Effect of refrigeration at -1°C on spermatozoa quality of domestic cats / Efeito da refrigeração a -1°C na qualidade de espermatozoides de gatos domésticos

The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%. In ejaculate samples, higher values of total morphological defects were observed after 24 and 48 hours of refrigeration at 4°C (P<0.022) compared to refrigeration at -1°C, using Friedman test. To quantify the decrease in sperm quality, parameter reductions were calculated among time points (F-24h/F-48h/24h-48h). In EPD samples, a greater reduction in sperm quality was detected after 24 hours of refrigeration at 4°C, both in motility and sperm kinetics and in the movement and velocity indices, compared to refrigeration at -1°C. Based on the results, it can be concluded that cooling of feline spermatozoa at -1°C for up to 48 hours was efficient in maintaining spermatic quality collected by EEJ and EPD, and it could be an alternative to spermatozoa cryopreservation in domestic felines.(AU)

O objetivo deste trabalho foi avaliar a qualidade espermática de gatos domésticos obtidos por eletroejaculação e recuperação da cauda do epidídimo após a refrigeração a -1°C e a 4°C por 24 e 48 horas. Vinte e nove gatos adultos (2 a 6kg) foram utilizados. A colheita de espermatozoides foi realizada por eletroejaculação (EEJ) e, após 48 horas, os gatos foram orquiectomizados, e as amostras espermáticas foram obtidas a partir do ducto deferente e da cauda do epidídimo (EPD). As amostras foram diluídas em ACP-117® e as características espermáticas foram avaliadas em três momentos distintos: fresco, 24 e 48 horas após a refrigeração. Para ser possível comparar as duas temperaturas de refrigeração, a primeira etapa foi analisar se havia diferença entre as técnicas de colheita. Após isto, dois experimentos foram conduzidos: no primeiro, espermatozoides de 14 gatos foram utilizados e a refrigeração foi realizada a -1°C; e no segundo, amostras de 15 gatos foram utilizados e os espermatozoides foram refrigerados a 4°C. A cinética espermática foi avaliada por análise computadorizada (CASA), a concentração por câmara de Neubauer, a morfologia espermática foi avaliada pela coloração de Karras modificada, e a integridade da membrana foi avaliada por eosina nigrosina. Os resultados obtidos foram analisados no software R, versão 3.2.5, utilizando o teste de Mann-Whitney para variáveis com distribuições anormais, considerando significância ao nível de 5%. No ejaculado, maiores valores de defeitos morfológicos totais foram observados após 24 e 48 horas de refrigeração a 4°C (P<0,022) em comparação com refrigeração a -1°C, usando o teste de Friedman. Para quantificar a diminuição na qualidade espermática, as reduções dos parâmetros foram calculadas entre os pontos de tempo (F-24h/F-48h/24h-48h). Na EPD, uma maior redução na qualidade espermática foi detectada após 24 horas de refrigeração a 4°C, tanto na motilidade e na cinética espermática quanto nos índices de movimento e velocidade, em comparação com a refrigeração a -1°C. Com base nos resultados, pode concluir-se que a refrigeração dos espermatozoides felino a -1°C, até 48 horas, foi eficaz na manutenção da qualidade espermático colhidos por EEJ e EPD, e pode ser uma alternativa para a criopreservação de espermatozoides em felinos domésticos.(AU)

Colheita farmacológica de sêmen de onças-pardas (Puma concolor: Mammalia: Carnivora: Felidae) / Pharmacological semen collection in cougars (Puma concolor: Mammalia: Carnivora: Felidae)

Objetivou-se, por meio do presente estudo, avaliar o método de colheita farmacológica de sêmen com sondagem uretral, em machos de onças-pardas (Puma concolor) mantidos em cativeiro. A técnica proposta (Cat; N=3) foi comparada com a eletroejaculação (EE; N=4). Para a colheita farmacológica, utilizou-se medetomidina para induzir a liberação de sêmen na uretra e sonda uretral para gatos, sem janela lateral, para colheita do sêmen por capilaridade. O método foi eficaz em todos os animais usados. Por meio dessa técnica, colheram-se amostras com menor volume (106,7±30,5aµL) e maior concentração (524,1±54,3b x 106 espermatozoides/mL) em relação à EE (450,0±0,1bµL e 205,0±141,8a x 106 espermatozoides/mL). As avaliações de vigor, motilidade e patologia espermática demonstraram que a técnica não afeta a qualidade do sêmen em relação à EE (P>0,05). Dessa forma, o método proposto consiste em uma técnica mais prática e eficiente para a colheita de sêmen com boa qualidade, dispensando o eletroejaculador.(AU)

The aim of this study was to evaluate the pharmacological semen collection method with urethral catheterization (CT) in captive cougar (Puma concolor) males. The pharmacological method (CT; N= 3) was compared to the electroejaculation technique (EE; N= 4). For CT collection, medetomidine was administrated to induce semen release using a tomcat catheter inserted into the urethra to collect by capillarity. The proposed method was efficacious on all animals used. Through the CT method, semen collected yielded smaller volume (106,7±30,5aµL) and higher concentration (524,1±54,3b x 106sperm/mL) compared to EE (450,0±0,1bµL and 205,0±141,8a x 106 sperm /mL). Evaluations of vigor, motility and sperm pathology demonstrated that CT does not affect semen quality when compared to EE (P> 0.05). Thus, the proposed method consists of a more practical and efficient technique for semen collection with good quality, eliminating the need for eletroejaculation.(AU)

Use of infrared thermography to evaluate the inflammatory reaction in cat testis after intratesticular injection of 0. 9% NaCl or 20% CaCl2 with 1% lidocaine / O uso da termografia infravermelha para avaliar a reação inflamatória após a injeção intratesticular de NaCL 0, 9 % e cloreto de cálcio 20% com lidocaína 1%

Due to the doubts and questions about the inflammatory reaction caused by chemical castration, this study aimed to use infrared thermography to detect, evaluate and monitor the inflammatory reaction caused by the intratesticular injection of calcium chloride (CaCl2) 20% with lidocaine 1%. For this, thermographic measurements were taken before (M0), 10 minutes (M1), 1 and 6 hours (M2 and M3), for 7 consecutive days (M4 to M10), at 15 (M11), 30 (M12) and 60 (M13) days after intratesticular injection. Additionally, changes to testicular tissue and effects over spermatogenesis were evaluated by andrological exam before (M0) and 60 days (M13) after intratesticular injection. All cats were orchiectomized at M13, and testicles were submitted to histological analysis. CaCl2 (20%) with lidocaine (1%) administration produced testicular tissue damage and interfered with the spermatogenesis in 70% of treated cats without exacerbating the inflammatory reaction or impairing the cat's welfare. It was concluded that thermographic evaluation is a useful, efficient, easy and quick method to diagnose and monitor cat testicular inflammatory reactions.(AU)

Devido a constantes dúvidas e questionamentos sobre a reação inflamatória ocasionada pela castração química, este estudo objetivou o uso da termografia infravermelha para detectar, avaliar e monitorar a reação inflamatória causada pela injeção intratesticular de cloreto de cálcio (CaCl 2 ) 20% associada à lidocaína 1%. Para isso, medidas termográficas foram aferidas antes (M0), 10 minutos (M1), uma e seis horas (M2 e M3), por sete dias consecutivos (M4 a M10), aos 15 (M11), 30 (M12), e 60 (M13) dias após injeção intratesticular, nos grupos tratado e controle. Todos os gatos foram orquiectomizados no M13, e os testículos submetidos à análise histológica. A injeção CaCl 2 a 20% associada com lidocaína a 1% produziu lesão testicular e interferiu na espermatogênese de 70% dos gatos tratados, sem exacerbar a reação inflamatória ou prejudicar o bem-estar do animal. Foi concluído que a avaliação termográfica é uma ferramenta útil, eficiente, rápida e fácil para o diagnóstico e o monitoramento da reação inflamatória em gatos.(AU)

Establishment of an anesthetic protocol for semen collection by electroejaculation in six-banded armadillos (Euphractus sexcinctus Linnaeus, 1758) / Estabelecimento de um protocolo anestésico para coleta de sêmen por eletroejaculação em tatus-peba (Euphractus sexcinctus Linnaeus, 1758)

The aim was to verify the effects of different anesthetic protocols used during electroejaculation (EEJ) in six-banded armadillos (Euphractus sexcinctus). Four sexually matured animals were physically restrained and subjected to semen collection by the EEJ following three treatments: The control group consisted of no use of anesthesia; in the others, the anesthetic combinations xylazine/ketamine/propofol or butorphanol/ ketamine/propofol were administered. For each group, twelve procedures were conducted for EEJ. Semen was evaluated for volume, color, aspect, motility, sperm concentration, morphology, viability, and functional membrane integrity. The highest efficiency (100% ejaculates) was achieved when the control group was used; the xylazine/ketamine/propofol association provided only 11 ejaculates from a total of 12 attempts (91.6% efficiency), while only 4 ejaculates (33% efficiency) were obtained with butorphanol/ketamine/propofol (P<0.05). Both protocols provided rapid induction and relaxation enough to perform the EEJ. In the use of butorphanol/ketamine/propofol, the animals recovered at 16.5±1.5min, a time shorter than in the use of xylazine/ketamine/propofol protocol, 20.7±1.0min (P>0.05). The semen volume and sperm concentration obtained in the use of xylazine/ketamine/propofol association were significantly higher than those verified for butorphanol/ketamine/propofol protocol. In conclusion, the xylazine/ketamine/propofol association is indicated for anesthesia of six-banded armadillos submitted to EEJ.(AU)

Objetivou-se verificar os efeitos de diferentes protocolos anestésicos usados durante a eletroejaculação (EEJ) em tatus-peba (Euphractus sexcinctus). Quatro animais sexualmente maduros foram contidos fisicamente e submetidos à coleta de sêmen por EEJ, seguindo três tratamentos: o grupo controle consistiu do não uso de anestesia; nos outros, foram administradas combinações anestésicas de xilazina/cetamina/propofol, ou butorfanol/cetamina/propofol. Para cada grupo, foram conduzidos 12 procedimentos de EEJ. O sêmen foi avaliado para volume, cor, aspecto, motilidade, concentração de espermatozoides, morfologia, viabilidade e integridade funcional da membrana. A mais alta eficiência (100% de ejaculados) foi alcançada quando o grupo controle foi utilizado; a associação de cetamina/xilazina/propofol forneceu apenas 11 ejaculados de um total de 12 tentativas (de eficiência 91,6%), enquanto apenas quatro ejaculados (eficiência de 33%) foram obtidos com butorfanol/cetamina/propofol (P<0,05). Ambos os protocolos forneceram rápida indução e relaxamento suficientes para executar a EEJ. Na utilização de butorfanol/cetamina/propofol, os animais se recuperaram em 16,5±1,5min, um tempo mais curto do que no uso de xilazina/cetamina/protocolo de propofol, 20,7±1,0min (P>0,05). O volume de sêmen e a concentração espermática obtidos no uso da associação xilazina/cetamina/propofol foram significativamente maiores do que os verificados para o protocolo butorfanol/cetamina/propofol. Em conclusão, a associação de cetamina/xilazina/propofol é indicada para anestesia de tatus-peba submetidos à EEJ.(AU)

Advances in the management of infertility in men with spinal cord injury / 亚洲男科学杂志(英文版)

Couples with a spinal cord injured male partner require assisted ejaculation techniques to collect semen that can then be further used in various assisted reproductive technology methods to achieve a pregnancy. The majority of men sustaining a spinal cord injury regardless of the cause or the level of injury cannot ejaculate during sexual intercourse. Only a small minority can ejaculate by masturbation. Penile vibratory stimulation and electroejaculation are the two most common methods used to retrieve sperm. Other techniques such as prostatic massage and the adjunct application of other medications can be used, but the results are inconsistent. Surgical sperm retrieval should be considered as a last resort if all other methods fail. Special attention must be paid to patients with T6 and rostral levels of injury due to the risk of autonomic dysreflexia resulting from stimulation below the level of injury. Bladder preparation should be performed before stimulation if retrograde ejaculation is anticipated. Erectile dysfunction is ubiquitous in the spinal cord injured population but is usually easily managed and does not pose a barrier to semen retrieval in these men. Semen analysis parameters of men with spinal cord injury are unique for this population regardless of the method of retrieval, generally presenting as normal sperm concentration but abnormally low sperm motility and viability. When sperm retrieval is desired in this population, emphasis should be placed on initially trying the simple methods of penile vibratory stimulation or electroejaculation before resorting to more advanced and invasive surgical procedures.

Reproductive evaluation of red deer (Cervus elaphus) stags under tropical conditions / Evaluación reproductiva del ciervo rojo (Cervus elaphus) macho bajo condiciones tropicales / Avaliação reprodutiva do cervo rojo (Cervus elaphus) macho sob condições tropicais

Background: evaluation of reproductive traits of red deer is important to understand its performance. Objective: to evaluate seminal traits of red deer (Cervus elaphus) at three different stages of the breeding season (beginning, middle, and end) and to relate semen quality traits with pregnancy rate of hinds. Methods: scrotal circumference, semen volume, mass motility, individual motility, sperm concentration, morphology, and intact acrosomes were evaluated in seven stags. After evaluation, each of five stags was bred to 23 to 30 hinds. Pregnancy diagnosis was carried out using ultrasonography 45 days after the breeding season. Data were analyzed using the Student's t and chi-square tests, and simple correlation procedures. Results: scrotal circumference was reduced 5.4 cm (p<0.05) from the beginning to the end of the reproductive season, although semen volume was similar at the three different stages of the season. Sperm concentration (194 vs. 622.7/10(6)), mass motility (1.6 vs. 2.8), individual motility (28.6 vs. 63.3%), and intact acrosome (52.7 vs. 75.5%) were greater (p<0.05) at the middle of the breeding season in comparison with values found at the beginning. Percentage of abnormal spermatozoa was similar at the beginning and middle of the breeding season (p>0.05). No spermatozoa were found in stags at the end of the breeding season. Pregnancy rates were similar among bucks (p>0.05), ranging from 80% to 91.3%, and there was no relationship between pregnancy rate and semen traits. Conclusions: there was a clear seasonality of semen traits of red deer and no relationship between semen traits and pregnancy rate.

Antecedentes: la evaluación reproductiva de los ciervos es de suma importancia para entender su comportamiento reproductivo bajo el sistema en el cual se producen. Objetivo: evaluar algunos rasgos seminales del venado rojo (Cervus elaphus) en diferentes momentos de la estación reproductiva (inicio, medio y final) y relacionar la calidad seminal con la tasa de preñez de las hembras. Métodos: se utilizaron siete machos para medir la circunferencia escrotal, volumen seminal, motilidad masal, motilidad individual, concentración de espermatozoides, morfología y acrosomas intactos. Después de la evaluación, cinco machos fueron apareados con 23 a 30 hembras cada uno. El diagnóstico de preñez se realizó por ultrasonografía, 45 días después de la estación de apareamiento. Los datos se analizaron mediante las pruebas de t de Student y chi cuadrado, y procedimientos de correlación simple. Resultados: la circunferencia escrotal se redujo 5,4 cm (p<0,05) del inicio al final de la estación reproductiva, aunque el volumen de semen fue similar en los tres momentos de la estación reproductiva. La concentración de espermatozoides (194 vs 622,7/106), motilidad masal (1,6 vs 2,8), motilidad individual (28,6 vs 63,3%) y acrosomas intactos (52,7 vs 75,5%) fueron mejores (p<0,05) en la mitad de la estación reproductiva que en comparación con los valores del inicio de la misma. El porcentaje de espermatozoides anormales fue similar al inicio y en la mitad de la estación reproductiva (p>0,05). No se encontraron espermatozoides al final de la estación reproductiva. Las tasas de preñez fueron similares entre machos (p>0,05) variando de 80 a 91,3%, y no se encontró relación entre la tasa de preñez y los rasgos seminales. Conclusiones: hubo una clara estacionalidad de los rasgos seminales del venado rojo bajo las condiciones tropicales y una falta de relación entre los rasgos seminales y la tasa de preñez.

Antecedentes: a avaliação reprodutiva do cervo rojo é muito importante para entender seu comportamento reprodutivo no sistema em que eles são produzidos. Objetivo: avaliar algumas caraterísticas do sêmen do cervo rojo (Cervus elaphus) em diferentes momentos da estação reprodutiva (inicio, meio e final) e relacionar a qualidade seminal com a taxa de prenhez das fêmeas. Métodos: foram utilizados sete machos para medir a circunferência escrotal, volume seminal, motilidade masal, motilidade individual, concentração de espermatozóides, morfologia e acrossomas intactos. Depois da avaliação, cinco machos foram acasalados com 23-30 fêmeas cada um. El diagnóstico de gestação foi realizado por ultrassonografia, 45 dias depois da estação de acasalamento. Os dados foram analisados usando provas de t de Student e chi-quadrado, e procedimentos de correlação simples. Resultados: a circunferência escrotal se reduz 5,4 cm (p<0,05) do inicio ao final da estação reprodutiva, ainda o volume de sêmen foi similar nos três momentos da estação reprodutiva. A concentração de espermatozoides (194 vs 622,7/10(6)), motilidade masal (1,6 vs 2,8), motilidade individual (28,6 vs 63,3%) e acrossomas intactos (52,7 vs 75,5%) foram melhores (p<0,05) na parte meia em comparação com os valores do inicio da estação reprodutiva. A porcentagem de espermatozoides anormais foi similar ao inicio e em meio da estação reprodutiva (p>0,05). No se encontraram espermatozoides ao final da estação reprodutiva. As taxas de prenhez foram similares entre machos (p>0,05) variando de 80 a 91,3%, e não se encontrou relação entre a taxa de prenhez e os rasgos seminais. Conclusões: houve uma clara estacionalidade dos rasgos seminais do cervo rojo sob as condiciones tropicais e uma falta de relação entre os rasgos seminais e a taxa de prenhez.

Contenção farmacológica da jaguatirica, Leopardus pardalis, para coleta de sêmen, pela associação de tiletamina-zolazepam e xilazina / Chemical restraint of ocelot, Leopardus Pardalis, for semen collection with the association of tiletamine-zolazepam and xylazine

Foram avaliados os efeitos anestésicos da associação de cloridrato de tiletamina (7 mg/kg), cloridrato de zolazepam (7 mg/kg) e cloridrato de xilazina (1 mg/kg), doses calculadas para peso estimado, aplicados por via intramuscular profunda para a contenção farmacológica de machos de jaguatirica, (Leopardus pardalis. Linnaeus, 1758) (Felidae), submetidos à coleta de sêmen por eletroejaculação. Quatro animais foram anestesiados em quatro a cinco ocasiões diferentes, com intervalo mínimo de 30 dias, totalizando 17 procedimentos. Em algumas ocasiões, quando a contenção farmacológica inicial não propiciava segurança para a manipulação do animal, foi necessária a aplicação complementar de anestésicos, sendo então administrada 1/3 da dose inicial, fato necessário em apenas três (17,64%) do total de procedimentos. Após a administração das drogas, foram monitorados durante 80 minutos parâmetros como frequência cardíaca, frequência respiratória, temperatura retal, miorrelaxamento e nocicepção. O período de latência foi de 3,56±0,55 minutos após a injeção e o tempo para recuperação foi de 169,60±12,37 minutos. Não foi evidenciada diferença estatística (p>0,05) entre os animais e os momentos avaliados para reações dolorosas, miorrelaxamento, bem como para os parâmetros: frequências respiratória e cardíaca e temperatura retal. A associação de tiletamina-zolazepam e xilazina, nas doses de 7mg/kg e 1mg/kg respectivamente, proporcionou uma boa qualidade anestésica, curto período de latência, bom período anestésico hábil e com segurança para a realização do procedimento de eletroejaculação nas jaguatiricas, com boa qualidade do sêmen obtido.

The anesthetic effects of tiletamine hydrochloride (7 mg/kg), zolazepam hydrochloride (7 mg/kg) and xylazine (1 mg/kg) association, doses calculated by the estimated weight, were evaluated after deep intramuscular injection for pharmacological restraint of male ocelot (Leopardus pardalis. Linnaeus, 1758) (Felidae), subjected to semen collection by electroejaculation. Four animals were anesthetized on four to five different occasions with an interval of 30 days, totaling 17 procedures. In some instances, when the initial pharmacological restraint was not propitiated safety for handling the animal, it was necessary to apply supplementary anesthetics, and then administered 1/3 of the initial dose, fact necessary only in three procedures (17.64%). After the injection, parameters like heart rate, respiratory rate, rectal temperature, muscle relaxation and nociception was monitored for 80 minutes. The latency period was 3.56±0.55 minutes after injection and the time for recovery was 169.60±12.37 minutes. There was no significant statistical difference (p>0.05) between the animals and moments evaluated for painful reactions, muscle relaxant, as well as for the parameters: respiratory and heart rates and rectal temperature. The combination of tiletamine-zolazepam and xylazine at doses of 1mg/kg and 7mg/kg respectively, provided a good quality of anesthesia, short latency period, good viable anesthetic period and safety to proceed the electroejaculation in ocelots, with good quality of obtained semen.

Comparação entre duas concentrações de glicerol para a criopreservação de sêmen de suçuarana (Puma concolor) / Comparison between two glycerol concentrations to cryopreservation of semen of mountain lions (Puma concolor)

O desenvolvimento de biotécnicas de reprodução é uma importante ferramenta para a conservação de animais silvestres ameaçados de extinção. Procedimentos de reprodução assistida em suçuarana, no entanto, são escassos na literatura, em especial aqueles relacionados à criopreservação de sêmen. Neste sentido, o presente trabalho objetivou avaliar a congelabilidade do sêmen de suçuaranas adultas mantidas em cativeiro, por meio da comparação entre duas concentrações de glicerol no meio de congelamento. Foram usados cinco machos adultos de suçuarana, mantidos no Centro de Reabilitação de Animais Silvestres do Mato Grosso do Sul (CRAS/MS). As amostras foram coletadas por eletroejaculação e avaliadas quanto ao seu aspecto físico, volume, vigor, motilidade, concentração e índice espermático. De cada ejaculado duas alíquotas foram diluídas em meio Tris-citrato-gema de ovo, em concentrações finais de 5 e 7,5% de glicerol, resfriadas a uma taxa de -0,55ºC/min e congeladas a uma taxa de -5,8ºC/min. Depois de descongeladas, as amostras foram reavaliadas e submetidas aos testes de termorresistência e hiposmótico. O protocolo de criopreservação e descongelamento de sêmen proposto se mostrou eficiente em ambas as concentrações de glicerol testadas, não havendo diferença (p>0,05) entre estas.

The development of biotechnologies of reproduction is an important tool for the conservation of wild animals threatened with extinction. Assisted reproduction procedures in mountain lions, however, are scarce, especially those related to sperm cryopreservation. In this context, this study aimed to evaluate the freezing capacity of semen from adult mountain lions in captivity through the comparison of two concentrations of glycerol in the freezing media. Five adult male mountain lions were used, held at the Rehabilitation Center for Wild Animals of Mato Grosso do Sul (CRAS/MS). Samples were collected by electroejaculation and evaluated for physical appearance, volume, sperm progressive status, sperm motility, sperm concentration and sperm motility index. Each ejaculate was spliced into two aliquots and diluted in Tris-citrate-half egg yolk, at final concentrations of 5 and 7.5% glycerol, cooled at a rate of -0.55ºC/min and frozen at a rate of -5.8ºC/min. Once thawed, the samples were re-evaluated and tested for thermoresistance and hypoosmotic swelling. The suggested protocol for cryopreservation and thawing of semen is efficient in both glycerol concentrations tested, with no difference (p>0.05) between them.

Colheita de sêmen por eletroejaculação em cutia-parda (Dasyprocta azarae) / Semen collection by electroejaculation in Azara´s agouti (Dasyprocta azarae)

As cutias são animais que vivem nas Américas Central e do Sul, com potencial zootécnico e que despertam crescente interesse científico. O objetivo deste estudo foi estabelecer um padrão para colheita de sêmen por eletroejaculação em cutia-parda (Dasyprocta azarae). Foram utilizados machos adultos (n=4) anestesiados e submetidos a quatro séries de 20 estímulos eletroejaculatórios, com 3 segundos de duração cada, com uma intensidade de 2, 4, 6 e 8 volts, com dois minutos de intervalo entre cada série. A colheita de sêmen foi obtida em todos os animais, com ereção parcial com 2 volts, ereção evidente com 4 volts, ejaculação com 6 volts e exacerbação da glande e exteriorização das espículas penianas com 8 volts, observando-se espermatozoides em 100% das amostras. A técnica descrita para colheita de sêmen em cutia-parda foi eficaz.

Agoutis are animals that live in South and Central America, present rearing potential and call increasing scientific interest. The aim of this paper was to establish a model for semen collection by electroejaculation in Azara´s agouti (Dasyprocta azarae). Anesthetized adult males (n=4) were submitted to four series of 20 electroejaculatory stimuli, during 3 seconds each and with 2, 4, 6 and 8 volts, with a 2 minutes rest among series. Semen collection was accomplished in every animal, with partial penis erection at 2 volts, makeable erection at 4 volts, ejaculation at 6 volts, glans and penis spines exposition at 8 volts, and sperm collected in all samples. The described technique for Azara's agouti semen collection was efficient.

Conservação do sêmen e liquefação do coágulo seminal de macaco-prego (Cebus apella) em água de coco em pó (ACP-118®), em diferentes temperaturas / Semen conservation and seminal clot liquefaction of capuchin monkey (Cebus apella) in powdered coconut water extender (PCW) at different temperatures

O objetivo do estudo foi avaliar a água de coco em pó (ACP) na conservação do sêmen e liquefação do coágulo seminal de Cebus apella. O sêmen de seis machos adultos foi coletado por eletroejaculação (EEJ), diluído em solução à base de ACP-118® e submetido à incubação em banho-maria a 33, 35 e 37°C, por 24 horas. Avaliou-se a integridade espermática por meio da coloração eosina-nigrosina a cada uma hora durante as seis horas iniciais e após 24 horas de incubação. Os volumes médios e as concentrações espermáticas das frações coagulada e líquida foram de 0,20±0,02 e 0,20±0,10mL; 1,1±0,3x10(8) e 1,3±0,9x10(7) espermatozoides mL-1, respectivamente. Somente em uma amostra da fração líquida foram verificados espermatozoides com motilidade (20 por cento) e vigor (4), perdurando por 40 minutos. A maior parte do coágulo liquefez em ACP-118® após 12 horas de incubação. O melhor tratamento observado foi sob 33°C, por manter até 47±12,8 por cento de espermatozoides vivos após 24 horas. Conclui-se que o diluente à base de ACP é eficiente na liquefação do coágulo seminal e na manutenção da integridade espermática até 24 horas após a EEJ, nas temperaturas de 33, 35 e 37°C.

The aim of this study was to evaluate the powdered coconut water (PCW) in the semen conservation and seminal clot liquefaction. The semen of six adult male Cebus apella was collected by electroejaculation (EEJ), diluted in ACP-118® extender and stayed in water bath at 33, 35 and 37°C for 24 hours. The sperm integrity was evaluated by eosin-nigrosine staining every one hour during the six initial hours and after 24 hours of incubation. The average volumes and sperm concentrations of clotted and liquid fractions were 0.20±0.02 and 0.20±0.10mL, 1.1±0.3x108 and 1.3±0.9x107 sperm mL-1, respectively. Immediately after collection, only in a sample of liquid fraction was observed 20 percent motility and vigor 4, which stopped after 40 minutes. Most of the clot was liquefied in ACP-118® after 12 hours of incubation. The best observed treatment was 33°C, because it kept 47±12.8 percent of sperm integrity after 24 hours. It was concluded that the PCW extender is effective in the liquefaction of seminal clot and maintenance of sperm viability 24 hours after the EEJ at 33, 35 and 37°C.

Improvement of Sperm Quality by Pseudoephedrine in Spinal Cord Injured Males

OBJECTIVE: To investigate the changes of sperm quality by taking adrenergic agonist in spinal cord injured (SCI) males treated by repeated electro-ejaculation. METHOD: Twenty-six SCI males were induced three times to ejaculate with rectal probe electrical stimulation, and then semen analysis for their antegrade and retrograde ejaculates was performed, at weekly interval, while they either took pseudoephedrine (Sudafed(R)) for 2 weeks or no medication as a control. RESULTS: Before pseudoephedrine medication, there was no significant difference in sperm quality between two groups. However, after 2 weeks' medication, SCI males taking pse-udoephedrine showed increased semen concentration, motile sperm count and sperm motility of the antegrade ejaculates compared to control group, although there was no difference in any parameter of the retrograde ejaculates. Moreover, sperm motility of the antegrade ejaculates significantly improved after 2 weeks' medication compared with those at the baseline. CONCLUSION: Pseudoephedrine, a sympathomimetic adrenergic agonist, may potentiate the improvement of sperm quality in SCI males undergoing repeated electro-ejaculation for short periods.

Semen parameters retrieved by electroejaculation in spinal cord injured Thai men: Preliminary report.

Semen characteristics from seven spinal injured Thai men retrieved by electroejaculation were studied at Spinal cord injury unit and Infertility unit, Siriraj hospital. The mean age of the subjects was 24.7 years. Three cases had sperm count within normal limit ( > 20x106/c.c. ) and oligospermia in three cases. One case had azoospermia. All had low motility or no motility at all. All had leukospermia ( > 1x106/c.c. ) except one case. This study shows the change in semen parameters and may help for planning the management of spinal cord injured men.

Clinical Experience with Electroejaculation in Anejaculatory Spinal Cord Injured Men Who Failed in Chemoejaculation Using Physostigmine / 대한비뇨기과학회지

About 85-97% of patients get to lose an ejaculatory function with a little difference according to the level of spinal cord injury. Fifty eight trials of electroejaculation stimulation were performed in total 17 patients that failed in chemoejaculation with physostigmine. Mean age was 36.9 years. Of the 17 spinal cord injured patients, 1 had cervical, 13 thoracic 3 lumbar paraplegia; 7 had complete and 10 had incomplete lesions. Semens could be obtained in 14 patients ( 82.4%) among total 17 patients and in 51 of total 58 trials( 87.9%). Average total sperm counts and motile sperm numbers were 35.98 x 106 and 6.09 x 106 respectively in the obtained semen. However, motility was very poor. We performed more than 4 trials in 6 patients, but there results were not predictable. Two pregnancies were achieved in their spouse after an artificial insemination. Side effects were trivial. and there were lower abdominal discomforts or pains in 7 patients among incomplete paraplegics. In conclusion we could expect high successful rates of ejaculation in anejaculatory spinal cord injured patients by electroejaculation stimulation in case of the failure in chemoejaculation. However, we suggest that in vitro fertilization with advanced recent technology should be tried with adequate preparation after an experimental electroejaculation, because a quality of sperm motility was very poor and sperm motility was not improved after several trials.

Successful combination therapy with electroejaculation and in vitro fertilization-embryo transfer in the treatment of a patient with neurogenic anejaculation / 대한비뇨기과학회지

Recent case reports of the successful application of electroejaculation techniques for seminal emission and subsequent pregnancies have offered promising fertility alternative for the men with neurogenic anejaculation. The success of his program requires multidisciplinary couple-oriented therapy combining both turologic and gynecologic infertility therapy. However, despite obtaining often normal numbers of spermatozoa. sperm motility rarely exceeds 10-20% in electroejaculation sample. Therefore, realistic potential for pregnancy with intrauterine insemination technique is extremely low. In this report we present the first domestic successful coordination of both electroejaculation and in vitro fertilization-embryo transfer in a neurogenic anejaculation patient with severe oligoasthenospermia.