrossoma foi maior no sêmen tratado com 100 pM de melatonina (P<0,05) do que nos demais tratamentos. A alta atividade mitocondrial foi maior nos espermatozoides tratados com 100 pM de melatonina (69,30%; P<0,05). A adição de 100 nM de melatonina reduziu a quantidade de TBARS após a criopreservação (2,84; P<0,05) quando comparado aos demais tratamentos. Após o descongelamento, o número de espermatozoides que se ligaram à membrana perivitelina foi maior nos tratados com 100 pM de melatonina (155,73; P<0,05). Portanto, a adição de melatonina no sêmen diluído pode ser útil para aperfeiçoar a criopreservação do sêmen de ovinos, melhorando as taxas de fertilização por meio da inseminação artificial.(AU)


The aim was to evaluate the effect of adding different concentrations of melatonin in ram semen diluted after cryopreservation. Ten ejaculates were collected0 from three adult ram (n=30) by means of artificial vagina for sheep. The collected samples were diluted in Tris-egg yolk, to a final concentration 200x106 sptz/mL kept in water bath at 32°C, and melatonin added as treatments: control; 100pM; 100nM; 100µM and 1mM melatonin. Then, the samples were cooled in a cold chamber at 5°C for two hours, in straws of 0.5mL and sealed. They were stored under the liquid nitrogen vapor for 15 minutes to 8cm of liquid blade and frozen with liquid nitrogen. Samples were analyzed for sperm motility, membrane integrity, acrosomal membrane, mitochondrial activity, oxidative stress and quantification of the binding capacity. The variables were subjected to analysis of variance and the means were compared by Tukey test at 5% probability. The total and progressive motility of thawed sperm were higher in samples treated with 100pM melatonin (62.99 and 45.07%, respectively; P<0.05) when compared to other treatments. The addition of different concentrations of melatonin in semen diluted with the exception of 1mM concentration, a higher percentage of cells with intact plasma membrane, as compared with the control (P<0.05). The percentage of sperm with acrosome membrane integrity was higher in the semen with 100pM melatonin (P<0.05) than the other treatments. The high mitochondrial activity was higher in spermatozoa treated with 100pM melatonin (69.30%; P<0.05). Addition of 100nM melatonin reduced the amount of TBARS after cryopreservation (2.84, P<0.05) when compared with the other treatments. After thawing, the number of sperm which bind to the perivitelline membrane was higher in the melatonin treated with 100pM (155,73; P<0.05). Therefore, melatonin addition the semen diluted can be useful to enhance the cryopreservation of sheep semen, improving fertilization rates through artificial insemination.(AU)


Subject(s)
Animals , Melatonin/administration & dosage , Melatonin/analysis , Oxidative Stress , Semen Analysis/veterinary , Sheep/physiology , Spermatozoa/physiology , Antioxidants/analysis , Cryopreservation/veterinary , Reactive Oxygen Species/analysis , Reproductive Techniques/veterinary
17.
Rev. bras. reumatol ; 56(3): 212-219, tab, graf
Article in English | LILACS | ID: lil-785756

ABSTRACT

Abstract Objective To investigate the deleterious effects of air pollutants exposure in the Sao Paulo metropolitan region on semen quality in systemic lupus erythematosus (SLE). Methods A seven-years longitudinal repeated-measures panel study was performed at the Laboratory of Experimental Air Pollution and Rheumatology Division. Two semen samples from 28 post-pubertal SLE patients were analyzed. Daily concentrations of air pollutants exposure: PM10, SO2, NO2, ozone, CO, and meteorological variables were evaluated on 90 days before each semen collection dates using generalized estimating equation models. Results Intravenous cyclophosphamide (IVCYC) and ozone had an association with a decrease in sperm quality of SLE patients. IVCYC was associated with decreases of 64.3 million of spermatozoa/mL (95% CI 39.01–89.65; p = 0.0001) and 149.14 million of spermatozoa/ejaculate (95% CI 81.93–216.38; p = 0.017). With regard to ozone, the most relevant adverse effects were observed from lags 80–88, when the exposure to an interquartile range increase in ozone 9-day moving average concentration led to decreases of 22.9 million of spermatozoa/mL (95% CI 5.8–40.0; p = 0.009) and 70.5 million of spermatozoa/ejaculate (95% CI 12.3–128.7; p = 0.016). Further analysis of 17 patients that never used IVCYC showed association between exposure to ozone (80–88 days) and decrease of 30.0 million of spermatozoa/mL (95% CI 7.0–53.0; p = 0.011) and 79.0 million of spermatozoa/ejaculate (95% CI 2.1–155.9; p = 0.044). Conclusion Ozone and IVCYC had a consistent adverse effect on semen quality of SLE patients during spermatogenesis. Minimizing exposure to air pollution should be taken into account, especially for patients with chronic systemic inflammatory diseases living in large cities.


Resumo Objetivo Investigar os efeitos deletérios da exposição aos poluentes do ar na Região Metropolitana de São Paulo sobre a qualidade do sêmen de pacientes com lúpus eritematoso sistêmico (LES). Métodos Foi feito um estudo longitudinal de painel com medidas repetidas de sete anos no Laboratório de Poluição Atmosférica Experimental e Reumatologia. Foram analisadas duas amostras de sêmen de 28 pacientes com LES pós‐púberes. Foram avaliadas as concentrações diárias de exposição aos poluentes do ar PM10, SO2, NO2, ozônio e CO e variáveis meteorológicas 90 dias antes de cada data de coleta de sêmen com o uso do método de equações de estimativas generalizadas. Resultados A ciclofosfamida intravenosa (CICIV) e o ozônio estiveram associados a uma diminuição na qualidade do sêmen dos pacientes com LES. A CICIV esteve associada a um decréscimo de 64,3 milhões de espermatozoides/mL (IC 95% 39,01‐89,65; p = 0,0001) e 149,14 milhões de espermatozoides/ejaculado (IC 95% 81,93‐216,38; p = 0,017). Em relação ao ozônio, os efeitos adversos mais relevantes foram observados entre os lags (intervalo de tempo) 80 e 88, quando a exposição a uma concentração média de ozônio um intervalo interquartil maior em nove dias móveis levou a um decréscimo de 22,9 milhões de espermatozoides/mL (IC 95% 5,8‐40; p = 0,009) e 70,5 milhões de espermatozoides/ejaculado (IC 95% 12,3‐128,7; p = 0,016). Uma análise mais aprofundada dos 17 pacientes que nunca usaram CICIV mostrou associação entre a exposição ao ozônio (80‐88 dias) e o decréscimo de 30 milhões de espermatozoides/mL (IC 95% 7‐53; p = 0,011) e 79 milhões de espermatozoides/ejaculado (IC 95% 2,1‐155,9; p = 0,044). Conclusão O ozônio e a CICIV tiveram um efeito adverso consistente sobre a qualidade do sêmen de pacientes com LES durante a espermatogênese. Deve‐se considerar a minimização da exposição à poluição do ar, especialmente para pacientes com doenças inflamatórias sistêmicas crônicas que vivem nas grandes cidades.


Subject(s)
Humans , Male , Ozone/adverse effects , Spermatozoa/drug effects , Air Pollution/adverse effects , Lupus Erythematosus, Systemic , Spermatozoa/physiology , Environmental Exposure/adverse effects , Semen Analysis
18.
Pesqui. vet. bras ; 36(supl.1): 58-64, June 2016. tab, graf, ilus
Article in English | LILACS, VETINDEX | ID: lil-798011

ABSTRACT

The aim was to study the effects of different gamete coincubation times on porcine in vitro fertilization (IVF), and to verify whether efficiency could be improved by reducing oocyte exposure time to spermatozoa during IVF. In groups of 50, a total of 508 immature cumulus-oocyte complexes (COCs) were matured in NCSU-37 medium. The COCs were cultured for 44 hours and then inseminated with in natura semen (2,000 spermatozoa/oocyte). The sperm and oocytes were coincubated according to the following treatments (T): T1 = oocytes exposed to spermatozoa for one hour (173 oocytes), T2 = oocytes exposed to spermatozoa for two hours (170 oocytes), and T3 = oocytes exposed to spermatozoa for three hours (165 oocytes). After these coincubation periods, the oocytes were washed in fertilization medium (TALP medium) to remove spermatozoa not bound to the zona pellucida and cultured in another similar medium (containing no sperm). Eighteen to twenty hours after fertilization, the putative zygotes were stained in Hoechst-33342 to evaluate the IVF results. The penetration rate was higher (P<0.05) after two hours of coincubation time than it was for one or three hours. Furthermore, 68.60% of the ova coincubated with the spermatozoa for two hours were monospermic. The oocytes exposed to spermatozoa for one hour (T1) presented a higher (P<0.01) rate of polyspermy than those in T2 and T3. Fertilization performance (%) did not differ (P>0.05) between oocytes exposed to spermatozoa for one (T1) and three hours (T3). However, optimum (P=0.048) results were obtained after two hours of coincubation, when the rate of fertilization performance was 50.16±8.52%. The number of penetrated sperm per oocyte, as well as male pronucleus formation, did not differ (P>0.05) between the treatments evaluated. Under these assay conditions, especially in relation to the sperm concentration used, gamete coincubation for a period of two hours appears to be optimal for monospermy and fertilization performance. Thus, it is the optimal time period for obtaining a large number of pig embryos capable of normal development.(AU)


Esse estudo foi realizado para avaliar os efeitos de diferentes tempos de coincubação dos gametas sobre a fertilização in vitro (FIV) de suínos e se a eficiência dessa técnica poderia ser melhorada pela redução no período que os ovócitos são expostos aos espermatozoides durante a FIV. Um total de 508 (em grupos de 50) complexos cumulus-ovócito (COCs) imaturos foram maturados no meio NCSU-37. Os COCs foram cultivados por 40-44 horas e então inseminados com sêmen in natura (2.000 espermatozoides/ovócito). Os espermatozoides e ovócitos foram coincubados de acordo com os seguintes tratamentos (T): T1 = ovócitos expostos aos espermatozoides por uma hora (173 ovócitos); T2 = ovócitos expostos aos espermatozoides por duas horas (170 ovócitos) e T3 = ovócitos expostos aos espermatozoides por três horas (165 ovócitos). Após esses períodos de coincubação, os ovócitos foram lavados em meio de fertilização (meio TALP) para remoção dos espermazoides não ligados a zona pelúcida e cultivados em outro mesmo meio (não contendo espermatozoides). Após 18-20 horas de fertilização, os prováveis zigotos foram corados com Hoechst-33342 para avaliação dos resultados da FIV. A taxa de penetração foi maior (P<0,05) após o tempo de coincubação de duas horas em comparação a uma e três horas. Além disso, 68,60% dos ovócitos coincubados com os espermatozoides por duas horas foram monospérmicos. Os ovócitos expostos aos espermatozoides por uma hora (T1) apresentaram elevada (P<0,01) taxa de polispermia em comparação com o T2 e T3. A eficiência da fertilização (%) não diferiu (P>0,05) entre os ovócitos expostos aos espermatozoides por uma (T1) e três horas (T3). Entretanto, ótimos (P=0,048) resultados foram obtidos após duas horas de coincubação, quando a taxa da eficiência da fertilização foi 50,16 ± 8,52%. O número de espermatozoides penetrados por ovócito e a formação de pro-núcleo masculino não diferiu (P>0,05) entre os tratamentos avaliados. Sob as condições de ensaio realizadas, especialmente em relação à concentração espermática utilizada, a coincubação dos gametas por um período de duas horas parece ser ótima para as taxas de monospermia e eficiência da fertilização. Portanto, um tempo provavelmente ótimo para obter um elevado número de embriões suínos capazes de ter um desenvolvimento normal.(AU)


Subject(s)
Animals , Fertilization in Vitro/veterinary , Oocytes/physiology , Spermatozoa/physiology , Swine , Reproductive Techniques/veterinary
19.
Pesqui. vet. bras ; 35(11): 925-932, nov. 2015. tab
Article in Portuguese | LILACS | ID: lil-767757

ABSTRACT

Este estudo descreveu as características seminais, da membrana plasmática e do acrossoma de espermatozoide congelado/descongelado de 19 ejaculados de garanhões da raça Nordestina. Os aspectos analisados incluíram os parâmetros físicos do sêmen fresco; a motilidade e a longevidade do sêmen diluído e descongelado; a morfologia espermática, integridade funcional e estrutural da membrana plasmática do espermatozoide e a habilidade de ligação do espermatozoide à membrana perivitelina da gema do ovo de galinha do sêmen descongelado. As variáveis foram avaliadas pela ANOVA com post hoc teste de Student Newman-Keuls (P<0,05). A MT e a MP foram maiores (P<0,05) no sêmen diluído do que no descongelado. A percentagem média de defeitos maiores, menores e totais foi muito inferior ao limite recomendado pelo CBRA. A porcentagem de reativos ao HOST foi de 14,21±1,12% e a porcentagem média de membranas íntegras detectadas pelo teste supravital de 62,22±9,06% e pela sonda SYBR-14 de 81,47±26,90. O número médio de espermatozoides ligados à MPV após a descongelação do sêmen foi de 230,39±57,09. A MT e MP no tempo 0 min do TTR foi superior (P<0,05) em relação a 150 min, não diferindo nos tempos 10 min e 30 min. Os resultados demonstram que a utilização dos testes laboratoriais adicionais ajudam no processo de avaliação das amostras, possibilitando a obtenção de informações mais confiáveis e precisas. Embora a criopreservação tenha provocado queda na motilidade seminal, o uso de diluidor contendo amidas minimizou os danos osmóticos nas células espermáticas e manteve a integridade morfológica, funcional e estrutural da membrana plasmática do espermatozoide. Estes resultados são um referencial em estudos futuros uma vez que, inexistem dados comparativos nesta raça...


This paper describes the seminal characteristics of the plasma membrane of frozen-thawed sperm. Nineteen ejaculates of Nordestino horse breed. The aspects analyzed in the physical parameters of fresh semen were total and progressive motility and your longevity after dilution or thawed; sperm morphology, functional and structural integrity of the plasma membrane of the sperm and the sperm-binding ability to the perivitelline membrane of the yolk (MPV) after thawed. The variables were assessed by ANOVA with post hoc test of Student Newman-Keuls test (P<0.05). The total and progressive motility were higher in diluted semen than thawed (P<0.05). The average percentage of the major, minor and total defects was lower than the limit recommended by the CBRA. The percentage of reactive to hypo-osmotic swelling test was 14.21±1.12%, the intact membrane detected by supravitally test was 62.22±9.06% and the SYBR-14 was 81.47±26.9. The ability of sperm to bind to the MPV after thawing semen was 230.39±57.09. The total and progressive motility at time 0 min of termo resistance test was higher than to 150 minutes (P<0.05), and no difference was observed in the times 10 and 30 minutes. The results demonstrate that the use of additional laboratory tests help in the process of evaluation of samples, making possible to obtain more reliable and accurate information. Although cryopreservation has caused decrease in sperm motility and was used diluents with amides to diluted and cryopreservation protocol and this minimized the osmotic damage to sperm cells and maintained the morphological, functional and structural integrity of the plasma membrane of the sperm. These results are a reference for future studies since there are no comparative data on this breed...


Subject(s)
Animals , Male , Horses/physiology , Spermatozoa/physiology , Semen Analysis/veterinary , Cryopreservation/veterinary , Sperm Motility/physiology , Semen Preservation/adverse effects , Semen Preservation/veterinary
20.
Int. j. morphol ; 33(3): 835-841, Sept. 2015. ilus
Article in Spanish | LILACS | ID: lil-762551

ABSTRACT

Conocer los aspectos moleculares que acontecen en el proceso de unión de los espermatozoides humanos a la zona pelúcida (ZP) humana es uno de los grandes retos de la biología de la Reproducción. Por otra parte conocer si el proceso de fecundación puede verse afectado por la criopreservación de los gametos femeninos sigue siendo otra cuestión debatida en la literatura. En base a esto, el objetivo principal de este trabajo fue conocer si la vitrificación ovocitaria puede alterar la interacción de los espermatozoides con el glicocáliz de la ZP y demostrar si la ZP de estos ovocitos pierde la capacidad de inducir la reacción acrosómica en los espermatozoides. Según nuestros resultados el método de vitrificación ovocitaria cerrado (S3) no altera la capacidad de unión de los espermatozoides a la zona pelúcida, ni la capacidad de ésta para inducir la reacción acrosómica.


To know the molecular aspects that occur in the process of human sperm binding to the human zona pellucida (ZP) is one of the great challenges of reproduction biology. Moreover knowing if the fertilization process may be affected by cryopreservation of female gametes is still another issue discussed in the literature. Based on this, the main objective of this study was to determine whether the oocyte vitrification may alter the interaction of sperm with the glycocalyx of ZP and show whether these oocytes lost the ability to induce the acrosome reaction in sperm. According to our results the oocyte closed vitrification method (S3) does not alter the ability of the sperm binding to the zona pellucida, and their ability to induce the acrosome reaction.


Subject(s)
Humans , Male , Female , Oocytes/physiology , Oocytes/ultrastructure , Spermatozoa/physiology , Spermatozoa/ultrastructure , Vitrification , Cryopreservation , Fertility , Microscopy, Electron , Sperm-Ovum Interactions , Zona Pellucida
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