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.

220. Equine growth hormone enhances motility and extends longevity of ram spermatozoa in vitro

2005 ◽  
Vol 17 (9) ◽  
pp. 85 ◽  
Author(s):  
B. J. King ◽  
V. V. Da Silva ◽  
J. D. Harper ◽  
C. J. Scott ◽  
M. N. Sillence
Keyword(s):  
Growth Hormone ◽  
Dilution Rate ◽  
Sperm Motility ◽  
Room Temperature ◽  
Final Concentration ◽  
Bovine Growth Hormone ◽  
Semen Preservation ◽  
Sperm Survival ◽  
Motility Of Sperm

Sperm survival in vitro decreases with time at room temperature, but may be improved by treatment with recombinant bovine growth hormone in rats1, bulls2 and horses3. Two experiments investigated the effect of equine growth hormone (eGH) on the longevity of ram spermatozoa in vitro. The aim of experiment 1 was to determine if the addition eGH would improve the motility of ram spermatozoa after 24 h and identify any interaction with semen dilution rates used for ram semen preservation. Semen was collected from five mature Merino rams. Ejaculates were assessed for good quality and were diluted 1+50, 1+4, 1+3, 1+2 (semen+diluent) with a Tris-based cryoprotectant. Aliquots from each ram were mixed with eGH to achieve a final concentration of 100 ng/mL eGH and stored at 20°C for 24 h. Motility of spermatozoa was then assessed manually. eGH improved the motility of spermatozoa at all dilution rates compared to controls (P < 0.0001) but most markedly in the 1 + 3 and 1 + 2 samples (42.6 ± 0.8% eGH v. 22.7 ± 2.6% control; 40.5 ± 1.4% eGH v. 22.7 ± 2.2% control, respectively, P < 0.01). The aim of experiment 2 was to determine the optimum eGH concentration for improving sperm motility. eGH was added to aliquots of diluted semen (1+3 dilution rate) to produce samples with final concentrations of 1000, 100, 10 and 1 ng eGH/mL. The samples were placed in a water bath at 20°C for 24 h at which time the motility of sperm was assessed as before. Sperm motility was higher in the 100 ng/mL eGH sample (P < 0.05; 39.6 ± 0.7%) compared to other concentrations (1000 ng/mL 11.8 ± 0.7%, 10 ng/mL 21.5 ± 0.7% and 1 ng/mL 11.3±0.7%). We conclude that growth hormone is effective in promoting the longevity in vitro of ram spermatozoa stored at room temperature, and that this effect is concentration dependent. (1)Breier et al. (1996). Endocrinology 137, 4061–4064.(2)Sauerwein et al. (2000). Dom. Anim. Endocrinol. 18, 145–158.(3)Champion et al. (2002). Theriogenology 57, 1793–1800.

50 Long-term Motility of Semen Following Breeding Soundness Exams for Yearling Beef Bulls

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 152-152
Author(s):  
Garrett R Seltzer ◽  
Ashley R Hartman ◽  
Sharon K Tucker ◽  
David M Grieger
Keyword(s):  
Gradual Decrease ◽  
Future Research ◽  
Motile Sperm ◽  
Biological Factors ◽  
Ph Values ◽  
Scrotal Circumference ◽  
Breeding Soundness ◽  
Over Time

Abstract To find an in vitro predictor of in vivoM/em&gt; semen motility prompted this study. Our objective was to evaluate semen motility for an 8-hour period immediately following a breeding soundness exam. Ejaculates from 52 Angus and 56 Charolais bulls were evaluated. Motility, morphology, scrotal circumference and pH of ejaculate were evaluated at the time of collection. Ejaculates were then extended using a one to one ratio and incubated in a water bath held at 37 degrees Celsius and evaluated hourly. Motility was evaluated hourly for 8 hours, or until motility of the sample reached zero. Data were analyzed for breed and hourly effects using the GLIMMIX procedure of SAS. There was statistical evidence for difference (P &lt; 0.0001) between breeds for motility over time. Angus ejaculates had higher pH values than Charolais ejaculates showing an association between breed and pH (6.82 vs 6.76, respectively). Primary spermatozoa abnormalities were greater (P &lt; 0.0001) for Angus bulls compared to Charolais bulls (13.33% vs. 10.91%, respectively). Scrotal circumference between breeds tended to be different (P &lt; 0.07), with Charolais bulls having a larger scrotal circumference compared to Angus bulls (38.29 vs. 38.03 centimeters, respectively). There was no difference (P &gt; 0.05) between breeds for secondary abnormalities. There was a significant interaction (P &lt; 0.01) between breed and time of motility measurement. Angus bull’s motility decreased drastically until hour 4, it then had a more gradual decrease until hour 8. Charolais bulls had a more gradual decrease in the percentage of motile sperm over time. In conclusion, there was evidence for difference between breeds for pH, primary spermatozoa abnormalities, and long-term motility, and a scrotal tendency. Understanding the effects of breed and individual biological factors may help producers adjust BSE expectations and lead to future research in long term semen motility.

Peculiarities of pregnancy and delivery course in women after the application of assisted reproductive technologies against obesity (Based on the retrospective analysis)

2018 ◽  
pp. 122-126
Author(s):  
I.A. Zhabchenko ◽  
◽  
O.R. Sudmak ◽  
Keyword(s):  
In Vitro Fertilization ◽  
Pregnant Women ◽  
Retrospective Analysis ◽  
Perinatal Outcomes ◽  
Reproductive Technologies ◽  
Ischemic Lesion ◽  
Vitro Fertilization ◽  
Pregnancy And Delivery

The objective: to study the structure and frequency of complications of pregnancy, deliveries and perinatal outcomes in three groups of women: women with infertility and obesity, treated by application of in vitro fertilization (hereinafter IVF), pregnant women after IVF application with normal body weight, and pregnant women on the background of obesity which did not have an infertility in past history. Materials and methods. A retrospective analysis of 221 case histories of pregnancies and labors in women who were treated and gave birth in the Pregnancy and delivery pathology Department of SI «Institute of Pediatrics, Obstetrics and Gynecology named after Acad. O. M. Lukyanova of NAMS of Ukraine» for 2012 – 2016 years was carried out. Results. The overwhelming majority of pregnant women after IVF on the background of obesity are primaparas, who have a complicated obstetric history, hormonal changes in the form of progesterone deficiency predominantly and chronic inflammatory processes. Pregnancy with a combination of infertility, treated by the means of IVF application, and obesity, in most cases is accompanied by a long-term threat of termination of pregnancy (48.8%), threatening preterm deliveries (56%), placental dysfunction (41.5%), premature rupture of the amniotic membranes (41.5%), other problems during pregnancy, at the same time, every second woman (58.5%) had a combination of several complications, and required a long-term and repeated inpatient treatment (53.7%). The specific gravity of surgical delivery was 90%, and 16.2% of such deliveries were complicated by pathological blood loss. The number of preterm deliveries was 17.1%, with perinatal losses up to 11.3‰. Among full-term newborns 21.3% of newborns had malnutrition of the I degree and 17% of them had hypoxic-ischemic lesion of CNS. Conclusion. The course of pregnancy, delivery and the postpartum period in the studied contingent of women has a significant frequency of complications, mainly the coinciding ones, which affects on the consequences of perinatal outcomes and requires further study of this problem and the development of differentiated algorithms for antenatal observation. Key words: pregnancy, obesity, in vitro fertilization, complications, delivery, newborn.

scholarly journals Evaluation of Two Supplemented Culture Media for Long-Term, Room-Temperature Preservation ofStreptococcus pneumoniaeStrains

2017 ◽  
Vol 2017 ◽  
pp. 1-9
Author(s):  
Beatriz Quintero Moreno ◽  
María Araque ◽  
Evelyn Mendoza
Keyword(s):  
Survival Time ◽  
Room Temperature ◽  
Culture Media ◽  
Skim Milk ◽  
Extended Period ◽  
Phenotypic Characteristics ◽  
Genetic Profiles ◽  
Sequence Types ◽  
Over Time

Objective. To produce two supplemented agar types in order to store pneumococci for several months at room temperature.Methods. Todd-Hewitt/Hemoglobin/Yeast/Charcoal/Agar (TH-HYC) and Todd-Hewitt/Skim-Milk/Yeast/Charcoal/Agar (TH-SYC) were used to prepare two supplemented agar types. Nineteen pneumococci isolated from patients or asymptomatic carriers displaying diverse serotypes and multilocus sequence types (MLST) were subcultured and stored onto supplemented agar types, in four different tests, at room temperature.Findings. At the end of all tests (4–6 months) all noncontaminated subcultures were viable and maintained all phenotypic characteristics. Survival-time curves revealed a slow decrease of viable CFU over time on agar types, but at the end the number of viable CFU was satisfactory (≥2+ of growth). Decreasing of CFU was significantly higher for clinical versus nasopharyngeal isolates. Subcultures contamination rates were 6.25% and 14.58% after 2 and 6 months of storage, respectively.Conclusion. TH-HYC and TH-SYC agar types allowed the viability of pneumococci with several serotypes, MLST, and genetic profiles, after 6 months of storage at room temperature. We consider that these agar types are a valid alternative to preserve pneumococci over an extended period, especially when methods as cryopreservation or lyophilization are not available, and are useful for transporting strains between laboratories.

Infertility Treatments and Long-Term Neurologic Morbidity of the Offspring

2018 ◽  
Vol 36 (09) ◽  
pp. 949-954 ◽  
Author(s):  
Shai Levin ◽  
Eyal Sheiner ◽  
Tamar Wainstock ◽  
Asnat Walfisch ◽  
Idit Segal ◽  
...  
Keyword(s):  
Medical Center ◽  
Cohort Analysis ◽  
Multivariable Analysis ◽  
Maternal Diabetes ◽  
Population Based ◽  
Vitro Fertilization ◽  
Increased Risk ◽  
Attention Deficit Hyperactivity Disorders

Objective To determine the risk of long-term neurologic morbidity among children (up to 18 years) born following in vitro fertilization (IVF) or ovulation induction (OI) treatments as compared with spontaneously conceived. Study Design A population-based cohort analysis was performed, including data from the perinatal computerized database on all singleton infants born at the Soroka University Medical Center (SUMC) between the years 1991 and 2014. This perinatal database was linked and cross-matched with the SUMC computerized dataset of all pediatric hospitalizations. Results Neurologic morbidity was significantly more common in IVF (3.7%) and OI (4.1%) offspring as compared with those following spontaneous pregnancies (3.1%; p = 0.017). In particular, attention deficit/hyperactivity disorders and headaches were more common in the OI group and sleep disorders in the IVF group, whereas autism and cerebral palsy were comparable between the groups. In the Weibull multivariable analysis, while controlling for maternal age, preterm delivery, birthweight centile, maternal diabetes, and hypertensive disorders, IVF (adjusted hazard ratio [HR]: 1.40; 95% confidence interval [CI]: 1.14–1.71; p = 0.001), but not OI (adjusted HR: 1.17' 95% CI: 0.92–1.48; p = 0.196), was noted as an independent risk factor for long-term pediatric neurologic morbidity. Conclusion IVF offspring appear to be at an increased risk of long-term neurologic morbidity up to 18 years of age.

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.

scholarly journals Sex-specific effects of in vitro fertilization on adult metabolic phenotypes and hepatic transcriptomic and proteomic pathways in mouse

2020 ◽  
Author(s):  
Laren Narapareddy ◽  
Eric A. Rhon-Calderon ◽  
Lisa A. Vrooman ◽  
Josue Baeza ◽  
Duy K. Nguyen ◽  
...  
Keyword(s):  
Regulatory Element ◽  
Perinatal Outcomes ◽  
Vitro Fertilization ◽  
Specific Effects ◽  
Cholesterol Levels ◽  
Element Binding Protein ◽  
Fat Composition ◽  
Sterol Regulatory Element

AbstractAlthough in vitro fertilization (IVF) is associated with adverse perinatal outcomes, an increasing concern is the long-term health implications. We augmented our IVF mouse model to longitudinally investigate cardiometabolic outcomes in offspring from optimal neonatal litter sizes. We found that IVF-conceived females had higher body weight and cholesterol levels compared to naturally-conceived females, whereas IVF-conceived males had higher levels of triglycerides and insulin, and increased body fat composition. Through transcriptomics and proteomics of adult liver, we identified sexually-dimorphic dysregulation of the sterol regulatory element binding protein (SREBP) pathways that are associated with the sex-specfic phenotypes. We also found that global loss of DNA methylation in placenta was linked to higher cholesterol levels in IVF-conceived females. Our findings indicate that IVF procedures have long-lasting sex-specific effects on metabolic health of offspring and lay the foundation to utilize the placenta as a predictor of long-term outcomes.

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