Effect of freezing and thawing rates on sperm motility in Bocachico Prochilodus magdalenae (Pisces, Characiformes)

Martínez, José Gregório; C, Sandra Pardo

doi:10.21897/rmvz.191

Cited by 6 publications

(6 citation statements)

References 23 publications

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“…Dry shippers are used in most studies on the cryopreservation of characiform fish sperm (Godinho et al, 2011), and the programmed freezing machine remains underutilized (Carneiro et al, 2012;Martínez and Pardo-Carrasco, 2013). The technique used to decrease seminal temperature during cryopreservation can also influence the results.…”

Section: Discussionmentioning

confidence: 99%

Use of glucose or BTS"! combined with DMSO or methylglycol under two different freezing protocols for the cryopreservation of sperm from the common curimatã (Prochilodus brevis)

et al. 2016

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This study evaluated the effect of glucose or Beltsville Thawing Solution (BTS™) combined with dimethyl sulfoxide (DMSO) or methylglycol (MG) under two different freezing protocols on the kinetics and morphology of cryopreserved Prochilodus brevis sperm. The semen samples were diluted using one of four different treatments (glucose+DMSO, glucose+MG, BTS™+DMSO, and BTS™+MG), loaded into 0.25-ml straws and subjected to two different freezing processes (programmed freezing machine and dry shipper). After 10 days, the semen samples were thawed, and the sperm morphology and kinetics were evaluated. The physicochemical parameters of the semen in natura were similar to those observed in other studies of Characiformes, indicating the feasibility of semen cryopreservation. Glucose, when used as a diluent with the cryoprotectant MG (glucose+MG), yielded higher percentages of mobile spermatozoa after freezing in a dry shipper (76.88 ± 4.84%) and in a programmed freezing machine (70.95 ± 1.76%) compared with the combination of glucose and DMSO. Moreover, the glucose+MG treatment yielded a higher sperm velocity (curvilinear velocity: 79.52 ± 2.88 µm s -1 ; straight-line velocity: 45.46 ± 3.01 µm s -1 ; average path velocity: 67.92 ± 3.08 µm s -1 ) than the other studied treatments, and a higher amount of normal sperm (74.56 ± 0.77%) was observed in the semen samples cryopreserved using a programmed freezing machine. The sperm abnormalities observed included a bent tail morphology. Therefore, the use of glucose+MG in combination with either a dry shipper or a programmed freezing machine is recommended for the cryopreservation of P. brevis sperm because these methods yielded high numbers of motile and morphologically normal spermatozoa.

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Section: Discussionmentioning

confidence: 99%

Use of glucose or BTS"! combined with DMSO or methylglycol under two different freezing protocols for the cryopreservation of sperm from the common curimatã (Prochilodus brevis)

et al. 2016

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“…Damage compromises the cell at the morphological level (membrane), affecting mitochondria, which are critical for energy production and post-thawing cell movement. Additionally, damage also occurs at the DNA level, causing the alteration of the genetic material despite the fact that cells can still move (Martínez and Pardo, 2013).…”

Section: Discussionmentioning

confidence: 99%

“…Additional advantages include broadening artificial fertilization processes in aquaculture (Watson and Holt, 2001) as well as increasing the availability of semen during periods of low sperm production observed in captive fish. Reports on advances in the development of protocols for bocachico semen cryopreservation include: a study on the effect of glucose concentration on sperm motility (Martínez et al, 2011), another study on the use of dimethylacetamide as cryoprotectant (Atencio-García et al, 2013), a further study into the use of different concentrations of dimethyl sulfoxide (DMSO 5, 10, 15%) and glucose (5.5, 6.0, 6.5%) as cryoprotectants (Martínez et al, 2012a), an assessment of membrane damage and DNA fragmentation of bocachico spermatozoa caused by cryopreservation with DMSO and glucose (Martínez et al, 2012b), and a report on the effects of freezing and thawing rates on sperm motility (Martínez et al, 2013). In spite of this, there is little information concerning effective spermatozoa/oocyte ratios for bocachico using fresh and cryopreserved semen to improve the efficiency of semen usage in artificial reproduction processes.…”

Section: Introductionmentioning

confidence: 99%

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Insemination of bocachico fish (Prochilodus magdalenae) with fresh or cryopreserved semen: effect of spermatozoa/oocyte ratio

García¹,

Espinosa²,

Martínez

et al. 2015

Rev Colomb Cienc Pecu

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Insemination of bocachico fish (Prochilodus magdalenae) with fresh or cryopreserved semen: effect of spermatozoa/oocyte ratio ¤ Inseminacion de bocachico (Prochilodus magdalenae) con semen fresco o crioconservado: efecto de la proporción espermatozoide/oocito Inseminação do bocachico (Prochilodus magdalenae) usando sêmen fresco o criopreservado: efeito da razão espermatozoide/oócitoVíctor J Atencio García 1* , MSc; José A Espinosa 1 , MSc; José G Martínez 2 , MSc; Sandra C Pardo Carrasco 3 , PhD. (Received: September 4, 2014; accepted: January 11, 2015) doi: 10.17533/udea.rccp.v28n4a07 Summary Background: cryopreservation is an important biotechnological tool in the conservation of biodiversity, particularly for endangered species. Objective: to evaluate six different spermatozoa/oocyte ratios using fresh and cryopreserved semen in bocachico fish (Prochilodus magdalenae). Methods: fresh semen was collected and its quality determined to verify cryopreservation feasibility. The semen was put in 5 mL straws and mixed with a solution (5.5% glucose, 12% egg yolk, and 10% dimethyl sulfoxide -DMSO-) in a 1:4 dilution (semen:solution). The semen was frozen in nitrogen vapor dry shipper for 30 min, rapidly transferred to storage thermos, and submerged directly into liquid nitrogen (LN;. Straws were thawed at 60 ºC for 45 seconds. Motility, velocity, and sperm progressivity of fresh and cryopreserved semen were assessed using Sperm Class Analyzer (SCA ® ) software. Each proportion of spermatozoa/oocyte was assessed with 2 g of eggs (1,630 ± 87 eggs/g) to evaluate fertility (F), hatching (H), and larval survival (LS) rates. Results: the best reproductive performance for fresh semen was obtained inseminating with 160,000 spermatozoa/oocyte (F = 75.0%, H = 67.7%, LS = 32.7%). Similarly, the best reproductive performance for cryopreserved semen was achieved with 320,000 spermatozoa/oocyte (F = 70.0%, H = 48.6%, LS = 19.5%). Conclusion: it is possible to achieve adequate reproductive performance in bocachico fish using cryopreserved sperm (10% DMSO, 5.5% glucose, and 12% egg yolk) at twice the spermatozoa/oocyte ratio used with fresh semen.Keywords: artificial reproduction, cryobiology, fertility, Prochilontidae. Atencio VJ et al. Fresh and cryopreserved semen in bocachico fishResumen Antecedentes: la crioconservación es una herramienta biotecnológica importante para la conservación de la biodiversidad, particularmente de especies en peligro. Objetivos: evaluar seis proporciones diferentes entre espermatozoides/ovocito en la fertilización de bocachico (Prochilodus magdalenae), usando semen fresco o crioconservado. Métodos: se colectó semen y se determinó su calidad para verificar su viabilidad de crioconservación. El semen fue colocado en pajillas de 5 mL y mezclado con una solución crioconservante (5,5% glucosa, 12% yema de huevo y 10% dimethyl sulfoxido -DMSO-) en una dilución 1:4 (semen:solución). El semen fue congelado en un termo de vapores de nitrógeno por 30 min y rápidamente se transfirió a termos de almacenami...

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“…It is also possible that the concentration of glucose affects the membrane through changes in osmotic pressure, considering that the damages increased significantly when reaching levels of 6% (360 mOsm kg -1 ) to a more hyperosmotic concentration of 6.5% (385 mOsm kg -1 ) respecting the seminal plasma of the species: 250 mOsm kg -1 (Martínez et al, 2011). The results of the present study permit the conclusion that cryopreservation originates significant damage to the DNA and plasmatic membrane of sperm cells of bocachico whose magnitude will depend on the variation of the concentration of the external cryoprotectant (glucose) as well as the internal (DMSO) and their interactions.…”

Section: Membrane Damage During Cryopreservationmentioning

confidence: 99%

DNA fragmentation and membrane damage of bocachico Prochilodus magdalenae (Ostariophysi: Prochilodontidae) sperm following cryopreservation with dimethylsulfoxide and glucose

2012

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The endangered bocachico Prochilodus magdalenae is a native freshwater fish of Colombia, the most captured species locally and one of the most important species for ex-situ conservation (germplasm banks). The aim of this study was to examine the effect of three concentrations of Dimethylsulfoxide (DMSO) (5%, 10%, 15%) and three of glucose (305, 333, 361 mM) in the extender on spermatic DNA fragmentation (F-DNA) (by Halomax®, Chromatin dispersion) and membrane damage (D-Me) (by eosin-nigrosin staining). After assessment of sperm quality by computer analysis of motility, one part of semen from males was diluted separately with three parts of extender and filled into 0.5 ml straws. Freezing was carried out in liquid nitrogen vapor dry shipper for 30 minutes and thawed at 60°C for 8 seconds in a water bath and evaluated for the percentage of cells found with F-DNA and D-Me. The results demonstrated that cryopreservation causes greater F-DNA (13.62 ± 1.6% to 28.91 ± 3.25) and D-Me (24.27 ± 1.1% to 58.33 ± 2.81%) when compared with pre-freezing semen (PFS) (6.71 ± 1.54% and 2.34 ± 0.5%, respectively for F-DNA and D-Me). A significant interaction was found between DMSO and glucose concentration in this experiment. Use of extender: 10% DMSO + 305 mM glucose + 12% chicken egg yolk and, 10% DMSO + 333 mM glucose + 12% chicken egg yolk, allow for lower F-DNA and D-Me during cryopreservation of bocachico semen. A high correlation between F-DNA and D-Me was found (r = 0.771).O curimba Prochilodus magdalenae, é uma espécie nativa de água doce da Colômbia ameaçada de extinção, sendo a mais capturada localmente e uma das mais importantes para a conservação ex-situ (bancos de germoplasma). O objetivo deste estudo foi avaliar o efeito de três concentrações de dimetilsulfóxido (DMSO) (5%, 10%, 15%) e três de glicose (305, 333, 361 mM) no diluente sobre a fragmentação do ADN espermático (F-DNA) (através de Halomax®, dispersão da cromatina) e danos em membrana (D-Me) (através da coloração eosina-nigrosina). Depois de avaliar a qualidade espermática por análise computacional da mobilidade, uma parte do sêmen dos machos foi diluída separadamente com três partes do diluente e colocado dentro de canudos de 0.5 ml. O congelamento foi feito em tanque de vapores secos de nitrogênio líquido por 30 minutos e descongelado a 60°C por 8 segundos em banho de água e avaliada a percentagem de células com F-DNA e D-Me. Os resultados demonstraram que a criopreservação causa grande F-DNA (13.62 ± 1.6% até 28.91 ± 3.25) e D-Me (24.27 ± 1.1% até 58.33 ± 2.81%) quando comparada com o sêmen pré-congelamento (PFS) (6.71 ± 1.54% e 2.34 ± 0.5%, respectivamente para F-DNA e D-Me). Uma interação significativa foi encontrada entre a concentração de DMSO e glicose neste experimento. O uso dos diluentes 10% DMSO + 305 mM glicose + 12% gema de ovo de galinha e 10% DMSO + 333 mM glicose + 12% gema de ovo de galinha permitem obter a menor F-DNA e D-Me durante a criopreservação do sêmen de curimba. Uma alta correlação entre F-DNA e D-Me foi encontrada (r = 0.771).

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Effect of freezing and thawing rates on sperm motility in Bocachico Prochilodus magdalenae (Pisces, Characiformes)

Cited by 6 publications

References 23 publications

Use of glucose or BTS"! combined with DMSO or methylglycol under two different freezing protocols for the cryopreservation of sperm from the common curimatã (Prochilodus brevis)

Use of glucose or BTS"! combined with DMSO or methylglycol under two different freezing protocols for the cryopreservation of sperm from the common curimatã (Prochilodus brevis)

Insemination of bocachico fish (Prochilodus magdalenae) with fresh or cryopreserved semen: effect of spermatozoa/oocyte ratio

DNA fragmentation and membrane damage of bocachico Prochilodus magdalenae (Ostariophysi: Prochilodontidae) sperm following cryopreservation with dimethylsulfoxide and glucose

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