Assessment of ram sperm membrane integrity following different thawing procedures

Söderquist, L.; Madrid-Bury, Ninoska; Rodriguez‐Martinez, Heriberto

doi:10.1016/s0093-691x(97)00344-0

Cited by 54 publications

(35 citation statements)

References 12 publications

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“…Para isso, uma amostra do sêmen descongelado (100 mL) foi acondicionada em tubo eppendorf com solução de formol salina tamponado (Hancock, 1957). Posteriormente, por ocasião do exame, confeccionou-se a preparação úmida da amostra de sêmen entre lâmina e lamínula, procedendo-se à avaliação acrossomal por meio de microscopia com contraste de fase em aumento de 1.000 vezes e imersão (Soderquist et al, 1997). O percentual de anormalidade de acrossoma foi feito sobre a contagem de 200 células espermáticas, computando-se os acrossomas intactos e anormais.…”

Section: Methodsunclassified

“…Entretanto, esses índices de danos acrossomais (edema, desprendimento parcial e até perda total de acrossoma) são aceitá-veis, tanto no pós-descongelamento como após o término do TTR. Em média, os valores encontrados foram de 50% de lesões acrossomais de 0 e 2 horas após o descongelamento, enquanto Tasseron et al (1977), Memon et al (1985), Soderquist et al (1997) obtiveram valores que de 35 a 40%.…”

Section: Alpina Saanenunclassified

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Uso de testes complementares para avaliação do congelamento do sêmen de bodes submetidos ao manejo de fotoperíodo artificial

Torres¹,

Fonseca

Borges³

et al. 2006

R. Bras. Zootec.

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RESUMO -O congelamento do sêmen de bodes das raças Alpina e Saanen submetidos ao manejo de fotoperíodo artificial foi avaliado por meio dos testes de termorresistência (TTR), hiposmótico (HOST) e de integridade do acrossoma. Foram utilizados oito machos caprinos (quatro da raça Alpina e quatro da raça Saanen) de duas idades diferentes (jovens e adultos). A qualidade do sêmen durante as etapas do congelamento foi superior em bodes jovens de ambas as raças. Avaliada pelo TTR, a motilidade do sêmen fresco apresentou longevidade pós-descongelamento. Os resultados de motilidade espermática, obtidos imediatamente após o descongelamento, tiveram reflexos positivos sobre o TTR, indicando que os sêmens que apresentaram maior motilidade pós-coleta (86,2% vs 79,3%) e pós-descongelamento (37,7% vs 32,0%) tiveram maior longevidade seminal. Os resultados do HOST, tanto para o sêmen fresco quanto para o congelado, não diferiram entre raças e idades. Houve redução na porcentagem de espermatozóides íntegros após o congelamento e descongelamento do sêmen dos animais da raça Alpina e dos adultos Saanen. Houve redução superior a 14% nos valores do HOST para o sêmen congelado em relação aos valores observados para o sêmen fresco (38,0 vs 52,0%, respectivamente). A motilidade espermática progressiva mostrou maior sensibilidade à criopreservação que a integridade da membrana espermática, indicando que a motilidade espermática é mais afetada pelo processo de congelamento que a membrana plasmática. A integridade do acrossoma não foi influenciada pelo descongelamento. Os índices de danos acrossomais (edema, desprendimento parcial e até perda total de acrossoma) mostraram-se dentro do padrão aceitável (44,5%), tanto no pós-descongelamento como após o término do TTR.Palavras-chave: caprino, congelamento, hiposmótico, integridade do acrossoma, sêmen, termorresistência Use of complementary tests to evaluate the freezing ability of semen from goats under artificial photoperiod exposureABSTRACT -The freezing ability of semen from Alpine and Saanen goats submitted to an artificial photoperiod exposure were evaluated by the thermo-resistance (TTR) and hypoosmotic (HOST) tests and integrity of the acrosome using eight male goats, four of each breed and two age groups (young and adults). The quality of semen during the freezing process was superior for the young animals of both breeds. Spermatic motility immediately after thawing showed a positive effect on TTR; animals with higher motility after collection (86.2% vs 79.3%) and after thawing (37.7% vs. 32.0%) had a higher seminal longevity. Both fresh and frozen semen did not differ between breeds and ages by the HOST test. There was a 14% decrease (P<0.05) in HOST values for frozen semen compared the fresh semen (38.0 vs. 52.0%, respectively). The spermatic progressive mobility was more sensible to cryopreservation than the integrity of the spermatic membrane, indicating that spermatic mobility is more affected by the frozen process than the plasmatic membrane. The level of acrosome damage (edema...

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Section: Methodsunclassified

Section: Alpina Saanenunclassified

Uso de testes complementares para avaliação do congelamento do sêmen de bodes submetidos ao manejo de fotoperíodo artificial

Torres¹,

Fonseca

Borges³

et al. 2006

R. Bras. Zootec.

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“…According to SODESQUIST et al (1997), fast thawing reduces the formation of intracellular ice crystals and decreases stress in the sperm cell. Conversely, slow thawing exposes the sperm cell to higher temperature oscillations, leading to an increase in lipid and protein deposition, and directly affecting the physical structure of the cell, thus reducing sperm vitality (PEÑA and LINDE-FORSBERG, 2000).…”

Section: Cryopreservation and Cold Storage Testsmentioning

confidence: 99%

Testes de toxicidade, congelamento e refrigeração em sêmen de espécie nativa

et al. 2017

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RESUMOA primeira estapa deste trabalho testou a toxicidade de quatro crioprotetores em espermatozoides de Macrobrachium acanthurus durante 10 e 20 min nas concentrações de 10 e 20%. A segunda etapa foi realizar a criopreservação aplicando o crioprotetor com menor grau de toxicidade, testanto dois mecanismos de congelamento, um automatizado (protocolos A e B) e outro convencional (protocolos C e D), durante 24 horas. O protocolo A apresentou velocidade de resfriamento de 0,5°C min -1 até alcançar -32°C, partindo de uma temperatura de -6°C e idem o protocolo B, com a diferença de partir de uma temperatura ambiente; os protocolos C e D apresentaram uma velocidade de resfriamento de 2 e 10°C min -1 , respectivamente, sendo as palhetas transferidas ao nitrogênio líquido. A terceira etapa foi verificar o tempo de vida do espermatozoide quando refrigerado a 5°C. A viabilidade espermática foi avaliada por meio do esfregaço de sêmen com eosina-nigrosina. Os crioprotetores que se apresentaram menos tóxicos foram o glicerol 10 e 20% e metanol 10%, num tempo de equilíbrio de 10 minutos. A melhor velocidade de congelamento foi a de 2°C min -1 para glicerol 10%, com 21,8% de sobrevivência espermática, sendo a refrigeração até três dias recomendada, com uma sobrevivência de 35,3%.Palavras-chave: caridea; criopreservação; crioprotetores; glicerol; metanol; pitú ABSTRACTIn the first part of this study, toxicity tests were performed on the sperm of Macrobrachium acanthurus using four cryoprotectants for periods of 10 and 20 min at concentrations of 10 and 20%. In the second part, cryopreservation was performed by applying the least toxic cryoprotectant, and two freezing methods were tested over 24 hours: automated (protocols A and B) and conventional (protocols C and D). Protocol A exhibited a cooling rate of 0.5°C min -1 from -6°C to -32°C; protocol B was similar to A except for the starting temperature, which was room temperature; whereas protocols C and D exhibited a cooling rate of 2 and 10°C min -1 , respectively. The third part of the study was conducted to assess the lifespan of the sperm when stored at 5°C, in which sperm viability was evaluated by a semen smear with eosin-nigrosin. The least toxic cryoprotectants were 10 and 20% glycerol, and 10% methanol, and the equilibrium time was 10 minutes. The optimal cooling rate was 2°C min -1 for 10% glycerol, which had a sperm survival rate of 21.8%. Cold storage for up to 3 days is recommended, presenting a sperm survival rate of 35.3%.

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“…The cooling process (during the initial stage of sperm cooling, prior to cryopreservation) causes a transition from fluid to gel phases and decreases membrane fluidity (Holt, 2000), which is probably responsible for thermal shock that may occur at this time. Furthermore, in addition to cooling, sperm must retain their potential viability at -196 o C and subsequently overcome stresses associated with thawing (Söderquist et al, 1997). It is believed that pre-freezing equilibration can help sperm reach an osmotic equilibrium following the addition of cryoprotectant (Emmens and Blackshaw, 1955).…”

Section: Introductionmentioning

confidence: 99%

Influence of catalase and pre-freezing equilibration on post-thaw semen quality and conception rate in ewes laparoscopically inseminated

Câmara

Pinto

et al. 2016

Anim Reprod

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The aim of this study was to investigate the effects of catalase and pre-freezing equilibration during ram sperm cryopreservation on motility and membrane and acrosomal integrity of frozen-thawed semen, as well as conception rate following laparoscopic timedinsemination. Semen was collected from four mature Dorper rams, pooled and diluted in Tris egg-yolk extender basic solution (CON), or this solution supplemented with catalase (CAT; 20 U/100 × 10 6 sperm). Extended semen was packaged in 0.25 ml mini straws (25 × 10 6 sperm/straw), chilled (to 5°C), and then either frozen immediately (CON and CAT) or maintained at 5°C for 12 h of pre-freezing equilibration (CON12 and CAT12). Immediately after thawing and at 1 h after incubation at 37°C, kinematic parameters (CASA), plasma membrane integrity (PI-FITC), and acrosomal status (FITC-PNA) of sperm were assessed. There were no significant differences among the four groups on sperm traits evaluated immediately postthaw. However, after 1 h of incubation, total motility (46.7 and 25.0%) and plasma membrane integrity (38.7 and 25.7%) were higher (P < 0.05) in CAT12 than CON. When these two treatments were used for laparoscopic timed artificial insemination of ewes (with synchronized ovulation), conception rate was similar for CAT12 and CON (32.8%, n = 61 vs. 27.3%, n = 55). In conclusion, the combination of catalase and pre-freezing equilibration resulted in significantly improved quality of post-thawed ram semen without affecting conception rate in fixed-time laparoscopically intrauterine inseminated ewes.

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Assessment of ram sperm membrane integrity following different thawing procedures

Cited by 54 publications

References 12 publications

Uso de testes complementares para avaliação do congelamento do sêmen de bodes submetidos ao manejo de fotoperíodo artificial

Uso de testes complementares para avaliação do congelamento do sêmen de bodes submetidos ao manejo de fotoperíodo artificial

Testes de toxicidade, congelamento e refrigeração em sêmen de espécie nativa

Influence of catalase and pre-freezing equilibration on post-thaw semen quality and conception rate in ewes laparoscopically inseminated

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