Sperm Cryopreservation in Live-Bearing <i>Xiphophorus</i> Fishes: Offspring Production from <i>Xiphophorus variatus</i> and Strategies for Establishment of Sperm Repositories

Yang, Huanming; Cuevas‐Uribe, Rafael; Savage, Markita; Walter, Ronald B.; Tiersch, Terrence R.

doi:10.1089/zeb.2012.0737

Cited by 11 publications

(13 citation statements)

References 42 publications

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“…Some counting chambers, like the haemocytometer, require relatively large volumes of sample (>10 μL), which is a problem when dealing with small fish where the total volume obtained is in the range 2–5 μL (Yang et al . 2007 a , 2012). In addition, this method is time consuming, requiring 10–15 min per sample, which may not be suitable for high-throughput applications (Tan et al .…”

Section: Sample Collection and Preparationmentioning

confidence: 99%

“…In addition, it is recommended for fertilisation testing to use spermatozoa of a different species to produce hybrids with distinguishable phenotypes to verify protocols, so the paternity can be readily verified (Yang et al . 2007 b , 2012). Furthermore, improper handling of the female during insemination can cause permanent injury or kill the fish.…”

Section: Egg Collection and Use Of Thawed Sperm Samplesmentioning

confidence: 99%

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Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development

Torres

Hu²,

Tiersch

2016

Reprod. Fertil. Dev.

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Cryopreservation in aquatic species in general has been constrained to research activities for more than 60 years. Although the need for application and commercialisation pathways has become clear, the lack of comprehensive quality assurance and quality control programs has impeded the progress of the field, delaying the establishment of germplasm repositories and commercial-scale applications. In this review we focus on the opportunities for standardisation in the practices involved in the four main stages of the cryopreservation process: (1) source, housing and conditioning of fish; (2) sample collection and preparation; (3) freezing and cryogenic storage of samples; and (4) egg collection and use of thawed sperm samples. In addition, we introduce some key factors that would assist the transition to commercial-scale, high-throughput application.

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Section: Sample Collection and Preparationmentioning

confidence: 99%

Section: Egg Collection and Use Of Thawed Sperm Samplesmentioning

confidence: 99%

Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development

Torres

Hu²,

Tiersch

2016

Reprod. Fertil. Dev.

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“…Repositories for livestock and other agricultural species have been long established, 26 but less has been done for aquatic species, in part due to the lack of comprehensive, standardized QC programs that would start with animal screening and address sperm quality in each step of the process. 27 The mission of germplasm repositories includes the commitment to conserve, maintain, and distribute genetic resources and the associated information regarding sample quality and inventory, genetics, phenotypes, locality, and other information about individual fish and populations. Given the myriad of cryopreservation protocols that have been developed for different aquatic species, 28 only through comprehensive QC and quality assurance strategies would it be possible to guarantee the quality of deposited sperm, and the reproducibility of cryopreservation protocols.…”

Section: Figmentioning

confidence: 99%

Challenges in Development of Sperm Repositories for Biomedical Fishes: Quality Control in Small-Bodied Species

et al. 2017

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Quality control (QC) is essential for reproducible and efficient functioning of germplasm repositories. However, many biomedical fish models present significant QC challenges due to small body sizes (<5 cm) and miniscule sperm volumes (<5 μL). Using minimal volumes of sperm, we used Zebrafish to evaluate common QC endpoints as surrogates for fertilization success along sequential steps of cryopreservation. First, concentrations of calibration bead suspensions were evaluated with a Makler counting chamber by using different sample volumes and mixing methods. For sperm analysis, samples were initially diluted at a 1:30 ratio with Hanks' balanced salt solution (HBSS). Motility was evaluated by using different ratios of sperm and activation medium, and membrane integrity was analyzed with flow cytometry at different concentrations. Concentration and sperm motility could be confidently estimated by using volumes as small as 1 μL, whereas membrane integrity required a minimum of 2 μL (at 1 × 10 cells/mL). Thus, <5 μL of sperm suspension (after dilution to 30-150 μL with HBSS) was required to evaluate sperm quality by using three endpoints. Sperm quality assessment using a combination of complementary endpoints enhances QC efforts during cryopreservation, increasing reliability and reproducibility, and reducing waste of time and resources.

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“…Poeciliids are popular ornamental species, important cancer research models, and have been used for mosquito control[24]. Sperm from Poeciliidae has been used in studies addressing reproductive behavior [25], evolution [26], toxicology [27], and establishment of germplasm repositories [28]. Goodeidae, the second largest freshwater livebearing family (about 38 livebearing species), is considered to be one of the most at-risk fish groups in the world [29].…”

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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Sperm Cryopreservation in Live-Bearing Xiphophorus Fishes: Offspring Production from Xiphophorus variatus and Strategies for Establishment of Sperm Repositories

Cited by 11 publications

References 42 publications

Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development

Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development

Challenges in Development of Sperm Repositories for Biomedical Fishes: Quality Control in Small-Bodied Species

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

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