27 Comparison of cryoprotectants and their combinations in the optimisation of a sperm cryopreservation protocol in the Argentine black and white tegu (Salvator merianae)

2022 ◽  
Vol 34 (2) ◽  
pp. 247
Author(s):  
C. Young ◽  
N. Ravida ◽  
M. Curtis ◽  
F. Mazzotti ◽  
B. Durrant
Keyword(s):  
Sperm Cryopreservation ◽  
Black And White ◽  
Cryopreservation Protocol

Development of a sperm cryopreservation protocol for the Argentine black and white tegu (Tupinambis merianae)

2017 ◽  
Vol 87 ◽  
pp. 55-63 ◽  
Author(s):  
Carly Young ◽  
Nicole Ravida ◽  
Michelle Curtis ◽  
Frank Mazzotti ◽  
Barbara Durrant
Keyword(s):  
Sperm Cryopreservation ◽  
Black And White ◽  
Tupinambis Merianae ◽  
Cryopreservation Protocol

scholarly journals Effects of Cryoprotective Medium Composition, Dilution Ratio, and Freezing Rates on Spotted Halibut (Verasper variegatus) Sperm Cryopreservation

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 2153
Author(s):  
Irfan Zidni ◽  
Yun Ho Lee ◽  
Jung Yeol Park ◽  
Hyo Bin Lee ◽  
Jun Wook Hur ◽  
...  
Keyword(s):  
Previous Experiment ◽  
Activity Index ◽  
Medium Composition ◽  
Freezing Rate ◽  
Dilution Ratio ◽  
Sperm Cryopreservation ◽  
Ringer’S Solution ◽  
Potential Market ◽  
Spotted Halibut ◽  
Cryopreservation Protocol

The spotted halibut is species that has a high potential market value in Korea, but the supply of seed is unstable because of the limited milt production of males. The objective of this research was to explore different aspects, such as CPAs, diluents, dilution ratio, and freezing rates, to develop an optimal sperm cryopreservation. The parameters assessed were movable sperm ratio, sperm activity index, survival rate, and DNA damage. The CPAs tested in this research were propylene glycol, dimethyl sulfoxide (DMSO), methanol, ethylene glycol, and glycerol. Different diluents, including 300 mM sucrose, 300 mM glucose, Stain’s solution, and Ringer’s solution, were investigated. The previous experiment showed that the optimal CPA for cryopreservation was DMSO with a concentration of 15% with 300 mM as diluent. To determine the effect of the dilution ratio, sperm was diluted to 1:1, 1:2, 1:10, 1:100, and 1:1000 with 300 mM sucrose containing DMSO at a final concentration of 15%. Lastly, the optimal freezing rate of the sperm was evaluated with four different freezing rates (−1, −5, −10, and −20 °C/min). Post-thaw sperm motility was higher with a dilution ratio lower than 1:2, and the freezing rate was less than −5 °C/min. In conclusion, these findings represent the development of a cryopreservation protocol for spotted halibut.

scholarly journals Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda

Reproduction ◽  
2004 ◽  
Vol 127 (5) ◽  
pp. 547-556 ◽  
Author(s):  
R E Spindler ◽  
Y Huang ◽  
J G Howard ◽  
P Wang ◽  
H Zhang ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Giant Panda ◽  
Calcium Ionophore ◽  
Sperm Cryopreservation ◽  
Management Tools ◽  
Giant Pandas ◽  
Before And After ◽  
Cryopreservation Protocol ◽  
Acrosomal Integrity

Sperm cryopreservation and artificial insemination are important management tools for giant panda breeding and the preservation of extant genetic diversity. This study examined the influence of freeze–thawing on sperm function, specifically capacitation. Sperm from nine giant pandas were assessed before and after rapid (− 40 and − 100 °C/min) cryopreservation by incubation in HEPES-buffered Ham’s F10 medium with and without the capacitation accelerators, 3-isobutyl-1-methylxanthine (IBMX) and dibutyryl cyclic AMP (dbcAMP). At 0, 3 and 6 h of exposure, aliquots were assessed for sperm motility traits and capacitation, defined as the proportion of sperm with intact acrosomes following exposure to solubilised zonae pellucidae (ursid or felid) or calcium ionophore subtracted from the proportion of sperm with intact acrosomes before exposure. Although mean±s.e.m. sperm motility post-thaw (56.1 ± 3.9% at 0 h) was less (P < 0.05) than pre-freeze (71.7 ± 6.0%), there was no difference (P > 0.05) in the proportion of acrosome-intact sperm (fresh, 93.0 ± 1.7% versus cryopreserved–thawed, 81.7 ± 4.7% at 0 h). Incidence of capacitation was greater (P < 0.05) in fresh sperm incubated with capacitation accelerators IBMX and dbcAMP (9 h: 50.9 ± 1.1) compared with fresh sperm incubated without accelerators (9 h: 41.2 ± 1.1%). Frozen–thawed sperm preincubated without accelerators underwent capacitation (49.6 ± 1.1%) to a greater extent (P < 0.05) compared with these fresh counterparts. Thawed samples with (9 h: 45.9 ± 1.4%) and without accelerators (9 h: 41.2 ± 1.1%) did not differ (P > 0.05) during the 9-h incubation. We conclude that giant panda spermatozoa (1) undergo capacitation in vitro with or without chemical accelerators and (2) withstand a rapid cryopreservation protocol, including retaining normal acrosomal integrity and functional capacitation ability.

Dimethylacetamide can be used as an alternative to glycerol for the successful cryopreservation of koala (Phascolarctos cinereus) spermatozoa

2008 ◽  
Vol 20 (6) ◽  
pp. 724 ◽  
Author(s):  
Yeng Peng Zee ◽  
William V. Holt ◽  
Jaime Gosalvez ◽  
Camryn D. Allen ◽  
Vere Nicolson ◽  
...  
Keyword(s):  
Disulfide Bonds ◽  
Membrane Integrity ◽  
Sperm Nucleus ◽  
Sperm Chromatin ◽  
Similar Degree ◽  
Sperm Cryopreservation ◽  
Phascolarctos Cinereus ◽  
Degree Of Protection ◽  
Fish Sperm ◽  
Cryopreservation Protocol

Swelling of koala sperm chromatin following cryopreservation has largely been attributed to the absence of intermolecular disulfide cross-linkages in the marsupial sperm nucleus. Fish spermatozoa also lack disulfide bonds within their chromatin, but have been successfully cryopreserved. The present study examined the hypothesis that the cryoprotectants used for fish sperm cryopreservation would confer a similar degree of protection on koala spermatozoa. Three concentrations each of five cryoprotectants (dimethyl sulfoxide, methanol, propylene glycol, ethylene glycol and dimethylacetamide (DMA)) were evaluated. Each treatment was compared against an established koala sperm cryopreservation protocol that uses 14% glycerol. Post-thaw assessment of progressive motility, plasma membrane integrity and mitochondrial membrane potential (MMP) revealed that protocols using 15% DMA achieved 62.2 ± 3.6% (P < 0.05) sperm survival, of which 79% (P < 0.05) had high MMP, an improvement of 32% and 40%, respectively, over sperm frozen in 14% glycerol. The percentage of spermatozoa with swollen nuclei was also lowest when frozen in 15% DMA, both immediately after thawing (18.0 ± 3.5%; P < 0.05) and after 2 h incubation at 35°C (35.8 ± 4.4%; P < 0.05). A second study was conducted to determine the optimal concentration of DMA for use in the cryopreservation of koala spermatozoa. High DMA concentrations (17.5% and 20%) resulted in significantly lower proportions of live spermatozoa showing high MMP immediately after thawing compared with spermatozoa frozen in the lower concentrations. The percentage of koala spermatozoa with swollen chromatin following cryopreservation was not affected by DMA concentration.

Development of sperm cryopreservation protocol of endangered spiny eel, Mastacembelus armatus (Lacepede 1800) for ex-situ conservation

Cryobiology ◽  
2016 ◽  
Vol 73 (3) ◽  
pp. 316-323 ◽  
Author(s):  
M. Mahfujur Rahman ◽  
M. Rahmat Ali ◽  
M. Rafiqul Islam Sarder ◽  
M. Fazlul Awal Mollah ◽  
Najmus Sakib Khan
Keyword(s):  
Ex Situ Conservation ◽  
Ex Situ ◽  
Sperm Cryopreservation ◽  
Mastacembelus Armatus ◽  
Cryopreservation Protocol

109 DEVELOPMENT OF A SPERM CRYOPRESERVATION PROTOCOL FOR THE ARGENTINE BLACK AND WHITE TEGU (TUPINAMBIS MERIANAE)

2014 ◽  
Vol 26 (1) ◽  
pp. 168 ◽  
Author(s):  
C. Young ◽  
M. Curtis ◽  
N. Ravida ◽  
F. Mazotti ◽  
B. Durrant
Keyword(s):  
Quality Index ◽  
Sperm Quality ◽  
Statistical Tests ◽  
Membrane Integrity ◽  
Monitoring Program ◽  
Florida Everglades ◽  
Equal Weight ◽  
Sperm Cryopreservation ◽  
Lizard Species ◽  
Black And White

Only 891 of the approximately 5600 lizard species have been evaluated by the International Union for Conservation of Nature (IUCN). Of those, at least one-third are threatened with extinction. However, there is no organised effort to preserve their genetic diversity through semen banking. As part of an invasive species monitoring program, Argentine black and white tegus were captured in the Florida Everglades. Following postmortem examination, sperm was collected by flushing the vas deferens and used as a model for the development of sperm cryopreservation protocols for related endangered lizards. Initial motility score (IMS; % motile × speed of progression2), plasma membrane integrity (IPL) and acrosome integrity (IAC) were recorded before freezing. Sperm was extended in TES and Tris (TEST)-yolk buffer with a final glycerol or dimethyl sulfoxide (DMSO) concentration of 8, 12, or 16%, and frozen in vials at 0.3, 1, or 6.3°C min–1. Vials were thawed at 37°C for 90 s. Cryoprotectant (CPA) was removed by centrifugation and resuspension of the sperm pellet in M199, at which time (T0) all variables were assessed and expressed as the percentage of initial (%IMS, %IPL, and %IAC). Statistical tests included multivariate ANOVA (MANOVA) and Student's t-test. Over all CPA concentrations and freeze methods, DMSO was significantly better than glycerol in maintaining %IMS (P = 0.01; 37.32 ± 3.5 and 25.44 ± 3.09, respectively) and %IAC (P < 0.01; 81.45 ± 3.45 and 22.99 ± 3.03, respectively). The 2 CPA were equally successful in protecting %IPL (P = 0.77; 56.61 ± 5.62 and 54.42 ± 4.93, respectively). The slowest freeze rate of 0.3°C min–1 was more successful than 1 and 6.3°C min–1 in preserving %IMS (P = 0.01; 37.85 ± 3.29, 26.03 ± 4.45, and 21.91 ± 4.45, respectively) and %IPL (P < 0.01; 77.43 ± 2.54, 27.99 ± 3.44, and 42.32 ± 3.44, respectively). The %IAC was not significantly affected by freeze rate (P = 0.14; 58.06 ± 6.89, 36.14 ± 9.33, and 42.99 ± 9.33, respectively). The interaction between CPA and freeze method affected %IMS (P < 0.01) and %IAC (P < 0.01), but did not affect %IPL (P = 0.28). All variables were affected (P < 0.05) by concentration of cryoprotectant as well as the interaction between freeze method and cryoprotectant concentration. To simplify these analyses and to determine the best overall freeze method for this species, a sperm quality index (SQI) was calculated, giving equal weight to each of the 3 measured indicators of cryosurvival. Table 1 depicts the product of %IMS, %IPL, and %IAC for each treatment. Because there were significant interactions between treatment parameters, each treatment was compared with all others (a–f). The SQI analysis revealed that tegu sperm frozen at 0.3°C min–1 in 8% DMSO exhibited a significantly higher post-thaw viability compared with all other treatments. Table 1.Sperm quality index (SQI) of thawed tegu sperm

Development of a Sperm Cryopreservation Protocol for Critically Endangered Mohashol, Tor tor (Hamilton)

2022 ◽  
Author(s):  
M. Salah Uddin Kabir ◽  
M. Rafiqul Islam Sarder ◽  
M. Matiur Rahman ◽  
M. Fazlul Awal Mollah ◽  
N. Binte Ryhan
Keyword(s):  
Critically Endangered ◽  
Sperm Cryopreservation ◽  
Cryopreservation Protocol ◽  
Tor Tor

scholarly journals Estudios preliminares de las características espermáticas y de criopreservación del coral formador de arrecifes Orbicella faveolata

2018 ◽  
Vol 47 (2) ◽  
Author(s):  
María Juliana Vanegas ◽  
Valeria Pizarro
Keyword(s):  
Sperm Morphology ◽  
Sperm Cryopreservation ◽  
Head Shape ◽  
First Report ◽  
Orbicella Faveolata ◽  
Cryopreservation Protocol ◽  
Mature Spermatozoa ◽  
Broad Scale

Cryopreservation has been recently applied to coral gametes and tissue with successful results that can be applied for different purposes on coral conservation and restoration. In this study, we decided to determine the sperm morphology of the coral Orbicella faveolata and assess the feasibility of sperm cryopreservation using a combination of intracellular (1,2-Propadiol) and extracellular (milk) cryoprotectants, and two frozen treatments for 24 h. Mature spermatozoa had a triangular-like head shape measuring 4.10 ± 0.69 μm(mean ± SD) and long flagellum (43.24 ± 7.99 μm). Fresh sperm remained viable and mobile for more than five hours after being released from the gamete bundles. After cryopreservation, all post-thaw sperm components assessed (morphology, motility and viability) showed no difference in contrast to fresh sperm. This study is the first report of cryopreservation of O. faveolata sperm, however further research is needed to increase the success of the cryopreservation protocol for broad-scale application.

scholarly journals Optimization of Sperm Cryopreservation Protocol for Peregrine Falcon (Falco peregrinus)

Animals ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 691
Author(s):  
Beatriz Cardoso ◽  
Irene Sánchez-Ajofrín ◽  
Cristina Castaño ◽  
Olga García-Álvarez ◽  
Milagros Cristina Esteso ◽  
...  
Keyword(s):  
Mitochondrial Activity ◽  
Peregrine Falcon ◽  
Sperm Cryopreservation ◽  
Freezing Method ◽  
Ultrarapid Freezing ◽  
Complex Process ◽  
Acrosome Integrity ◽  
Cryopreservation Protocol ◽  
Motility Parameters ◽  
Important Progress

Sperm cryopreservation is a complex process that needs to be adapted to wild and domestic avian species to ensure proper efficiency. Because of its accessibility, the peregrine falcon may be used as a good model for studying other raptor species. To find the most optimal cryopreservation protocol for peregrine falcon ejaculates, sperm parameters such as motility, viability, DNA fragmentation, acrosome integrity, and mitochondrial activity were analyzed under different conditions by varying the freezing method (slow freezing in straws vs. ultrarapid freezing in pellets), thawing conditions (37 °C for 30 s vs. 5 °C for 1 min), type of cryoprotectant (DMA vs. DMSO), and concentration of DMSO (4% vs. 8%). Results show that slow cryopreservation in straws yielded greater percentages (p < 0.05) of motile spermatozoa (22.5% ± 4.4% vs. 0.0% ± 4.1%), viable spermatozoa with intact acrosomes (84.6% ± 4.3% vs. 77.4% ± 4.3%), and spermatozoa with active mitochondria (41.0% ± 6.7% vs.12.8% ± 6.7%), compared with those obtained by the ultrarapid freezing in pellets. However, no differences were found between different thawing conditions. Moreover, all sperm motility parameters were greater (p < 0.05) when DMSO was used during freezing compared with DMA, although the use of 3% and 8% DMSO produced similar results. In conclusion, these results represent important progress in the study of falcon semen cryopreservation protocol, highlighting the crucial steps of the process and the most suitable conditions.

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