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Post-thaw Motility (post-thaw + motility)
Selected AbstractsThe Effects of Osmolality, Cryoprotectant and Equilibration Time on Striped Bass Morone saxatilis Sperm MotilityJOURNAL OF THE WORLD AQUACULTURE SOCIETY, Issue 3 2003Shuyang He Four experiments were designed to evaluate the effects of osmolality, cryoprotectant, and equilibration time on striped bass sperm motility. In the first experiment, solutions of NaCI or KCI with osmolalities ranging from 0 to 700 mmol/kg were tested on sperm activation. Over 60% of the sperm were activated by isotonic NaCI and KCI solutions with a treatment osmolality of 350 mmol/kg. Sperm remained motile until osmolality increased to 600 mmol/ kg. In the second and third experiments, Extenders 1, 2 and 3 with osmolalities of 350, 500, and 600 mmol/kg, respectively, were tested. Sperm samples stored in Extender 2 showed significantly higher (P 0.01) sperm motility after 10 min of exposure as well as greater (P < 0.01) post-thaw motility when compared to samples stored in Extenders 1 and 3. In the fourth experiment, two trials were carried out to evaluate the effects of cryoprotectant and equilibration time. In the first trial, methanol with a concentration of 5% and 10% yielded the highest (P < 0.05) sperm motility prior to freezing at all equilibration times examined. However, 5% DMSO yielded the highest (P < 0.01) post-thaw motility (38 ± 3.6%). DMSO with concentrations of 10% and 15% resulted in 17 ± 2.3% and 6 ± 1.0% post-thaw motility, respectively. Both methanol and DMA, at all concentrations tested, resulted in less than 10% post-thaw motility. In the second trial, four DMSO concentrations with three different equilibration times were examined. We observed a significant (P < 0.001) interaction effect between DMSO concentration and equilibration time. Post-thaw motility was significantly greater (P < 0.01) with a concentration of 5% DMSO at all equilibration times examined, compared to 1.25, 2.5, and 10% DMSO. An average post-thaw motility of 40 ± 2.9% was achieved after 10 min equilibration using 5% DMSO. [source] Evaluation of Fertilizing Potential of Frozen-thawed dog Spermatozoa Diluted in ACP-106® using an In Vitro Sperm,Oocyte Interaction AssayREPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2007RCS Cardoso Contents The aim of present study was to evaluate frozen canine semen with ACP-106® (Powder Coconut Water) using an in vitro sperm,oocyte interaction assay (SOIA). Ten ejaculates from five stud dogs were diluted in ACP-106® containing 20% egg yolk, submitted to cooling in a thermal box for 40 min and in a refrigerator for 30 min. After this period, a second dilution was performed using ACP-106® containing 20% egg yolk and 12% glycerol. Samples were thawed at 38°C for 1 min. Post-thaw motility was evaluated by light microscopy and by using a computer aided semen analysis (CASA). Plasma membrane integrity and sperm morphology/acrosomal status were evaluated by fluorescent probes (C-FDA/PI) and Bengal Rose respectively. Moreover, frozen-thawed semen was analysed by a SOIA. Subjective post-thaw motility was 52.0 ± 14.8% and it was significant higher than the total motility estimated by CASA (23.0 ± 14.8%) because this system considered the egg yolk debris as immotile spermatozoa. Although normal sperm rate and acrosomal integrity evaluated by Bengal Rose stain was 89.6 ± 3.1% and 94.3 ± 3.1%, respectively, post-thaw percentage of intact plasma membrane was only 35.1 ± 14.3%. Regarding SOIA, the percentage of interacted oocytes (bound, penetrated and bound and/or penetrated) was 75.3%. Using regression analysis, it was found significant relations between some CASA patterns and data for SOIA. In conclusion, the freezing-thawing procedure using ACP-106® was efficient for maintain the in vitro fertility potential of dog spermatozoa. [source] Effect of pentoxifylline on motility and membrane integrity of cryopreserved human spermatozoaINTERNATIONAL JOURNAL OF ANDROLOGY, Issue 3 2002P. Stanic The purpose of this study was to examine the effects of pentoxifylline used before and after semen cryopreservation,thawing on sperm motility and membrane integrity. Twenty-four semen samples were split into four equal aliquots. Aliquots were incubated at 37 °C for 30 min, followed by cryopreservation with TEST-yolk freezing medium using slow programmable freezing protocol. After 2 weeks the sperm samples were thawed, washed twice in Quinn's Sperm Washing Medium (modified HTF with 5.0 mg/mL Human Albumin) and incubated at 37 °C for 30 min. Aliquots were treated by adding 3 mmol/L pentoxifylline to: (1) fresh sperm samples during incubation period prior to cryopreservation, (2) sperm samples as a supplement to the cryoprotectant prior to cryopreservation, and (3) thawed sperm samples during incubation period. One aliquot received no treatment (control group). The addition of 3 mmol/L pentoxifylline to fresh semen during incubation period prior to cryopreservation significantly decreased progressive and total motility compared with controls. However, the addition of 3 mmol/L pentoxifylline to cryopreserved semen after thawing significantly increased progressive and total motility compared with controls. After post-thaw, no differences in motion characteristics between sperm samples treated by adding 3 mmol/L pentoxifylline as a supplement to the cryoprotectant and control groups were observed. Post-thaw hypoosmotic swelling (HOS) test scores did not improve with the addition of pentoxifylline compared with the control group. It is concluded that pentoxifylline enhanced post-thaw motility of cryopreserved human spermatozoa when added after thawing. No improvement was found by freezing sperm with pentoxifylline. [source] The development of haddock and Atlantic cod sperm cryopreservation techniques and the effect of sperm age on cryopreservation successJOURNAL OF FISH BIOLOGY, Issue 2 2004R. M. Rideout Three cryoprotectants [dimethyl sulphoxide (DMSO), propylene glycol (PG) and glycerol], two diluents (sucrose- and saline-based), two sperm collection times, two freezing rates and three times between thaw and activation (0, 30 and 60 min) were tested in order to develop a protocol for the cryopreservation of sperm of haddock Melanogrammus aeglefinus and Atlantic cod Gadus morhua. The faster freezing rate resulted in extremely low post-thaw motility in comparison to the slower freezing rate, which was successful for sperm from both gadids. In both cases, the use of PG resulted in significantly higher post-thaw sperm motility-recovery indices than with DMSO or glycerol, which did not differ significantly from one another. Diluent had no effect on post-thaw sperm motility for Atlantic cod or haddock. Sperm collected at the end of the spawning season tended to have reduced post-thaw motility compared to that collected 2 weeks after the start of spawning. A 30 min delay between thaw and activation of haddock and Atlantic cod sperm resulted in a significant decrease in sperm motility. When PG was used as cryoprotectant, sperm motility continued to decrease between 30 and 60 min post-thaw. With DMSO or glycerol as cryoprotectant, motilities were already very low after 30 min post-thaw and did not decrease any further after 60 min. Cryoprotectant, diluent and time between thaw and activation had no effect on mean or maximum sperm swimming speeds for either Atlantic cod or haddock sperm. Fertilization success for haddock eggs, like sperm motility, was higher with PG-frozen sperm than DMSO- or glycerol-frozen sperm. These results constitute the first reported successful cryopreservation of haddock sperm and improve on previous methods used to cryopreserve sperm from Atlantic cod. [source] The Effects of Osmolality, Cryoprotectant and Equilibration Time on Striped Bass Morone saxatilis Sperm MotilityJOURNAL OF THE WORLD AQUACULTURE SOCIETY, Issue 3 2003Shuyang He Four experiments were designed to evaluate the effects of osmolality, cryoprotectant, and equilibration time on striped bass sperm motility. In the first experiment, solutions of NaCI or KCI with osmolalities ranging from 0 to 700 mmol/kg were tested on sperm activation. Over 60% of the sperm were activated by isotonic NaCI and KCI solutions with a treatment osmolality of 350 mmol/kg. Sperm remained motile until osmolality increased to 600 mmol/ kg. In the second and third experiments, Extenders 1, 2 and 3 with osmolalities of 350, 500, and 600 mmol/kg, respectively, were tested. Sperm samples stored in Extender 2 showed significantly higher (P 0.01) sperm motility after 10 min of exposure as well as greater (P < 0.01) post-thaw motility when compared to samples stored in Extenders 1 and 3. In the fourth experiment, two trials were carried out to evaluate the effects of cryoprotectant and equilibration time. In the first trial, methanol with a concentration of 5% and 10% yielded the highest (P < 0.05) sperm motility prior to freezing at all equilibration times examined. However, 5% DMSO yielded the highest (P < 0.01) post-thaw motility (38 ± 3.6%). DMSO with concentrations of 10% and 15% resulted in 17 ± 2.3% and 6 ± 1.0% post-thaw motility, respectively. Both methanol and DMA, at all concentrations tested, resulted in less than 10% post-thaw motility. In the second trial, four DMSO concentrations with three different equilibration times were examined. We observed a significant (P < 0.001) interaction effect between DMSO concentration and equilibration time. Post-thaw motility was significantly greater (P < 0.01) with a concentration of 5% DMSO at all equilibration times examined, compared to 1.25, 2.5, and 10% DMSO. An average post-thaw motility of 40 ± 2.9% was achieved after 10 min equilibration using 5% DMSO. [source] Testing Usability of Butylated Hydroxytoluene in Conservation of Goat SemenREPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2008TAA Khalifa Contents The objective of this study was to investigate whether butylated hydroxytoluene (BHT) could be used as a suitable supporter or alternative of egg yolk during preservation of goat spermatozoa. Three in vitro experiments and a fertility test were conducted to evaluate the effect of BHT on viability of chilled-stored semen as well as motility and kidding rate of frozen-thawed spermatozoa. In the first two experiments, ejaculates (n = 30/experiment) were collected from 10 bucks, split, diluted with egg yolk-based and egg yolk-free extenders supplemented with or without 0.3, 0.6, 2, 5 and 8 mm BHT and stored at 5°C for 168 h. In the third experiment, 30 ejaculates were collected from the above-mentioned bucks, split and diluted with egg yolk-free extenders supplemented with or without 0.3, 0.6 and 0.9 mm BHT and egg yolk-based extenders supplemented with or without 5 mm BHT. Diluted semen was cooled to 5°C over a period of 4 h, frozen and thawed in the form of 0.3-ml pellets. In the fertility test, 75 ejaculates were collected from two proven fertile bucks, split, diluted with egg yolk-free extenders containing 0.6 mm BHT and egg yolk-based extenders supplemented with or without 5 mm BHT, frozen and thawed as described above. An insemination volume of 0.6 ml containing 120,140 × 106 progressively motile spermatozoa was used for a single cervical insemination of cloprostenol-synchronized does (n = 230). The results showed that addition of 5 mm BHT to egg yolk-deficient (2.5%) extenders significantly improved viability of chilled-stored semen together with motility (48.5%) and fertility (62.5%) of frozen-thawed spermatozoa. Replacement of egg yolk in semen extenders by 0.6 mm BHT could sustain not only viability of chilled-stored semen but also post-thaw motility (47.5%) and fertility (53.75%) of frozen-thawed spermatozoa. In conclusion, supplementation of semen diluents with BHT can ameliorate preservability of goat sperm. [source] Evaluation of Fertilizing Potential of Frozen-thawed dog Spermatozoa Diluted in ACP-106® using an In Vitro Sperm,Oocyte Interaction AssayREPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2007RCS Cardoso Contents The aim of present study was to evaluate frozen canine semen with ACP-106® (Powder Coconut Water) using an in vitro sperm,oocyte interaction assay (SOIA). Ten ejaculates from five stud dogs were diluted in ACP-106® containing 20% egg yolk, submitted to cooling in a thermal box for 40 min and in a refrigerator for 30 min. After this period, a second dilution was performed using ACP-106® containing 20% egg yolk and 12% glycerol. Samples were thawed at 38°C for 1 min. Post-thaw motility was evaluated by light microscopy and by using a computer aided semen analysis (CASA). Plasma membrane integrity and sperm morphology/acrosomal status were evaluated by fluorescent probes (C-FDA/PI) and Bengal Rose respectively. Moreover, frozen-thawed semen was analysed by a SOIA. Subjective post-thaw motility was 52.0 ± 14.8% and it was significant higher than the total motility estimated by CASA (23.0 ± 14.8%) because this system considered the egg yolk debris as immotile spermatozoa. Although normal sperm rate and acrosomal integrity evaluated by Bengal Rose stain was 89.6 ± 3.1% and 94.3 ± 3.1%, respectively, post-thaw percentage of intact plasma membrane was only 35.1 ± 14.3%. Regarding SOIA, the percentage of interacted oocytes (bound, penetrated and bound and/or penetrated) was 75.3%. Using regression analysis, it was found significant relations between some CASA patterns and data for SOIA. In conclusion, the freezing-thawing procedure using ACP-106® was efficient for maintain the in vitro fertility potential of dog spermatozoa. [source] Studies on the Effect of Supplementing Boar Semen Cryopreservation Media with Different Avian Egg Yolk Types on in Vitro Post-thaw Sperm QualityREPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2006R Bathgate Contents Fertility after insemination of cryopreserved boar semen is currently below that of fresh semen. In an attempt to improve the post-thaw motility and acrosome integrity of boar sperm, semen was frozen using an adapted Westendorf method in which the chicken egg yolk was replaced by either duck or quail egg yolk. The different composition of the yolk types, particularly the amount of cholesterol, fatty acids and phospholipids, were thought to potentially afford a greater level of protection to sperm against damage during freezing and thawing. Sperm frozen in medium containing chicken egg yolk displayed higher motility immediately after thawing, but there was no difference in the motility of sperm frozen with different types of egg yolk 3 or 6 h after thawing and maintenance at 37°C. Sperm frozen in media containing chicken or duck egg yolk had a higher proportion of intact acrosomes immediately after thawing than sperm frozen in medium containing quail egg yolk, but 6 h after thawing and maintenance at 37°C the sperm that had been frozen in medium containing chicken egg yolk had a higher proportion of intact acrosomes than the sperm frozen in media containing duck or quail egg yolk. Analysis of the composition of the different yolk types showed that the basic components of the yolks were similar, but the ratios of fatty acids and phospholipid classes differed. Duck egg yolk had more monounsaturated fatty acids (MUFA) than chicken egg yolk, which had more MUFA than quail egg yolk. Duck egg yolk contained more phosphotidylinositol (PI) than chicken or quail egg yolks and quail egg yolk contained more phosphotidylserine than either chicken or duck egg yolks. The differences in post-thaw motility and acrosome integrity of boar sperm when frozen in media containing the different types of egg yolk may be due to the variation in composition. [source] Relationship between Sperm Response to Glycosaminoglycans in vitro and Non-return Rates of Swedish Dairy AI BullsREPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2000A Januskauskas Contents In this study, the relations between fertility (56-day non-return rates, 56-day NRR) after artificial insemination (AI) and bull sperm characteristics post-thaw, after swim-up and after co-incubation with heparin (Hep) and hyaluronan (HA), respectively, were determined, attempting to determine if such a procedure could be of value to evaluate the potential fertilizing ability of frozen-thawed AI bull spermatozoa. Spermatozoa from 20 semen batches derived from 20 Swedish Red and White AI bulls ranging widely in their field fertility after AI (55,79% 56-day NRRs) were evaluated with regards to post-thaw motility, membrane integrity, and migration through a simple swim-up procedure. Sperm viability and capacitation status were evaluated by two different vital staining procedures and chlortetracycline hydrochloride staining. Sperm motility and membrane integrity post-thaw (e.g. indicators of sperm viability) were significantly correlated (r = 0.53, p < 0.05 and r = 0.59, p < 0.01, respectively) with fertility. Heparin (5 µg/ml) significantly (p lt; 0.001) increased the frequencies of capacitation and acrosome-reaction (AR) among swim-up separated spermatozoa, whereas HA at a concentration of 50 ng/ml did not have any significant capacitating effect. The incidences of capacitated or AR-spermatozoa following Hep-treatment were not correlated with fertility. On the other hand, the percentage of viable spermatozoa was significantly (p < 0.001) lower in Hep-treated samples than in control and HA-treated samples and was significantly (r = 0.49, p < 0.05) correlated with fertility after AI (56-day NRR). The results indicate that the percentage of viable spermatozoa after swim-up separation and heparin-exposure from a selected population of AI bulls were significantly and positively related to the AI fertility of the donors and thus could be used as a parameter to determine the fertilizing ability of frozen,thawed AI bull spermatozoa. [source] Cryopreservation technique: comparison of Test yolk buffer versus SpermCryo and vapour versus computerised freezingANDROLOGIA, Issue 1 2008L. Paras Summary Semen cryopreservation offers the possibility to maintain fertility over a long time period e.g. for male cancer patients. Although its use expands worldwide, there is no established method that can be referred to as an entrenched standard for routine laboratory use. Cryodamage is still a general phenomenon and the success of cryopreservation is affected on one side by the cryoprotective agent and on the other side by the technique of freezing. In this methodological study, we compared the newly offered SpermCryo (SC) with the standard used cryoprotectant Test yolk buffer (TYB). We could show that TYB is superior to SC. In addition, we compared the two mainly used techniques for cryopreservation: computerised slow-stage freezing versus nitrogen vapour fast freezing. Regarding the sperm post-thaw motility and viability, no significant difference was found between these two methods. In conclusion, TYB can be recommended as a cryomedium of first choice and the appropriate freezing technique can be selected according to the local facilities of the institution. [source] Hormone injections enhance the tolerance of land-locked ayu spermatozoa to cryopreservationAQUACULTURE RESEARCH, Issue 16 2009Ken-ichi Yokoi Abstract We evaluated the effects of maturation-stimulating hormones on the post-thaw motility of land-locked form ayu (Plecoglossus altivelis) spermatozoa. Male ayu were administered three intraperitoneal injections of either salmon pituitary extract (SPE; 0.2 or 0.6 mg g,1 BW day,1) or of 17, 20,-dihydroxy-4-pregnen-3-one (DHP; 2 or 10 ,g g,1 BW day,1), the maturation-inducing steroid (MIS) in ayu. Before cryopreservation, the motility of spermatozoa of the SPE- and DHP-treated groups was significantly higher than that of the control group. Similarly, the comparative post-thaw motility (presented as a percentage of the motility obtained before cryopreservation) was significantly higher in the SPE group than in the control; however, there was no significant difference between the DHP group and the control. The effect of SPE and DHP on pre- and post-cryopreservation motility was not dose dependent. Our results suggest that the hormone(s) present in salmon pituitary are effective in enhancing the tolerance of ayu sperm cells to cryopreservation and that the MIS (DHP) is not involved in this process. [source] Effect of extender composition and freezing rate on post-thaw motility and fertility of Arctic char, Salvelinus alpinus (L.), spermatozoaAQUACULTURE RESEARCH, Issue 9 2006Nabil Mansour Abstract The effects of extender composition and freezing rate on motility and fertility of frozen-thawed Arctic char, Salvelinus alpinus, spermatozoa were investigated. Three freezing rates, two semen diluents and three cryoprotectants were tested. Semen frozen in 0.3 mol L,1 glucose diluent with 10% methanol as a cryoprotectant or in a diluent described by Lahnsteiner with 10%N,N- dimethylacetamide (DMA) resulted in the highest sperm motility. Fertility was the highest for semen frozen in a glucose,methanol extender but was not significantly different than that for semen frozen in Lahnsteiner's diluent with 10% DMA. Dimethyl sulphoxide (DMSO) at 10% was a relatively ineffective cryoprotectant with either semen diluent. Semen frozen at 6 cm above the surface of liquid nitrogen resulted in a higher post-thaw sperm motility and fertility than semen frozen at 5 cm. The addition of 7% fresh egg yolk to glucose diluent containing methanol or DMSO did not improve the fertility of frozen-thawed spermatozoa. However, the addition of 7% fresh egg yolk to glucose,DMA extender significantly improved the fertilization percentages of frozen-thawed spermatozoa. In conclusion, dilution of semen 1:3 in 0.3 mol L,1 glucose with 10% methanol and freezing 6 cm above the surface of liquid nitrogen (freezing rate of 40±8°C min,1, mean±SD from ,5 to ,55°C) is a promising protocol for cryopreservation of Arctic char semen. [source] Freezing equine semen: the effect of combinations of semen extenders and glycerol on post-thaw motilityAUSTRALIAN VETERINARY JOURNAL, Issue 7 2009J Scherzer Objective We evaluated combinations of two commercial semen extenders and three concentrations of glycerol to determine the combination that yielded the highest post-thaw sperm motility. Design A randomised 2 × 3 block design was used. Procedure Semen was collected from four stallions (6 collections per stallion). The sample was diluted with either a dried skim-milk glucose extender (EZ Mixin Original Formula) or a chemically defined, milk-free diluent (INRA 96), and each was used in combination with 2%, 3% or 4% glycerol in standard commercial freezing medium. Sperm motility was assessed by microscopy in fresh and post-thaw semen. Results There was a significant difference between the two extenders in the motility of spermatozoa after cryopreservation (48.9% for INRA 96; 38.6% for EZ Mixin OF; P < 0.0001). Glycerol at 4% in freezing medium yielded the highest post-thaw motility, significantly better than 2% (P < 0.05). Three of four stallions had significantly higher post-thaw motility using INRA 96 relative to EZ Mixin OF (P < 0.01), and two of four stallions had significantly higher post-thaw motility using 4% glycerol (P < 0.05). The combination of INRA 96 and 4% glycerol in freezing medium gave the highest average post-thaw motility of 51.5%. Conclusion In this study, INRA 96 combined with 4% glycerol yielded an average recovery of progressively motile sperm consistently above the 35% target. [source] | |||||||||||||