Artificial fertilisation in a terrestrial toadlet (Pseudophryne guentheri): effect of medium osmolality, sperm concentration and gamete storage

2013 ◽  
Vol 25 (8) ◽  
pp. 1134 ◽  
Author(s):  
Aimee J. Silla

Anurans exhibit a greater reproductive diversity than any other vertebrate order. However, studies investigating the effects of the external fertilisation environment on fertilisation success are limited to aquatic-breeding species. This study investigated the effects of fertilisation medium osmolality, sperm concentration and short-term oocyte storage on fertilisation success in a terrestrial-breeding anuran, Pseudophryne guentheri. Split-clutch experimental designs were used to determine optimal fertilisation conditions. To determine the effect of short-term sperm storage, sperm viability was assessed using fluorescence microscopy and percentage sperm motility and velocity quantified with a computer-assisted sperm analysis system. Fertilisation success was highest in media ranging in osmolality from 25 mOsm kg–1 to 100 mOsm kg–1, representing a broader range and higher optimal osmolality than previously reported for aquatic breeders. High rates of fertilisation (>75%) were achieved in relatively low sperm concentrations (2.5 × 104 mL–1). Oocytes stored in isotonic solutions (200 mOsm kg–1) retained fertilisation capacity (32%) after 8 h of storage, while sperm suspensions maintained motility (≥26%) for 13 days. Additional studies on terrestrial-breeding anurans will be required to ascertain whether the optimal fertilisation conditions reported reflect adaptations to achieve fertilisation in a terrestrial environment.

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1885
Author(s):  
José Néstor Caamaño ◽  
Carolina Tamargo ◽  
Inmaculada Parrilla ◽  
Felipe Martínez-Pastor ◽  
Lorena Padilla ◽  
...  

Genetic resource banks (GRB) preserve the genetic material of endangered, valuable individuals or genetically relevant breeds. Semen cryopreservation is a crucial technique to reach these goals. Thus, we aimed to assess the sperm parameters of semen doses from the native pig breed Gochu Asturcelta stored at the GRB of Principado de Asturias (GRB-PA, Gijón, Spain), focusing on intrinsic and extrinsic (boar, season) factors. Two straws per boar (n = 18, 8–71 months of age) were thawed, pooled, and assessed after 30 and 150 min at 37 °C by CASA (computer-assisted sperm analysis system; motility and kinematic parameters) and flow cytometry (viability, acrosomal status, mitochondrial activity, apoptosis, reactive oxygen species, and chromatin status). The effects of age, incubation, and season on post-thawing quality were determined using linear mixed-effects models. Parameters were on the range for commercial boar breeds, with chromatin status (SCSA: fragmentation and immaturity) being excellent. Incubation decreased sperm quality and functionality. The boar age did not have a significant effect (p > 0.05), but the between-boar variability was significant (p < 0.001). The season significantly affected many parameters (motility, kinematics, viability, acrosomal status, mitochondrial activity), especially after 150 min of incubation. In general, samples collected in spring and summer showed higher quality post-thawing, the lowest in winter. In conclusion, the sperm doses from the Gochu Asturcelta breed stored at the GRB-PA showed excellent chromatin status and acceptable characteristics after thawing. Therefore, boar and seasonal variability in this autochthonous breed could be relevant for cryobank management.


2001 ◽  
Vol 26 (2) ◽  
pp. 75-83 ◽  
Author(s):  
Hashihiro HIGUCHI ◽  
Masanao NAKAOKA ◽  
Satoshi KAWAMURA ◽  
Yusuke KAMITA ◽  
Akira KOHDA ◽  
...  

Author(s):  
Bernadett Pataki ◽  
Berta Izabella Roberta ◽  
Gyöngyi Gazsi ◽  
Béla Urbányi ◽  
Tímea Kollár ◽  
...  

Abstract The effect of age on the sensitivity of zebrafish sperm against mercury exposure was investigated in the present study. Although results of the use of sperm from mature individuals for toxicity tests have been published, there is no information about the exact age of the fish in some cases, which can affect the results. During the experiments, pooled sperm was stripped from males of 7, 12, or 18 months of age, divided into 5 sub-groups, diluted with different concentrations of Hg (0, 0.5, 1, 2.5, and 5 mg/L Hg), and incubated for 240 min. The motility parameters of sperm (progressive motility (%), curvilinear velocity (VCL)) were measured by a computer-assisted sperm analysis system, at 30, 120, and 240 min of exposure. Regarding the age, significant differences were found in PMOT (p = 0.0267) as well as in VCL (p = 0.0004) among the three different age groups. The different concentrations of Hg also caused significant differences. The most significant differences in PMOT were between the 7- and 18-month-old groups; these differences were observed at 0.5, 1 and 2.5 mg/L Hg at 30 min, at 0.5 and 1 mg/L at 120 min, as well as at 0.5 mg/L at 240 min. In VCL the most significant differences were found between the 7- and 12-month-old groups; significant differences were found at each tested concentration at 30 min as well as at 0.5 and 2.5 mg/L at 240 min. According to the results, the age of zebrafish negatively influences the sensitivity of its sperm. This may concern not only toxicology tests but many techniques in fish breeding where the sperm is treated before use (cryopreservation, pressure shock, etc.).


2020 ◽  
Vol 68 (2) ◽  
pp. 197-199
Author(s):  
Eszter Balogh ◽  
Anna Boglárka Dálnoki ◽  
László Rózsa ◽  
Viktória Johanna Debnár ◽  
Orsolya Varga-Balogh ◽  
...  

AbstractWhen using artificial insemination in porcine reproduction, one of the most important requirements is the suitable quality of semen regarding its total motility (TM) and progressive motility (PM). Computer-assisted sperm analysis (CASA) is an appropriate method to analyse the quality of semen. Recently a portable instrument has been developed to help specialists in their everyday field work. In our study, semen quality was measured simultaneously by the portable device (Ongo) and a laboratory CASA system (Microptic) to compare TM and PM values obtained by these appliances at a concentration of 50 × 106 spermatozoa/mL. Agreement between measurements was evaluated with a Bland-Altman plot. Strong correlation was found between the investigated instruments for all the three parameters, i.e. sperm concentration, TM and PM. However, a few measurements fell outside the defined range of acceptance.


Biology ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 207
Author(s):  
Jesús Yániz ◽  
Carlos Alquézar-Baeta ◽  
Jorge Yagüe-Martínez ◽  
Jesús Alastruey-Benedé ◽  
Inmaculada Palacín ◽  
...  

Computer assisted sperm analysis (CASA) systems can reduce errors occurring in manual analysis. However, commercial CASA systems are frequently not applicable at the forefront of challenging research endeavors. The development of open source software may offer important solutions for researchers working in related areas. Here, we present an example of this, with the development of three new modules for the OpenCASA software (hosted at Github). The first is the Chemotactic Sperm Accumulation Module, a powerful tool for studying sperm chemotactic behavior, analyzing the sperm accumulation in the direct vicinity of the stimuli. This module was validated by comparing fish sperm accumulation, with or without the influence of an attractant. The analysis clearly indicated cell accumulation in the treatment group, while the distribution of sperm was random in the control group. The second is the Sperm Functionality Module, based on the ability to recognize five sperm subpopulations according to their fluorescence patterns associated with the plasma membrane and acrosomal status. The last module is the Sperm Concentration Module, which expands the utilities of OpenCASA. These last two modules were validated, using bull sperm, by comparing them with visual counting by an observer. A high level of correlation was achieved in almost all the data, and a good agreement between both methods was obtained. With these newly developed modules, OpenCASA is consolidated as a powerful free and open-source tool that allows different aspects of sperm quality to be evaluated, with many potential applications for researchers.


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