Anormalidades espermáticas de Piaractus mesopotamicus após descongelamento utilizando diferentes metodologias

Paulino, Michelle Sampaio; Murgas, Luís David Solis; Felizardo, V. O.; Freitas, Rilke Tadeu Fonseca de

doi:10.1590/s0102-09352012000600027

Cited by 4 publications

(6 citation statements)

References 8 publications

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“…As for the morphological analysis, the highest percentage of normal sperm was obtained using the thaw rates of 25 °C for 30 sec and 40 °C for 12 sec, regardless of the freezing media; these results are similar to those achieved in the thawing of pacu semen (PAULINO et al, 2012) which was performed at the second thawing rate. In addition, the best result was observed using glucose and MG, corroborating the findings of Lopes et al (2014).…”

Section: Sperm Cryopreservation and Thawingsupporting

confidence: 72%

Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates

Nunes

Oliveira

Lopes

et al. 2016

SCA

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Prochilodus brevis is a migratory fish, an important component of its river ecosystem and an appreciated animal in northeastern cuisine. However, human activities have threatened its survival. Thus, researchers have become interested in developing genetic material storage protocols, such as seminal cryopreservation. Therefore, determination of the appropriate freezing media and thawing rate is a fundamental step toward the use of this biotechnology in the production of common curimatã and for reducing risks to the species' survival. As such, the aim of the current study was to evaluate the effect of different freezing media and thawing rates on the quality of cryopreserved semen from P. brevis. For this study, males received a single dose of pituitary extract of carp 18 hours before semen collection. The semen samples were diluted in 5% glucose + 10% methyl glycol (MG), 5% glucose + 10% dimethyl sulfoxide (DMSO), 0.9% NaCl + 10% methyl glycol, and 0.9% NaCl + 10% dimethyl sulfoxide, loaded into 0.25-mL straws and frozen in liquid nitrogen vapor. The semen from each treatment was thawed at three different thawing rates: 25 °C for 30 sec, 30 °C for 16 sec and 40 °C for 12 sec. Motility, vitality and morphology analyses were performed by computer-assisted sperm analysis (CASA). The characteristics of the fresh sperm mostly resembled those found in the literature. For the parameters analyzed, fresh sperm presented higher sperm quality in comparison to all treatments with cryopreserved sperm (p<0.05), except for the characteristic of normal morphology, for which the sperm cryopreserved in glucose and MG did not differ statistically from the fresh sperm. For the cryopreserved semen, the greatest results of total motility and curvilinear velocity (VCL) were obtained using glucose and DMSO, regardless of the thawing rate employed. For the straight-line velocity (VSL) and average path velocity (VAP), DMSO showed the best results, regardless of the diluent and thawing rate. With regard to vitality, the highest values were achieved when DMSO and thawing rates of 30 °C for 16 sec or 40 °C for 12 sec were used. In the morphological analysis, the greatest percentage of normal sperm cells was obtained using thawing rates of 25 °C for 30 sec and 40 °C for 12 sec, regardless of the freezing media. Sperm quality was found to suffer interference from the freezing media, as well as from interaction between its components (diluent and cryoprotectant) and the thawing rate used. Under the methodological conditions employed, the use of 5% glucose + 10% DMSO and a thawing rate of 30 °C for 16 seconds or 40 °C for 12 seconds is recommended for P. brevis semen cryopreservation. ResumoO Prochilodus brevis é um peixe reofílico, importante componente do ecossistema fluvial e apreciado na culinária nordestina. No entanto, ações antrópicas têm ameaçado sua sobrevivência. Desta forma, surge, nos pesquisadores, o interesse no desenvolvimento de protocolos de conservação do material genético, como a criopreservação seminal. Logo, a deter...

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Section: Sperm Cryopreservation and Thawingsupporting

confidence: 72%

Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates

Nunes

Oliveira

Lopes

et al. 2016

SCA

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“…After 10 days, the individual straws were thawed by immersion in water (water bath) to 60 °C for 8 sec (MARIA et al, 2006;PAULINO et al, 2012). A 10 μL aliquot of semen was deposited on a histological slide and homogenized with 40 μL of distilled water for activation of the sperm cells.…”

Section: Cryopreservation and Thawing Of Semenmentioning

confidence: 99%

Cryopreservation of Prochilodus lineatus semen: effect of cryoprotectants combination

et al. 2015

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The study was conducted with the objective of comparing the toxicity and the effect of the combination of intra and extracellular cryoprotectants in curimba Prochilodus lineatus sperm cells subjected to cryopreservation. Semen from 19 males were analyzed and diluted in four solutions comprise of intra and extracellular cryoprotectants in the following combinations: methanol+lactose, methanol+egg yolk, DMSO+lactose and DMSO+egg yolk. A portion of the diluted semen was frozen while the remaining fraction was kept in repose and evaluated after 10 min. For freezing, the diluted samples were packaged in 0.50 mL straws and placed into liquid nitrogen vapor for 24 h and, after this time, submerged into liquid nitrogen for 10 days. The combination of DMSO+lactose was less toxic to the diluted semen, resulting in higher motility rates and durations when compared to the other treatments. After thawing, the highest motility rate and duration were obtained using lactose as extracellular cryoprotectant, regardless of its combination. There was no significant difference between treatments when analyzing sperm morphology after thawing. Considering the effects of the tested treatments, the use of lactose as an extracellular cryoprotectant added with DMSO or methanol is the most suitable, since these combinations presented the highest motility rates and durations and low morphological change rate after thawing.Keywords: genetic preservation; cryobiology; sperm analysis; freezing protocol; toxicity; curimba CRIOPRESERVAÇÃO DO SÊMEN DO Prochilodus lineatus: EFEITO DA COMBINAÇÃO DE CRIOPROTETORES RESUMOO estudo foi realizado com o objetivo de comparar a toxicidade e o efeito da combinação de crioprotetoresintra e extracelulares em espermatozóides de curimba Prochilodus lineatus submetidos à criopreservação. O sêmen de 19 machos foi analisado e diluído em quatro soluções com crioprotetores intra e extracelulares nas seguintes combinações: metanol+lactose, metanol+gema de ovo, DMSO+lactose e DMSO+gema de ovo. Uma porção do sêmen diluído foi congelada enquanto que a fração remanescente foi mantida em repouso e avaliada após 10 min. Para o congelamento, as amostras diluídas foram envasadas em palhetas de 0,50 mL e colocadas em vapor de nitrogênio líquido durante 24 h e, após este tempo, submersas em nitrogênio líquido durante 10 dias. A combinação de DMSO+lactose se mostrou menos tóxica para o sêmen diluído, resultando em taxas mais elevadas de motilidade e duração espermática quando comparadas com os outros tratamentos. Após descongelamento, as maiores taxas de motilidade e duração foram obtidas usando lactose como crioprotector extracelular, independentemente da sua combinação. Não houve diferença significativa entre os tratamentos ao analisar a morfologia espermática após a descongemento. Considerando-se os efeitos dos tratamentos utilizados, o uso de lactose como crioprotector extracelular, adicionada ao DMSO ou metanol, é o mais adequado, uma vez que estas combinações apresentaram maiores taxas de motilidade e duração espe...

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“…This requires an understanding of both male and female gametes to succeed. Although there has been a growing body of research on male gametes, as exemplified by Dreanno et al (1998), Streit Jr et al (2002; Paulino et al (2012), Silva et al (2014), Lenz et al (2018), Caldas and Godoy (2019) and Kowalski and Cejko (2019), very little information is known about the semen of many native species, including the fire-eye tetra, M. oligolepis.…”

Section: Introductionmentioning

confidence: 99%

“…Studies related to sperm morphology have become an important tool in seminal evaluation. This is especially true with regards to cryopreservation, as some morphological abnormalities can be caused by cryoprotectant, the storage temperature and thawing time (Baulny et al, 1997;Paulino et al, 2012). Plasma membrane damage, like that observed with the sperm cells of this species, results in mitochondrial injuries that affect ATP production and, consequently, sperm motility (Baulny et al, 1997).…”

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confidence: 99%

“…As a result, the rate established for mammals is normally used as a benchmark. According to Paulino et al (2012), the Brazilian College of Animal Reproduction does not recommend the use of semen with abnormal sperm counts higher than 30% for cattle and horses, or 20% for pigs. Given this information, it can be inferred that the amount of abnormal sperm found in M. oligolepis would not result in unsuccessful fertilization.…”

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Seminal characterization of the Amazonian fire-eye tetra Moenkhausia oligolepis (Günther, 1864)

Rodrigues

Santos

Cordeiro

et al. 2020

Zygote

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Summary The seminal characteristics of Moenkhausia oligolepis are described. Three males were induced with a single dose of carp pituitary. Semen was collected 6 h after induction, and diluted in dibasic sodium phosphate extender solution. For motility analysis, 1 µl of diluted semen was added to 10 µl of distilled water to achieve gamete activation. The average duration of total motility was 76.67 s; while the average sperm motility rate at intervals of 15 s was 95.3, 85.3, 59.6, 31.7, 13.0, 4.6 and 1.2%. To determine sperm concentration in samples, 0.5 μl of semen was diluted with 500 μl of glutaraldehyde. An aliquot of 10 μl of this dilution was utilized for cell counting. An average count of 4.97 × 109 ± 3.46 sperm/ml was obtained. Morphological analyses were performed using eosin–nigrosine dye; 20.33% of the sperm were observed to be dead. Live sperm, comprising the other 79.67%, had an average length of approximately 30 µm, with a head diameter of 4.488 ± 0.7 µm; and a flagella plus mid-piece length of 26.071 ± 12.4 µm. Of those sperm, 69% had a normal morphology, while 31% had primary and secondary abnormalities. The observed abnormality rate did not have a detrimental effect on artificial fertilization potential for the species. The description of the seminal characteristics of a species is one of the most important sets of information required for artificial reproduction of fish in captivity. It also contributes significantly to the total biological knowledge of the studied species.

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Anormalidades espermáticas de Piaractus mesopotamicus após descongelamento utilizando diferentes metodologias

Abstract: Anormalidades espermáticas de Piaractus mesopotamicus após descongelamento utilizando diferentes metodologias

Cited by 4 publications

References 8 publications

Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates

Cryopreservation of Prochilodus brevis semen: freezing media and thawing rates

Cryopreservation of Prochilodus lineatus semen: effect of cryoprotectants combination

Seminal characterization of the Amazonian fire-eye tetra Moenkhausia oligolepis (Günther, 1864)

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