scholarly journals Cryopreservation of tambaqui semen using a dry shipper and a programmed freezing machine

2016 ◽  
Vol 37 (4) ◽  
pp. 2167 ◽  
Author(s):  
Mayara Setúbal Oliveira ◽  
Priscila Silva de Almeida-Monteiro ◽  
Larissa Teixeira Nunes ◽  
Francisco Renan Aragão Linhares ◽  
João Paulo Silva Pinheiro ◽  
...  

Tambaqui (Colossoma macropomum) is a native freshwater fish that is of great importance for Brazilian aquaculture. Because of this importance, several techniques have been developed to improve the reproduction of this species in captivity. One of these techniques is the cryopreservation of sperm. In an effort to increase the efficiency of cryopreservation protocols, researchers have tried to determine suitable diluting solutions and freezing methods, which will provide a better post-thaw sperm quality. Thus, this study aimed to evaluate the efficiency of different diluents and freezing methods for the cryopreservation of tambaqui (C. macropomum) sperm. Samples of fresh semen were diluted in different treatments (Glucose 5% + 10% Dimethyl sulfoxide – DMSO, Glucose 5% + 10% Methyl glycol – MG, BTS + 10% DMSO and BTS + 10% MG) at a 1:9 dilution rate and frozen in a programmed freezing machine and a dry shipper. The semen samples were thawed and evaluated for vitality, sperm morphology and kinetics. Cryopreserved semen with DMSO and using the programmed freezing machine provided a greater percentage of motile sperm (15.44 ± 1.04%) after thawing compared to the dry shipper (3.99 ± 0.55%), regardless of the diluent. Additionally, DMSO showed better sperm velocities than MG regardless of the freezing method and the extender employed. A higher percentage of living spermatozoa was observed when glucose (37.28 ± 1.32%) (regardless of the freezing method and cryoprotectant) and DMSO (37.98 ± 1.25%) was used in the programmed freezing machine. For morphology, a greater amount of normal spermatozoa (46.10 ± 1.82%) was observed when the semen was cryopreserved using a freezing machine programmed with DMSO as the cryoprotectant and Glucose or BTS (38.16 ± 1.9% and 39.26 ± 1.87%, respectively) as extenders. Therefore, we suggest the use of the DMSO (10%) cryoprotectant in association with the Glucose (5%) extended in the programmed freezing machine for cryopreservation of C. macropomum semen.

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Monika Trzcińska ◽  
Magdalena Bryła

AbstractThis study aimed to define potential markers that could determine the suitability of ejaculate for cryopreservation. Fresh semen from eleven boars (4–7 ejaculates/boar), regardless of their sperm motility, was subjected to a cryopreservation procedure. The sperm quality before and after freezing was assessed based on the sperm membrane permeability and acrosome integrity. The results showed that it was possible to effectively cryopreserve ejaculates below the accepted standards of 70–80% of fresh motile sperm and still obtain a high cryosurvival rate. Moreover, a significant correlation was found between the percentage of viable sperm with apoptotic-like changes, viable sperm with reacted acrosomes, and the cryosurvival rate. The proposed markers for assessing the quality of fresh semen could be used to predict the success of cryopreservation procedures.


2019 ◽  
Vol 15 (01) ◽  
pp. 9-13
Author(s):  
P K Pathak ◽  
A J Dhami ◽  
D V Chaudhari

This investigation was carried out on semen of three healthy mature breeding bulls of Gir breed to evaluate the interrelationships among sperm quality attributes of fresh and frozen-thawed semen assessed by Biovis CASA. The ejaculates (n = 24) having >75% initial motility were diluted @80 million sperm/mL using TFYG extender, filled in French mini straws, and were frozen using a programmable bio freezer after 4 hours of equilibration. The straws were thawed in a water bath at 37°C for 30 sec. The freshly diluted and frozenthawed samples were assessed for routine subjective tests and various motion characteristics/kinematics by Biovis CASA. The Pearson’s correlations for sperm motility and velocity/kinematic parameters of total motile sperm as well as of progressively motile sperm were studied in freshly diluted and frozen-thawed semen. In fresh semen, total motile sperm assessed by CASA had significant (p less than 0.05, 01) correlations with rapid progressive motile sperm (r = 0.46), wobbling index (r = 0.52) and dancing frequency (r = -0.43) in fresh semen. In frozen-thawed semen, it was significantly correlated only with linearity (r = 0.46). The rapid progressive motile sperm in both fresh (r = 0.41 to 0.92) and frozen-thawed (r = 044 to 0.88) semen, however, had significant correlations with most of their velocity traits. Further, the average path velocity (VAP), curvilinear velocity (VCL), straight line velocity (VSL), linearity (LIN), straightness (STR), wobbling (WOB), beat-cross frequency (BCF), amplitude of lateral head displacement (ALH), and dancing mean (DNM) of sperm showed significant positive or negative interrelationships among each other in both fresh (r = 0.41 to 0.91) as well as post-thawed (r = 0.44 to 0.90) semen. Moreover, the correlations of motility and kinematics parameters of total motile sperm in both fresh and frozen-thawed semen were highly significant with velocity/kinematics traits of only progressively motile sperm, and the velocity traits among only motile sperm were highly significantly interrelated in both fresh (r = 0.46 to 0.98) and frozen-thawed (r = 0.43 to 0.93) semen of Girbulls, though the magnitudes of correlations were lower in frozen-thawed semen as compared to fresh semen. Thus, CASA analysis offresh semen for motility and velocity traits could predict the post-thawed sperm motility and velocity/kinematics of bovine semen.


2014 ◽  
Vol 74 (4) ◽  
pp. 933-938 ◽  
Author(s):  
JM Galo ◽  
DP Streit Jr. ◽  
JA Povh ◽  
DC Fornari ◽  
EK Resende ◽  
...  

This study aimed at assessing the sperm quality of the Amazon catfish, Leiarius marmoratus ¸ after refrigeration without extenders. After capturing the animals and stripping of semen, the following parameters were analyzed: progressive motility, motility quality score, duration of motility and sperm morphology. An aliquot of fresh semen from each male was kept at room temperature (28 ± 2°C) as a control, for further comparison with cooled semen. The semen from each animal was stored in extenders-free individual syringes. The syringes were kept in ice within polystyrene boxes at 13 ± 2°C. For both fresh and cooled semen, seminal parameters were evaluated every one-hour interval, reaching seven hours of analysis. Fresh semen showed a significant decrease in motility, motility quality score and duration of motility remaining viable only for three hours. Progressive motility of the cooled semen displayed a negative linear pattern (P<0.05). The duration of motility increased (P<0.05), reaching its peak after three hours of storage. The motility quality score showed a quadratic pattern. No statistical differences were observed when sperm morphology was assessed (P>0.05), even though the mean values of total abnormalities have increased over the storage time. Further studies focusing on the application of this technique should be performed, including the addition of extenders and cryoprotectants for preservation of the sperm over longer periods.


2021 ◽  
Vol 63 (1) ◽  
Author(s):  
Maja Zakošek Pipan ◽  
Petra Zrimšek ◽  
Breda Jakovac Strajn ◽  
Katarina Pavšič Vrtač ◽  
Tanja Knific ◽  
...  

ABSTRACT Background Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.


2018 ◽  
Vol 110 (7) ◽  
pp. 1277-1284 ◽  
Author(s):  
Ashok Agarwal ◽  
Manesh Kumar Panner Selvam ◽  
Rakesh Sharma ◽  
Kruyanshi Master ◽  
Aditi Sharma ◽  
...  

2017 ◽  
Vol 38 (5) ◽  
pp. 3069 ◽  
Author(s):  
Ana Carolina Rodrigues ◽  
Camila Montanari Ruiz ◽  
Carla Daniela Dan De Nardo ◽  
Gabriele Barros Mothé ◽  
Fabiano Martinez Rossi ◽  
...  

For years, fatty acids have been recommended as a dietary supplement to improve canine hair. For animal reproduction, supplementation with omegas has been used to increase the reproductive efficiency and conception rate, but few studies have been conducted in dogs. The aim of this study was to evaluate the effects of daily dietary supplementation with omega-3 and -6 on the quality of fresh and frozen/thawed semen in canines. Semen was collected from seven dogs and evaluated for sperm motility, vigor, concentration, and morphology. The 17-week study included 119 ejaculates and was divided according to oral supplementation with omega-3 and -6: M1 (1st-5th week) or pre-supplementation; M2 (6th-9th week) and M3 (10th-13th week) or during supplementation; and M4 (14th-17th week) or post-supplementation. After analysis, the semen was frozen and then revaluated both immediately and 30 minutes (at 37° C) after thawing. Supplementation with omegas increased sperm motility, vigor, and concentration; however, supplementation had no influence on semen freezability. In addition, there was no improvement in sperm motility after supplementation when the thawed cells were maintained at 37° C for 30 minutes. We concluded that dietary supplementation with omega-3 and -6 for 4 to 8 weeks can improve the quality of fresh semen, although it has no effect on the freezability of canine semen.


2020 ◽  
Vol 3 (2) ◽  
pp. 99-106
Author(s):  
Sara Mahmood Qureshi ◽  
Salma Kafeel ◽  
Riffat Bibi ◽  
Jawad Mohmand

Introduction: The unrestricted use of intracytoplasmic sperm injection (ICSI) for non-male factor infertility is associated with adverse outcomes. Post-wash total motile sperm count (PW-TMSC) offers prognostic value to assess sperm quality and aid in the decision to perform in vitro fertilization (IVF) or ICSI. Objectives: The aim of this study was to identify the effect of PW-TMSC on fertilization rates in patients undergoing IVF cycles exclusively with non-male factor infertility. It also aimed to identify whether unnecessary ICSI could be avoided in such cases, thus maximizing optimal outcomes. Materials & Methods: We retrospectively analyzed age, semen volume, prewash TMSC, and PW-TMSC in 68 conventional IVF cycles of infertile couples with non-male factor infertility. Clinical characteristics including female age, number of follicles, level of estradiol on trigger day, mature cumulus-oocyte complexes (COCs) collected, were also included. Results: Incidence of <30% fertilization was significantly higher in the 4-<10 Million group compared with the ≥20 Million post-wash TMSC group (P<0.001). Furthermore, Receiver operating characteristics (ROC) analysis revealed post-wash TMSC as a significant predictor (P<0.05) of total failed fertilization (TFF) and of ≥30% fertilization (P<0.05) with area under curve (AUC) of 0. 79 and 0.77, respectively, with a deemed cutoff of 10.89 Million. Conclusion: Post-wash TMSC is a good predictor of fertilization; it can help in avoiding potentially low or even total fertilization failure (TFF). A cut-off point of 10.89 Million or less should warrant the use of ICSI.


2021 ◽  
Vol 10 (19) ◽  
pp. 1417-1421
Author(s):  
Jyothi A. Raj ◽  
Heera Sankar ◽  
Sagarika Mahapatra ◽  
Ashima Binny

BACKGROUND Semen analysis is an integral part of work up for infertility in men, with sperm morphology being an important qualitative parameter. Qualitative defects can affect any part of the sperm and are classified as defects in the head, middle piece, and tail, based on morphology. The focus of the study was to assess qualitative defects in sperms by light microscopy, in semen with normal sperm counts. METHODS This study is hospital based, descriptive, retrospective study. Of the semen samples received in the clinical laboratory, fifty with normal sperm counts were included in the study and processed according to standard protocol. For evaluation of qualitative defects by sperm morphology, smears were fixed in ethanol, stained with Papanicolaou stain [PAP], and assessed under light microscope. RESULTS The 50 semen samples included in the study had sperm counts ranging from 15 to 80 million / ml. Thirty samples had less than 10 % abnormal forms, fourteen samples had 11 - 20 % abnormal forms, five samples had 21 - 30 % abnormal forms and one sample had 40 % abnormal sperms. Qualitative defects were classified as morphological abnormalities in head, neck, and tail. Of the fifty cases, most defects were found in the head, followed by those in the neck and tail. Common defects noted were double head (44 %), abnormal sized heads, and bent neck (48 %). Coiling was a common defect noted in the tail (10 %). Most sperms showed a combination of defects. CONCLUSIONS Qualitative defects in sperm morphology are often seen in samples with normal sperm counts. Assessment of microscopic characteristics of human spermatozoa is as important as count and motility in the complete evaluation and work-up of semen samples in cases of infertility. KEY WORDS Semen, Sperm, Quality, Microscopy, Morphology


2010 ◽  
Vol 90 (3) ◽  
pp. 389-392 ◽  
Author(s):  
N. Am-in ◽  
R N Kirkwood ◽  
M. Techakumphu ◽  
W. Tantasuparuk

Boars having normal (71.1 ± 1.2%; n = 10) or low (35.12 &plusmn 3.9%; n = 10) sperm motility 24 h after collection were used, and semen was evaluated following storage in Beltsville Thawing Solution (BTS) for 24 h at 18°C. Sperm lipids were extracted and lipid peroxidation quantified. No differences were evident in fresh semen, but after 24 h, sperm motility, viability and membrane permeability in the low motility group were lower (P < 0.001) compared with the normal motility group. Sperm membrane lipid peroxidation was greater (P < 0.001) in the low motility group. A factor influencing sperm storability is membrane lipid peroxidation, which can be accurately assayed using a commercial kit.Key words: Boars, sperm motility, sperm quality, lipid peroxidation


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