scholarly journals Application of computer-assisted sperm analysis system to elucidate lack of effects of cyclophosphamide on rat epididymal sperm motion.

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


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.


2008 ◽  
Vol 20 (1) ◽  
pp. 86
Author(s):  
M. Filliers ◽  
T. Rijsselaere ◽  
P. Bossaert ◽  
V. De Causmaecker ◽  
J. Dewulf ◽  
...  

Feline epididymal sperm is commonly used for in vitro fertilization. It also yields the opportunity to conserve genetic material from valuable males that suddenly die. Epididymal sperm quality parameters vary considerably among laboratories, implicating the need for objective evaluation methods. The aim of the present study was to describe reference values of computer-assisted sperm analysis (CASA) parameters of fresh epididymal cat sperm and to assess the effect of prolonged cooled storage (4�C) on various sample characteristics. Epididymides obtained from tomcats after routine orchiectomy (2–4 pairs/replicate) were sliced to release spermatozoa. The sperm suspension was placed on a 2-layer gradient and, after centrifugation, the sperm pellet was recovered. In Experiment 1 (20 replicates), sperm motility parameters were assessed immediately after retrieval (T0) using the Hamilton Thorne analyzer Ceros 12.1 (HTR; Hamilton Thorne Biosciences, Beverly, MA, USA). In Experiment 2, fresh (T0) sperm samples (4 replicates) were evaluated for motility parameters (HTR), acrosomal status (FITC-Pisum sativum agglutinin staining), morphology (eosin/nigrosin (E/N) staining), and membrane integrity (E/N and SYBR�-14-propidium iodide staining; Molecular Probes, Inc., Eugene, OR, USA). After addition (1:2) of a Tris-glucose-citrate diluent containing 20% egg yolk, samples were cooled and reassessed on Days 1 (T1), 3 (T3), 5 (T5), 7 (T7), and 10 (T10). Results were analyzed in a mixed linear model, with replicate as random factor and time as fixed effect (S-PLUS 7.0; Insightful Corp., Seattle, WA, USA). Results of Experiment 1 were as follows (mean � SD): motility (MOT): 80.8% � 23.5; progressive motility (PMOT): 69.9% � 23.2; velocity average pathway (VAP): 98.7 µm s–1 � 24.2; velocity straight line (VSL): 89.3 µm s–1 � 25.4; velocity curved line (VCL): 134.8 µm s–1 � 31.9; amplitude lateral head (ALH): 4.3 µm � 2.0; beat cross frequency (BCF): 34.6 Hz � 7.0; and straightness (STR): 89.6% � 6.6. In Experiment 2, MOT, PMOT, VAP, VSL, VCL, BCF, and the percentage of normal spermatozoa showed a decrease over time (P < 0.05) compared to fresh samples, starting from T1, T3, T5, T7, T5, T3, and T1, respectively. In contrast, STR, ALH, membrane integrity, and the percentage of acrosome-intact spermatozoa were not affected (P > 0.05) by cooled storage. To summarize, we have presented a set of reference values for CASA-parameters of fresh, epididymal cat spermatozoa. Cooled storage impaired most motility parameters and lowered the percentage of normal spermatozoa, but did not influence membrane integrity or acrosomal status. The effect of cooled storage on DNA fragmentation of sperm and its subsequent influence on in vitro embryo development require further investigation.


1994 ◽  
Vol 61 (1) ◽  
pp. 75-80
Author(s):  
G. Bonanni ◽  
A. Colonna ◽  
S. Masiero ◽  
I. Mastrogiacomo

The recent introduction of various devices for computer-assisted sperm analysis allows us to have not only objective measurements of the seminal parameters but also to determine some characteristics of sperm motion such as curvilinear and linear velocity and amplitude of lateral head displacement that cannot be obtained by routine analysis. These new seminal parameters seem to be very useful in the diagnosis of male infertility and, among them, ALH seems to play a more important role; in one of our studies we found a significant correlation between this parameter and the results of the bovine cervical mucus penetration test (Penetrak). Furthermore it is possible to determine sperm subpopulations for each parameter and so it will be possible to express a more accurate judgement not only on the fertilizing capacity of semen but also to know the effects on spermatozoa of different activating and capacitating treatments.


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.).


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