scholarly journals Short-term storage of tiger salamander (Ambystoma tigrinum) spermatozoa: The effect of collection type, temperature and time

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245047
Author(s):  
Amanda B. Gillis ◽  
Emmet L. Guy ◽  
Andrew J. Kouba ◽  
Peter J. Allen ◽  
Ruth M. Marcec-Greaves ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Hormone Treatment ◽  
Effect Of Temperature ◽  
Initial Assessment ◽  
Ambystoma Tigrinum ◽  
Tiger Salamander ◽  
Storage Duration ◽  
Spermatozoa Motility ◽  
Over Time

The aims of this project were to characterize tiger salamander (Ambystoma tigrinum) spermatozoa motility over time, when excreted as either milt or spermic urine prior to packaging into a spermatophore, and to determine the effect of temperature on sperm motility. A split-plot design was utilized to assess the motility of the two pre-spermatophore sample types at two temperatures, 0°C and 20°C (n = 10 for each treatment). Spermiation was induced through exogenous hormone treatment of luteinizing hormone releasing hormone analog in order to collect both milt and spermic urine, which were evaluated for motility, divided into two separate aliquots, and subsequently stored in either an ice-bath (0°C) or on the benchtop (20°C). The decay rate of sperm motility was assessed by reevaluating subsamples at 0.5, 1, 2, 3, 5, 7, and 24 hours following the initial assessment. Results showed that sperm stored at 0°C had significantly higher progressive, non-progressive, and total motility for both sperm collection types over time. An interaction was found between collection type and time, with milt exhibiting lower initial motility that was more sustainable over time, compared to spermic urine. For both milt and spermic urine, motility decreased rapidly with storage duration, indicating samples should be used as soon as possible to maximize motility for in-vitro fertilization and cryopreservation. This is the first study to describe the differences in sperm motility between milt and spermic urine from an internally fertilizing caudate and demonstrates the benefits of near freezing temperatures on sperm longevity.

271 THE EFFECT OF TEMPERATURE ON HUMAN TESTICULAR TISSUE TO OPTIMIZE THE IN VITRO MATURATION OF PRE-FREEZE MOTILITY

2009 ◽  
Vol 21 (1) ◽  
pp. 233
Author(s):  
M. C. Schiewe ◽  
A. Spitz ◽  
R. E. Anderson
Keyword(s):  
Sperm Motility ◽  
Seminiferous Tubules ◽  
Type I ◽  
Obstructive Azoospermia ◽  
Percentage Increase ◽  
Testicular Sperm ◽  
Incubation Condition ◽  
Progressive Motility ◽  
Over Time

The development of intracytoplasmic sperm injection has made the use of testicular sperm a viable option for infertile men with obstructive and nonobstructive azoospermia. Over the past decade, testicular biopsies have been handled and processed using a variety of different methods. Whole biopsy pieces can be effectively cryopreserved in a 10% glycerol diluent (Schiewe et al. 1997 67, S115 abst); however, the ability to find viable, motile sperm post-thaw is improved when prefreeze motility exists. The purpose of this study was to comparatively document in vitro sperm motility enhancement over time at different temperatures, and to prove that an intermediate temperature (28 to 30°C) would optimize sperm longevity for up to 1 week. In this study, 10 men with obstructive azoospermia underwent a surgical, open testicular biopsy procedure. Each biopsy was placed in HEPES buffered-human tubal fluid (mHTF) medium supplemented with 5% human serum albumin (HSA; Irvine Sci., Santa Ana, CA, USA) and transported to the laboratory at room temperature. Each testis biopsy (TBx) was dissected into 8 equal pieces (approximately 2 × 2 × 1 mm). Five intact pieces of TBx were cryopreserved in separate cryovials for future use. The remaining TBx tissue was subdivided into 1 of 3 temperature treatment groups (24, 30, or 37°C) for extended IVM. The 30°C incubation condition was achieved by placing the dish(es) in a Styrofoam box placed on a 37°C warming plate. Each TBx was placed into a separate 100 × 35 mm Falcon dish in 150-μL droplets of mHTF under oil and shredded by needle dissection to disperse the contents of the seminiferous tubules. Reminant tissue was placed into another droplet for additional dissection, as needed. Sperm were analyzed for motility (graded as Type I = twitching, II = undulating, III = slow progression, and IV = rapid progression) at 0, 24, 96, and 144 h without replacing the IVM medium. The percentage increase in motility, compared with 0 h, was statistically contrasted across temperature treatments by chi-squared analysis. Testicular sperm motility ranged from 5 to 25% at 0 h and increased in all groups at 24 h. There was no difference in total motility at 24 h, but progressive motility was higher (P < 0.05) at 37°C compared with 24°C. Significant differences (P < 0.05) were observed in total percentage of motility/percentage of progressive motility at 96 h with treatment differences (*) being 30°C (66%*/44%*) > 24°C (42%*/14%) > 37°C (18%*/10%). This statistical trend continued at 144 h with 30°C (42%*/23%*) > 24°C (24%*/8%) > 37°C (9%*/5%). During this study, a viable pregnancy was achieved using a 30°C sample 8 days post biopsy, exceeding 2 previous healthy triplet pregnancies, which were successful using 4-day-old TBx specimens in 1996 and 2004. This study confirms that an intermediate IVM temperature of 30°C is optimal to enhance TBx cryopreservation, which in our laboratory is generally performed at 24 to 72 h of IVM. Our routine pregnancy success in applying ICSI with IVM, thawed TBx sperm discounts the concern some individuals express regarding DNA fragmentation of sperm over time.

scholarly journals IN VITRO EFFECTS OF ACINETOBACTER BAUMANNII AND SELECTED NATURAL BIOMOLECULES ON RABBIT SPERMATOZOA MOTILITY

AGROFOR ◽  
2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Eva TVRDÁ ◽  
Michal ĎURAČKA ◽  
Attila KÁNTOR ◽  
Marek HALENÁR ◽  
Lukáš HLEBA
Keyword(s):  
Sperm Motility ◽  
Negative Impact ◽  
White Male ◽  
Selective Advantage ◽  
Rapid Decline ◽  
Sperm Analysis ◽  
Spermatozoa Motility ◽  
In Vitro Effects ◽  
Rabbit Spermatozoa

The aim of this study was to assess the potential efficiency of selected biologicallyactive substances on the motility behaviour of rabbit spermatozoa subjected to invitro induced A. baumannii contamination. The semen samples used for A.baumannii detection were collected from 10 New Zealand white male rabbits andthe presence of the bacterium was confirmed using MALDI-TOF MassSpectrometry. For the in vitro experiments rabbit spermatozoa were re-suspendedin PBS, containing mineral supplements, BSA and glucose in the presence of 3x105CFU A. baumannii and diverse concentrations of selected biomolecules (resveratrol- RES, quercetin - QUE, curcumin - CUR, epicatechin - EPI, isoquercitrin - ISO).The sperm motility was assessed using the computer-aided sperm analysis at 0h,2h, 4h and 6h. A. baumannii significantly decreased the sperm motility (P<0.001)at Time 2h and maintained this negative impact throughout the in vitro culture.Meanwhile, the motility at Time 2h was significantly higher in the samples subjectedto A. baumannii together with 10 μmol/L RES (P<0.01); 5, 10 and 50 μmol/L QUE(P<0.001); 1 μmol/L CUR (P<0.05); 10, 50 and 100 μmol/L EPI (P<0.01) as well as 50μmol/L (P<0.05) and 100 μmol/L ISO (P<0.001) in comparison to the control exposedto the bacterium exclusively. After 4h, the motility remained significantly higher in thegroups co-treated with the inoculum and 10 μmol/L RES (P<0.05), 50 μmol/L QUE(P<0.05) as well as 50 μmol/L EPI (P<0.05) when compared to the positive control.Nevertheless, none of the biomolecules was effective against the rapid decline ofsperm motility caused by A. baumannii during later stages of the experiment (Time6h). Based on these results, one can conclude that RES, QUE and EPI exhibitantibacterial properties providing a selective advantage to spermatozoa in thepresence of A. baumannii, particularly during short-term rabbit semen handling.

scholarly journals Mouse sperm delivery via cauda epididymis without transport medium

2016 ◽  
Author(s):  
Man-Xi Jiang ◽  
Mei-Shan Wang ◽  
Xiang-Hong Ou ◽  
Xue-Jin Chen ◽  
Yan Zhu
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Room Temperature ◽  
Sperm Quality ◽  
Mouse Sperm ◽  
Spermatozoa Motility ◽  
Transport Medium ◽  
Fertilization Rates ◽  
Progressive Motility

This study was aimed to investigate the effects of room temperature (RT, 20-25 oC) and absence of medium during cauda epididymis transport on spermatozoa quality, fertility and embryo development. In the first experiment, fresh sperm from one side of cauda epididymis was used for in vitro fertilization, and another side was delivered at RT or 4-8 oC either with or without M2. In the second experiment, each side of cauda epididymis obtained from the same mouse was individually delivered at RT or 4 oC with or without M2. Finally, sperm motility, progressive motility scores and fertility of fresh spermatozoa or those from transported cauda epididymis, and IVF embryo development were evaluated. Progressive motility scores and fertilization rates were higher in fresh spermatozoa than transported sperm; sperm motility of transported cauda epididymis at 4-8 oC was comparable to fresh spermatozoa, but spermatozoa motility of transported cauda epididymis at RT was inferior to fresh spermatozoa. Spermatozoa motillty of transported cauda epididymis at 4-8 oC with transport medium was much higher than that without transport medium; absence of transport medium did not affect sperm motility of transported cauda epididymis at 4-8 oC but affected sperm motility of transported cauda epididymis at RT. Sperm quality from transported cauda epididymis can be efficiently kept at 4-8 oC, and cauda epididymis transport at 4-8 oC without M2 is more beneficial on keeping their fertility. Moreover cauda epididymis transport at RT without medium could sufficiently produce embryos for obtainning live offsprings.

scholarly journals In Vitro Effects of Selected Trichothecenes on the Rabbit Spermatozoa Motility Behavior – A Comparative Study

2016 ◽  
Vol 65 (3-4) ◽  
pp. 21-26
Author(s):  
Eva Tvrdá ◽  
Michal Ďuračka ◽  
Marek Halenár ◽  
Norbert Lukáč ◽  
Adriana Kolesárová
Keyword(s):  
Sperm Motility ◽  
Toxic Substance ◽  
Reproductive Capacity ◽  
Rapid Decline ◽  
Sperm Analysis ◽  
Spermatozoa Motility ◽  
In Vitro Effects ◽  
Rabbit Spermatozoa ◽  
Motility Behavior

Summary This study was designed to describe and compare the time- and dose-dependent in vitro effects of selected trichothecenes (deoxynivalenol-DON, zearalenone-ZEA and T-2 toxin) on the motility behavior of rabbit spermatozoa. The rabbit semen was diluted in PBS supplemented with different concentrations (1, 5, 10, 50, 100 μmol/L) of DON, ZEA or T-2 while the Control carried no mycotoxin. At culture times of 0h, 2h, 4h and 8h, the spermatozoa motility was assessed using the computer-aided sperm analysis (CASA) with the help of the IDENT stain and fluorescent illumination. The motility assessment revealed different behavior patterns, specific and unique to each of the studied mycotoxins. DON exhibited the ability to temporarily increase the sperm motility, followed by its rapid decline at later stages of the experiment (P<0.001). ZEA proved to act as a highly toxic substance on the spermatozoa, causing a rapid decline of the motion and resulting in a fast and complete sperm motility inhibition (P<0.001). Lastly, T-2 revealed to be highly detrimental to the sperm activity even at small concentrations (P<0.001). Our data suggest that further experiments are needed due to the lack of evidence emphasizing the toxinogenic effects of trichothecenes on male reproductive capacity.

scholarly journals Mouse sperm delivery via cauda epididymis without transport medium

2016 ◽  
Author(s):  
Man-Xi Jiang ◽  
Mei-Shan Wang ◽  
Xiang-Hong Ou ◽  
Xue-Jin Chen ◽  
Yan Zhu
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Room Temperature ◽  
Sperm Quality ◽  
Mouse Sperm ◽  
Spermatozoa Motility ◽  
Transport Medium ◽  
Fertilization Rates ◽  
Progressive Motility

This study was aimed to investigate the effects of room temperature (RT, 20-25 oC) and absence of medium during cauda epididymis transport on spermatozoa quality, fertility and embryo development. In the first experiment, fresh sperm from one side of cauda epididymis was used for in vitro fertilization, and another side was delivered at RT or 4-8 oC either with or without M2. In the second experiment, each side of cauda epididymis obtained from the same mouse was individually delivered at RT or 4 oC with or without M2. Finally, sperm motility, progressive motility scores and fertility of fresh spermatozoa or those from transported cauda epididymis, and IVF embryo development were evaluated. Progressive motility scores and fertilization rates were higher in fresh spermatozoa than transported sperm; sperm motility of transported cauda epididymis at 4-8 oC was comparable to fresh spermatozoa, but spermatozoa motility of transported cauda epididymis at RT was inferior to fresh spermatozoa. Spermatozoa motillty of transported cauda epididymis at 4-8 oC with transport medium was much higher than that without transport medium; absence of transport medium did not affect sperm motility of transported cauda epididymis at 4-8 oC but affected sperm motility of transported cauda epididymis at RT. Sperm quality from transported cauda epididymis can be efficiently kept at 4-8 oC, and cauda epididymis transport at 4-8 oC without M2 is more beneficial on keeping their fertility. Moreover cauda epididymis transport at RT without medium could sufficiently produce embryos for obtainning live offsprings.

107 COMPARISON OF PRIMATE SPERM CRYOPRESERVATION PROTOCOLS: POST-THAW SPERM RECOVERY AND HYPERACTIVATION IN CULTURE

2006 ◽  
Vol 18 (2) ◽  
pp. 161
Author(s):  
S. Nichols ◽  
B. Bavister
Keyword(s):  
New York ◽  
Sperm Motility ◽  
Room Temperature ◽  
Vitro Fertilization ◽  
Treatment Groups ◽  
Motility Of Sperm ◽  
Fertilizing Capability ◽  
Over Time

Cryopreservation of spermatozoa provides material for gene banking of genetically valuable males and offers convenience for in vitro fertilization (IVF). In addition, cryobanking of spermatozoa allows less frequent ejaculation collections from males. The present study compared the effectiveness of several published techniques in non-human primates to find the most efficient one for rhesus macaque (Macaca mulatta) semen cryopreservation. The effectiveness of each method was assessed by longevity (post-thaw motility % and duration) and ability to hyperactivate in culture in response to chemical activators (caffeine, dbcAMP) needed for rhesus sperm capacitation (Boatman and Bavister 1984 J. Reprod. Fertil. 71, 357-366). The ability to hyperactivate provides a reasonable assessment of the fertilizing capability of spermatozoa prior to performing IVF; the latter was impractical for this study, given the large number of treatments and endpoints. Spermatozoa were collected via electroejaculation from one male on three occasions to avoid confounding treatments with male effects. Each ejaculate was divided into one of four treatment groups for cryopreservation: Method A (Seier et al. 1993 J. Med. Primatol. 22, 355-359); Method B (Wei et al. 2000); Method C (Sanchez-Partida et al. 2000, Biol. Reprod. 63, 1092-1097); and Method D (Isachenko et al. 2005 Reprod. Biomed. Online 10, 350-354). Protocols were followed according to each published technique. Upon thawing, each sample was split into different incubation conditions: 37�C, 5% CO2 in air or room temperature for 0-24 h. One dose of activators was used according to standard protocol. Statistical analyses of motility rates were performed using 2 � 2 G tests (Sokal and Rohlf 1981 Biometry. New York: W. H. Freeman Co.) to determine significance. Samples cryopreserved using method D did not survive the method (motility = 0) and were not included in the statistical analysis. Methods A-C all demonstrated reasonable post-thaw motility recovery rates (68%, 73%, and 62%, respectively) and underwent capacitation within 30 min of exposure to activators. Sperm motility decreased over time in culture within each treatment at 37�C. However, spermatozoa in Method A were significantly less motile at 4 and 24 h than those in Methods B and C, and Method B spermatozoa were significantly less motile at 24 h than those in Method C. Sperm motility also decreased over time in samples incubated at room temperature, with motility of sperm in Method A motility being significantly less at 24 h than that of sperm in Methods B and C. Method C best preserved motility over time regardless of temperature of incubation upon thawing. Overall, incubation at room temperature preserved motility better than incubation at 37�C. Methods A-C yielded satisfactory post-thaw recovery of progressively motile spermatozoa despite the various differences among their protocols. For long-term use of each sample, however, it would be beneficial to incubate spermatozoa at room temperature after using Method C. This technique appears to be more appropriate for gene banking rhesus semen, and applying this protocol would allow more efficient usage of each semen sample, potentially providing for multiple IVF cases over a 24-h period. This work was supported by NIH Grant RR15395.

The development of dormancy in bulblets of Lilium speciosum generated in vitro. II. The effect of temperature

1990 ◽  
Vol 80 (3) ◽  
pp. 431-436 ◽  
Author(s):  
Isabelle Delvallee ◽  
Annie Paffen ◽  
Geert-Jan De Klerk
Keyword(s):  
Effect Of Temperature ◽  
Lilium Speciosum

Effect of Temperature on ADP-Induced Platelet Aggregation

1973 ◽  
Vol 29 (01) ◽  
pp. 183-189
Author(s):  
C. A Praga ◽  
E. M Pogliani
Keyword(s):  
Platelet Aggregation ◽  
Incubation Period ◽  
Room Temperature ◽  
Important Variable ◽  
Practical Significance ◽  
Effect Of Temperature ◽  
Induce Platelet Aggregation ◽  
Cuvette Holder ◽  
Exact Temperature

SummaryTemperature represents a very important variable in ADP-induced platelet aggregation.When low doses of ADP ( < 1 (μM) are used to induce platelet aggregation, the length of the incubation period of PRP in the cuvette holder of the aggregometer, thermostatted at 37° C, is very critical. Samples of the same PRP previously kept at room temperature, were incubated for increasing periods of time in the cuvette of the aggregometer before adding ADP, and a significant decrease of aggregation, proportional to the length of incubation, was observed. Stirring of the PRP during the incubation period made these changes more evident.To measure the exact temperature of the PRP during incubation in the aggre- gometer, a thermocouple device was used. While the temperature of the cuvette holder was stable at 37° C, the PRP temperature itself increased exponentially, taking about ten minutes from the beginning of the incubation to reach the value of 37° C. The above results have a practical significance in the reproducibility of the platelet aggregation test in vitro and acquire particular value when the effect of inhibitors of ADP induced platelet aggregation is studied.Experiments carried out with three anti-aggregating agents (acetyl salicyclic acid, dipyridamole and metergoline) have shown that the incubation conditions which influence both the effect of the drugs on platelets and the ADP breakdown in plasma must be strictly controlled.

The Development of Homologous (Canine/Anti-Canine) Antibodies in Dogs with Haemophilia A (Factor VIII Deficiency): A Ten-Year Longitudinal Study

1993 ◽  
Vol 69 (01) ◽  
pp. 021-024 ◽  
Author(s):  
Shawn Tinlin ◽  
Sandra Webster ◽  
Alan R Giles
Keyword(s):  
Factor Viii ◽  
Canine Model ◽  
Significant Inhibition ◽  
Human Condition ◽  
Haemophilia A ◽  
Variable Expression ◽  
The Family ◽  
Over Time

SummaryThe development of inhibitors to factor VIII in patients with haemophilia A remains as a serious complication of replacement therapy. An apparently analogous condition has been described in a canine model of haemophilia A (Giles et al., Blood 1984; 63:451). These animals and their relatives have now been followed for 10 years. The observation that the propensity for inhibitor development was not related to the ancestral factor VIII gene has been confirmed by the demonstration of vertical transmission through three generations of the segment of the family related to a normal (non-carrier) female that was introduced for breeding purposes. Haemophilic animals unrelated to this animal have not developed functionally significant factor VIII inhibitors despite intensive factor VIII replacement. Two animals have shown occasional laboratory evidence of factor VIII inhibition but this has not been translated into clinical significant inhibition in vivo as assessed by clinical response and F.VIII recovery and survival characteristics. Substantial heterogeneity of inhibitor expression both in vitro and in vivo has been observed between animals and in individual animals over time. Spontaneous loss of inhibitors has been observed without any therapies designed to induce tolerance, etc., being instituted. There is also phenotypic evidence of polyclonality of the immune response with variable expression over time in a given animal. These observations may have relevance to the human condition both in determining the pathogenetic factors involved in this condition and in highlighting the heterogeneity of its expression which suggests the need for caution in the interpretation of the outcome of interventions designed to modulate inhibitor activity.

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