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OBJECTIVE: To demonstrate nanoscale motion tracing of spermatozoa and present analysis of the motion traces to characterize the consistency of motion of spermatozoa as a complement to progressive motility analysis. DESIGN: Anonymized sperm samples were videographed under a quantitative phase microscope, followed by generating and analyzing superresolution motion traces of individual spermatozoa. SETTING: Not applicable. PATIENT(S): Centrifuged human sperm samples. INTERVENTION(S): Not applicable. MAIN OUTCOME MEASURE(S): Precision of motion trace of individual sperms, presence of a helical pattern in the motion trace, mean and standard deviations of helical periods and radii of sperm motion traces, speed of progression. RESULT(S): Spatially sensitive quantitative phase imaging with a superresolution computational technique MUltiple SIgnal Classification ALgorithm allowed achieving motion precision of 340 nm using ×10, 0.25 numerical aperture lens whereas the diffraction-limited resolution at this setting was 1,320 nm. The motion traces thus derived facilitated new kinematic features of sperm, namely the statistics of helix period and radii per sperm. Through the analysis, 47 sperms with a speed >25 µm/s were randomly selected from the same healthy donor semen sample, it is seen that the kinematic features did not correlate with the speed of the sperms. In addition, it is noted that spermatozoa may experience changes in the periodicity and radius of the helical path over time. Further, some very fast sperms (e.g., >70 µm/s) may demonstrate irregular motion and need further investigation. Presented computational analysis can be used directly for sperm samples from both fertility patients with normal and abnormal sperm cell conditions. We note that MUltiple SIgnal Classification ALgorithm is an image analysis technique that may vaguely fall under the machine learning category, but the conventional metrics for reporting found in Enhancing the QUAlity and Transparency Of health Research network do not apply. Alternative suitable metrics are reported, and bias is avoided through random selection of regions for analysis. Detailed methods are included for reproducibility. CONCLUSION(S): Kinematic features derived from nanoscale motion traces of spermatozoa contain information complementary to the speed of the sperms, allowing further distinction among the progressively motile sperms. Some highly progressive spermatozoa may have irregular motion patterns, and whether irregularity of motion indicates poor quality regarding artificial insemination needs further investigation. The presented technique can be generalized for sperm analysis for a variety of fertility conditions.

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Purpose: This study aimed to evaluate the effects of swim-up and density gradient centrifugation methods on sperm DNA fragmentation. Methods: Nineteen normozoospermic patient samples with ≥100 × 106 motile sperms were included in this study. Sperm DNA fragmentation, progressive motility, and progressive motile sperm number were measured before and after the swim-up method or density gradient centrifugation. Results: Sperm DNA fragmentation was not statistically different between swim-up-(14.4 ± 2.1%, p = 0.32) and density gradient centrifugation-processed (25.0 ± 3.0%, p = 0.20) and unprocessed semen samples (19.2 ± 1.9%). Sperm DNA fragmentation was significantly lower in swim-up-than in density gradient centrifugation-processed samples (p < 0.05). Sperm progressive motility was significantly higher (p < 0.05) in swim-up-(92.9 ± 1.0%) and density gradient centrifugation-processed (81.3 ± 2.0%) samples, with the former being higher, than in unprocessed semen samples (53.1 ± 3.7%). The recovery rate of progressive motile sperms was significantly lower in swim-up-(9.7 ± 1.4%) than in density gradient centrifugation-processed samples (17.2 ± 1.8%, p < 0.05). Conclusions: The swim-up method is superior to density gradient centrifugation, evidenced by less sperm DNA fragmentation and higher sperm progressive motility. The recovery rate of progressive motile sperms was better after density gradient centrifugation than after swim-up.

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Boar seminal plasma (SP) proteins were associated with differences on sperm resistance to cooling at 17°C. However, information about seminal plasma proteins in boars classified by capacity of semen preservation and in vivo fertility remains lacking. Thus, the objective was to evaluate the SP proteome in boars classified by capacity of semen preservation and putative biomarkers for fertility. The ejaculates from high-preservation (HP) showed higher progressive motility during all 5 days than the low-preservation (LP) boars. There was no difference for farrowing rate between ejaculates from LP (89.7%) and HP boars (88.4%). The LP boars presented lower total piglets born (14.0 ± 0.2) than HP (14.8 ± 0.2; p < 0.01). A total of 257 proteins were identified, where 184 were present in both classes of boar, and 41 and 32 were identified only in LP and HP boars, respectively. Nine proteins were differently expressed: five were more abundant in HP (SPMI, ZPBP1, FN1, HPX, and C3) and four in LP boars (B2M, COL1A1, NKX3-2, and MPZL1). The HP boars had an increased abundance of SP proteins related to sperm resistance and fecundation process which explains the better TPB. LP boars had a higher abundance of SP proteins associated with impaired spermatogenesis.

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BACKGROUND: Accumulating evidence has linked exposure to ambient air pollution to a reduction in semen quality; however, the exposure-response associations are yet to be synthesized. OBJECTIVE: To summarize the exposure-response associations between air pollution and semen quality. METHODS: We systematically searched PubMed, Embase, and Web of Science for relevant studies published before April 20, 2022. Studies investigating the exposure-response association of PM2.5, PM10, SO2, NO2, CO, and O3 with semen quality written in English were included. Semen quality parameters included semen volume, sperm concentration, total sperm number, total motility, progressive motility, and normal forms. Random-effects and fixed-effects models were performed to synthesize associations in the meta-analysis. RESULTS: The search returned 850 studies, 11 of which were eligible for meta-analysis. Each 10 µg/m3 increase of exposure to PM10 and SO2 was respectively associated with a 2.18 % (95 % confidence interval [CI]: 0.10 %-4.21 %) and 8.61 % (1.00 %-15.63 %) reduction in sperm concentration, and a 2.76 % (0.10 %-5.35 %) and 9.52 % (5.82 %-13.93 %) reduction in total sperm number. Each 10 µg/m3 increase of exposure to PM2.5 and PM10 was respectively associated with a 1.06 % (95 % CI: 0.31 %-1.82 %) and 0.75 % (0.43 %-1.08 %) reduction in total motility, and a 0.55 % (0.09 %-1.01 %) and 0.31 % (0.06 %-0.56 %) reduction in progressive motility. No association was observed for PM2.5 or PM10 with semen volume; PM2.5, NO2, CO, or O3 with sperm concentration or total sperm number; and gaseous air pollutants with total or progressive motility. The association between air pollution and normal forms was not summarized due to insufficient number of studies. No significant publication bias was detected. CONCLUSIONS: Exposure to ambient PM2.5, PM10, and SO2 was inversely associated with sperm concentration, total sperm number, total motility, and/or progressive motility. Our findings add to the evidence that air pollution may lead to adverse effects on male reproductive system and suggest that reducing exposure to air pollution may help maintain better semen quality.

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Previous studies have indicated that outdoor air pollution has a negative impact on semen quality; however, few studies have examined whether living in a recently renovated residence is one of the factors influencing semen parameters. We aimed to examine the association between household renovation and semen parameters among infertile men. Our study was conducted at the Reproductive Medicine Center, The First Hospital of Jilin University (Changchun, China) from July 2018 to April 2020. A total of 2267 participants were enrolled in the study. The participants completed the questionnaire and provided a semen sample. Univariate and multiple logistic regression models were used to estimate the association between household renovations and semen parameters. Of the participants, about one-fifth (n = 523, 23.1%) had undergone renovations in the last 24 months. The median progressive motility was 34.50%. There was a significant difference between participants whose residences had been renovated in the last 24 months and those whose residences had not been recently renovated (z = -2.114, p = .035). Compared with participants whose residences were not recently renovated, participants who moved into the residence within 3 months after renovation had a higher risk of abnormal progressive motility after adjusting for age and abstinence time (odds ratio [OR] = 1.537, 95% confidence interval [CI]: 1.088-2.172). Our findings indicated that progressive motility was significantly associated with household renovations.

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Objective To investigate the effects of high concentration of semen samples in detection by fully automated sperm quality analyzer for sperm concentration,progressive motility(PR),non-progressive motility(NP)and percentage of immotile sperm(IM).Methods The semen samples from the patients who visited the reproductive clinic in our hospital from February 2023 to May 2023 were analyzed using the fully automated sperm quality analyzer.A total of 155 semen samples with initial sperm concentration ≥100× 106/mL and semen volume ≥1.5 mL were set as the high concentration semen group(group A).The semen samples with self seminal plasma dilution group(group B)and 37 ℃ normal saline dilution group(group C)were prepared with their own seminal plasma and normal saline at 1∶2 dilution ratio respectively.The sperm concentration of group A was detected by the manual method as the control.The sperm concentration and motility parameters of each group were detected.The differences of sperm concentration and the values of PR,NP and IM among the group A,B and C were analyzed.The correlations between sperm concentration and PR,NP and IM were also assessed.Results The sperm concentration of group A was significantly lower than that of the control group and groups B and C(all P＜0.05).There was no statistically significant difference for the results between group B and group C(P＞0.05).The PR in group C was significantly lower than that in groups A and B(all P＜0.05),and there was no significant difference of PR between group A and group B(P＞0.05).The values of NP in group A was significantly higher than that in groups B and C(all P＜0.05),and there was no significant difference in NP between group B and group C(P＞0.05).The IM in group A was significantly lower than that in groups B and C(all P＜0.05),and there was no statistical difference of IM values between group B ang group C(P＞0.05).Conclusion The direct analysis of undiluted semen samples with high sperm concentration may lead to lower results of sperm concentration,increased percentage of NP sperm and decreased percentage of IM sperm.The dilution of semen samples may improve the accuracy of the detec-tion results.The dilution with 37 ℃ normal saline can lead to decrease of the percentage of PR sperm.The self seminal plasma should be recommended as the first choice of diluent for the semen samples with high sperm concentration.

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Objective: This proof-of-principle aims to develop an index to aid the differential diagnosis of disorders affecting testicular and/or epididymis. A total of 202 subject data were evaluated in two groups: fertile men with children naturally conceived within 1 year of unprotected intercourse (n = 36) and infertile men (n = 166) who had attempted a pregnancy more than 1 year with unprotected intercourse. Materials and methods: Semen parameters (sperm count, vitality, motility, morphology, and hypoosmotic swelling test [HOST]) were evaluated. The index was calculated by dividing the percentage HOST by the percentage of sperm progressive motility in the fertile group (n = 36). Results: A normal range from 1.23 to 1.53 was determined. Using this index, the outcomes of semen analysis from infertile men were grouped in three study groups: below 1.23 (n = 24), normal (n = 44), and higher than 1.53 (n = 98). These parameters were significantly decreased in semen with normal range (p < 0.01) and in indexes higher than 1.53 (p < 0.0001). Receiver operating characteristic curves compared progressive motility and morphology in infertile men with indexes higher than 1.53 shows that semen samples with normal sperm progressive motility and morphology did not suggest dysfunctions in testis and epididymis. Semen samples with asthenozoospermia suggested epididymal dysfunction (area under the curve [AUC] 0.889, confidence interval [CI] 0.783-1), whereas semen samples with teratoasthenozoospermia suggested dysfunction in both testicles and epididymis (AUC 0.891, CI 0.77-1). Conclusions: The current index proof-of-principle of the success of such a strategy provides valuable information about whether a disorder individually affects testicular and/or epididymal function.

Objetivo: Esta prueba de principio tiene como objetivo desarrollar un índice que ayude al diagnóstico diferencial de los trastornos testiculares y/o epidídimales. Métodos: Se evaluaron 202 individuos divididos en dos grupos: hombres fértiles con hijos concebidos de forma natural en el plazo no mayor a un año (n = 36) y hombres infértiles (n = 166), los cuales habían intentado un embarazo por más de un año. Se evaluaron los parámetros seminales (concentración, viabilidad, movilidad, morfología y prueba de hinchazón hipoosmótica [HOST]). El índice se calculó dividiendo el porcentaje de HOST por el porcentaje de movilidad espermática progresiva en el grupo fértil (n = 36). Resultados: Se determinó un rango normal de 1,23 a 1,53. Utilizando este índice, los resultados del análisis del semen de los hombres infértiles se agruparon en tres grupos de estudio: por debajo de 1,23 (n = 24), normal (n = 44) y superior a 1,53 (n = 98). En contraste, estos parámetros disminuyeron significativamente en el semen de rango normal (p < 0,01) y en los índices superiores a 1,53 (p < 0,0001). Las curvas ROC comparadas con la movilidad espermática progresiva y la morfología en los hombres infértiles con índices superiores a 1,53 muestran que las muestras de semen con movilidad progresiva y morfología normales no sugieren disfunciones en los testículos y epidídimos. Las muestras de semen con astenozoospermia sugerirían una disfunción del epidídimo (AUC 0,889, IC 0,783-1), mientras que las muestra de semen que presentaban teratoastenozoospermia sugerirían una disfunción tanto en los testículos como en el epidídimo (AUC 0,891, IC 0,77-1).

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Testis-specifically expressed genes are important for male reproduction according to their unique expression patterns. However, the functions of most of these genes in reproduction are unclear. Here, we showed that mouse 4930590J08Rik was a testis-specifically expressed gene. 4930590J08Rik knockout mice exhibited a delay in the first wave of spermatogenesis and a reduction of cauda epididymal sperm. Furthermore, knockout spermatozoa exhibited defective acrosome reactions and decreased progressive motility, which led to impaired in vivo fertilization. Transcriptome analysis of testes revealed that most of the differentially expressed genes in knockout testes were associated with metabolic processes. 4930590J08Rik knockout sperm exhibited oxidative phosphorylation deficiency and were highly dependent on increased anaerobic glycolysis to compensate for ATP demands. Taken together, the 4930590J08Rik-disrupted mouse partially mimics the phenotypes of human asthenospermia and oligozoospermia, which provides a new model for further understanding the pathogenesis of idiopathic male infertility.

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Male fertility, as manifest by the quantity and progressive motility of spermatozoa, is negatively impacted by obesity, dyslipidaemia and metabolic disease. However, the relative distribution of lipids in spermatozoa and the two compartments which supply lipids for spermatogenesis (seminal fluid and blood serum) has not been studied. We hypothesised that altered availability of lipids in blood serum and seminal fluid may affect the lipid composition and progressive motility of sperm. 60 men of age 35 years (median (range 20-45) and BMI 30.4 kg/m2 (24-36.5) under preliminary investigation for subfertility were recruited at an NHS clinic. Men provided samples of serum and semen, subject to strict acceptance criteria, for analysis of spermatozoa count and motility. Blood serum (n = 60), spermatozoa (n = 26) and seminal fluid (n = 60) were frozen for batch lipidomics analysis. Spermatozoa and seminal fluid had comparable lipid composition but showed marked differences with the serum lipidome. Spermatozoa demonstrated high abundance of ceramides, very-long-chain fatty acids (C20-22), and certain phospholipids (sphingomyelins, plasmalogens, phosphatidylethanolamines) with low abundance of phosphatidylcholines, cholesterol and triglycerides. Men with spermatozoa of low progressive motility had evidence of fewer concentration gradients for many lipid species between blood serum and spermatozoa compartments. Spermatozoa are abundant in multiple lipid species which are likely to contribute to key cellular functions. Lipid metabolism shows reduced regulation between compartments in men with spermatozoa with reduced progressive motility.

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OBJECTIVE: To elucidate the mechanism underlying how Ureaplasma urealyticum (UU) affects sperm quality and identify a therapeutic target. METHODS: In this prospective observational study, the differences in and relationships among semen volume, pH, viscosity, liquefaction time, sperm concentration, sperm motility [progressive motility (PR)], and seminal polymorphonuclear (PMN) elastase were analyzed in 198 normal semen samples (control group) and 198 UU-infected semen samples (observation group). The UU-infected samples were treated and the above parameters were compared between the two groups. RESULTS: The semen volume, viscosity, liquefaction time, and seminal PMN elastase were significantly higher in the observation than control group, but the pH and PR were significantly lower. In the observation group, the pH and PR were significantly higher after than before treatment, whereas the semen volume, PMN elastase, viscosity, and liquefaction time were lower. UU was closely related to semen volume, pH, viscosity, liquefaction time, sperm motility (PR), and PMN elastase. PMN elastase had significant negative effects on semen pH and sperm motility (PR) but positive effects on viscosity and liquefaction time. CONCLUSION: UU might induce PMN elastase to increase the liquefaction time and viscosity of semen, eventually decreasing PR. PMN elastase might be a therapeutic target of UU.

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Sperm motility and viability of cryopreserved semen vary between boars and straws, which influences the outcomes of in vitro embryo production (IVEP). However, progressive motility is usually not considered during IVEP. The aim of this study was to assess fertilization with a 500:1 and 250:1 'progressively motile sperm to oocyte' ratio on IVEP outcomes using semen from three Duroc and three Landrace boars. Frozen-thawed sperm was centrifuged through a 45/90% Percoll® density gradient and sperm quality parameters were assessed. In vitro matured oocytes were fertilized at the two ratios, a portion was stained 10-12 h after start of fertilization to analyze fertilization and polyspermy, while the remaining zygotes were cultured up to day 7. The 500:1 ratio resulted in a higher fertilization and blastocyst yield on day 6 compared with the 250:1 ratio, but no effect of ratio was observed for polyspermy, cleavage rate or blastocyst cell number. Individual differences between boars were observed for fertilization, cleavage and blastocyst rates, but not for the other IVEP outcomes. In conclusion, a higher fertilization and blastocyst yield was obtained with the 500:1 ratio compared with the 250:1 ratio, while polyspermy level was consistent across ratios. Differences in IVEP outcomes were still observed between the individual boars although adjusted for progressive motility. Promising blastocyst yields and high total blastocyst cell counts were obtained with sperm from both breeds.

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Objective: To summarize the currently available phase I and II clinical trials of the effects of nonoxynol-9 (N-9) on human sperm structure and functions. Methods: A systematic review and meta-analysis aiming to evaluate the spermicidal activity of N-9 on motility, was conducted in PubMed, EMBASE, and Cochrane databases by 10 March 2021. The counted numbers of progressive motile (PR) sperm in cervical mucus and the vanguard sperm penetration distances were analyzed. Other effects on sperm structures and physiological activities were reviewed as well. Results: In the pooled results, percentages or counted numbers of PR sperm decreased after the treatment of N-9. Vanguard sperm penetration distance was shortened in treated groups. N-9 has been confirmed to damage the structures of sperm, as well as other organelles like acrosome and mitochondria. The physiological activities such as generation of reactive oxygen species, superoxide dismutase activity, acrosin activity, and hemizona binding were all inhibited in the reviewed studies. Conclusions: N-9 has several impacts on sperm owing to its potency in reducing sperm motility and cervical mucus penetration, as well as other functional competencies. Lay summary: Nonoxynol-9 (N-9) has been used worldwide as a spermicide to kill sperm for more than 60 years but can cause side effects including vaginal irritation and can increase the rate of contraceptive failure. A detailed analysis of published literature aiming to evaluate the spermicidal activity of N-9 on sperm was carried out. In the pooled results, N-9 reduced the number of active sperm and the distance they traveled. It also caused damage to the structures of sperm and to the way the sperm acted and interacted with the egg. In conclusion, N-9 impacts on sperm in a number of ways that lead to sperm death and dysfunction.

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Semen quality has a direct relation to male fertility. Whether sperm variables in humans have decreased over the last years is still uncertain, with some studies showing a decline and others reporting no changes. In this regard, previous research has suggested that lifestyle and environmental conditions may contribute to this variability, calling for regional studies. The present work is a retrospective, unicentric study that includes semen samples analyzed between 1997 and 2017 at the Parc Taulí Hospital (Barcelona metropolitan area). First, a multivariate analysis including the age as a confounding factor showed a statistically significant decrease in semen volume, pH, progressive motility, morphology and total motile sperm over time. Contrarily, no significant variation in sperm count or concentration was observed. Mean reductions per year were -0.02 mL for volume, -0.57% for progressively motile sperm and -0.72% for sperm with normal morphology. Interestingly, the average annual temperature registered by the Spanish Meteorology Agency negatively correlated to sperm morphology and sperm count (Rs = -0.642; p = 0.002 and Rs = -0.435; p = 0.049, respectively). In conclusion, the present study based on infertile patients from the Barcelona area found a decline in sperm motility and morphology, without effects on sperm count. Changes in temperature appeared to be associated to this decline, but further studies are needed to address the mechanisms linked to the observed variations.

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Fatty acid-based lipidomic analysis has been widely used to evaluate health status in human medicine as well as in the veterinary field. In equine species, there has been a developing interest in fertility and sperm quality. Fatty acids, being the principal components of the membranes, play an active role in the regulation of the metabolic activities, and their role on spermiogenesis seems to be of great importance for the resulting quality of the sperm and, thus, fertility. With the application of widely used lipidomic techniques, the aim of this study was to evaluate: (a) the fatty acid content of the spermatozoa's membranes of 26 healthy male Martina Franca donkeys and its possible correlation with sperm parameters, and (b) the evaluation of the composition of the red blood cells' membrane. PUFA omega-6 are the principal components (40.38%) of the total PUFA content (47.79%) in both types of cells; however, DPA is the predominant one on the spermatozoa's membrane (27.57%) but is not present in the erythrocyte's membrane. Spermatozoa's motility (%) is positively correlated with stearic acid and EPA, and progressive motility (%), with oleic acid. These findings offer information on the composition of both types of cells' membranes in healthy male MF donkeys and reflect the metabolic transformations of the spermatozoa's membrane during the maturation period, providing a better perception of the role of fatty acids in sperm parameters and fertility.

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Over the past five decades, there has been increasing evidence to indicate global declines in human semen quality. Parallel adverse trends measured in male animals indicate a potential environmental aetiology. This study evaluated the progressive motility (PM) of stallion ejaculate through a systematic review and meta-analysis. A total of 696 estimates of equine PM from 280 studies, which collected semen samples between the years 1990 and 2018, were collated for meta-analysis. The method of motility analysis, breed, season of collection, and geographical location were extracted. Simple linear regression determined temporal trends in stallion PM. Studies using microscopy estimated PM to be significantly greater compared to computer-automated methods (p ≤ 0.001). For Arabian breeds, PM was consistently higher than other breeds. Over time, there was a significant decline in PM for studies from Europe (n = 267) but a significant increase for studies from North America (n = 259). Temporal trends indicate the fertilising capacity of equine ejaculate has remained consistently high in the last three decades. That being so, variations observed suggest methodological, geographical, and individual stallion differences may significantly influence actual and reported stallion fertility potential.

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An association between progressive motility (PM) and spermatozoa fertility competence has been suggested. However, the mechanism that underlies PM is not clear enough. We examined physiological characteristics and fatty acid composition of fresh spermatozoa with high and low PM. Additional analysis of fatty acid composition and structural characteristics was performed on spermatozoa samples with high and low progressively motile spermatozoa's survival (PMSS), i.e., the ratio between the proportion of progressively motile spermatozoa after and before cryopreservation. Finally, a fertility field trial was conducted to examine the association between the number of PM spermatozoa within the insemination straw post thawing and conception rate. Analysis of fresh spermatozoa revealed a higher omega-6 to omega-3 ratio in ejaculates with low PM relative to those with high PM (p < 0.01). The proportion of polyunsaturated fatty acids was higher in low-PMSS fresh samples (p < 0.05) relative to their high-PMSS counterparts. Fresh samples with high-PMSS expressed a higher mitochondrial membrane potential (p < 0.05) and a higher proportion of viable cells that expressed reactive oxygen species (ROS; p < 0.05). Post-thawing evaluation revealed a reduced proportion of progressively motile sperm, with a prominent effect in samples with high PM relative to low PM, defined before freezing (p < 0.01). No differences in spermatozoa mitochondrial membrane potential or ROS level were found post-thawing. A fertility study revealed a positive correlation between the number of progressively motile spermatozoa within a standard insemination straw and conception rate (p < 0.05). Considering these, the bull PMSS is suggested to be taken into account at the time of straw preparation.

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CONTEXT: Some vaginal lubricants and ultrasound gels are known to be detrimental to sperm function and therefore could negatively affect fertility. AIMS: The aim of the current study was to develop a sperm motility index (SMI) to test the sperm toxicity of ultrasound gels and vaginal lubricants used in reproductive medicine. SETTINGS AND DESIGN: Two ultrasound gels (Aquasonic® and Kefus®) and five vaginal lubricants (Vaginesil™, Velastisa®, K-Y Jelly®, Control®, and Durex®) were studied. Three different concentrations (1%, 5%, and 10%) of each lubricant were tested. SUBJECTS AND METHODS: SMI was calculated dividing the percentage of progressively motile sperm in each tested gel by that in the control at 0.5, 1, 2, and 24 h of incubation at 5% of CO2 and 37°C. SMI values <0.75 indicate sperm toxicity. STATISTICAL ANALYSIS USED: The main outcome measured was SMI for each concentration and time of incubation. RESULTS: Only Durex® did not show any deleterious effect on sperm quality. The rest of lubricants presented different degrees of toxicity. Vaginesil™ resulted in toxic for all concentrations and incubation periods (SMI < 0.12). Control® and Velastisa® presented toxicity at 10% after 2 h, while K-Y Jelly® showed toxicity at 10% from 1 h of incubation. Regarding ultrasound gels, Aquasonic® showed toxic effects after only 0.5 h (SMI = 0.70 ± 0.15), while Kefus® showed slightly toxic effects after 2 h (SMI 0.69 ± 0.07). CONCLUSIONS: SMI is an accurate tool to evaluate sperm toxicity. One of the main strengths of the article is the inclusion of representative semen samples and known products used worldwide. This study has a relevant clinical translation since it highlights the importance of evaluating the possible sperm toxicity of simple products used in reproductive medicine.

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OBJECTIVE: To evaluate the changes of conventional sperm parameters in men who referred to an andrology reference center in Catania (Eastern Sicily, Italy) in the decade 2011-2020. METHODS: For this purpose, we selected-retrospectively and randomly-the reports of 1409 semen analyses performed according to the 2010 WHO criteria. Data on sperm concentration, total sperm count, progressive sperm motility, and percentage of normal forms were analyzed using linear regression of the raw and logarithmic-transformed data. The sperm parameters were subsequently pooled in two five-year periods (2011-2015 and 2016-2020) and compared with each other. Finally, the influence of the city of residence was assessed on five-year pooled data. MAIN RESULTS: A slight but non-significant decline of total sperm count (-2.26 million/year; p = 0.065) and the percentage of spermatozoa with normal morphology (-0.08%/year; p = 0.057) was observed. In contrast, a significant increase of progressive sperm motility (+0.28%/year; p = 0.008) over time was found. The total sperm count of the quinquennium 2016-2020 was significantly lower. and an upward trend of progressive sperm motility was found. compared to the years 2011-2015. No changes in sperm concentration and morphology occurred in the years 2011-2015 vs. 2016-2020. Sperm conventional parameters did not differ when the five-year pooled data were analyzed according to the town of residence. CONCLUSIONS: Divergent trends of total sperm count and progressive sperm motility over time were found in patients from Eastern Sicily. This may point out the need of assessing whether a time-dependent change of biofunctional sperm parameters occurs to really understand the trend of sperm quality over time.

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BACKGROUND: Conventional cryopreservation methods induce chemical and mechanical damage to the sperm membranes. The cryoprotectant potential of phospholipids of vegetal origin as soybean lecithin has been investigated as a substitute for egg yolk in diluents used for the cryopreservation of human spermatozoa. Therefore, the objective of this study was comparing the efficacy of a synthetic cryoprotectant supplemented with L-α-phosphatidylcholine (PC) and L-acetyl-carnitine (ANTIOX-PC) and the standard egg-based TEST-yolk buffer (TYB) in preserving sperm motility and chromatin quality in cryopreserved semen samples. METHODS: Prospective experimental study in which semen samples from 63 men with normal sperm motility and 58 men with low sperm motility were included and analyzed both before and after cryopreservation using ANTIOX-PC or TYB freezing media. Sperm quality was evaluated by routine semen analysis and DNA fragmentation index using the Terminal deoxynucleotidyl transferase dUTP nick end labeling assay. RESULTS: Differences in the post-thaw progressive motility and DNA fragmentation index were not detected between TYB and ANTIOX-PC cryoprotectants in both normal and low sperm motility groups (P>0.05). However, ANTIOX-PC medium retained higher non-progressive motility and lower percentage of immotile sperm when compared to TYB medium, resulting in a greater total motile sperm count (P<0.05), regardless baseline values of motility characteristic of the normospermic or asthenozoospermic samples. CONCLUSIONS: ANTIOX-PC medium was effective to protect human sperm during a freeze-thaw cycle compared to the TYB medium. A clinically relevant advantage in better preserving kinetic parameters as higher total motility and lower immotile post-thawed sperm from ANTIOX-PC, in normal and low motility semen samples, demonstrated the positive impact of phospholipid and antioxidant treatment on sperm cryotolerance with high potential for egg yolk lipids replacement and biosafety.