Linear Motility Research Articles

Quercetin Protects Goat Sperm Motility by Inhibiting Neutrophil Extracellular Traps and Maintaining Plasma Membrane and Acrosome Integrity

Enhancing or protecting sperm motility has always been a pivotal approach to improving the ewe pregnancy rate. Sperm motility is highly susceptible to the immune status of the reproductive tract. Neutrophil extracellular traps (NETs) have been demonstrated to capture sperm and impair its motility in human, swine, and goat species. Quercetin is a flavonoid derived from Cuscuta Chinensis Lam., which can protect sperm from oxidative damage. In this study, we investigated whether inflammation decreases sperm motility and tried to clarify the potential protective mechanism of quercetin on goat sperm motility. Sperm-triggered NETs were analyzed by immunofluorescence analysis. Sperm acrosome integrity was detected by using giemsa staining. Quercetin exhibited no cytotoxicity towards sperm and PMNs within the concentration range of 20–80 μM. PMNs impaired both the survival rate and rapid linear motility of sperm, while quercetin significantly enhanced these parameters. PMNs captured sperm through NETs composed of DNA, citrullinated histone 3 (citH3), and neutrophil elastase (NE); however, quercetin effectively inhibited the release of sperm-stimulated NETs. The stimulation of PMNs with sperm resulted in a significant increase in levels of ROS and MDA, which decreased by quercetin. Moreover, PMNs caused integrity violation to both the plasma membrane and acrosome in sperm; this effect was significantly alleviated by quercetin. In conclusion, quercetin effectively ameliorated PMN-reduced sperm motility through the inhibition of NETs and oxidative stress, and preserving sperm plasma membrane and acrosome integrity, thereby providing preliminary insights into the underlying mechanisms and theoretical support for the development of potential sperm protectors.

Coordinated linear and rotational movements of endothelial cells compartmentalized by VE-cadherin drive angiogenic sprouting

Angiogenesis is a sequential process to extend new blood vessels from preexisting ones by sprouting and branching. During angiogenesis, endothelial cells (ECs) exhibit inhomogeneous multicellular behaviors referred to as "cell mixing," in which ECs repetitively exchange their relative positions, but the underlying mechanism remains elusive. Here we identified the coordinated linear and rotational movements potentiated by cell-cell contact as drivers of sprouting angiogenesis using invitro and in silico approaches. VE-cadherin confers the coordinated linear motility that facilitated forward sprout elongation, although it is dispensable for rotational movement, which was synchronous without VE-cadherin. Mathematical modeling recapitulated the EC motility in the two-cell state and angiogenic morphogenesis with the effects of VE-cadherin-knockout. Finally, we found that VE-cadherin-dependent EC compartmentalization potentiated branch elongations, and confirmed this by mathematical simulation. Collectively, we propose a way to understand angiogenesis, based on unique EC behavioral properties that are partially dependent on VE-cadherin function.

Glucose Starvation Inhibits Ferroptosis by Activating the LKB1/AMPK Signaling Pathway and Promotes the High Speed Linear Motility of Dairy Goat Sperm.

In mammals, sperm acquire fertilization ability after capacitation in vitro or when in the female reproductive tract. The motility patterns of sperm undergo continuous changes from the moment of ejaculation until fertilization in the female reproductive tract. In vitro, hyperactivated motility can be induced through high glucose mediums, while in vivo, it is induced by oviduct fluids. Conversely, sperm maintain linear motility in seminal plasma or uterine fluids that contain low glucose levels. In dairy goat sperm, energy metabolism associated with capacitation depends on the energy sources in vitro, seminal plasma, or the female reproductive tract, especially the glucose levels. However, there is little experimental knowledge that glucose levels affect sperm energy metabolism in dairy goats. To clarify these hypotheses, we incubated dairy goat spermatozoa with different concentrations of rotenone-glucose (ROT), carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), and tigecycline (TIG) in vitro. Sperm motility attributes, ATP content, pyruvate and lactate levels, mitochondrial permeability transition pore fluorescence intensity, mitochondrial membrane potential (MMP), and protein synthesis were analyzed. Sperm motility patterns changed from circular to linear under low glucose conditions compared with those in high glucose conditions and showed a significant improvement in progressive motility and straight line speed, whereas lactate and pyruvate levels and MMP decreased remarkably. Incubation of spermatozoa with ROT, FCCP, and TIG inhibited sperm mitochondrial activity, protein synthesis, oxidative phosphorylation, and ATP levels, thereby reducing sperm motility, including the progressive motility, straight line speed, and total motility. Simultaneously, incubation of spermatozoa with Compound C under low glucose conditions significantly decreased the ATP levels and MMP, as well as liver kinase B1 and AMPK protein expression. Under low glucose conditions, sperm mainly produce ATP through mitochondrial OXPHOS to achieve high speed linear movement, inhibit ferroptosis through the LKB1/AMPK signaling pathway, and further maintain energy metabolism homeostasis.

P–074 Chromatin Maturity Index (CMI) in unfixed and live spermatozoa and Aniline Blue (AB) stained as an additional evaluation parameter in idiopathic male infertility

Abstract Study question To investigate whether idiopathic male infertility may be due to the presence of histones in motile spermatozoa using a modified AB staining protocol. Summary answer No correlation between CMI in live motile spermatozoa, DNA Fragmentation Index (DFI) and other conventional seminal parameters were found in male infertile patients. What is known already The AB stain discriminates between lysine-rich histones and arginine/cysteine-rich protamines. Transition from histones to protamines during spermatogenesis remodels chromatin packaging and abnormalities in the substitution of those proteins maybe interfere with seminal parameters and affect male infertility. The correlation between CMI and seminal parameters is known, but little is knowledge about live and motile spermatozoa associated to CMI because literature report only spermatozoa fixation before staining. Sperm chromatin carries half of the genomic material to offspring. Spermatozoa nuclear status is crucial for balanced transmission to future generations, and histones modifications are directly involved in epigenetic mutations. Study design, size, duration Retrospective observational study of 77 men underwent to standard semen analysis, including the evaluation of CMI and DFI, enrolled from January to December 2020. Mean age of the men was 36.63±8.26 years old, sperm concentration 46.69±37.23 mill/mL, linear progressive motility 39.35±15.31%, normal morphology 6.42±3.40%, DFI 25.91±10.29%. 200 spermatozoa for evaluation of CMI and 300 for DFI were analyzed respectively. Participants/materials, setting, methods Semen samples of 77 patients were collected and analyzed according to 5th edition of WHO guidelines (2010) for examination of human semen. For the evaluation of CMI we performed a new modified protocol for AB stain directly in live spermatozoa. Dilution 1:1 fresh semen and Aniline Blue colorant were mixed and placed on a slide and examined in bright field microscopy x1000 magnification. DFI was evaluated using Sperm Chromatin Dispersion (SCD) test. Main results and the role of chance Of all spermatozoa analyzed, 82.58±29.98% were white, 17.17±17.21% were pale blue, and 28.53±21.09% were dark blue. By our modified protocol, directly in live spermatozoa, we correlated AB staining with motility and , surprisingly, all motile spermatozoa observed were not stained (white), while pale or dark blue spermatozoa resulted always immotile. For this reason, we have considered pale blue spermatozoa as AB positive, in disagreement with some authors. So, maybe, we should reconsider pale blue stained spermatozoa as abnormal. We also observed AB negative spermatozoa with morphological head, neck and tail defects, underlining the independence of these two parameters: nuclear status and morphology. We have observed no statistically significant differences between conventional semen parameters, DFI and CMI, so nuclear analysis seems to be independent parameters. The statistical analysis was performed by Matlab statistical toolbox version 2008 (MathWorks, Natick, MA, USA) for Windows at 32 bit; finally all tests with p-value (p) < 0.05 were considered significant. Attention should be paid to the evaluation of CMI not only in astenozoospermic patients, where a lower CMI is known, but also in normozoospermic infertile patients. Limitations, reasons for caution This is a preliminary observational study on a small number of normozoospermic or mild asthenozoospermic patients. The study should be considered as a pilot study. Future studies with higher number of samples are necessary in order to confirm the results obtained. Wider implications of the findings: This is the first study that reports AB staining on unfixed live spermatozoa with a modified protocol. Our study underlines the necessity of classify pale blue spermatozoa as AB positive. Further investigations are necessary. This is a starting point for future analysis to be carried out under the project EcoFoodFertility. Trial registration number Not applicable

Proteins involved in mitochondrial metabolic functions and fertilization predominate in stallions with better motility

Even in stallions with sperm quality within normal reference ranges at ejaculation, subtle differences in sperm quality exist that in many cases lead to reduced time frames for conservation of the ejaculate and/or reduced fertility. The spermatozoon is a cell highly suitable for proteomics studies, and the use of this technique is allowing rapid advances in the understanding of sperm biology. The aim of the present study was to investigate differences among stallions of variable sperm quality (based on motility and sperm velocities), although all horses had sperm characteristics within normal ranges. The proteome was studied using UHPLC/MS/MS and posterior bioinformatic and enrichment analysis; data are available via ProteomeXchange with identifier PXD025807. Sperm motility, linear motility and circular, straight line and average velocities (VCL, VSL, VAP) were measured using computer assisted sperm analysis (CASA). In stallions showing better percentages of motility, circular and average velocity predominated mitochondrial proteins with roles in the Citric acid cycle, pyruvate metabolism and oxidative phosphorylation. Interestingly, in stallions with better percentages of total motility, sperm proteins were also enriched in proteins within the gene ontology (G0) terms, single fertilization (G0: 0007338), fertilization (G0: 0009566), and zona pellucida receptor complex (GO:0002199). The enrichment of this proteins in samples with better percentages of total motility may offer a molecular explanation for the link between this parameter and fertility. SignificanceProteomic analysis identified a high degree of specificity of stallion sperm proteins with discriminant power for motility, linear motility, and sperm velocities (VCL, VAP and VSL). These findings may represent an interesting outcome in relation to the molecular biology regulating the movement of the spermatozoa, and the biological meaning of the measurements that computer assisted sperm analysis (CASA) provide. Of a total of 903 proteins identified in stallion spermatozoa, 24 were related to the percentage of total motility in the sample; interestingly, gene ontology (G0) analysis revealed that these proteins were enriched in terms like single fertilization and fertilization, providing a molecular link between motility and fertility. Field studies indicate that the percentage of total motility is the CASA derived parameter with the best correlation with fertility in stallions.

Saturated fatty acids accelerate linear motility through mitochondrial ATP production in bull sperm.

PurposeThe present study was undertaken to clarify whether bovine sperm could take up fatty acids (FAs) and produce ATP to maintain linear motility.MethodsFrozen bovine semen was thawed in media containing either lipid mixture (LM) or FAs, and sperm motility was analyzed. The kinetic changes in FA levels in sperm were detected using gas chromatography‐mass spectrometry. The mitochondrial activity of sperm thawed in media containing LM or FAs was analyzed based on the fluorescence intensity of JC‐1 staining and the oxygen consumption rate. FA transporters were observed using whole‐mounted immunofluorescence.ResultsSperm linear motility was significantly (P < .05) increased after thawing in media with LM and FA. Moreover, saturated fatty acids were predominant in sperm thawed in media with LM. Notably, our study revealed that frozen bovine sperm possessed FA transporters in the midpiece where the fluorescence signals were detected after treatment with fluorescence‐tagged FA. Treatment with FA activated electron transport in mitochondria through β‐oxidation.ConclusionsSperm linear motility is facilitated by FAs in the thawing media used for frozen bovine sperm. This might provide a new approach for upgrading the artificial insemination technique used in both livestock animals and human infertility care.

Development and validation of a novel mail-in semen analysis system and the correlation between one hour and delayed semen analysis testing

To develop and validate a novel, mail-in semen analysis (SA) system. Prospective cohort. Not applicable. Ejaculates from normospermic men. One-hour SA, then repeat SAs (on same ejaculate) over 52 hours using a novel technique for maintaining sperm viability. World Health Organization SA parameters. One-hour SA on 104 ejaculates in the validation phase of the study demonstrated normal semen parameters. With up to 52 hours of observation and four subsequent SA measurements/ejaculate, concentration remained stable, motility decreased by 0.39%/h, and normal morphology decreased by 0.1%/h. Measured 1-hour and calculated motility (correlation coefficients 0.87) and morphology (correlation coefficients 0.82) strongly were correlated. This novel, mail-in, Clinical Laboratory Improvement Amendments-approved SA testing system demonstrates a strong degree of correlation between 1-hour and delayed SA testing. Given the linear motility and morphology decrease and stability of sperm concentration, this test may be used in clinical practice to evaluate semen quality for fertility evaluations. Furthermore, this approach significantly improves the ease, comfort, and efficiency of obtaining a SA, likely breaking down early barriers to accessing successfully a male fertility evaluation.

Retrospective Analysis of Semen Quality Using Sperm Enhancing Technique for Intrauterine Insemination

Objective: To use sperm wash swim up technique as a diagnostic tool in Intra Uterine Insemination. Design: Male semen data was recorded for 181 couples who chose to undergo IUI procedures at the Bangladesh Institute of Research &amp; Rehabilitation in Diabetes, Endocrine &amp; Metabolic Disorders (BIRDEM) Centre for Assisted Reproduction (CARE) from 2013- 2016. Couples were selected for IUI based on initial complaints of primary or secondary subfertility. The female partner had at least patent one tube based on hysterosalphingogram or laparoscopy results. Polycystic ovarian syndrome and minimal to mild endometriosis cases were included. Male factor includes sexual dysfunction and moderate sperm abnormalities. Statistical Analysis: Baseline and stratified analyses were conducted using semen data form. Results: There were 181 IUI cycles done in total. 126 couples (70%) suffered from primary sub fertility while 55 couples (30%) suffered from secondary subfertility. The baseline findings show that morphology of samples is improved by nearly 28%, while rapid linear motility improves by 50% after the first wash and has less variation after the second wash. Concentration, rapid linear motility, and morphology, all have strengthened parameters with each progressive wash process (p&lt; .0001, p&lt; .0001, and p = .0004, respectively). Conclusions: The result of our study provides some interesting observations in the treatment modalities for male infertility. Relatively low cost, less time consuming Swim up sperm wash technique can improve the quality of semen in a great extent. Bangladesh Journal of Urology, Vol. 20, No. 1, January 2017 p.36-41

Itaconate regulates the glycolysis/pentose phosphate pathway transition to maintain boar sperm linear motility by regulating redox homeostasis

Mammalian cells improve redox homeostasis under reactive oxygen species (ROS) stress conditions via the enhancement of the pentose phosphate pathway (PPP). However, it is not clear how the cell reprograms glucose metabolism from glycolysis to the PPP. Hence, in the present study, we used boar sperm as a model to elucidate the mechanism by which the glycolysis/PPP transition occurs under ROS stress. The boar sperm treated with moderate glucose levels for 3 h exhibited increased sperm linear motility patterns, ATP levels and GSH/GSSG ratios and decreased ROS levels compared to the boar sperm treated without glucose. In addition, the hexokinase activity, glucose-6-phosphate dehydrogenase (G6PD) activity, NADPH level, NADPH/NADP+ ratio and mitochondrial activity were higher in the sperm treated with moderate glucose than in those not treated with glucose. Interestingly, the enzyme activity of fructose-1,6-bisphosphate aldolase (ALDOA) was not significantly changed during the incubation. The sperm linear motility patterns were decreased by treatment with the G6PD inhibitor 6-aminonicotinamide. Moreover, moderate glucose treatment significantly increased the itaconate levels in sperm. Both endogenous and exogenous itaconate increased the total itaconate modifications and the itaconate-modified ALDOA levels in sperm, suggesting that under moderate-glucose conditions, glycolysis in the sperm was suppressed by an increase in the itaconate levels. Furthermore, the addition of itaconate improved the sperm linear motility patterns by suppressing glycolysis and enhancing oxidative phosphorylation (OXPHOS). Therefore, the itaconate generated from OXPHOS regulates the glycolysis/PPP transition to maintain redox homeostasis. In sperm, this itaconate-dependent mechanism plays an important role in maintaining their high linear motility.

Effect of low-frequency electric field screening on motility of human sperm.

The human body is constantly exposed to an extremely low electromagnetic field (ELF-EMF), in particular at 50Hz, emitted by power lines, domestic distribution lines, electrical appliances, etc. It is assumed that the increase in electromagnetic exposure may cause adverse effects upon human health, as well as raising concerns regarding the impact on human fertility. The aim of this in vitro study was to investigate the influence of ELF-EMF with a frequency of 50 Hz on the motility of human sperm. At the same time, the effectiveness of the dielectric screen constructed by ADR Technology ® in absorbing the emitted radiation was examined. Semen samples of 20 patients were exposed to the influence of an extremely low electromagnetic field. After 5, 15 and 30 min., spermatozoa motility was analysed using a computer-assisted spermatozoa motility analysis system. The following sperm motility parameters were examined: 1) velocity straight linear motility; 2) cross-beat frequency; 3) lateral head displacement; 4) homogeneity of progressive motility velocity. It was found that the ELF-EMF presented a negative effect on the motility of human spermatozoa. A significant decrease in spermatozoa motility speed and a significant increase in lateral head deviation values were observed under the influence of the electromagnetic field. ELF-EMF did not show an effect on either lateral head displacement or homogeneity of progressive motility velocity. A positive effect of the dielectric screen ADR Technology® was found. This effect compensated spermatozoa motility changes induced with ELF-EMF.

Negative effects of ROS generated during linear sperm motility on gene expression and ATP generation in boar sperm mitochondria

Mitochondrial oxidative phosphorylation (OXPHOS) is essential for ATP production to maintain sperm linear motility during migration from the uterus to the oviduct. However, ROS are generated as by-products of OXPHOS, causing stress and damaging the sperm quality. This study aimed to clarify the ROS targets in sperm mitochondria that decrease linear motility and to investigate whether mitochondria-target antioxidants (PQQ and CoQ10) affect mitochondrial activity and sperm motility. Sperm linear motility pattern, ATP production, and mitochondrial activity were decreased with increasing ROS levels during incubation in the low-glucose medium. However, sperm motility patterns and ROS levels were not significantly changed in the high-glucose medium. Moreover, the gene expression system (mt-DNA, mitochondrial transcription factor-A (TFAM) and RNA polymerase (POLRMT)) in sperm mitochondria was damaged during incubation in the low-glucose medium. Interestingly, PQQ treatment increased the mt-DNA stability and decreased the damage to TFAM and POLRMT, which resulted in high expression of mitochondrial genes. Furthermore, the antioxidants increased mitochondrial activity and maintained sperm linear motility under the low glucose condition. These results revealed that both ATP production and the mitochondrial transcription system are damaged with increasing ROS levels in sperm that show a linear motility pattern. Treatment with antioxidants, such as PQQ and CoQ10, is beneficial tool to maintain sperm linear motility.

Gene Expression and Protein Synthesis in Mitochondria Enhance the Duration of High-Speed Linear Motility in Boar Sperm.

Sperm motility patterns are continuously changed after ejaculation to fertilization in the female tract. Hyperactivated motility is induced with high glucose medium in vitro or the oviduct fluids in vivo, whereas sperm maintain linear motility in the seminal plasma or the uterine fluids containing low glucose. Therefore, it is estimated that sperm motility patterns are dependent on the energy sources, and the mitochondrial oxidative phosphorylation is activated to produce ATP in low glucose condition. To elucidate these hypotheses, boar sperm was incubated in different energy conditions with the transcription and translation inhibitors in vitro. Sperm motility parameters, mitochondrial activity, ATP level, gene expression and protein synthesis were analyzed. Sperm progressive motility and straight-line velocity were significantly increased with decreasing glucose level in the incubation medium. Moreover, the mitochondrial protein turnover meaning transcription and translation from mitochondrial genome in sperm is activated during incubation. Incubation of sperm with mitochondrial translation inhibitor (D-chloramphenicol) suppressed mitochondrial protein synthesis, mitochondrial activity and ATP level in sperm and consequently reduced the linear motility speed, but not the motility. Thus, it is revealed that the mitochondrial central dogma is active in sperm, and the high-speed linear motility is induced in low glucose condition via activating the mitochondrial activity for ATP generation.

Manipulation of bicarbonate concentration in sperm capacitation media improves in vitro fertilisation output in porcine species

BackgroundThe in vivo concentration of bicarbonate (HCO3−), one of the essential sperm capacitating effectors, varies greatly in the different environments sperm go through from cauda epididymis to the fertilisation site. On the contrary, porcine in vitro sperm capacitation and fertilisation media usually contains a standard concentration of 25 mmol/L, and one of the main problems presented is the unacceptable high incidence of polyspermy. This work hypothesised that by modifying the HCO3− concentration of the medium, the output of in vitro sperm capacitation and fertilisation could be increased.ResultsOnce exposed to the capacitation medium, the intracellular pH (pHi) of spermatozoa increased immediately even at low concentrations of HCO3−, but only extracellular concentrations of and above 15 mmol/L increased the substrates protein kinase A phosphorylation (pPKAs). Although with a significant delay, 15 mmol/L of HCO3− stimulated sperm linear motility and increased other late events in capacitation such as tyrosine phosphorylation (Tyr-P) to levels similar to those obtained with 25 mmol/L. This information allowed the establishment of a new in vitro fertilisation (IVF) system based on the optimization of HCO3− concentration to 15 mmol/L, which led to a 25.3% increment of the viable zygotes (8.6% in the standard system vs. 33.9%).ConclusionsOptimising HCO3− concentrations allows for establishing an IVF method that significantly reduced porcine polyspermy and increased the production of viable zygotes. A concentration of 15 mmol/L of HCO3− in the medium is sufficient to trigger the in vitro sperm capacitation and increase the fertilisation efficiency in porcine.

Fresh and post thaw quality characteristics of Holstein Friesian bull semen maintained at semen production unit Quetta Baluchistan, Pakistan

Abstract The study was conducted to determine the quality characteristics of fresh and post-thaw semen of Holstein Friesian bull, maintained at Semen Production Unit, Quetta Baluchistan, Pakistan. The semen were evaluated for volume, pH, mass activity, sperm concentration, sperm motility and integrity of sperm cell membrane. The semen qualifying these criteria were processed for freezing. The frozen semen after thawing was assessed for progressive linear motility and intact cell membrane. The mean ejaculate volume of semen was recorded as 6.93, 6.59 and 6.61 ml, pH; 6.58, 6.59 and 6.61, mass activity of sperms 3.87, 3.75 and 3.5 and sperm concentration 1349.12x106, 1509.37x106 and 1364.37x106 /ml during the month of May, June and July respectively. The color of semen was not consistent pattern, it varied as creamy white and milky white. The motility of fresh and post thaw semen was assessed as 73.9 vs 48.75%; 75 vs. 53.75%; 75 vs. 53.12% for the month of May, June and July respectively. The membrane integrity of fresh semen samples over a period of three month were 58.37% against post-thaw membrane integrity of 44.79%. It was concluded that the semen of Holstein Friesian bull tolerates freezing and thawing stress and maintained reasonable good fertility. The summer season has no significant adverse effect on quality of semen. Keywords: Bull; Fresh; Holstein Friesian; Post-thaw; Quality; Semen http://dx.doi.org/10.19045/bspab.2019.80020

The Effect of Suboptimal Semen Parameters on Male Partner's Ability to Conceive. Is He Really Subfertile Because the Test Says So?

Context:In many developing countries, subfertility treatment is not covered by government-funded institutions. It is observed that healthcare providers incriminate male factor for subfertility even when only a slight deviation from presumed normal criteria is observed.Aim:This study aims to provide scientific evidence that pregnancies are possible at semen parameters that are below the generally accepted lower limits of normal.Setting and Design:This was a retrospective cohort study conducted from January 2014 to December 2018.Materials and Methods:During the study period, couples who conceived without any treatment of male partner were included. The World Health Organization (WHO) reference values for semen analysis were utilized to assess the reports. The primary outcome measure was conception despite abnormal semen parameters.Statistical Analysis Used:Data were analyzed using the SPSS software program, version 15.0 (IBM, Armonk, USA).Results:Of the 332 couples included, 233 (70.1%) couples conceived despite suboptimal semen parameters, The most common criterion not satisfied was rapid linear motility –200 (85.8%), 87 (37.3%) men were oligozoospermic, 94 (40.3%) were asthenozoospermic, and 21 (9%) were teratozoospermic. The abnormalities were more common in men having primary subfertility (71.7% vs. 28.3%, P = 0.002). The abnormalities were most common in the age group 40–44 years (n = 91, 39.1%) and those who were overweight (n = 110, 47%).Conclusions:A consensus for defining poor semen criteria is the need of the hour so that these males can be counseled satisfactorily. WHO criteria are a standard commonly employed, but they do not necessarily predict the fertility potential.

Semen Proile of Infertile Males Attending Private Infertility Clinic of Metropolitan City of Pakistan

Objective: To determine the semen profile of male partner of infertile couples coming for treatment of infertility in a private infertility clinic of metropolitan city of Pakistan. Methods: A retrospective study was conducted in Australian concept infertility medical center, Karachi from January 2017 to August 2017. Data of married males unable to conceive after 12 months of regular unprotected intercourse were retrieved from the medical record. The semen analysis was the baseline investigation that was carried out to analyze the different sperm abnormalities. WHO criteria published in 2010 was used to assess the motility and morphology of the sperms. Results: A total of 120 couples were screened to _ind out the reason behind infertility. Amongst these, 61 (50.83%) males were found to have infertility. Oligoasthenospermia was found higher, i.e. 26 (42.6%). Total sperm count was found less than the normal values in 70.50% men, live count in 98.40% men, rapid linear motility in 95.10% men and normal morphology in 60.70% men. A signi_icant association of total sperm count was found with type of infertility (p-value 0.002) and male infertility type (p-value 0.032). Conclusion: Oligoteratozoospermia was observed as predominant cause of infertility among males. Semen analysis could be used as a baseline investigation to exclude male causes before going for extensive and invasive investigation of females.

Evaluation of the impact of marijuana use on semen quality: a prospective analysis from the university of washington

The use of marijuana has been increasing among U.S. men of reproductive age and its recreational consumption was legalized in Washington State (WS) on November 12th, 2012. Marijuana and its active metabolite tetrahydrocannabiol (THC) can alter the signaling system within spermatozoa which may result in negative effects on spermatogenesis and male fertility. We aimed to characterized differences in semen quality among men with variable consumption of marijuana. Prospective cohort study. We prospectively evaluated semen analyses (SA) from men who presented for infertility evaluation at a single male fertility laboratory. Semen analyses were performed from July 2017 to April 2018. All participants completed a reproductive health questionnaire which included specific queries on past and present marijuana consumption. Questionnaire data included age, ethnicity, marijuana use (≤1 or >1 time per week), duration (months), and tobacco smoking. SA was performed in accordance with World Health Organization (WHO) 2010 criteria. SA parameters included volume (mL), concentration (million/mL), motility (%), progressive motility (%), rapid and linear motility (%) and strict normal morphology (%). A total of 409 patients underwent SA and completed the questionnaire of which 174 (43%) men reported marijuana use (ever users.) Among the ever users, current and past users compromised 71 (18%) and 103 (25%) respectively. Compared to non-user, semen quality was significantly decreased in volume, concentration, morphology, total motile count (TMC) and total progressive motile count (TPMC) compared to never users (Table 1). In multivariate logistic regression analyses, controlling for age, marijuana use was associated with increased odds of abnormal morphology (OR 2.28 (95% confidence interval (CI): 1.52-3.43)), volume (OR 2.02 (1.04-3.94)), and TPMC (OR 1.8 (1.05-3.06)). There was no significant association between marijuana use and sperm concentration, motility or progressive motility as solitary parameters. Marijuana use is common among men presenting for fertility evaluation in our cohort and may have a detrimental effect on semen quality, particularly morphology, volume, and TPMC. Given these findings, large, prospective studies of both semen quality and fertility in this growing, at-risk population are warranted.Table 1Percentages of normal and abnormal semen analysis [WHO 2010] among three groupsParametersGroupNormal Semen Analysis N (%)Abnormal Semen Analysis N (%)P valueVolumeNever216 (92%)19 (8%)0.044Past93 (90%)10 (10%)Current58 (82%)13 (18%)ConcentrationNever209 (89%)26 (11%)0.086Past87 (85%)16 (15%)Current56 (79%)15 (21%)% MotilityNever145 (62%)90 (38%)0.011Past75 (73%)28 (27%)Current56 (79%)15 (21%)% Progressive motilityNever162 (69%)73 (31%)0.513Past73 (71%)30 (29%)Current54 (77%)17 (23%)% Strict morphologyNever156 (67%)79 (33%)0.000Past48 (47%)55 (53%)Current35 (49%)36 (51%)Total motile countNever199 (85%)36 (15%)0.608Past84 (82%)19 (18%)Current57 (80%)14 (20%)Total progressive motile countNever204 (87%)31 (13%)0.111Past83 (81%)20 (19%)Current55 (76%)16 (23%) Open table in a new tab

Effect of Extender Supplementation with Low-molecular-weight Antioxidants on Selected Quality Parameters of Cryopreserved Canine Spermatozoa.

Introduction The addition of low-molecular-weight antioxidants during the freezing process improves post-thaw sperm quality. The high antioxidant potential of cryopreserved semen could have a positive effect on the motility, viability, and energy status of sperm cells and their ability to bind to the zona pellucida of oocytes. The aim of the study was to determine the effects of different concentrations and combinations of vitamins E and C in a semen extender on selected quality parameters of frozen-thawed canine spermatozoa.Material and Methods The experimental material was the semen of four mixed-breed dogs. Sperm viability (motility, plasma membrane integrity, and mitochondrial function) was examined at 0, 60, and 120 min in semen samples supplemented with the extender and in the controls.Results Combined supplementation with vitamins C + E at a concentration of 200 + 200 μM /1 × 109 spermatozoa had the most profound effect on total sperm motility, linear motility, and the percentage of spermatozoa with intact plasma membrane and active mitochondria.Conclusion The synergistic activity of vitamins E and C had a more beneficial influence on the quality of frozen–thawed sperm than these non-enzymatic antioxidants applied separately.

Impact of Seminal Plasma Zinc and Serum Zinc Level on Semen Parameter of Fertile and Infertile Males

Background: Despite Zinc (Zn) deficiency being prevalent in humans, less emphasis has been given on the understanding of its impact on male reproduction. Spermatogenesis has a strict requirement for zinc. The relationship of seminal plasma zinc level and semen parameter until now is controversial.Objective : The study was done to find out the impact of seminal plasma zinc and serum zinc level on semen parameter of fertile and infertile males.Subjects and methods: The study was done in Center for Assisted Reproduction, a tertiary Infertility center in Dhaka and in the Biochemistry Department of Bangabandhu Sheik Mujib Medical University (BSMMU), Dhaka. Sixteen fertile males were taken as control and sixty nine infertile males were taken as cases. Semen analysis was done according to WHO criteria (2004). Serum zinc and seminal plasma zinc levels were measured in the Biochemistry Department of BSMMU by Graphite Furnace Atomic Absorption Spectrophotometer.Result: Seminal parameters between fertile and infertile men showed significantly high sperm count, sperm motility, rapid linear motility, and morphology in fertile group. In fertile men, serum zinc level was lower than the infertile group, whereas the seminal plasma zinc level was higher in the fertile than the infertile group which was not statistically significant. In fertile men except for serum zinc, all parameters showed positive relationship; and in infertile men, except sperm morphology, all other parameters showed negative relationship. Seminal plasma zinc shows positive correlation with all semen parameters in fertile group and negative correlation in infertile group except for sperm morphology. There is negative correlation of serum zinc and seminal zinc-in both groups. Regarding other parameters, semen zinc shows positive correlation with all parameter except sperm morphology in fertile men and negative correlation with all parameter except sperm count in infertile men.Conclusion: Seminal zinc levels in fertile men are higher than those in the infertile patients although the serum zinc level is lower. Zinc levels in seminal plasma has a direct relationship with semen parameters. Zinc deficiency may be an important risk factor for low semen parameters and idiopathic male infertility.J Bangladesh Coll Phys Surg 2017; 35(1): 15-19

Dietary polyunsaturated fatty acid supplementation of young post-pubertal dairy bulls alters the fatty acid composition of seminal plasma and spermatozoa but has no effect on semen volume or sperm quality

The aim of this study was to examine the effects of dietary supplementation with rumen protected n-6 or n-3 polyunsaturated fatty acids (PUFA) on the quantity and quality of semen from young post-pubertal dairy bulls. Pubertal Holstein-Friesian (n = 43) and Jersey (n = 7) bulls with a mean ± s.e.m. age and bodyweight of 420.1 ± 5.86 days and 382 ± 8.94 kg, respectively, were blocked on breed, weight, age and semen quality (based on the outcomes of two pre-trial ejaculates) and randomly assigned to one of three treatments: (i) a non-supplemented control (CTL, n = 15), (ii) rumen-protected safflower (SO, n = 15), (iii) rumen-protected n-3 PUFA-enriched fish oil (FO, n = 20). Bulls were fed their respective diets, ad libitum for 12 weeks; individual intakes were recorded using an electronic feeding system for the initial 6 weeks of the feeding period. Semen was collected via electro-ejaculation at weeks −2, −1, 0, 7, 10, 11 and 12 relative to the beginning of the trial period (week 0). On collection, semen volume, sperm concentration and progressive linear motility (PLM) were assessed. On weeks −2, −1, 0, 10, 11, 12, semen was packaged into 0.25 mL straws and frozen using a programmable freezer. On weeks −1, 7 and 11; a sub-sample of semen was separated into sperm and seminal plasma, by centrifugation and stored at – 20 °C until analysis of lipid composition. Semen from 10 bulls per treatment were used for post-thaw analysis at weeks 10, 11 and 12 (3 straws per ejaculate). Sperm motility was analysed by computer assisted semen analysis (CASA). In addition, membrane fluidity, acrosome reaction and oxidative stress were assessed using flow cytometry. Sperm from bulls fed SO had a 1.2 fold higher total n-6 PUFA content at week 11 compared to week −1 (P < 0.01) while bulls fed FO had a 1.3 fold higher total n-3 PUFA content, in sperm by week 11 (P < 0.01). There was no effect of diet on semen volume, concentration or PLM of sperm when assessed either immediately following collection or post-thawing. Membrane fluidity and oxidative stress of sperm were also not affected by diet. The percentage of sperm with intact-acrosomes was lower in CTL bulls compared to those fed SO (P < 0.01). In conclusion, while the lipid composition of semen was altered following dietary supplementation with either n-6 or n-3 based PUFA, this did not lead to measurable improvements in the quantity or quality of semen produced by young post-pubertal dairy bulls.