Ejaculated Spermatozoa Research Articles

Effect of Management and Environmental Factors on Semen Production Potential of Indigenous Cattle Breeds

Background: The production of higher number of quality semen doses from the indigenous bulls is required to mitigate the issue of declining population of indigenous cattle breeds. To make the semen production more efficient, the knowledge of factors affecting the semen production is of utmost importance. However, studies on different indigenous breeds are scarcely available and very little information is available on effect of ejaculate number, collection interval, semen collector etc. The present investigation was hence focused to study the effect of different management and environmental factors affecting semen production traits in indigenous breeds. Methods: Semen data comprising of 15,435 ejaculates from 60 bulls of 10 indigenous cattle breeds were analysed for semen production traits. Prior to the analysis, data on initial motility was transformed using the arcsine transformation. Least square analysis was carried out using “lm” function while ANOVA (Type III sum of square) and Duncan’s multiple range test were computed using “car” and “agricolae” packages from R statistical software. Result: The overall means for the semen production traits: ejaculate volume, sperm concentration, total sperm, initial motility and total motile sperm were 5.62±0.041 ml, 1.42±0.51× 109/ml, 7.96±0.25× 109/ejaculate, 74.77±0.002% and 5.92± 0.11 × 109/ejaculate, respectively. All traits were significantly affected by order of ejaculate, collection interval, age at collection, breed, semen collector, season and year of collection except collection interval which had no effect on motility. Except initial motility, semen characters showed quadratic pattern over the age. Results of the study would help in suggesting suitable management and breeding plans for semen production traits.

Cтворення інформаційної бази про кнурів миргородської породи, кріоконсервована сперма яких зберігається у банку генетичних ресурсів тварин

Goal. To provide information about boars of Myrgorod breed, which sperm production is stored in deep-chilled state in the Bank of animal genetic resources (Institute of genetics and animal breeding named after M.V. Zubets of NAAS) in the context of the formation of the structured information on animal genetic resources at the national level and possible recovery of the breed using modern methods. Methods. The study was conducted on boars of Myrgorod breed belonging to the lines Veselyi, Dnipro, Komysh, and Lovchyk. The study of immunogenetic characteristics of boars was carried out by 9 genetic systems of blood groups: EAA, EAB, EAD, EAE, EAF, EAG, EAH, EAK, and EAL. The processing involved data from all boars of these lines that were in the breed. Assessment of boar semen was carried out on the ejaculate volume and mobility of sperm cells in the laboratory of artificial insemination of breeding plant SE «SF named after Decembrists of the Institute of pig breeding and agro-industrial production of NAAS». They took semen from boars during the year in the mode of the sexual use — one cage in 3 days. They determined the average volume of ejaculate and motility of sperm cells. The characteristic of boars as to the productivity was made on the indicators of live weight, body length, age of reaching a live weight of 100 kg during growth, and multiple pregnancies in sows inseminated by them. Results. It was found that among the examined boars the most common allelic variants were A, Ba, Db, Eedg, Eaeg, Fb, Gb, H, Kb, Lb with a frequency of 0.233–1.000. To the category of relatively rare were read alleles Eaef, Ka, Ld with a frequency of 0.033–0.067. The boars of Myrgorod breed did not differ significantly among themselves for the most of the genetic systems of blood groups, besides systems of EAE, EAK. and EAL. The obtained results give the possibility to realize a homogeneous or heterogeneous selection and to create desirable genotypes in the breed in case it is subjected to recovery. The largest volume of ejaculate sperm was obtained from boars of lines Veselyi and Dnipro (223.8 and 227.4 ml respectively), the smallest – from line Komysh (139.6 ml), which was probably due to the individual characteristics of animals and their heredity. The sperm of the studied boars were not significantly different in the mobility of sperm cells (9.0–9.5 points). Conclusions. The characteristics of the boars-manufacturers on the genetic systems of blood groups, sperm quality, and performance are given. This information can be used to retrieve ancestors of lines or families in the recovery process of the Myrgorod breed of pigs and the creation of other genotypes. It will also help to solve the complex problems of preservation and use of local endangered breeds of agricultural animals of Ukraine under the recommendations of the Commission on animal genetic resources of FAO.

Spermatozoa retrieved by electroejaculation: Should we prefer fresh or cryopreserved spermatozoa for intracytoplasmic sperm injection?

We aim to evaluate our experience, comparing intracytoplasmic sperm injection (ICSI) outcomes of cycle using fresh versus thawed electroejaculated spermatozoa. All consecutive couples undergoing ICSI cycles using electroejaculated spermatozoa, during a 16-year period, were evaluated. Embryological/laboratory variables of the ICSI cycles were assessed and compared between those utilising fresh (fresh group) versus thawed (thawed group) electroejaculated spermatozoa. Fifty-seven couples were evaluated, 30 used a fresh electroejaculated spermatozoa in 55 ICSI cycles, while 27 used a thawed sperm sample in 41 ICSI cycles. There were no in-between group differences in the mean numbers of oocytes retrieved per oocyte retrieval nor the percentage of MII oocytes. The fresh group demonstrated significantly higher fertilisation (71.5% vs. 64.1%, respectively, p<.05), top-quality embryos (66.5% vs. 54.9%, respectively, p<.02), clinical pregnancy per transfer (41.3% and 21.2%, respectively, p<.05) and cumulative clinical pregnancy (58.2% vs. 26.8%, respectively, p<.001) rates, as compared to the thawed group. Independent of the source of spermatozoa used, no pregnancy was achieved following ICSI utilising immotile spermatozoa. In conclusion, ICSI cycles using ejaculated spermatozoa of patients suffering from neurologic or psychogenic anejaculation are reassuring. The use of fresh ejaculated spermatozoa retrieved on the day of the female spouse oocyte retrieval might improve outcome. Whenever a thawed electroejaculated spermatozoa yield no motile spermatozoa, emergency electroejaculation is mandatory.

Effect of thermal stress on basic seminal characteristics of Gyr bulls.

The objective of this study was to evaluate the influence of thermal stress indices and minimum temperature on the semen quality of dairy Gyr bulls and to determine whether the minimum night temperature is able to balance the thermal stress that occurred during the day. A total of 922 semen samples were collected from 391 bulls aged between 15 and 49months, which participated in progeny pretest trials, from December 2011 to April 2017. Semen characteristics (ejaculate volume, gross motility, sperm motility, sperm vigor, sperm concentration, and percentage of total defects) and scrotal circumference were analyzed. Meteorological data were recorded daily by the National Institute of Meteorology. The analyses were carried out using mixed models. The following fixed effects were included in the model: test, month of sample collection, scrotal circumference, the age of the bull at sampling as covariate (linear regression), and the climatic variables (equivalent temperature index [ETI], temperature and humidity index [THI], or minimum temperature). The effect of the bull nested within the test was included in the model as a random effect. Two periods were considered to evaluate the effect of the climatic variables on sperm quality: 7days before collection (period 1) and the day of collection (period 2). There was an effect of ETI on sperm vigor and concentration in both periods and on volume and percentage of total defects only in the second period. The THI exerted a significant effect on volume in both periods and on sperm motility and concentration only in the second period. There was an effect of the minimum temperature on ejaculate volume, sperm motility, sperm concentration, and percentage of total defects only in the second period, which corresponds to the day of semen collection. Taken together, the results show that there was sufficient thermal stress to negatively affect semen quality. However, the minimum temperature during the night was a significant factor that balanced these negative effects of THI and ETI on semen traits of Gyr bulls.

Effects of digoxin on full-type hyperactivation in bovine ejaculated spermatozoa with relatively lower survivability for incubation with stimulators of cAMP signaling cascades

Previous researches of our laboratory reported that addition of cAMP analog cBiMPS and protein phosphatase inhibitor calyculin A (stimulators of cAMP signaling cascades) improved the capacity of incubation medium to induce full-type hyperactivation in bovine ejaculated spermatozoa. However, this modified medium was valid only for samples with relatively good survivability for incubation with stimulators of cAMP signaling cascades. Thus, it is necessary to make further modified medium for evaluation of potentials to exhibit full-type hyperactivation in bovine sperm samples with relatively lower survivability. Na+/K+-ATPase is an integral membrane protein and involved with the regulation of rodent sperm motility. To make further modification of the medium, we examined effects of Na+/K+-ATPase inhibition with digoxin on motility, full-type hyperactivation and protein tyrosine phosphorylation in bovine ejaculated spermatozoa with relatively lower survivability for incubation with stimulators of cAMP signaling cascades and also performed the immunodetection of bovine sperm Na+/K+-ATPase. The addition of Na+/K+-ATPase inhibitor digoxin to the incubation medium containing cBiMPS and calyculin A had the tendency to lessen the decreases in the percentages of motile spermatozoa in all of 12 samples after the incubation for 1–3 h and significantly increased the percentages of full-type hyperactivation in one group of 4 samples (Sample-A1) and another group of 4 samples (Sample-A2) after 1 and 2 h respectively, though it had no significant effects on full-type hyperactivation in the other group of 4 samples (Sample-B). In addition, incubation time-related changes in the sperm protein tyrosine phosphorylation (a good marker for sperm capacitation) were correlated with those in the percentages of full-type hyperactivation in Sample-A1 containing digoxin. Immunodetection showed that Na+/K+-ATPase is present in the middle and principal pieces of the flagella, indicating that Na+/K+-ATPase has possible relations with sperm motility. These results obtained with bull ejaculated spermatozoa with relatively lower survivability indicate that incubation method using digoxin is useful to evaluate potentials of sperm samples to exhibit full-type hyperactivation, that digoxin has effects on suppressing reduction of sperm motility, and that prolonged incubation with digoxin induces reduction of capacitation state which may suppress the maintenance of full-type hyperactivation.

Fibronectin induces capacitation-associated events through the endocannabinoid system in bull sperm

Mammalian ejaculated spermatozoa must undergo a series of changes in the female reproductive tract, collectively called capacitation, in order to fertilize the oocyte. We reported that fibronectin (Fn), a glycoprotein from the extracellular matrix, and anandamide (AEA), one of the major members of the endocannabinoid family, are present in the bovine oviductal fluid and regulate bull sperm function. Also, AEA induces bovine sperm capacitation, through CB1 and TRPV1 receptors.In this work, we investigated if Fn induces bovine sperm capacitation thought the activation of the endocannabinoid system in this process. We incubated sperm with Fn (100 μg/ml) and/or capsazepine, a TRPV1 antagonist (0.1 μM) and some events related to sperm capacitation such as LPC-induced acrosome reaction, sperm-release from the oviduct, induction of PKA phosphorylated substrates (pPKAs) and protein tyrosine phosphorylation (pY) and nitric oxide (NO) production were assessed. Also, we studied the activity of fatty acid amide hydrolase (FAAH), the enzyme that degrades AEA. We found that Fn, via α5β1 integrin, induced capacitation-associated events. Also, Fn stimulated signaling pathways associated to capacitation as cAMP/PKA and NO/NO synthase. Moreover, Fn decreased the FAAH activity and this correlated with sperm capacitation. Capsazepine reversed fibronectin-induced capacitation, and pPKAs and NO levels. The incubation of spermatozoa with R-methanandamide (1.4 nM), a stable analogue of AEA, increased cAMP and pPKAs levels. The presence of H89 (50 μM) or KT5720 (100 nM) (PKA inhibitors) prevented AEA-induced capacitation. In addition, R-methanandamide and capsaicin (0.01 μM), a TRPV1 agonist, increased NO production via the PKA pathway. These results indicate that Fn, through α5β1, supports capacitation in bovine spermatozoa. This effect is dependent on the activation of TRPV1 through cAMP/PKA and NO signaling pathways. We propose that Fn could be considered as a new agent that promotes sperm capacitation in bull sperm. Our findings contribute to better understand the significance of Fn signaling in the capacitating events that lead to successful fertilization and embryo development in mammals including humans.

REPRODUCING QUALITIES OF RAMS OF THE MOLDOVAN TYPE KARAKUL BREED

Issues related to the reproduction of animals have been and remain one of the most complex and relevant problems of biology and are constantly finding a direct and effective way out into livestock farming practice. The rational use of breeding sheep as producers is limited by the lack of standard, objective methods and methods for the timely assessment of their reproductive ability. The article presents the results of evaluating the sperm production of rams-producers. For the first time, the freshly obtained sperm of rams of Moldavian type producers of the Karakul breed was evaluated using the macroscopic method - ejaculate volume, color and smell, as well as the microscopic method - motility, sperm concentration in the ejaculate, total sperm count in the ejaculate sperm movements (VAP; VSL and VCL) as well as the percentage of abnormal forms of sperm. The experiments were carried out on sheep producers grown on a pedigree farm of the experimental farm of the Moldavian Scientific and Practical Institute of Biotechnology in Animal Husbandry and Veterinary Medicine. As a result of the studies, it was found that the ejaculate volume on average was 0.99 ± 0.04 ml, motility was 0.95 ± 0.02 and sperm concentration 1.51 ± 0.14 billion/ml. The percentage of pathological forms in sperm averaged 13.72 ± 0.61, an indicator that characterizes the high quality of sperm. The average quality indicator of ejaculates obtained from ram-producers of the Moldavian type of the Karakul breed corresponds to physiological standards for the Karakul breed. The average percentage of pathological forms of sperm found in ejaculates indicates that the rams were in good conditions of feeding and keeping. According to research, we can say that the Moldovan type of Karakul rams can be successfully used to obtain high quality ejaculates suitable for freezing sperm at 196°C.

Modified permeable cryoprotectant-free vitrification method for three or fewer ejaculated spermatozoa from cryptozoospermic men and 7-year follow-up study of 14 children born from this method.

What technique can be used to successfully cryopreserve three or fewer ejaculated spermatozoa from cryptozoospermic men and is the physical and cognitive development of children born after this technique normal? The modified cryopreservation method for three or fewer human spermatozoa from cryptozoospermic men showed a recovery rate above 95% and a survival rate just under 90%, and the physical and cognitive abilities of the children born after ICSI were comparable to those born after natural conception. Clinical outcomes of ICSI using cryptozoospermic men's ejaculated spermatozoa are considered to be inferior to that using testicular spermatozoa from microsurgical testicular sperm extraction (Micro-TESE), possibly because the DNA fragmentation rate is higher in ejaculated spermatozoa than in testicular spermatozoa from Micro-TESE. Evaluation of the efficiency of cryopreservation of three or fewer spermatozoa was conducted retrospectively at St. Mother Clinic. The physical and cognitive development of children born after this method was studied between 2011 and 2018. This study included 28 cryptozoospermic men who had three or fewer morphologically normal and motile spermatozoa in their ejaculate after centrifugation and who preferred using cryopreserved spermatozoa to Micro-TESE. Control subjects were 31 cryptozoospermic patients using fresh spermatozoa from their ejaculates and 20 non-obstructive azoospermic patients with fewer than 10 spermatozoa obtained by TESE and vitrified. Clinical outcomes among three groups, vitrified spermatozoa from the ejaculate, fresh spermatozoa from the ejaculate and vitrified spermatozoa from the testis, were statistically analysed. For the 7-year follow up study of the 14 children born after ICSI using the ejaculated vitrified spermatozoa, the Japanese government-issued Boshi Kenko Techo (Mother-Child Handbook) and Kinder Infant Development Scale (KIDS scale) were used to determine whether their physical and cognitive development was comparable to that of naturally conceived children. Recovery and survival rates were 97.8% (510/521) and 87.1% (444/510) for vitrified spermatozoa from the ejaculate and 92.7% (152/164) and 60.5% (92/152) for vitrified spermatozoa from the testis. Clinical pregnancies (%), miscarriages (%) and live birth rates (%), respectively, among the three groups were as follows: vitrified spermatozoa from the ejaculate: 15(25.0), 2(13.3), 13(21.7); fresh spermatozoa from the ejaculate: 26(24.3), 5(19.2), 20(18.7); and vitrified spermatozoa from the testis: 3(16.7), 0(0.0), 3(16.7). Among the groups, there were no statistically significant differences except for the sperm survival rate and the oocyte fertilisation rate, which were lower for vitrified spermatozoa from the testis compared with vitrified spermatozoa from the ejaculate. The 7-year follow-up study showed that the physical and cognitive development of 14 children born after ICSI using vitrified ejaculated spermatozoa from the ejaculate was comparable to that of naturally conceived children. The maximum number of spermatozoa to which this method can be applied successfully is about 10. When the number of aspirated spermatozoa is over 10, some of them change direction after colliding with each other inside the aspiration pipette and reach the mineral oil, and once this happens, they cannot be expelled out of the pipette. Even though we did not find evidence of DNA fragmentation, further studies with larger participant numbers and longer time periods are necessary. This technique is very useful for the cryopreservation of very small numbers of testicular spermatozoa (fewer than 10) in order to avoid or reduce Micro-TESE interventions. No external funding was received to undertake this study. There are no competing interests. N/A.

Influencia de la procedencia de los espermatozoides (testículo o eyaculado) en ciclos de ICSI de parejas estériles por factor masculino severo

BackgroundIt is estimated that around a 3% of the couples are affected by severe male factor infertility -oligozoospermia, cryptozoospermia and non-obstructive azoospermia-. It is suggested the existence of postesticular sperm damage due to the oxidative stress that sperm suffers through the male genital tract. Base on this, different authors have propose the generalised use of testicular sperm, surgically retrieved (TESE), instead of ejaculated sperm. ObjectivesAnalysing the influence of the sperm source in couples affected by severe male factor infertility in the ICSI results, when DNA ejaculated sperm fragmentation has not been tested before. Material and methodsA retrospective study was conducted based on a specific severe male factor ICSI cycle database, according to the selected sperm source -testicle or ejaculate-. ResultsWe analysed a total amount of 152 ICSI cycles which were divided into two groups, 107 cycles conducted with fresh ejaculated spermatozoa and 45 cycles conducted with criopreserved testicular biopsy spermatozoa. The results showed no significant differences in social-demographical or ovarian stimulation variables. ICSI cycles results were not different either; we obtained similar fertilization rates (54.3% vs. 48.6%), implantation rates (23.1% vs. 22.8%) and pregnancy per embryo transfer rates (35.5% vs. 37.1%). We observed a trend towards greater miscarriage rate between the testicular sperm cycles (18.5% vs. 46.1%), as well as a reduced live birth rate (28.9% vs. 20.0%) in this group. However, no significance differences were found after the analyses. ConclusionNo evidences were found in favour of mass testicular spermatozoa vs. ejaculated spermatozoa use in ICSI cycles, when DNA ejaculated sperm fragmentation has not been tested before.

Does Use of Testicular Sperm Improve Outcomes in Nonazoospermic Couples With Previous IVF Failure Using Ejaculated Sperm? [OP02-2B

INTRODUCTION: Due to controversial evidence that testicular sperm is associated with lower sperm DNA fragmentation (SDF) and improved outcomes compared to ejaculated sperm (ES), this study evaluates intracytoplasmic sperm injection (ICSI) outcomes using testicular sperm in nonazoospermic couples with prior IVF failure using ES. METHODS: From Jan 2015-Aug 2018, nonazoospermic couples with ≥1 prior failed ART cycles using ES underwent testicular sperm extraction (TESE) for IVF/ICSI. Outcomes using TESE sperm were compared to the mean values of couples’ prior cycles using ES. Primary outcomes: clinical pregnancy and live birth rates (CPR &amp; LBR). Secondary outcomes: fertilization and blastocyst conversion. RESULTS: 64 couples were evaluated with an average of 2.5 prior failed ART cycles using ES. There were 76 total transfers including fresh/frozen transfers and supernumerary embryo transfers. Compared to prior failed cycles with ES, TESE sperm resulted equivalent fertilization rates (61.4% vs. 59.0%, p=0.66), but significantly improved blastocyst conversion rates (61.9% vs. 42.9%, p&lt;0.001) and blastocysts available for vitrification (1.6 vs. 0.6, p&lt;0.003). CPR and LBR were improved (42.1% and 35.5%, respectively), but not statistically compared to the prior failed cycles due to regression of the mean. A comparison of TESE-ICSI cycles in couples with ≥2 prior failed ART cycles using ES did not change the findings. CONCLUSION: In nonazoospermic couples with failed ART using ES, ICSI using TESE sperm may improve blastocyst development, number of embryos available for vitrification, CPRs, and LBRs. Testicular sperm may bypass adverse effects of elevated SDF from ES and improve pregnancy outcomes.

Testicular Sperm Extraction Versus Ejaculated Sperm Use for Non-azoospermic Male Factor Infertility [OP02-7B

INTRODUCTION: Testicular sperm extraction (TESE) has traditionally been reserved for couples with azoospermia. However, recent data suggests TESE may be beneficial for men with abnormal semen parameters without azoospermia. The objective of this study was to compare in vitro fertilization (IVF) outcomes between TESE and ejaculated sperm (ES) for non-azoospermic male factor infertility. METHODS: Following IRB exemption, couples undergoing IVF for non-azoospermic male factor infertility with total motile sperm (TMS) &lt;25 million motile/ejaculate from 1/2016 to 1/2019 were included. Men were grouped by whether TESE or ES was utilized for insemination. Couple characteristics and cycle outcomes were analyzed. Wilcoxon rank sum and Fisher exact tests were utilized as appropriate. Values are reported in medians. RESULTS: The analysis included 36 men who utilized ES and 23 men who utilized TESE. The median female age was 32 while the median male age was 35 with ES and 37 with TESE. TMS (6.1 vs 3.9 million motile/ejaculate, P=.46), motility (40% vs 30%, P=.07), and strict morphology (2.5% vs 1.0%, P=.15) were similar between ES and TESE. The total mature oocytes retrieved was similar between the groups but compared to ES, TESE was associated with 20% decrease in fertilization (80.6% vs 60.0%, P=.02) and half the number of blastocyst embryos (4 vs 2 embryos, P=.02). Although not statistically significant, TESE also had &gt;10% decrease in live birth (48.5% vs 36.8%, P=.56). CONCLUSION: TESE is associated with lower fertilization and blastocyst embryos as well as decreased live birth rates compared to ES use for non-azoospermic male factor infertility.

PD25-01 DOES THE USE OF TESTICULAR SPERM IMPROVE OUTCOMES IN NON-AZOOSPERMIC COUPLES WITH PREVIOUS IN-VITRO FERTILIZATION (IVF) FAILURE USING EJACULATED SPERM?

You have accessJournal of UrologyInfertility: Epidemiology & Evaluation II (PD25)1 Apr 2020PD25-01 DOES THE USE OF TESTICULAR SPERM IMPROVE OUTCOMES IN NON-AZOOSPERMIC COUPLES WITH PREVIOUS IN-VITRO FERTILIZATION (IVF) FAILURE USING EJACULATED SPERM? M. Blake Evans, Jessica Marinaro*, Allison Eubanks, Kate Devine, Micah Hill, Alan DeCherney, Russell Hayden, Paul Shin, and Cigdem Tanrikut M. Blake EvansM. Blake Evans More articles by this author , Jessica Marinaro*Jessica Marinaro* More articles by this author , Allison EubanksAllison Eubanks More articles by this author , Kate DevineKate Devine More articles by this author , Micah HillMicah Hill More articles by this author , Alan DeCherneyAlan DeCherney More articles by this author , Russell HaydenRussell Hayden More articles by this author , Paul ShinPaul Shin More articles by this author , and Cigdem TanrikutCigdem Tanrikut More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000882.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The use of testicular sperm for prior in-vitro fertilization (IVF) failure has been shown to have higher clinical pregnancy rates (CPR), live birth rates (LBR), and reduced spontaneous abortion (SAB) rates compared to ejaculated sperm (ES). However, some argue that this treatment is an invasive surgical procedure without clear benefit. This study evaluates the outcomes of IVF-intracytoplasmic sperm injection (ICSI) cycles using testicular sperm in non-azoospermic couples with prior IVF failure(s) using ES. METHODS: From 1/2015 to 10/2019, 116 non-azoospermic couples with ≥1 prior IVF cycle using ES underwent testicular sperm extraction (TESE) for IVF/ICSI. Failed cycles with ES were defined as those not progressing to clinical pregnancy. Outcomes using TESE sperm were compared to the mean values of couples’ prior cycles using ES. Primary outcomes included clinical pregnancy rates (CPR) and live birth rates (LBR). Secondary outcomes included fertilization and blastocyst conversion. RESULTS: Among 116 couples, the average number of prior failed IVF cycles using ES was 2.3 (range: 1-8). A total of 140 IVF/ICSI cycles were performed using TESE sperm (116 fresh TESE sperm, 24 frozen/thawed TESE sperm). Of these, there were 71 fresh blastocyst transfers, 31 frozen blastocyst transfers (FET), and 38 cycles without transfer. There were also 9 FETs using supernumerary embryos, for a total of 111 embryo transfers. Compared to ES, IVF/ICSI using TESE sperm significantly improved blastocyst development (p=0.003), blastocyst conversion (p=0.003) and number of embryos available for vitrification (p=0.003). A subset of men (n= 66) had a sperm DNA fragmentation (SDF) assessment of ES. Mean SDF was 45% (7- 90%). Of these 66 men, 55 (83%) had an SDF >25%. The outcomes for patients with elevated SDF did not differ significantly from the overall cohort. CONCLUSIONS: In non-azoospermic couples with prior failed IVF cycles using ES, IVF/ICSI using TESE sperm may improve embryo development, CPR and LBR. Testicular sperm may avoid the adverse effects of elevated SDF from ES and improve pregnancy outcomes in some patients. Further randomized studies are needed to determine if such a benefit exists. Source of Funding: None © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e538-e538 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information M. Blake Evans More articles by this author Jessica Marinaro* More articles by this author Allison Eubanks More articles by this author Kate Devine More articles by this author Micah Hill More articles by this author Alan DeCherney More articles by this author Russell Hayden More articles by this author Paul Shin More articles by this author Cigdem Tanrikut More articles by this author Expand All Advertisement PDF downloadLoading ...

Combined cues of male competition influence spermatozoal investment in a moth

Abstract Male animals usually raise their sperm allocation after detecting sperm competition risk. To date, only a few studies have investigated the cues used by males to sense and respond to rivals. Yet, it is still largely unknown whether males respond to single or combined cues and whether they can increase their lifetime spermatozoal investment after a perception of rival cue(s). Here we postulate that males increase ejaculation and production of sperm after detecting combined cues from rivals, but such response quickly diminishes after the cues are removed. We exposed newly emerged and virgin focal males of the moth Ephestia kuehniella to various rival cues and then permanently removed the cues. We introduced a virgin female to an exposed focal male and an unexposed focal male, respectively, per day and counted the number of sperm transferred by the focal males in each mating and recovered in their body after death. We demonstrate that males significantly increased their lifetime sperm allocation and production after premating detection of either single (acoustic or chemical) or combined cues (acoustic + chemical or acoustic + chemical + tactile) from their rivals with combined cues (acoustic + chemical + tactile) somewhat strengthening such response in eupyrene production. The number of sperm ejaculated by males significantly decreased over successive matings in their lifetime regardless of whether they were exposed to rival cues, but the decline was significantly faster in rival‐cue‐exposed males than in unexposed ones. This suggests that the increase in spermatogenesis cannot fully compensate for that of sperm expenditure in response to rival cues. We show that 10‐hr premating rival exposure was sufficient to maximize males' response in sperm ejaculation and production. The impact of the rival perception on sperm transfer persisted for most of males' reproductive life, suggesting that the moth males have a long‐term memory of sperm competition risk experienced in the early adulthood. A free Plain Language Summary can be found within the Supporting Information of this article.

Vasovasostomy: kinetics and predictors of patency

To assess the timing of patency and late failure (secondary azoospermia) after vasovasostomy (VV) using standardized kinetics definitions. Retrospective cohort study. University-affiliated hospital. Patients with obstructive azoospermia. Vasovasostomy. Univariate and multivariate logistic regression assessed predictors of patency and late failure. Patency was defined as any sperm return to the ejaculate; and >2 million total motile sperm (TMS) in ejaculate. Late failure after VV was defined as azoospermia; or <2 million TMS in ejaculate. 429 men underwent VV, with median follow up of 242 days. Mean time to patency was 3.25 months versus 5.29 months in the "any sperm" versus ">2 million TMS" groups. Finding sperm intraoperatively during VV significantly improved patency rates in multivariable analysis (odds ratio [OR] 4.22). This association was further boosted when sperm was found bilaterally (OR 6.70). Late failure rate (azoospermia) was 10.6% at mean time of 14.1 months and 23% for <2 million, at mean time of 15.7 months. When assessing predictors of late failure, intraoperative motile sperm bilaterally was a statistically significant protective factor on multivariate analysis (hazard ratio 0.22). Vasovasostomy remains highly efficacious in treating obstructive azoospermia. Young patients, shorter obstructive intervals, and sperm identified intraoperatively predict improved outcomes. Clinicians can expect VV patency in 3 months and late failure within the first 2 years after surgery. However, patency rates, late failure rates, and kinetics vary by definition.

Comparison of the structure of chinchilla sperm isolated from semen and from the tail of the epididymis.

Sperm cells isolated from the tail of the epididymis and from the semen of the same individuals were analysed. The use of silver nitrate to stain sperm cells isolated from the tail of the epididymis made it possible to identify structures that were not visible in the sperm from semen. Silver nitrate very clearly distinguished the acrosomal and distal parts of the sperm head. Following silver nitrate staining, the sperm isolated from the tail of the epididymis were characterized by dark 'collars' in the distal part of the head. These 'collars' are not visible in the sperm cells isolated from semen. The results of the study indicate differences in the dimensions of sperm isolated from the tail of the epididymis and sperm in semen. Sperm isolated from the tail of the epididymis had smaller heads, despite their longer length, and had longer midpieces and tails than ejaculate sperm. Silver nitrate staining is a simple and fast technique. Silver nitrate makes it possible to identify the acrosome and post-acrosomal region of the sperm head and to clearly identify the midpiece. Therefore, it can be successfully used to supplement routine techniques for evaluating sperm morphology or as an independent technique.

Investigation of serum vitamin D and ischaemia-modified albumin levels in infertile Turkish men.

Our aim was to explore the existence of a possible relationship of sperm motility with serum 25-hydroxyvitamin D3 (25-OH VD) levels and with ischaemia-modified albumin (IMA) levels in infertile Turkish men. A total of 30 men with nonobstructive azoospermia (no spermatozoa in ejaculate), 30 men with oligospermia (total progressive motile sperm count (TPMSC) <15×106 /ml) and 33 fertile men with normospermia (with at least one child, as the control group) were enrolled in the study. The mean 25-OH VD levels for groups 1, 2 and 3 were 9.31±6.46, 19.71±12.80 and 30.52±12.49 respectively (p<.05). There was a statistically significant difference in serum IMA levels among the groups (479.32±307.56 vs. 296.37±127.27 vs. 150.04±81.05, respectively; p<.05). A positive correlation between serum 25-OH VD levels and TPMSC, and a negative correlation between TPMSC and serum IMA levels were determined. Infertile men had lower serum 25-OH VD and higher IMA levels than fertile men, with a positive correlation between serum 25-OH VD levels and TPMSC, and a negative correlation between TPMSC and serum IMA levels. Vitamin D supplementation may increase the sperm motility.

Missing Information from the Estrogen Receptor Puzzle: Where Are They Localized in Bull Reproductive Tissues and Spermatozoa?

Estrogens are steroid hormones that affect a wide range of physiological functions. The effect of estrogens on male reproductive tissues and sperm cells through specific receptors is essential for sperm development, maturation, and function. Although estrogen receptors (ERs) have been studied in several mammalian species, including humans, they have not yet been described in bull spermatozoa and reproductive tissues. In this study, we analyzed the presence of all types of ERs (ESR1, ESR2, and GPER1) in bull testicular and epididymal tissues and epididymal and ejaculated spermatozoa, and we characterize them here for the first time. We observed different localizations of each type of ER in the sperm head by immunofluorescent microscopy. Additionally, using a selected polyclonal antibody, we found that each type of ER in bull sperm extracts had two isoforms with different molecular masses. The detailed detection of ERs is a prerequisite not only for understanding the effect of estrogen on all reproductive events but also for further studying the negative effect of environmental estrogens (endocrine disruptors) on processes that lead to fertilization.

Study on the role of calmodulin in sperm function through the enrichment and identification of calmodulin-binding proteins in bovine ejaculated spermatozoa.

Calmodulin is a small, highly conserved acidic protein present at high levels in spermatozoa that mediates numerous intracellular Ca2+ -dependent events. Sperm motility and fertilizing ability results from an array of biochemical pathways under Ca2+ control, in which the importance of calmodulin is not fully understood. The role of calmodulin in sperm function has been mostly assessed using antagonists. Nevertheless, few known calmodulin-regulated enzymes have been described in spermatozoa regarding their involvement in sperm function. To further understand the role of this important Ca2+ mediator in spermatozoa, different studies were also undertaken to investigate and to identify sperm calmodulin-binding proteins and determine their localization and subcellular distribution as an attempt to elucidate the role of this important Ca2+ mediator. In the present study, sperm calmodulin-binding proteins were identified by mass spectrometry after Ca2+ -dependent biotinylated-calmodulin binding on sperm head proteins subjected to 2D electrophoresis and transferred on a polyvinylidene difluoride membrane. Calmodulin binding protein identification was also done on detergent extracted whole sperm proteins pulled down in a Ca2+ -dependent manner by calmodulin-conjugated sepharose beads. In this latter group, 300 proteins were identified in at least two experiments out of three, and those identified in the three independent experiments were analyzed for overrepresented biological processes using the Bos taurus Gene Ontology database. Proteins with known function in reproductive processes, fertilization, sperm-egg recognition, sperm binding to the zona pellucida, regulation of sperm capacitation, and sperm motility were identified and further emphasize the importance of calmodulin in sperm function.

The ABCA1 blocking agent probucol decreases capacitation in ejaculated dog spermatozoa

BackgroundThe ATP binding cassette (ABC) transporters participate in the cholesterol and phospholipid transport within and through cell membranes of many cells including spermatozoa. Cholesterol efflux is important for capacitation of spermatozoa. ABCA1 expression has been assessed in canine spermatozoa previously but its role in capacitation still has to be determined. The aim of the study was to test whether inhibition of ABCA1 (1) decreases capacitation in ejaculated and epididymal canine sperm samples and (2) decreases cholesterol efflux in the same samples. Twenty-one ejaculates and sperm from 22 epididymal tails were collected from healthy dogs. Motility was measured by CASA and viability assessed after staining with SYBR-14/PI. Samples from ejaculated sperm and sperm from epididymal tails were aliquoted. One part was incubated with the ABCA1 inhibitor probucol, the other served as a negative control. In all samples, capacitation was evaluated by chlortetracyclin (CTC) assay and cholesterol was measured by cholesterol efflux assay and colorimetric enzymatic assay.ResultsIn ejaculated sperm, blockade of ABCA1 with 100 µM of probucol/mL of sample resulted in a significantly higher percentage of uncapacitated and acrosome reacted spermatozoa (P < 0.001 and P = 0.031), capacitation was significantly decreased (35% in probucol samples vs 54.2% in controls, P < 0.001). In probucol inhibited sperm samples from epididymal tails, the percentage of capacitated spermatozoa did not differ between groups but the percentage of acrosome reacted spermatozoa increased significantly (P = 0.014). The cholesterol measurement revealed significantly lower cholesterol concentration in the probucol group when compared to the controls (P = 0.035), however only in ejaculated sperm samples.ConclusionsCTC assay and cholesterol measurement revealed significant differences between groups; we conclude that inhibition of ABCA1 significantly decreased capacitation and cholesterol efflux in ejaculated canine spermatozoa. The inhibition was not complete but ABCA1 is supposed to contribute to capacitation in canine ejaculated spermatozoa. ABCA1 is probably not important for capacitation of epididymal spermatozoa but might exert other functions during spermatozoa ripening.

Determinant factors for sperm transfer and sperm storage within Ceratitis capitata (Diptera: Tephritidae) and impact on Sterile Insect Technique

The Sterile Insect Technique (SIT) is a method of pest control used in the management of Ceratitis capitata (Medfly). This technique is based on the mating and transfer of ejaculate sperm to the wi...