Sperm Motility Index Research Articles

Evaluation of duck egg yolk for the cryopreservation of Nili-Ravi buffalo bull semen

This study was carried out to investigate if the substitution of chicken egg yolk (CEY) with duck egg yolk (DEY) in extenders can improve the quality of frozen-thawed semen of Nili-Ravi buffalo bulls and to study if reducing DEY level in extender affects the freezability results. Thirty semen samples collected from three buffalo bulls were diluted in extenders A, B, C, D and E containing tris, citric acid, fructose, egg yolk, glycerol and antibiotics. Extender A contained 20% CEY (control), while extenders B, C, D and E contained 5, 10, 15 and 20% DEY, respectively. After freezing and storage for 24h in liquid nitrogen, samples were evaluated for post-thaw quality.The post extension sperm motility did not differ between extenders A (control) and E (20% DEY). The same was true for post-thaw percentage of sperm with functional plasma membrane and percentage of sperm with abnormal heads or mid pieces. However, extender E showed higher (P<0.05) values for post-thaw sperm motility, livability and absolute index of livability of spermatozoa at 37°C compared to extender A. Spermatozoa with abnormal tail were lower (P<0.05) in extender E compared to extender A. Values of these parameters of post-thaw semen quality were highest for extender E containing 20% DEY and decreased significantly with decrease in the concentration of DEY, except sperm abnormalities (head, mid-piece and tail) which increased with decrease in DEY level. These results showed that replacement of 20% CEY with 20% DEY in extenders significantly improved post-thaw sperm motility, livability and absolute index of livability of spermatozoa and reduced tail abnormalities. Reduction in the level of DEY in extenders from 20% adversely affected post-thaw semen quality of Nili-Ravi buffalo bulls.

Relationship among fluctuating asymmetry, morphological traits, and sperm quality in layers

The aim of the present study was to analyze the relationship between fluctuating asymmetry and the mean value of several bilateral traits (toe, leg, wing, and wattle lengths, and leg width) and sperm quality variables (volume, appearance, concentration, motility, morphological abnormalities, and acrosome and membrane integrities) in different breeds of layers. In total, 165 roosters were sampled at 36 wk of age. There was no significant correlation between fluctuating asymmetry and sperm quality variables. There was a positive association (P < 0.01) between sperm motility and concentration, and between sperm concentration and volume. Sperm motility was positively correlated (P < 0.001) with morphologically normal sperm. The membrane integrity did not correlate with motility. This study shows different results for the association between sperm quality variables and morphological traits. There was a negative association (P < 0.05) between toe length and membrane integrity and between leg length and sperm motility. In disagreement with the expected results, wattle length did not correlate with any sperm quality parameter. In general, roosters with longer legs had longer toes and wings, whereas those with longer wattles had longer toes, legs, and wings. Breed effect was significant for the sperm motility index (P < 0.001), concentration and membrane integrity (P < 0.05), and morphologically normal sperm (P < 0.01). Spanish breeds had similar sperm quality variables to the White Leghorn population, although the Blue Andaluza (a Spanish breed) showed smaller values for sperm motility index and concentration and membrane integrity. Breed effect was not significant for any fluctuating asymmetry, with values ranging from 1 to 6% of the trait value; however, breed effect was significant for all of the morphological traits (P < 0.001). The CV was higher for ejaculate volume and concentration, intermediate for semen appearance, motilities, and morphologically normal sperm, and smaller for membrane and acrosome integrity. In conclusion, the results suggest that fluctuating asymmetry is of limited use for assessment of rooster sperm quality. Some morphological traits (toe length, leg length, and leg width) might be associated with some sperm quality variables (motility and membrane integrity).

Testicular apoptosis after dietary zinc deficiency: Ultrastructural and TUNEL studies

The present study was conducted in Wistar rats to determine whether prepubertal dietary zinc deficiency causes apoptotic changes in testes. Prepubertal male Wistar rats (40–50 gm) were divided into 3 groups: zinc control (ZC), pairfed (PF), and zinc deficient (ZD). Control and pairfed groups were given a 100 ppm zinc diet while the deficient groups received 1 ppm zinc diet for 2 and 4 weeks (w), respectively. Ultrastructural studies revealed several apoptotic features such as wavy basement membrane, displaced nuclei, chromatin condensation, plasma membrane blebbing, nuclear membrane dissolution, loss of inter-Sertoli cell junctional complexes, and intercellular bridges and deformed mitochondria. A variable spectrum of sperm defects had also been visualized e.g., acrosomal deformities such as decapitation and a ring of condensed chromatin around the nuclear periphery, deformed sperm heads with a condensed nucleus, tail-elements with superfluous cytoplasm, and damage to the mitochondrial sheath and aggregation of spermatozoa within the membrane. This was further supported by TUNEL studies. Apoptotic index, epididymal sperm concentration, motility, and fertility index also revealed a significant (P < 0.05) decrease in zinc deficient groups (2 and 4 w) when compared with their respective control and pairfed groups. All the above findings are indicative that changes observed in the testes after dietary zinc deficiency are due to the onset of apoptosis. Increased apoptotic degeneration in testes may cause irreversible changes in the germ cells associated with decreased epididymal sperm concentration, motility, and fertility index which contributes to the low efficiency of spermatogenesis thereby indicating a possible role of zinc in fertility.

Characterization of epididymal spermatozoa motility rate, morphology and longevity of springbok ( Antidorcas marsupialis), impala ( Aepyceros melampus) and blesbok ( Damaliscus dorcus phillipsi): Pre- and post-cryopreservation in South Africa

Cryopreservation of antelope epididymal spermatozoa could play a vital role in future breeding by developing a successful protocol for cryo-conserving them. The aim of this study was to characterize morphology, motility rates and longevity of epididymal spermatozoa from springbok, impala and blesbok. Cauda epididymal spermatozoa were collected post-mortem from both testicles of free-ranging springbok ( n = 18), impala ( n = 21) and blesbok ( n = 21), and divided into two groups (pre- and post-cryopreservation). Spermatozoa were cryopreserved in Biladyl supplemented with 20% egg yolk and 7% glycerol under field conditions. Pre-freeze and post-thaw sperm quality was evaluated. The longevity of thawed spermatozoa was evaluated under culture conditions that support domestic cattle in vitro fertilization. There was a significant difference between pre-freeze and post-thaw sperm motility index (SMI) ( p < 0.05), plasma membrane integrity ( p < 0.05) and acrosome integrity ( p < 0.05) for all species. Post-thaw SMI and plasma membrane integrity were comparable between species ( p > 0.05). The effects of cryopreservation on sperm cell morphology differed between species and between specific abnormal morphology. Blesbok had the least abnormalities in post-thaw spermatozoa. Cryopreservation substantially reduced the survivability and motility rates of antelope species. Blesbok spermatozoa tolerated cryopreservation and thawing process better than impala and springbok. The antelope cauda epididymal sperm maintained viability and acrosome integrity for at least 4 h following incubation under conditions that support domestic cattle in vitro fertilization (IVF) with a decline in longevity over time across species however; species responded differently over time in terms of plasma membrane integrity and acrosome integrity. The antelope species may have different in vitro culture requirements, indicating differences in sperm physiology between the species. This research could contribute species-specific protocol development for IVF thus promoting ex-situ conservation strategies of African antelope species in South Africa.

2185 THE INCIDENCE AND EFFECT OF BACTERIOSPERMIA AND ELEVATED SEMINAL WHITE BLOOD CELLS ON SEMEN PARAMETERS

You have accessJournal of UrologyInfertility: Evaluation1 Apr 20112185 THE INCIDENCE AND EFFECT OF BACTERIOSPERMIA AND ELEVATED SEMINAL WHITE BLOOD CELLS ON SEMEN PARAMETERS Trustin Domes, Kirk Lo, Ethan Grober, Brendan Mullen, Tony Mazzulli, and Keith Jarvi Trustin DomesTrustin Domes Toronto, Canada More articles by this author , Kirk LoKirk Lo Toronto, Canada More articles by this author , Ethan GroberEthan Grober Toronto, Canada More articles by this author , Brendan MullenBrendan Mullen Toronto, Canada More articles by this author , Tony MazzulliTony Mazzulli Toronto, Canada More articles by this author , and Keith JarviKeith Jarvi Toronto, Canada More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.2423AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES In the infertile male, the effect of bacteriospermia and elevated seminal white blood cells (ESWBC) on semen parameters remains controversial. Despite this, the majority of men with bacteriospermia are treated with antibiotics. The objectives of this study are to determine the incidence of ESWBC and bacteriospermia (specific bacterial isolates) in a large non-azoospermic male infertility population, correlate these results with semen parameters and to determine which bacterial isolates negatively affect semen parameters preferentially. METHODS Non-azoospermic male patients attending a tertiary-level infertility clinic over nine years (2001–2010) that had a semen analysis and semen culture performed concurrently were included in the study. ESWBC was defined as ≥ 1 WBC in semen per 100 sperm. The prospectively collected microbiology results were merged with a prospectively collected andrology database for analysis. RESULTS During the study period, 8158 semen analysis and concurrent semen culture pairs were identified in 5088 patients with a median age of 37 years. The rate of bacteriospermia was 15%, incorporating 22 bacterial species. The four most common species accounted for 90% of bacterial isolates: E. fecalis (56%), E. coli (16%), Group B Streptococcus (13%) and S. aureus (5%). The rate of ESWBC was 19% and there was no correlation between bacteriospermia and ESWBC (18% ESWBC associated with bacteriospermia versus 19% ESWBC not associated with bacteriospermia, p= 0.383). There were statistically significant differences in sperm motility (24.5 vs. 25.5%, p=0.045) and DNA fragmentation index (DFI) (24.9 vs. 22.6%, p=0.024) in the bacteriospermia versus no bacteriospermia group. ESWBC alone was associated with more profound deterioration in semen parameters, including sperm concentration (20.6 vs. 55.9 x 106/cc, p<0.001), motility (21.5 vs. 26.8%, p<0.001), normal morphology (12.2 vs. 17.4%, p<0.001) and DFI (26.7 vs. 22.2%, p<0.001) with no additional deterioration identified with bacteriospermia and ESWBC together. No specific bacterial isolate preferentially deteriorated any semen parameter more than the other. CONCLUSIONS This large series demonstrates that both bacteriospermia and ESWBC are prevalent in men with infertility, however bacteriospermia was not statistically associated with ESWBC. While both are associated with diminished semen parameters, ESWBC is associated with the most profound deterioration in semen parameters. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e876 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Trustin Domes Toronto, Canada More articles by this author Kirk Lo Toronto, Canada More articles by this author Ethan Grober Toronto, Canada More articles by this author Brendan Mullen Toronto, Canada More articles by this author Tony Mazzulli Toronto, Canada More articles by this author Keith Jarvi Toronto, Canada More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Pentoxifylline induces capacitation and acrosome reaction and improves quality of motility in canine ejaculated spermatozoa

To evaluate effects of different concentrations of pentoxifylline, as phosphodiesterase inhibitor, on quality of motility, capacitation and acrosome reaction, Ejaculated spermatozoa were collected from crossbred dogs. The sperm were incubated at concentrations of 0.1, 1, 10 and 100 mM pentoxifylline for 2 h. Conventional assessment was also made on the percentage of motility and quality of motility of spermatozoa; values were expressed as sperm motility index (SMI). Capacitation and acrosome reaction were also evaluated by chlortetracycline fluorescence staining. SMI as quality index of sperm was significantly increased in concentrations of 10 and 100 mM pentoxifylline during 1 and 2 h compared to control. The number of capacitated or acrosome reacted spermatozoa significantly ( P < 0.05) were higher than controls at high concentrations of pentoxifylline (10 and 100 mM) during 1 and 2 h. In conclusion, high concentration of pentoxifylline is able to induce capacitation and acrosome reaction and improves quality of motility in canine ejaculated spermatozoa.

Effects of Radiographic Contrast Dye on Domestic Cat Spermatozoa.

Laparoscopic artificial insemination has played an important role in felid conservation, but requires minor surgery. The less invasive technique of transcervical artificial insemination has not been heavily pursued because of concerns that sperm is not adequately transported through the uterine horns. The use of radiographic contrast dyes to visualize and verify gamete placement within the uterine horns could ensure the success rate of this less invasive procedure. Two radiographic dyes (Omnipaque and MD76) were diluted 1:1.1 (Omnipaque) and 1:4.4 (MD76) with ultra-purified water to a final osmolarity of 320-330±3 mOsm, consistent with the osmolarity of domestic cat spermatozoa. Each diluted dye was tested for radio-opacity by injecting aliquots into uterine horns recovered from spayed cats and radiographing the tissue. Gamete rescued freshly collected and frozen-thawed spermatozoa were extended 1:1 with feline in vitro fertilization media (control) or with diluted radiographic dyes. Motility, forward progression, and acrosomal integrity were recorded every 30 minutes for four hours to analyze felid sperm activity after the addition of the experimental media, while morphology data was taken to determine if radiographic dye induced abnormalities in the sperm cells. To assess sperm penetration abilities, spermatozoa (fresh: 2×105cells/100µl droplets; frozen: 5×105cells/100µl droplets) extended with treatment media were coincubated with conspecific in vitro matured oocytes for 18-20h. All presumptive embryos were fixed and stained three days post insemination to determine fertilization and cleavage rates based on DNA content. Morphology analysis confirmed radiographic dyes did not induce abnormalities in felid spermatozoa. Motility and forward progression were converted into sperm motility indices, and results demonstrated no significant differences between media for freshly collected spermatozoa. For frozen-thawed spermatozoa, a lower sperm motility index was calculated for Omnipaque vs control, but not compared with MD76. Motility index decreased over time for all treatments for both freshly collected and frozen-thawed sperm. Acrosomal integrity for freshly collected spermatozoa was significantly reduced when treated with Omnipaque vs MD76 but not compared with control media. Differences in acrosomal integrity were not seen within frozen-thawed treatment groups. Acrosomal integrity for freshly collected and frozen-thawed sperm cells also significantly decreased with time. Preliminary data also indicate sperm exposed to radiographic contrast media maintained their ability to penetrate and fertilize conspecific oocytes. These findings propose radiographic dye can be added to domestic cat spermatozoa with minimal reduction to sperm fitness, thus creating a foundation for developing improved transcervical insemination techniques for non-domestic felids. (poster)

A sperm viability test using SYBR-14/propidium iodide flow cytometry as a tool for rapid screening of primary ciliary dyskinesia patients and for choosing sperm sources for intracytoplasmic sperm injection

Spermatozoa viability tests based on dual-color flow cytometry after staining with Sybr-14/propidium iodide were performed on 44 men with complete asthenospermia for primary ciliary dyskinesia (PCD) screening, and seven were identified with PCD by electron microscopy of ultrastructural ciliary defects. Six PCD patients underwent eight intracytoplasmic sperm injection therapy cycles using ejaculated sperm or testicular sperm, obtaining a mean fertilization rate of 46.6%, with three healthy babies born and one in utero at the time of writing.

Analysis of sperm concentration and motility in a microfluidic device

A home-use device that allows rapid and quantitative sperm quality analysis is desirable but not yet fully realized. To aid this effort, this article presents a microfluidic device capable of quantifying sperm quality in terms of two critical fertility-related parameters—motile sperm concentration and motility. The microdevice produces flow field for sperms to swim against, and sperms that overcame the flow within a specified time are propelled along in a separate channel and counted via resistive pulse technique. Data are compared to two control methods clinically utilized for sperm quality exam—hemocytometer and the sperm quality analyzer. Results reveal the numbers of pulses generated by passage of sperms correlates strongly with the two control methods: pulse number from 0 to 335 corresponds to progressively motile sperm concentrations from 0 to 19 × 106/ml (hemocytometer) and Sperm Motility Index from 0 to 204 (sperm quality analyzer). The microdevice should be applicable to facilitate self-assessment of sperm quality at home.

Long-term ofloxacin testicular toxicity: an experimental study

The aim of this study was to assess the long-term toxic effect of ofloxacin on the testes and epididymides of 72 adult male albino rats. The rats were divided into group A and group B. Group A, which received ofloxacin for 14 days, was subdivided into two subgroups; LD-14 received low dose 72 mg KBW(-1) daily and HD-14 received high dose 216 mg KBW(-1) daily. Group B, which received ofloxacin for 28 days, was subdivided into two subgroups; LD-28 received 72 mg KBW(-1) and HD-28 received 216 mg KBW(-1) daily. Two matched control groups were followed up for 14 and 28 days respectively. The animals were evaluated for body weight, testicular weight, relative testicular weight, serum testosterone (T), epididymal sperm analysis (sperm count, motility, morphology, curvilinear velocity, linear velocity and linearity index) and testicular histopathology. The adverse effects of ofloxacin were correlated with increased treatment duration and/or dose. It is concluded that long-term ofloxacin has a direct detrimental effect on the testicles of albino rats at the studied doses and durations.

Effect of storage of domestic cat ( Felis catus) epididymides at 5 °C on sperm quality and cryopreservation

Postmortem sperm recovery from the epididymides may constitute a powerful tool for the conservation of valuable genetic material. The domestic cat ( Felis catus) is a good model for wild felids and, using this model, we have explored the effect of epididymides storage time on sperm motility and percentage of intact acrosomes upon sperm recovery and after cryopreservation. We also examined the effect of time of sperm equilibration with glycerol before freezing on sperm motility and the percentage of intact acrosomes. Motility varied between sperm recovered from epididymides that were stored for different times. Significant differences were seen in the sperm motility index (SMI) before freezing (55.91 ± 2.02, 48.21 ± 1.47, and 43.03 ± 1.32) and after thawing (51.81 ± 3.02, 41.90 ± 2.14, and 42.35 ± 1.95) of sperm recovered from epididymides stored for 0, 48, or 72 h, respectively. The percentage of intact acrosomes did not vary significantly with storage time (average 60.33 ± 1.38% before and 52.50 ± 1.91% after freezing, respectively). The percentage of normal sperm after different storage times did not differ (average 19.22 ± 1.25% normal sperm after recovery). When epididymides were stored for 72 h, time of sperm equilibration with glycerol (30 vs. 120 min) resulted in significant differences in both motility (SMI = 39.17 ± 2.76 and 45.00 ± 2.65, respectively) and the percentage of intact acrosomes (45.76 ± 4.91% and 60.67 ± 3.64%, respectively) after thawing. In conclusion, best results are achieved when sperm are recovered from epididymides within 24 h of cool storage and when they are equilibrated with glycerol during 120 min before freezing. The current results should be useful in the further development of techniques for the rescue and cryostorage of epididymal spermatozoa of endangered felids.

Retrospective Assessment of Potential Negative Synergistic Effects of Varicocele and Tobacco Use on Ultrastructural Sperm Morphology

To investigate, in a retrospective study, whether smoking cigarettes increases the effect of varicocele on sperm morphology. The semen quality of 2 groups of patients with varicocele were compared, those who smoked (n = 121) and those who did not (n = 158). The semen parameters were evaluated, and sperm morphology was assessed using transmission electron microscopy and quantitatively elaborated (fertility index, immaturity, necrosis, and apoptosis percentages). In the smoker and nonsmoker varicocele-associated cases, sperm motility and the results from transmission electron microscopy analysis were significantly impaired compared with controls. However, a nonsignificant difference was detected when the semen parameters were compared. Subsequently, we divided the patients into 4 groups: mild (> or = 1 but < or = 10 cigarettes/d), moderate (>10 but <20 cigarettes/d), and heavy (> or = 20 cigarettes/d) smokers and a group of randomly chosen nonsmoker patients with varicocele. The sperm motility, sperm concentration, and fertility index decreased and the percentage of sperm pathologic features increased as the number of cigarettes smoked daily increased. A detrimental effect of cigarette smoking (>10 cigarettes/d) associated with varicocele on sperm motility and morphology was observed. Because much of reduced fecundity associated with smoking may be reversed within 1 year of cessation, as reported in published studies, effective interventions targeted at helping patients quit smoking should be addressed for the benefit of general health and fertility.

Ghrelin enhances viability of rat spermatozoa during incubation at 37°C

Summary Antioxidant properties of ghrelin have been demonstrated in recent studies. In the present study, the effects of chronic administration of ghrelin on the motility and plasma membrane integrity of rat spermatozoa during incubation at 37oC were investigated. Thirty 45-day-old male Wistar rats were divided into control and treatment groups. Rats in the treatment group were daily injected subcutaneously with 1 nmol of ghrelin for 10 consecutive days and the control rats received normal saline. Sperm was collected after killing of rats on days 5, 15 and 40 after the last injection, and sperm characteristics were examined at 0, 3 and 5 h after incubation at 37oC. Mass motility and forward progressive movement of spermatozoa were significantly higher in ghrelin-treated animals at 3 and 5 h of incubation on day 5 (P<0.05). After 3 h of incubation on day 15, only mass motility was greater than that of the control group. Plasma membrane integrity was assessed by hypoosmotic swelling (HOS) “water test”. The mean value of HOS reacted spermatozoa was higher in the treatment group on days 5 and 15 during 0, 3 and 5 h of incubation (P<0.05). However, the percentage of HOS-positive spermatozoa was not significantly different on day 40 between groups. There was a high correlation at 3 and 5 h of day 5 between the forward progressive movement (r = 0.92 and 0.94, P<0.0001) as well as overall sperm motility (r = 0.78 and 0.81, P<0.01) with HOS test in the ghrelin-treated animals. These results can be attributed to the antioxidative effects of ghrelin on the rat sperm especially on its plasma membrane which probably protects the sperm plasma membrane against oxidative damage during incubation and causes subsequent significant increase in the HOS test results. This may result in higher sperm motility index during 5 h of incubation.

Lycopene supplementation in vitro can protect human sperm deoxyribonucleic acid from oxidative damage

To examine the effects of lycopene on human sperm motility and DNA damage. Prospective study. Andrology research laboratory. Twelve fertile donors. Preincubation of washed sperm suspensions with or without lycopene (2 or 5 micromol/L) followed by a 2-hour incubation with or without hydrogen peroxide (H2O2, 50 micromol/L). Assessments of sperm motility (percentage) and DNA fragmentation index (percent DNA fragmentation index) before and after incubation. Sperm motility (percentage) and sperm percent DNA fragmentation index. Incubation of spermatozoa with H2O2 resulted in a significant decline in mean (+/-SD) percent sperm motility (28%+/-13% vs. 73%+/-4%, respectively) and a significant increase in percent DNA fragmentation index compared with samples incubated without H2O2 (29.8%+/-39.4% vs. 11.1%+/-14.6%, respectively). Pretreatment of samples with 5 micromol/L lycopene resulted in a significantly lower percent DNA fragmentation index than samples incubated without lycopene (8.0%+/-7.9% vs. 29.8%+/-39.4%, respectively). However, lycopene did not protect spermatozoa from the decline in motility after H2O2 treatment. The data suggest that preincubation of spermatozoa with lycopene offers protection against oxidative DNA damage in vitro. These data also highlight the differential protective effects of lycopene on sperm motility and sperm DNA integrity.

Improved Quality of Cryopreserved Cheetah (Acinonyx jubatus) Spermatozoa After Centrifugation Through Accudenz

Sperm cryopreservation, in combination with assisted reproductive techniques, is a valuable tool for the genetic management of endangered felids. However, the acrosome of the cheetah spermatozoon is especially sensitive to cryopreservation, with approximately 40% of spermatozoa experiencing acrosomal damage immediately after thawing and then another approximately 15% loss during the next 4 hours in vitro. Additionally, thawing causes a reduction in sperm motility by approximately 20% with another decrease of approximately 12% during subsequent incubation in vitro. We hypothesized that slow removal of glycerol from cryopreserved cheetah spermatozoa using an Accudenz gradient would improve acrosomal integrity, sperm motility longevity, and structural morphology. Accudenz was compared with traditional cheetah sperm processing methods for glycerol removal that involves washing, multistep resuspension, and swim-up processing. Electroejaculates (n = 21 total from 8 males) were washed in Ham F10 medium, and sperm pellets were resuspended in TEST-yolk buffer with 0% glycerol. Samples were cryopreserved in straws in 4% final glycerol, thawed, and assessed for percent intact acrosomes (% IA), percent motility (% M), and forward progressive status (FPS; scale, 0-5). Sperm motility index (SMI) was calculated as (% M + [FPS x 20]) / 2. In study 1, glycerol removal by centrifugation through an Accudenz gradient (4%, 10%) was compared with traditional sperm washing (control) and multistep resuspension protocols. At each time after centrifugation (hourly for 4 hours), % IA was improved (P < .05) for Accudenz (range, 36%-39%) compared with control (30%-33%) and multistep (29%-33%) treatments. In study 2, a modified Accudenz protocol was compared with traditional washing and was found to improve (P < .05) SMI (range, 52-64) compared with controls (range, 41-52) at each time postthaw after centrifugation. In study 3, swim-up processed sperm were compared with those treated by centrifugation through Accudenz and traditional sperm washing for improving sperm morphology. The percentage of structurally-normal sperm recovered postthawing increased (P < .05) for both the Accudenz (38%) and swim-up (33%) treatments compared with controls (21%). Percent IA and SMI also were improved (P < .05) for Accudenz (range, 39%-47% and 46-59, respectively) compared with controls (range, 26%-33% and 40-53, respectively). Results indicate that using Accudenz for glycerol removal from cryopreserved cheetah sperm mitigates the significant loss in sperm quality that occurs after freeze-thawing. This alleviation of cellular damage resulting from cryopreservation contributes to a more than 10% improvement in overall sperm motility and, more importantly, allows retention of 40% or more of sperm with intact acrosomes.

Survival of Human Sperm Following Controlled- and Noncontrolled-Rate Cryopreservation/Die Überlebensrate menschlicher Spermatozoen nach einer kontrollierten und nicht kontrollierten Kryokonservierung

The purpose of this study was to develop a noncontrolled-rate (NCR) method of freezing semen in cryovials which could be used as a backup to controlled-rate (CR) freezing when the CR freezer was being utilized for freezing of human embryos or oocytes. Semen was frozen with three concentrations of glycerol (0%, 3.75%, 7.5%) in cryoprotective media in a CR freezer or using a newly developed NCR method of freezing. Semen was thawed in 40 degrees C water or in air at room temperature (RT). Controlled-rate freezing and 40 degrees C thawing resulted in significantly greater postthaw sperm motility and motility index compared with NCR freezing and RT thawing. Sperm velocity and linearity were not significantly effected by rate of freezing or thawing except when there was no glycerol present in the cryoprotective media. Postthaw motilities following CR-40 degrees C, NCR-40 degrees C, CR-RT and NCR-RT freezing-thawing were 25.6%, 24.8%, 21.6% and 18.3%, respectively. Because NCR freezing resulted in only a 0.8% decline in motility compared with CR freezing when semen was thawed at 40 degrees C it appears that this new NCR method of freezing can be successfully used as a backup to CR freezing when the CR freezer is being used for embryo or oocyte cryopreservation.

Assessment of Palomo's operative method for infertile varicocele.

Palomo's operative method was employed in the surgical treatment of 58 cases with infertile varicocele, and the operative method and therapeutic results were assessed. With the lapse of time after operation, statistically significant improvement was achieved in the sperm density and sperm motility. The post-operative impregnation rate was 55.2%. When the pre-operative sperm motility index had been 20% or less, the post-operative impregnation rate was 23%, which was significantly lower than the overall rate of 55.2%. Thus, the pre-operative sperm motility index is a good indicator of the post-operative capacity for impregnation. Even in patients with severe oligozoospermia (< 5 x 10(6) ml-1), a post-operative impregnation rate of 45.5% was obtained. Thus, their prognosis is by no means hopeless. Study of the cross-sectional profile of the internal spermatic vessels revealed the possibility of overlooking small internal spermatic venous branches. Therefore, Palomo's method was concluded to be excellent as an approach performed under direct visual observation. Palomo's method did not cause any complication of testicular atrophy and improved the fertility potential.

Associations between dietary patterns and semen quality in men undergoing IVF/ICSI treatment

BACKGROUND This study investigates whether dietary patterns, substantiated by biomarkers, are associated with semen quality. METHODS In 161 men of subfertile couples undergoing in vitro fertilization treatment in a tertiary referral clinic in Rotterdam, the Netherlands, we assessed nutrient intakes and performed principal component factor analysis to identify dietary patterns. Total homocysteine (tHcy), folate, vitamin B12 and B6 were measured in blood and seminal plasma. Semen quality was assessed by sperm volume, concentration, motility, morphology and DNA fragmentation index (DFI). Linear regression models analyzed associations between dietary patterns, biomarkers and sperm parameters, adjusted for age, body mass index (BMI), smoking, vitamins and varicocele. RESULTS The 'Health Conscious' dietary pattern shows high intakes of fruits, vegetables, fish and whole grains. The 'Traditional Dutch' dietary pattern is characterized by high intakes of meat, potatoes and whole grains and low intakes of beverages and sweets. The 'Health Conscious' diet was inversely correlated with tHcy in blood (beta = -0.07, P = 0.02) and seminal plasma (beta = -1.34, P = 0.02) and positively with vitamin B6 in blood (beta = 0.217, P = 0.01). An inverse association was demonstrated between the 'Health Conscious' diet and DFI (beta = -2.81, P = 0.05). The 'Traditional Dutch' diet was positively correlated with red blood cell folate (beta = 0.06, P = 0.04) and sperm concentration (beta = 13.25, P = 0.01). CONCLUSIONS The 'Health Conscious' and 'Traditional Dutch' dietary pattern seem to be associated with semen quality in men of subfertile couples.

Influence of the Prostatic Fluid from the First and Third Fractions of the Ejaculates on the Cryosurvival of Poodle Dog Sperm

Problem statement: Two experiments were conducted to determine the effect of the prostatic fluid from the first and third fractions of the ejaculates on the freezability of poodle dog sperm. Approach: The ejaculates from 2 trained poodle dogs were collected. Effect of prostatic fluid from the first and third fractions of ejaculates on cryosurvival of dog sperm was examined. SDS-PAGE analyses were performed to verify whether there were any modifications in the protein patterns of the first and third fractions. Results: When second fractions of Dog A sperm were combined with the prostatic fluid from the first and third fractions of the ejaculates, sperm motility index was decreased compared to second fractions were collected alone. The post-thaw motility of Dog A sperm was significantly reduced when prostatic fluid from the first and third fractions of the ejaculates was added. The motility of frozen-thawed Dog B sperm did not appear to be altered when it was diluted with either of the prostatic fluids. The major bands present in both prostatic fluids corresponded to proteins with molecular masses of 30 and 75 kDa. The amount of proteins in the prostatic fluid from the third fraction was twice-three times as much protein present compared to the first fraction. Conclusion: Collection of the sperm-rich fraction along with either the prostatic fluid from the 1st and 3rd fractions of the ejaculates during sperm processing has varying detrimental effects on the motility characteristics of frozen-thawed sperm.

Influence of the Prostatic Fluid from the First and Third Fractions of the Ejaculates on the Cryosurvival of Poodle Dog Sperm

Problem statement: Two experiments were conducted to determine the effect of the prostatic fluid from the first and third fractions of the ejaculates on the freezability of poodle dog sperm. Approach: The ejaculates from 2 trained poodle dogs were collected. Effect of prostatic fluid from the first and third fractions of ejaculates on cryosurvival of dog sperm was examined. SDS-PAGE analyses were performed to verify whether there were any modifications in the protein patterns of the first and third fractions. Results: When second fractions of Dog A sperm were combined with the prostatic fluid from the first and third fractions of the ejaculates, sperm motility index was decreased compared to second fractions were collected alone. The post-thaw motility of Dog A sperm was significantly reduced when prostatic fluid from the first and third fractions of the ejaculates was added. The motility of frozen-thawed Dog B sperm did not appear to be altered when it was diluted with either of the prostatic fluids. The major bands present in both prostatic fluids corresponded to proteins with molecular masses of 30 and 75 kDa. The amount of proteins in the prostatic fluid from the third fraction was twice-three times as much protein present compared to the first fraction. Conclusion: Collection of the sperm-rich fraction along with either the prostatic fluid from the 1st and 3rd fractions of the ejaculates during sperm processing has varying detrimental effects on the motility characteristics of frozen-thawed sperm.