Standardization of spermatozoa concentration for cryopreservation of rainbow trout semen using a glucose-methanol extender

Nynca, Joanna; Judycka, Sylwia; Liszewska, Ewa; Dobosz, Stefan; Ciereszko, Andrzej

doi:10.1016/j.aquaculture.2017.04.036

Cited by 39 publications

(26 citation statements)

References 28 publications

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“…An effective semen cryopreservation method was recently developed by a Polish research group for several cultivated salmonid species [8][9][10][11]13]. The high post-thaw semen quality (total motility above 85%) reported in brown trout (Salmo trutta m. fario), rainbow trout (Oncorhynchus mykiss), sea trout (Salmo trutta m. trutta), brook trout (Salvelinus fontinalis), Atlantic salmon (Salmo salar) and whitefish, as well as the remarkable fertilizing ability (eyed embryos and hatching rates above 80%) recorded in brown trout, attracted our interest, and prompted us to test the effectiveness of this semen freezing protocol on the wild salmonid of the Mediterranean area (S. macrostigma = S. cettii).…”

Section: Discussionmentioning

confidence: 99%

“…The semen cryopreservation protocol based on the above mentioned procedure [10], which uses a simple glucose-methanol (GM) extender, was used. Briefly, the semen of five individual males was diluted to a final extender concentration of 0.15 M glucose and 7.5% methanol and loaded into 0.25 mL plastic straws, obtaining a final sperm concentration of 3.0 × 10 9 sperm/mL.…”

Section: Sperm Cryopreservationmentioning

confidence: 99%

“…They were then placed in liquid nitrogen. The straws were then thawed by immersion in a water bath at two different thawing rates: at 40 • C for 5 s, as reported in the original protocol by Nynca et al [10], or at 10 • C for 30 s, as already used in our previous work [7].…”

Section: Sperm Cryopreservationmentioning

confidence: 99%

“…However, despite our efforts and promising results obtained so far, we still aim to obtain a freezing procedure reporting a fertilization rate as similar as possible to that of fresh semen. Recently, a simple and efficient semen cryopreservation method for several cultivated salmonid species was developed by some Polish researchers, which attracted our interest [8][9][10][11]. These authors suggested that this freezing method could be "universal" for the semen cryopreservation of salmonids species, attributing its effectiveness to the composition of the freezing medium (glucose-methanol extender).…”

Section: Introductionmentioning

confidence: 99%

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A Simple and Efficient Semen Cryopreservation Method to Increase the Genetic Variability of Endangered Mediterranean Brown Trout Inhabiting Molise Rivers

Rusco

Iorio

Iampietro

et al. 2020

Animals

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The aim of our study was to test the effectiveness of a simple semen cryopreservation procedure, developed for cultivated salmonid, on the wild salmonid of the Mediterranean area and to evaluate the effect of different thawing rates and sperm-to-egg ratios. The semen of five individual males was diluted into a final extender concentration of 0.15 M glucose and 7.5% methanol and loaded into 0.25 mL plastic straws, and a final sperm concentration of 3.0 × 109 sperm/mL was obtained. After equilibration, the straws were frozen by exposure to liquid nitrogen vapor at 3 cm above the liquid nitrogen level for 5 min. The semen was thawed at 40 °C/5 s or 10 °C/30 s. The sperm cryosurvival was evaluated by examining in vitro the sperm motility parameters using the CASA system, followed by fertilization trials in vivo, using three different sperm-to-egg ratios 6 × 105, 4.5 × 105 and 3 × 105:1. The applied cryopreservation procedure resulted in remarkably high (85.6%) post-thaw sperm total motility, when the semen was thawed at 40 °C/5 s, whilst the highest fertilization rate (53.1%) was recorded for a sperm-to-egg ratio of 4.5 × 105:1. According to these outcomes, the cryopreservation procedure that was tested turned out to be effective for the wild population of Mediterranean brown trout and practical for the creation of the first European semen cryobank foreseen as part of our “LIFE” Nat.Sal.Mo. project.

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Section: Discussionmentioning

confidence: 99%

Section: Sperm Cryopreservationmentioning

confidence: 99%

Section: Sperm Cryopreservationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A Simple and Efficient Semen Cryopreservation Method to Increase the Genetic Variability of Endangered Mediterranean Brown Trout Inhabiting Molise Rivers

Rusco

Iorio

Iampietro

et al. 2020

Animals

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“…Various procedures of motility evaluation from different sources make it difficult or impossible to directly compare research results, and insufficient checkpoints during commercial-scale cryopreservation make the quality of final products unpredictable. Sperm concentration is another important indicator of initial sperm quality upon collection, and it can significantly affect motility and the level of agglutination of thawed sperm and fertilization rates [8–10], but sperm concentration is often not adjusted or reported in standardized methods in publications regarding sperm cryopreservation for aquatic species.…”

Section: Introductionmentioning

confidence: 99%

Quality evaluation of sperm from livebearing fishes: Standardized assessment of sperm bundles (spermatozeugmata) from Xenotoca eiseni (Goodeidae)

2018

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Standardized evaluation of sperm quality is essential for research, commercial-scale cryopreservation, and induced spawning. However, standardized methods for evaluation of sperm bundles (spermatozeugmata or spermatophores) have not been established. The purpose of the present study was to use Redtail Splitfin (Xenotoca eiseni) as a model for freshwater livebearing fishes to establish initial standardized methods to collect sperm bundles, and quantitatively and qualitatively evaluate quality-related attributes. No sperm or sperm bundles were able to be collected by stripping. Testes were removed, rinsed, weighed, placed in 50 μL of buffer solution on a glass slide, and crushed gently 3-5 times with angled spade-tip forceps. Sperm bundles were released into the buffer solution and collected with a pipette into 1.5-mL centrifuge tubes. To quantify size and shape, images of bundles were captured with a CCD camera connected to a microscope, and measured with computer software. There was no significant correlation between body wet weight and major bundle axis length (P = 0.6759), minor axis length (P = 0.5658), average axis length (P = 0.5869), aspect ratio (P = 0.7839), and observed area (P = 0.5727). The concentrations of sperm bundles, estimated with the three methods (Makler counting chamber, a hemocytometer, and direct counting) were significantly different (P < 0.0001). Hemocytometers were suitable for estimation of bundles from X. eiseni. To evaluate activation of sperm, bundles were viewed with a microscope, and classified into one of five phases by evaluating morphology of the bundles and motion of sperm within the bundles as Phase 0 through Phase 4 that represented early through late activation stages. The frequencies and duration of each activation phase were used to evaluate dissociation of sperm bundles and motility capability of sperm within the bundles. Within 180 min of activation, all five phases were observed. Overall, this study for the first time established standardized methods to collect and evaluate quality-related attributes of sperm bundles. These standardized evaluations provide a basis for further modification, standardization, and generalization, which are useful in research on livebearing fishes involving male gametes, such as studies on cryopreservation, artificial insemination, and in development of germplasm repositories for imperiled species including goodeids.

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Chapter 15 Sperm Cryopreservation of Aquatic Species

Horváth

Urbányi

2020

Reproduction in Aquatic Animals

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Standardization of spermatozoa concentration for cryopreservation of rainbow trout semen using a glucose-methanol extender

Cited by 39 publications

References 28 publications

A Simple and Efficient Semen Cryopreservation Method to Increase the Genetic Variability of Endangered Mediterranean Brown Trout Inhabiting Molise Rivers

A Simple and Efficient Semen Cryopreservation Method to Increase the Genetic Variability of Endangered Mediterranean Brown Trout Inhabiting Molise Rivers

Quality evaluation of sperm from livebearing fishes: Standardized assessment of sperm bundles (spermatozeugmata) from Xenotoca eiseni (Goodeidae)

Chapter 15 Sperm Cryopreservation of Aquatic Species

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