Sperm Motility Characteristics Research Articles

Effect of Quercetin and Mirtazapine on Spermatogenesis and Testis Structure in Phenylhydrazine-Induced Hemolytic Anemia Mice: An Experimental Study

Anemia poses a significant healthcare challenge across different socioeconomic groups and can result in reproductive system damage through the generation of free radicals and lipid peroxidation. This study examines the protective effects of quercetin (QUE) and mirtazapine (MIR) against the reproductive damage caused by phenylhydrazine (PHZ) in mice. Fifty NMRI mice, aged 8 to 10 weeks with an average weight of 27.0 ± 2.0 g, were randomly divided into five groups. The control group (Group 1) received oral administration of 10 mL/kg/day of normal saline. Group 2 (PHZ group) received an initial intraperitoneal dose of 8 mg/100 g body weight of PHZ, followed by subsequent doses of 6 mg/100 g every 48 hr. Group 3 received PHZ along with oral QUE at a dosage of 50 mg/kg/day. Group 4 received PHZ along with oral MIR at a dosage of 30 mg/kg/day. Group 5 received PHZ along with oral QUE at a dosage of 50 mg/kg/day and MIR at a dosage of 30 mg/kg/day. The treatment duration was 35 days. Sperm samples were collected from the caudal region of the epididymis post-euthanasia to assess the total mean sperm count, sperm viability, motility, DNA damage, and morphology. Testicular tissue was employed to quantify total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) concentrations, while serum levels of testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were analyzed. Additionally, various aspects, including testicular histopathology, oxidative enzyme levels, gene expression related to apoptosis and antiapoptotic pathways, and in vivo fertility index, were evaluated after 35 days. The QUE, MIR, and QUE+MIR groups showed less abnormal morphology and DNA damage, as well as better total and progressive sperm motility, motility characteristics, viability, and plasma membrane function compared to the PHZ group. QUE, MIR, and QUE+MIR administration increased TAC, SOD, and GPx activities in testicular tissue, while reducing MDA levels compared to the PHZ group. Furthermore, QUE, MIR, and QUE+MIR significantly reduced Bax, and caspase-3 expression levels, and increased Bcl-2 expression levels, compared to the PHZ group. Mice treated with QUE, MIR, and QUE+MIR exhibited an increased in vivo fertility index and plasma sex hormone levels compared to the PHZ group. These results show that QUE, MIR, and QUE+MIR might be able to improve the fertility index, boost the testicular antioxidant defense system, and control the death of germ cells. This could mean that they could be used to treat mice with PHZ-induced testicular damage.

Factors affecting the analysis and interpretation of sperm quality in frozen/thawed stallion semen

In the current study, we examined: 1) the agreement (bias) between fluorescence-based methods (NucleoCounter-SP100 [NC] vs. flow cytometry [FC]) for determining the viability (VIAB) of frozen/thawed stallion sperm; 2) the agreement between post-thaw sperm total motility (TMOT) and VIAB; 3) whether a difference between TMOT and VIAB [VIAB – TMOT] in frozen/thawed stallion sperm could be explained by the level of lipid peroxidation in viable sperm (VLPP); 4) the repeatability of post-thaw analysis of sperm quality; and 5) the effect of final post-thaw semen dilution (10, 30, or 50 x 106 sperm/mL) on sperm motion characteristics. Post-thaw VIAB was similar between NC and FC (P > 0.05), and the agreement between these two methods was high (bias: 1 to −3). The agreement between post-thaw TMOT and VIAB decreased as the pre-freeze percentages of morphologically normal sperm and DNA quality decreased: bias – 4 to – 25. The bias between [VIAB – TMOT] and VLPP ranged from – 5 to 7. Differences in post-thaw sperm quality (TMOT, PMOT, VIAB, and sperm concentration) were not observed when analyzing one or three straws per ejaculate (P > 0.05). There was no effect of post-thaw sperm concentration (i.e., 10 vs. 30 vs. 50 x 106 sperm/mL) on sperm motion characteristics (P > 0.05). This study reports factors other than post-thaw sperm motility that warrant further consideration when analyzing frozen/thawed stallion sperm.

Effect of anti-adhesive supplement doses on sperm motility characteristics in the European whitefish (Coregonus lavaretus)

Sperm adhering to glass slides is one of the main problems during fish sperm motility analyses with CASA systems. To mitigate this, albumin is the supplement added most frequently to activating solutions. However, there is no data on the use of supplements other than albumin (in various concentrations) in analyses of European whitefish (Coregonus lavaretus) sperm motility. This issue was investigated in the presented research using three anti-adhesive supplements (albumin, casein, Pluronic F-127) that were added to Billard solution (BS: 20 mM Tris, 1 mM CaCl2, 154 mM NaCl, 30 mM glycine at pH 9.0) at different concentrations (0.0; 0.1; 0.2; 0.5; 1.0; 2.0%). It was noted that the addition of the lowest concentration (0.1%) of albumin, casein, or the pluronic to BS had a significant effect on the motility and kinetic parameters of whitefish sperm compared to pure BS. BS supplemented with 0.2–0.5% albumin was the most appropriate variant used for whitefish sperm motility activation in the present experiment. BS supplemented with the pluronic at 1.0–2.0% concentrations resulted in significantly higher values of almost all CASA parameters compared to casein at the same concentrations. Moreover, CASA parameters determined in this variant of the pluronic (1.0–2.0%) were similar to those when BS was supplemented with the same albumin concentrations. This indicated that instead of albumin, the pluronic at higher concentrations in BS might be used to analyze whitefish sperm motility.

Timing of puberty in F1-generation hatchery-produced greater amberjack (Seriola dumerili)

We evaluated the onset of puberty of first-generation (F1) hatchery-produced greater amberjack (Seriola dumerili) reared in sea cages for 5 years. Fish were sampled every year in June, at the expected peak of the spawning period in the Mediterranean Sea. No sexual dimorphism in body weight was observed in the study. The ovaries of 1 and 2-year-old (yo) females consisted of primary oocytes only, while at the age of 3-yo early vitellogenic (Vg) oocytes were also identified, but with extensive follicular atresia. At the age of 4-yo, late Vg oocytes were observed, but again extensive follicular atresia characterized the ovaries of 50 % of females. At the age of 5-yo, follicular atresia of Vg oocytes was very limited. In males, gametogenesis was evident already in 1- and 2-yo fish, and 100 % of sampled 3-yo males produced collectable viable sperm. Plasma testosterone (T), 17β-estradiol (E2), and 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) remained similar in 3 – 5-yo females, with T and E2 levels being highest in females in advanced vitellogenesis or with significant follicular atresia, compared to immature females. In males, plasma T declined over the years, while 11-ketotestosterone (11-KT) and 17,20β-P were highest in 4 and 5-yo males, with spermatozoa motility characteristics being improved from the 4th year onwards. The administration of GnRHa implants to 5-yo fish induced only two spawns, albeit no fertilized eggs were obtained. The results indicate that hatchery-produced greater amberjack males mature well and within the same age observed in the wild, however with smaller gonad size. On the contrary, females mature later than in the wild, also with a smaller gonad size. Spawning in response to GnRHa treatment was not effective, suggesting that Mediterranean hatchery-produced broodstocks may be dysfunctional, and further research is needed to document any improvement as the fish get older, or to determine if the results may be related to the specific stock of fish.

Improving the quality of chilled semen from Thai native chicken using phosphorus and vitamin B12 supplementation in semen extender

This study aimed to determine phosphorus and vitamin B12 supplementation effect in semen extender on the quality and fertility ability of chilled Thai native rooster semen. Eighty-four ejaculates of semen from 26 Thai native roosters (Burmese x Vietnam crossbreed) were included. Semen was collected by applying dorsal–abdominal massage once a week, pooled, diluted to 500 million sperms per dose, and divided into six groups. The semen samples used for control group were diluted with modified Beltsville poultry semen extender (BPSE). For the treatment groups 2–6: semen samples were diluted with modified BPSE and enriched with phosphorus and vitamin B12 (Octafos® Octa Memorial Co., Ltd., Bangkok, Thailand) at concentrations 0.02%, 0.04%, 0.06%, 0.08%, and 0.10%. Semen fertility ability was tested in six replications by inseminating layer hens. Thirty-six Thai native hens were randomly assigned to three groups (control, 0.04%, and 0.08%) of 12 hens and were inseminated with a dose of 0.2 mL on collecting day. Sperm motion characteristics (i.e., sperm motility, sperm progressive motility, and sperm kinetic parameters) were measured using a computer-assisted sperm analysis system (SCA®, Proiser S.L., Valencia, Spain). Sperm viability, mitochondrial activity, acrosome integrity, plasma membrane integrity, and malondialdehyde (MDA) concentration were also evaluated. The sperm motion characteristics were the highest in the 0.04% supplementation group on all days of collection, especially the VCL and VAP (P < 0.05). The viability, mitochondrial activity, plasma membrane and acrosome integrity of spermatozoa were greater in the 0.04% supplementation group than in the control groups (P < 0.05). The 0.04% supplementation group had the lowest MDA concentration in all days of collection. The 0.04% supplementation group were higher both fertility (66.59% vs. 48.50%: P < 0.05) and hatching rates (58.80% vs. 43.18%: P < 0.05) than in the control group. In conclusion, 0.04% phosphorus and vitamin B12 concentrations supplementation in semen extender improved rooster semen quality and fertility in chilled rooster semen.

Effect of feeding dried olive (Olea europaea) leaves on the reproductive parameters of Awassi rams

Abstract The aim of this study was to estimate the effect of feeding adult Awassi rams different ratio of dried olive leaves on the reproductive parameters during seven weeks during the breeding season. Three groups of Awassi rams were fed for twelve weeks either a conventional wheat straw-concentrate ration (Control) or experimental rations where 30% (G1) and 60% (G2) of the wheat straw were replaced with dried olive leaves. Animals were fed the same amounts of crude protein and metabolizable energy daily. Results indicated that with the exception of scrotal circumference, no significant (p&gt;0.05) responses were noted for testosterone level, body weight, semen volume, sperm concentration or semen pH values between the tested groups. Analysis of motility characteristics of sperm by computer-assisted semen analysis (CASA) technique showed no significant differences (p&gt;0.05) between the three groups in the percent motility, percent of sperm showing progressive motility, average path velocity, straight line velocity, curvilinear velocity, percent straightness and percent linearity. It could be concluded that feeding Awassi rams dried olive leaves even at the 60% level did not affect the reproductive characteristics of the Awassi rams and therefore could be used as supplement sources of feeds for small ruminants.

P-090 The in vitro motility characteristics of human sperm collected by microfluidic device

Abstract Study question Do the in vitro motility characteristics of sperm collected using a microfluidic device (ZyMot) differ from those of sperm collected by density gradient centrifugation (DGC)? Summary answer ZyMot-collected sperm had longer-lasting motility than those collected by DGC, but the head beat-cross frequency was inferior due to the absence of washing in vitro. What is known already ZyMot (DxNow, Inc., USA) can sort highly motile sperm with low reactive oxygen species (ROS), and DNA damage, without centrifugation. In IUI cycles, although the total number of ZyMot collected sperm was lower than that of DGC, it was reported that the pregnancy rate of ZyMot cycles was higher than that of DGC cycles. Washing sperm with a medium is essential for in vitro capacitation; however, the protocol for microfluidic devices lacks this procedure. The aim of this study was to clarify the motility characteristics of ZyMot-collected sperm. We also examined how washing ZyMot-collected sperm affected motility. Study design, size, duration The specimens were provided by patients who underwent semen analysis between October 2022 and January 2023, with written consent. We used 12 samples with a semen volume of 2.5 ml or more, or a motile sperm concentration of 4 million/ml or more. The same donor samples were divided into three groups (DGC, ZyMot, and ZyMot with wash). Dunnett’s test was performed with the DGC group as the control group. Participants/materials, setting, methods In the DGC group, 2-layer DGC (400G, 20 min) was performed, followed by washing (400G, 5 min). In the ZyMot group, ZyMot was used according to the manufacturer’s protocol. In the ZyMot with wash group, ZyMot-collected sperm was centrifugally washed (300G, 5 min). All samples were cultured at 30 °C until motility analysis. Sperm motility was analyzed using a computer-assisted sperm analysis system (SMAS) after 0, 4, and 24 h of semen preparation. Main results and the role of chance There was no significant difference in the motility rate of the two ZyMot groups compared to that of the DGC group at 0 and 4h after sperm preparation. On the other hand, the motility of the ZyMot group tended to be higher (53.3%, p = 0.0787), and the motility of the ZyMot with wash group was significantly higher (85.4%, p &amp;lt; 0.001) than that of the DGC group (33.5%), after 24h of sperm preparation. The straight-line velocity, curvilinear velocity, and average speed of the ZyMot and ZyMot with wash group did not differ from those of the DGC group at 0 and 4h. All speed parameters of the ZyMot with wash group were significantly higher than those of the DGC group, after 24h of culturing. Regarding sperm head movement, there was no significant difference in the amplitude of lateral head displacement between the two ZyMot groups compared to that of the DGC group at 0 and 4h of culture. In contrast, the sperm head beat-cross frequency of the ZyMot group was lower than that of the DGC group (9.18±0.31 Hz vs 8.71±0.49 Hz, p = 0.0409). Both head movement parameters of the ZyMot with wash group were significantly higher than those of the DGC group. Limitations, reasons for caution We chose semen samples with a high number of motile sperm because we divided the sperm into three experimental groups. Therefore, it is unclear whether our findings are applicable to patients with poor semen parameters. Wider implications of the findings ZyMot may be advantageous for IUI cycles because ZyMot-collected sperm maintains sustainable motility for extended periods. Alternately, ZyMot may be unfavorable compared to DGC in conventional IVF cycles due to inferior sperm head movement, which is a marker of capacitation. However, this disadvantage can be overcome by washing. Trial registration number not applicable

The Difference in Subpopulation Structures in Sex-Sorted and Non-Sorted Semen by Discriminant Analysis of Bull Sperm Motility Data Collected by a Computer-Assisted Sperm Analysis System

We examined the motility characteristics of sperm from sex-sorted bull semen as a model for low-quality (expected low conception rate) sperm compared to that of sperm from non-sorted semen (expected average conception rate) using cluster analysis followed by discriminant analysis. The results indicated that sex-sorted semen contained sperm with hyperactivation-like motility as the main subpopulation immediately after thawing, and this subpopulation decreased after 2 h of incubation. The main subpopulation in the non-sorted semen had progressive motility that was maintained during incubation. A conventional comparison of the mean values of kinematic parameters could not distinguish the samples. In conclusion, discriminant analysis using data from a cluster analysis of motile sperm may accurately describe differences in the structures of sperm motility subpopulations between low and normal fertility semen.

Assessment of semen quality and anti-oxidative enzyme activity between bovine sex-sorted and non-sex-sorted frozen-thawed semen.

In the current study, the difference between the sex-sorted and non-sex-sorted frozen semen of Holstein Friesian breed cattle was investigated. Significant variation (p < .05) was found in the semen quality parameters such as motility; vitality; acrosome integrity rate; the anti-oxidative enzyme activity including GSH (glutathione); SOD (superoxide dismutase); CAT (catalase); GSH-Px (glutathione peroxidase) and the rate of fertilization. The results showed that the sperm acrosome integrity and motility of the non-sorted sperm were higher compared to sex-sorted sperm (p < .05). The linearity index and mean coefficient analysis revealed that the percentage of 'grade a' in sex-sorted sperm were significantly (p < .05) lower than non-sorted sperm. Interestingly, low SOD level and high CAT level was found in the non-sexed semen than in the sexed semen (p < .05). Furthermore, the GSH and GSH-Px activity in the sexed semen was found lower than the non-sexed semen (p < .05). In conclusion, sperm motility characteristics were lower in sex-sorted semen than in non-sex-sorted semen. This might be related to the complex process of sexed semen production, which could reduce sperm motility and movement characteristics, acrosomal integrity, CAT, SOD, GSH and GSH-Px, and finally lead to the decline in the fertilization rate.

Effect of BNT162b2 mRNA COVID-19 vaccine on sperm morphokinetics and DNA integrity: A prospective observational study in Japan

Objective: To assess whether the coronavirus disease 2019 (COVID-19) mRNA vaccine affects sperm morphokinetics using a computer-assisted semen analyzer and other semen parameters using a sperm chromatin structure assay.Methods: Healthy male volunteers in two Japanese clinics between May 2021 and December 2021 were prospectively analyzed. Participants donated sperm twice, two days apart, in the following phases: before vaccination, 2 weeks after the first vaccine dose, and 2, 4, and 12 weeks after the second dose. Basic sperm parameters, sperm motility characteristics, and the percentage of DNA-damaged sperm were compared among the different phases.Results: Ninety-six semen samples from ten volunteers, who were vaccinated with the BNT162b2 mRNA vaccine, were evaluated. There were no significant differences between any phases in basic semen findings and parameters of the sperm chromatin structure assays. Regarding sperm motion characteristics, the average linear velocity, beat-cross frequency, and sperm motility index significantly decreased after the second vaccine dose (P=0.018, P=0.003, and P=0.027, respectively), with no significant differences between any two phases by post-hoc pairwise comparisons.Conclusions: After COVID-19 mRNA vaccination, while sperm motion characteristics might fluctuate, no apparent deterioration of basic sperm parameters or sperm DNA integrity was observed. Given the adverse effects of COVID-19 on sperm, our findings suggest that there might be no reason to refrain from vaccination for healthy individuals.

Evaluation of the in vitro cryopreservative performance of Juniper berry oil (Juniperus communis) on frozen-thawed bull semen

This study aimed to reveal the effects of juniper (Juniperus communis) berry oil (JB) when added to the Tris extender, on bull (Holstein) sperm motility characteristics, abnormal sperm ratio, DNA fragmentation and some oxidative stress markers. The pooled ejaculate was divided into five equal parts as control (C) and Tris diluent containing 25, 50, 100, 200 μg/ml JB. Diluted semen was incubated at 4 °C for 4 h and then frozen in liquid nitrogen (−196 °C). After freezing-thawing, motility characteristics were determined using a sperm analyzer system. The results showed that the 25 μg/ml dose of JB had an improving effect on overall sperm motility (P &lt; 0.05). The total abnormal spermatozoa ratio was not significantly different between the groups (P &gt; 0.05). The JB25 and JB50 groups had higher DNA integrity than the other groups (P &lt; 0.05). This finding was supported by low malondialdehyde levels (P &lt; 0.05). Especially the 25 μg/ml dose of JB was found to have a remedial effect on DNA integrity, as well as increasing progressive and total motility rates. This study has revealed new information on the effects of JB on bull semen cryopreservation. Juniper berry oil may be used as an antioxidant in bull semen freezing.

PROTECTIVE ENVIRONMENTS FOR ANIMAL SPERM: EVOLUTION OF METHODS AND TOPICAL ASPECTS (REVIEW ARTICLE)

A retrospective review of domestic and foreign sources of literature is presented, as well as data of published own research on cryopreservation of animal sperm. The main historical stages of the creation of protective environments for deep freezing of sperm are given. In the 30s of the last century, a phenomenon characterized by the death of spermatozoa upon sharp cooling in the range of positive temperatures was discovered. It is called temperature shock of sperm. To prevent it, it is proposed to add substances containing phospholipids to the composition of diluents. Such environments can contain both simple components - native chicken egg yolk or milk, and high-tech - lipoproteins, isolated phospholipids of various origins. To stabilize protein-lipid complexes of plasma membranes and acrosomes of sperm during the cooling process, carbohydrates are added to the diluents. Sugars are components of energy supply for sperm and, along with salts, they are the main osmotic regulators. A combination of two or three carbohydrates in the medium was traditionally considered necessary. However, the Kharkiv school of reproductive specialists has proven the possibility of creating effective protective environments using only one sugar - sucrose or lactose - based on considerable practical experience. The effectiveness of germ cell freezing is shown depending on the cryoprotectants used. Glycerin is the first known endocellular cryoprotectant, which is still unsurpassed in sperm cryopreservation. Our own experimental data on the effect of combinations of glycerol with substances from the amide group on the main biological indicators of sperm after deconservation are presented. Cryoprotectants dimethylacetamide (DMAC) and dimethylformamide (DMF) were tested in own experiments on stallion semen. The experiments studied the effect of different concentrations of the above-mentioned penetrating cryoprotectants both on the main physiological characteristics of stallion sperm (motility, survival), and on the degree of damage to the membrane apparatus of sperm. The effectiveness of certain combinations of these substances has been proven. Methods of preventing the negative impact of oxygen and the development of lipid peroxidation processes in sperm during cryopreservation are presented. The concept of using additional hormonal components in diluents, in particular prostaglandin F2a, is revealed. The materials related to the effect on the quality of reproductive cells of healing preparations are displayed. Keywords: artificial insemination, environments, semen, animals, bulls, stallions, cryoprotectants, freezing.

Tribulus terrestris ekstraktının ilavesiyle gökkuşağı alabalığının (Oncorhynchus mykiss) sperm motilitesinin arttırılması

Tribulus terrestris (TT) is a famous traditional plant of family Zygophyllaceae and widely distributes around the world. TT has potential to elevate the testosterone, testosterone precursor and luteinizing hormone levels. In this study, trials were conducted to assessment the impact of Tribulus terrestris addition on sperm motility of rainbow trout (Oncorhynchus mykiss) for the first time. In the trial, we used to different concentrations [0 µg L-1 (Control), 200 µg L-1, 400 µg L-1, 600 µg L-1, 800 µg L-1 and 1000 µg L-1] of T. terrestris extract. Sperm motility characteristics and longevity were determined. In addition, protodioscin content of T. terrestris extract was assessed. The present study revealed that the presence of T. terrestris caused to increase in sperm motility. The increases in duration (49.00±3.61 s) and motility rate (91.67±2.89%) at 400 µg L-1 were statistically significant (p&amp;lt;0.05). Overall, addition of T. terrestris to activation solution can increase the sperm motility of rainbow trout.

Predicting the outcome of Thoroughbred stallion matings on the basis of dismount semen sample analyses.

A capacity to predict the likelihood of pregnancy following natural matings would be of considerable benefit to the Thoroughbred horse breeding industry. In this article, we describe a strategy for achieving this outcome through the analysis of dismount samples, that achieved an overall accuracy of 94.6%. The purpose of this study was to determine whether the analysis of dismount semen samples from Thoroughbred stallions could be used to predict whether a given mating would result in a pregnancy. The analysis was based on 143 matings of 141 mares by a cohort of 7 Thoroughbred stallions over a 4-week period at an Australian Stud. The criteria of semen quality utilized in this analysis involved a preliminary assessment of the raw dismount sample in terms of semen volume, sperm number, and sperm movement characteristics using an iSperm® Equine portable device. Following this initial assessment, a subpopulation of progressively motile spermatozoa was isolated by virtue of the cells ability to migrate across a 5 µm polycarbonate filter in a Samson™ isolation chamber over a 15-minute period. These isolated cells were again evaluated for their number and quality of movement using the iSperm® system and, in addition, assessed for their ability to reduce WST-1, a membrane impermeant tetrazolium salt. These data were then combined with additional information describing the ages of the animals used in this study, their historical breeding records, and mating frequency during the breeding season. The total data set was then used to predict the occurrence of pregnancy, as confirmed by ultrasound at ~14 days post mating. The criteria used to predict fertility in the ensuing multivariate discriminant analysis were optimized for each individual stallion. Using this strategy, we were able to successfully predict the outcome of a cover with an overall accuracy of 94.6%.

Post-Thaw Characteristics of the Simmental Sperm Function in Different Ages of Bulls

This study aims to determine the effect of the age difference of Simmental bulls on motion characteristics, capacitation status, and DNA fragmentation of post-thawing sperms. The frozen semen used was collected from twelve bulls, which were divided into four groups of age, which include a group of two, four, ≥ 10 years old with high semen rejection (≥ 10 HR), and ≥ 10 years old with low semen rejection (≥ 10 LR). Computer Assisted Sperm Analysis (CASA) was used to determine sperm motion characteristics, capacitation status by chlortetracycline (CTC) staining, plasma membrane integrity, and viability using eosin-nigrosine staining. In contrast, the DNA fragmentation index was determined using the Sperm-Bos-halomax kit. The results showed that the four year old group had a higher total and progressive motility percentage than the others (p<0.05). In all groups, there was no significant difference among sperm kinematics such as VAP, VSL, VCL, STR, and ALH. However, the LIN, WOB, and BCF of the ≥ 10 HR year old groups were significantly lower (p<0.05) than those of the other groups. However, un-capacitated sperm was higher (p<0.05) in the two years and four years old groups compared to the ≥ 10 years old, while the four years old group had lower capacitated and acrosome-reacted (p<0.05) than the other groups. Furthermore, the sperm membrane integrity, viability, and DNA fragmentation index of the ≥ 10 years old groups were significantly higher (p<0.05) than those of the other groups. The research concludes that aging in the Simmental bull affects motion characteristics, capacitation status, and DNA fragmentation of post-thawing sperm. However, the semen rejection rate in the older bull did not directly affect the post-thawing sperm quality.

LACTATIONAL EXPOSURE OF METHOXYCHLOR IMPAIRS SPERM QUALITY IN BALB/C MICE

Numerous pesticides derived from human activity are now disseminated in the environment that have been identified as an estrogenic activity that mainly affects the male reproductive system. The objective study was to determine the dose of methoxychlor (MXC) pesticides administered to the mother during the lactation period that would affect the sperm quality of the offspring in mice. Twenty-four BALB/c strains were divided into control and treatment groups. MXC was administered once daily by intraperitoneal to female mice for days 1 to 21 of lactation periods, with 0.14 mg/g, 0.28 mg/g, and 0,42 mg/g doses of MXC. Male litters were killed at postnatal day (PND) 60, and the cauda epididymis was used to observe sperm motility and morphology characteristics. Parameters observed were the number of spermatozoa abnormal and the percentage of progressive spermatozoa. ANOVA showed that the effect of MXC was significantly different in the case of the number of abnormal sperm and the rate of progressive sperm parameters. Duncan's Multiple Range Test (DMRT) showed that D3 (0.42 mg/g) gave the highest effect of MXC on increasing abnormal spermatozoa morphology and decreasing the percentage of motile sperm. These results indicate that exposure to MXC, which showed a physiological response in the lactation period, reduces the sperm quality of offspring

Spirulina (Arthrospira platensis) Extract Promotes Motility, Microscopic, and Antioxidative Parameters of Ram Semen during Refrigerated Storage

This study investigated the effect of spirulina ethanolic extract (SEE) on the quality of ram semen during low-temperature liquid storage and the relationship between sperm features. Ejaculates were collected from five Djallonké rams, pooled, extended with Tris-egg yolk (TEY) enriched with 0 (control), 20 (SEE20), 40 (SEE40), and 80 µg/mL (SEE80) of SEE to reach the concentration of 200×106 spz/mL, and stored at 4 °C for 72 h. Extended semen samples were assessed for total motility, progressive motility, sperm motion characteristics, viability, membrane integrity, and morphology at 6, 24, 48 and 72 h of storage. Moreover, malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase, (SOD) and catalase (CAT) levels were measured at 72 h of storage. The enrichment of TEY with SEE at 40 and 80 µg/mL, improved sperm total motility at 48 and 72 h of storage (P

P-022 Relationship between sperm movement characteristics assessed by a sperm motility analysis system (SMAS) and normal fertilization rates in vitro

Abstract Study question Is there a relationship between movement characteristics of human sperm assessed by SMAS and normal fertilization rates in vitro? Summary answer The curvilinear path wobble of motile sperm before insemination is predictive of normal fertilization rates after in vitro fertilization (IVF). What is known already Assessment of semen parameters is very important in both the diagnosis of male infertility and the selection of methods for treatment with conventional-IVF (C-IVF) or intracytoplasmic sperm injection (ICSI). Several studies have reported that objectively movement characteristics of motile sperm assessed by SMAS are related to the functional capacity of human sperm. However, there are no established the objective indicators and criteria for sperm motility characteristics assessed by SMAS. In this study, we investigated the utility of the movement characteristics of motile sperm measured by SMAS as an indicator of normal fertilization rates. Study design, size, duration Retrospective experimental trial conducted in a single IVF center between January 2018 and February 2020. Sperm movement characteristic analysis was performed using the SMAS in 300 IVF cycles using sperm from 270 men. At least three metaphase II oocytes were inseminated in C-IVF. The SMAS estimates were measured in semen and before insemination. This study was approved by the Ethics Committee of the Institutional Review Board (IRB) of Kinutani Women’s Clinic, Hiroshima, Japan. Participants/materials, setting, methods Motile sperm from at least 3 different fields were observed in each sample and their parameters were examined. First, we investigated the correlation between the normal fertilization rates and sperm motility characteristics. Next, the group comprising 300 cycles was classified into a group of a ≥ 60% normal fertilization (group 1: 229 cycles) and a group with &amp;lt;60% normal fertilization (group 2: 71 cycles) to assess the sperm characteristics described below. Main results and the role of chance The following indicators were evaluated: amplitude lateral head displacement (ALH), best cross frequency (BCF), curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), linearity (LIN=VSL/VCL), straightness (STR=VSL/VAP), and curvilinear path wobble (WOB=VAP/VCL).There were significant correlations between normal fertilization rates and semen parameters, including VCL (r = 0.014; P &amp;lt; 0.05) and ALH (r = 0.177; P &amp;lt; 0.05). However, there were no significant correlations for other parameters. Moreover, there were no significant correlations between normal fertilization rates and sperm parameters before insemination. The SMAS estimates in semen and sperm before insemination were compared between group 1 and group 2. There were no significant differences in most baseline characteristics across groups, except for the number of normally fertilized oocytes. However, there were significant differences in sperm WOB before insemination between group 1 and group 2 (0.589 vs. 0.572; P &amp;lt; 0.05). Sperm motility before insemination was more curvilinear in group 1 than in group 2. The optimal cut-off value for WOB of motile sperm before insemination for predicting≥60% normal fertilization was 0.587 (AUC was 0.60; P &amp;lt; 0.05). These results were suggested that the WOB of sperm before insemination is useful as an indicator in determining the selection of methods for fertilization in vitro. Limitations, reasons for caution The main limitations of this study are its retrospective design and small sample size. A prospective study is necessary to further clarify the prediction criteria. Wider implications of the findings This study suggests that the WOB of motile sperm before insemination may be used as one of the indicators to predict good fertilization rates after C-IVF. Trial registration number N.A.

P-094 Fundamental understanding of sperm motion in viscoelastic media

Abstract Study question How does viscosity influence the flagellar beating behaviour of free-swimming bull, mouse, and human sperm? Summary answer Sperm flagellar beating behaviour exhibits a transition from an irregular three-dimensional (3D) beating at 5 mPa·s to an organized two-dimensional (2D) waveform at 20 mPa·s. What is known already Sperm migrate in a complex viscoelastic environment through the female reproductive tract. The viscoelastic properties of the oviductal fluid significantly influence the progressive motility of sperm, acting as one of the key guidance mechanisms in vivo. However, the biomechanics of sperm flagellar activity in response to varying viscosity of the oviductal fluid is poorly understood. Understanding sperm flagellar behaviour in physiologically relevant environments is crucial to understanding reproduction and may help to describe unknown causes of infertility. Lack of high-speed high-resolution imaging techniques and automated image-analysis capabilities have been the main barriers to fully describe the flagellar beating behaviour. Study design, size, duration We used a custom-built high-speed high-resolution dark-field microscopy platform to resolve the flagellar dynamics of human, bull, and mouse sperm near surfaces in viscoelastic media ranging in viscosity from 1 to 250 mPa·s. The imaging system includes an automated image analysis algorithm to quantify sperm flagellar waveform and motility characteristics by extracting the flagellar centreline, reconstructing the waveform and calculating tangent-angle profiles. 20 sperm from 3 different bull, mice and humans were analyzed. Participants/materials, setting, methods Bull, mouse, and human sperm were used in this study. In each experiment, a diluted sperm sample in a buffer supplemented with methylcellulose was used and free-swimming sperm were imaged using dark-field microscopy at 200 frames per second. An automated image analysis algorithm was used to extract sperm flagellar centreline and Proper Orthogonal Decomposition (POD) was then used to study sperm flagellar waveform. Statistical analysis was performed using one-way ANOVA. Main results and the role of chance The reconstructed flagellar beating pattern was different for sperm swimming in low and high-viscosity media. Bull sperm exhibited a lower flagellar beating amplitude along the end piece when swimming in a high-viscosity media, a potential energy-efficient strategy to navigate a high viscosity fluid. The first two dominant POD modes (shape modes) describe more than 90% of the beating pattern for all species. Bull sperm exhibited a transition mode with irregular loops in 5 mPa·s buffer, but the flagellar shape cycle created an organised repetitive circular cycle in 1 mPa.s (3D beating) buffer and at viscosities higher than 5 mPa·s (2D beating). Human sperm also indicated a similar behaviour but with the transition happening at higher viscosities. Mouse sperm in high-viscosity media had a lower flagellar beating amplitude across the principal piece and higher beating amplitude across the end piece. The flagellar shape cycle in mouse sperm showed a periodic flagellar beating behaviour at high-viscosities (&amp;gt;20 mPa·s), but a shape cycle with distorted loops at lower viscosities. Our results showed in quantitative detail that increasing viscosity alters sperm flagellar beating pattern, and how sperm migration behaviour in low viscosity media can be distinct from their swimming behaviour in vivo. Limitations, reasons for caution A more comprehensive study of sperm motility parameters such as curvilinear velocity, average path velocity and straight line velocity with a larger sample size is required to fully characterise sperm swimming behaviour as a function of viscosity. Wider implications of the findings The increasing viscosity of the oviductal fluid regulates the sperm flagellar beating behaviour to switch from a 3D swimming behaviour with irregular shape cycles at lower viscosities to a 2D slithering mode with repetitive circular shape cycles at higher viscosities to achieve a more energy-efficient beating pattern for navigation. Trial registration number Not applicable

Heat shock protein family D member 1 in boar spermatozoa is strongly related to the litter size of inseminated sows

BackgroundSperm quality evaluation is the logical first step in increasing field fertility. Spermatozoa contain cytoplasmic organelles and biomolecules known as sperm-intrinsic factors, which play key roles in sperm maturation, sperm-oocyte fusion, and embryo development. In particular, sperm membrane proteins [e.g., arginine vasopressin receptor 2, beta-actin, prohibitin, and heat shock protein family D member 1 (HSPD1)] and RNA could be used as functional indicators of male fertility. We sought to clarify the effects of differential mRNA expression of selected genes on several fertilisation parameters, including sperm motility, motion kinematics, capacitation, and litter size, in a porcine model.ResultsOur results demonstrated that HSPD1 expression was significantly correlated with male fertility, as measured by the litter size of inseminated sows. The expression of HSPD1 mRNA was linked to sperm motility and other motion kinematic characteristics. Furthermore, HSPD1 had a 66.7% overall accuracy in detecting male fertility, and the high-litter size group which was selected with the HSPD1 marker had a 1.34 greater litter size than the low-litter size group.ConclusionsOur findings indicate that HSPD1 might be a helpful biomarker for superior boar selection for artificial insemination, which could boost field fertility.