ResumenEl objetivo del presente estudio fue evaluar la cualidad espermática del semen refrigerado de carneros, en cajas térmicas con diferentes temperaturas. Fueron utilizados 6 eyaculados, de 2 carneros colectado con vagina artificial. En el semen se analizó volumen, movimiento en masa, motilidad, vigor, concentración espermática, test hipo-osmótico y coloración supra-vital. Las muestras fueron divididas en 2 alícuotas y diluidas en solución NaCl 0,9% o en un medio (Botubov ® , Botupharma, Botucatu, SP, Brasil); después, la dilución fue mantenida a temperatura ambiente, nevera (5ºC), caja Botubox ® (15ºC, Botupharma, Botucatu, SP, Brasil), caja Botutainer ® (5ºC, Botupharma, Botucatu, SP, Brasil) y caja MaxSemen ® (15ºC, EHG Agrofarma, Campinas, SP, Brasil). Todas las muestras fueron analizadas cada 24 horas, siguiendo los mismos parámetros. De acuerdo con los resultados, las muestras mantenidas en el diluyente presentaron viabilidad hasta 48 horas de refrigeración en las cajas térmicas y en la nevera, y la motilidad se mantuvo entorno de 30% hasta las 72 horas. Las muestras diluidas en solución NaCl 0,9% conservaron la motilidad en 30% hasta las 24 horas de refrigeración. Basado en los resultados se concluye que las tres cajas pueden ser utilizadas para transporte de semen ovino, diluido y refrigerado por un período de 24 a 48 horas.Palabras clave: carnero, crio-preservación, espermatozoide. AbstractThe aim of this study was to evaluate the ovine sperm quality colled in thermal boxs. Six ejaculates from 2 rams were collected using artificial vagina. Samples were analyzed to volume, mass movement, motility, vigor, sperm cell concentration, hypo-osmotic swelling test and supravital stain. Ejaculates were divided into 2 aliquots and diluted in 0.9% NaCl or in the extender (Botubov ® , Botupharma, Botucatu, SP, Brazil), and was kept at room temperature, refrigerator (5°C), Botubox ® (15°C, Botupharma, Botucatu, SP, Brazil), Botutainer ® (5ºC, Botupharma, Botucatu, SP, Brazil) and MaxSemen ® (15°C, EHG Agrofarma, Campinas, SP, Brazil). All samples were analyzed each 24 hours to the same parameters. According to the results the samples diluted and cooled in extender preserved sperm viability until 48 hours in thermal boxes and in refrigerator; and the samples extended preserved in environment maintained motility 30% up to 72 hours. Samples diluted in 0.9% saline retained motility around 30% until 24 hours of cooling. Based on the results it is concluded that the three boxes may be used for transport of ram semen, diluted and refrigerated for a period of 24 to 48 hours.Keywords: cryopreservation, ram, spermatozoa. IntroducciónLa criación de ovinos en los últimos años ha crecido y el mercado internacional de carne ovina se mantiene en alta, debido al menor costo de producción. Este crecimiento ha impulsado y estimulado la aplicación de biotécnicas de reproducción en la especie (Oliveira y Oliveira, 2008) con el objetivo de aumentar la productividad y rentabilidad del rebaño.Dentro de las biotecnologías se debe resalta...
Pesq. Vet. Bras. 31(Supl.1):33-38, dezembro 2011 33 RESUMO.-A principal barreira para a aplicação da inseminação artiϐicial transcervical é a anatomia cervical aliada à baixa viabilidade e sobrevida do sêmen ovino congelado. Assim, este experimento teve como objetivo estudar a morfologia da cérvice de ovelhas. Para tal, foram adquiridas, em matadouro, 81 peças do trato reprodutor de ovelhas, nas quais se avaliou a morfologia cervical, segundo as seguintes características: tipo de óstio cervical, mensuração do tamanho da cérvice, integralidade e interdigitação entre os anéis das pregas cervicais, tamanho e característi-cas macroscópicas dos ovários (folículos e corpo lúteo) e tempo da passagem do aplicador de sêmen pela cérvice. Foi identiϐicada maior frequência do tipo liso de abertura da cérvice e integralidade e interdigitação dos anéis grau II. O tempo de passagem do aplicador pela cérvice foi em The major barrier to accomplishment of artiϐicial insemination in ewes is the anatomy of the cervix combined with low viability and survival of frozen ram semen. Thus, the aim of the present study was to evaluate the morphology of the cervix in ewes. Eighty-one specimens were obtained from a slaughterhouse and the following characteristics were evaluated: type of external cervical os, cervical size, integrity and interdigitation of cervical rings, gross characteristics and size of the ovaries (follicles and corpus luteum), and the time spent to pass an insemination catheter through the lumen of the cervix. The most frequent was the slit type cervical opening and grade II internal ring arrangement. The time spent to pass the insemination catheter through the cervix was 6 minutes and 15 seconds, and the dye was spread throughout the cervical lumen reaching the uterus in most sheep. The average values of cervical opening diameter and cervical length were 0.68cm and 4.4cm respectively. Ovarian follicular activity was found in 75% of the ewes. A positive correlation was established between some of the variables. We conclude that cervical opening size is inϐluenced by estrogen, slit type cervical os and grade III cervical ring arrangement; also, the greater length of the cervix was associated with greater difϐiculty to pass the insemination catheter. INTRODUÇÃOInúmeras tentativas já foram realizadas para superar a barreira cervical em ovelhas e alcançar o útero, tendo como objetivo a inseminação artiϐicial (IA). Isto se deve a anatomia da cérvice, a qual é o principal fator que diϐiculta a IA
ResumoO objetivo deste estudo foi avaliar o efeito da refrigeração do epidídimo, sobre a viabilidade dos espermatozoides congelados. Foram colhidos dez pares de testículos/epidídimos de um abatedouro comercial. Um dos epidídimos foi refrigerado a 4°C durante 12 horas e o contralateral foi imediatamente processado. O epidídimo foi isolado, as células espermáticas extraídas e analisadas quanto à motilidade, vigor, concentração, morfologia e integridade da membrana. Após a análise, os espermatozoides foram congelados em diluente Botubov ® (Botupharma Biotecnologia Animal, Botucatu, SP, Brasil) e descongelados para análise. O mesmo procedimento foi realizado com o testículo/epidídimo refrigerado. Os resultados evidenciaram maior viabilidade (p≤0,05) das células pré-congelação para os parâmetros, número total de espermatozides (1,9 ± 1,2 versus 0,9 ± 0,9 x 10 9 espermatozoides), motilidade (74,0 ± 15,1 versus 20,5 ± 13,8%) e vigor (3,7 ± 0,5 versus 1,7 ± 0,8), e pós-congelação, motilidade (23,5 ± 16,7 versus 8,0 ± 7,9%) e vigor (2,0 ± 0,8 versus 0,8 ± 0,8) quando os espermatozoides foram colhidos a partir de epidídimos processados imediatamente após o abate quando comparados aos mantidos 12 horas sob refrigeração, respectivamente. Conclui-se que 12 horas de refrigeração do epidídimo após o abate, prejudica a qualidade das células espermáticas, impossibilitando a congelação do sêmen.Palavras-chave: bovino, cauda-epidídimo, célula-espermática, congelação. AbstractThe purpose of this study was to evaluate effect of epididymis cooling on bovine frozen sperm viability. Ten pairs of testes/epididymes were collected of a commercial slaughterhouse; one epididymis from each pair was refrigerated at 4ºC for 12 hours and the other immediately proceeded. Epididymis was isolated, sperm cells collected after epididymal slicing and then analyzed regarding to motility, vigor, total number of sperm, morphology and membrane integrity. Sperm cells were frozen in Botubov® extender (Botupharma Biotecnologia Animal, Botucatu, SP, Brazil) and thawed for semen analysis. The same procedure was performed with cooled testis/epididymis. Results demonstrated higher viability (p≤0,05) of fresh cells to the total number of spermatozoa (1,9 ± 1,2 versus 0,9 ± 0,9 x 10 9 spermatozoa), sperm motility (74,0 ± 15,1 versus 20,5 ± 13,8%) and vigor (3,7 ± 0,5 versus 1,7 ± 0,8), and for pos-thawing motility (23,5 ± 16,7 versus 8,0 ± 7,9%) and vigor (2,0 ± 0,8 versus 0,8 ± 0,8) when spermatozoa were collected immediately post-slaughter than maintained cooling 12 hours, respectively. We conclude that 12 hours of epididymis cooling after slaughter decreases sperm cells quality.
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