Midpiece Defects Research Articles

Is male factor infertility rising? Another side of the equation from an in vitro fertilization clinic in southwestern Nigeria

Background: Male factors account for about half of the causes of infertility and a high premium is placed on childbirth and rearing in developing countries. With the introduction of assisted conception, the burden of care for managing male infertility has shifted to the female partner. Male infertility is generally evaluated via seminal fluid analysis which includes parameters such as concentration, motility, and morphology. The etiology of male infertility is multifactorial, and majority of cases are known to be idiopathic. Materials and Methods: A descriptive cross-sectional study of 77 male partners of infertile couples presenting to an in vitro fertilization clinic in Ibadan, Nigeria, between January 1 and June 30, 2019. Participants were selected consecutively at their first consultation visit. A self-administered questionnaire was completed by each participant comprising socio-demographic information, results of laboratory screening, and seminal fluid analysis. Data was analyzed using the Statistical Package for Social Sciences (SPSS, IBM, NY), version 22. Descriptive statistics were generated, and presented with the aid of a bar chart, pie chart, and frequency tables. Results: The average age of the participants was 43.64 ± 3.1 years and 97.4% (75) were in a stable relationship. Duration of infertility ranged between 3 and 17 years with an average of 7.6 ± 1.8 years. Human immunodeficiency virus, syphilis, hepatitis B, and hepatitis C were diagnosed in 3%, 4%, 5%, and 1%, respectively. Over half of the participants had semen volume and liquefaction time of 1-2 mL and 15-30 min, respectively. 34 men (44.1%) had normal semen parameters while the remaining 43 (55.9%) had various forms of abnormalities. Mid-piece defects were the most prevalent morphological defects and were found in 70 men (90.9%) while immature spermatozoa were found in 38 men (49.4%). Only one participant had azoospermia (1.3%). Conclusion: The impact of male factor infertility cannot be over-emphasized. Male contribution to infertility is thought to be rising and this study from male partners of infertile couples revealed semen abnormalities in over 50% of the participants. Addressing preventable causes of male infertility through health awareness and lifestyle modifications are important steps towards engendering optimal reproductive health.

Feeding yearling Angus bulls low-level ergot daily for 9 weeks decreased serum prolactin concentrations and had subtle effects on sperm end points

Our objective was to determine whether feeding yearling bulls with the higher recommended Canadian limit of ergot alkaloids (∼3 mg/kg dry matter intake, DMI) would affect sperm characteristics and plasma prolactin concentrations. Aberdeen Angus bulls (12–13 mo old, n = 7/group) allocated by blocking for sperm concentration and body weight, were fed placebo or ergot alkaloids in gelatin capsules (60 μg/kg body weight daily, 3.4 mg/kg of DMI) for 9 wk. Semen samples were collected weekly by electroejaculation and examined with a computer assisted semen analyzer (CASA) and flow cytometry, for the intervals 5 wk before (Pre-exposure period), 9 wk during (Exposure period) and 9 wk after (Post-exposure period) treatment. Weekly plasma samples were analyzed for prolactin by radioimmunoassay. Plasma prolactin concentrations decreased markedly (mean ± SEM, 16.74 ± 3.70 in Exposure and 33.42 ± 3.08 ng/mL in Post-Exposure periods; P < 0.01) compared to Control (67.54 ± 21.47 and 42.59 ± 15.06 ng/mL). Treatment did not affect (P ≥ 0.17) body weight gain, sperm concentration, sperm count/ejaculate, motility or percent live sperm. Averaged over the exposure and post-exposure durations, the scrotal circumference was smaller (P = 0.02) by 2.7% in the Ergot group. Progressive motility remained unchanged from 59.92 ± 2.31% in Exposure to 59.61 ± 2.59% in Post-Exposure periods, compared to marked increase in Control (61.42 ± 1.60% to 67.52 ± 1.47%; P = 0.02). Straight-line sperm velocity decreased (−3.15 ± 1.53 μm/s) from exposure to post-exposure periods in Ergot group (P = 0.04) versus an increase (2.96 ± 2.17 μm/s) in Control. Midpiece defects decreased from Exposure to Post-exposure periods in Control group but remained unchanged in Ergot group (trt∗age, P < 0.01). Ergot feeding resulted in a smaller proportion of sperm with medium mitochondrial potential (Ergot: 22.65 ± 0.98%, Control: 24.35 ± 1.05%, P = 0.04). In conclusion, feeding ergot at Canadian permissible limit for 9-wk resulted in a 4-fold decrease in plasma prolactin concentrations. Semen end points were not significantly affected, although there were subtle effects on progressive motility, midpiece defects and mitochondrial membrane potential. Clinical relevance of observed changes requires further evaluation. Results supported our hypothesis that prolonged low-level ergot will adversely affect plasma prolactin. However, semen parameters were partially affected, supporting similar work on fescue toxicosis.

Acute mild heat stress alters gene expression in testes and reduces sperm quality in mice

Heat stress is a major concern in animal reproduction, as testicular temperature must be 3–5 °C below body core temperature for production of motile and fertile sperm in mammals. Although recent studies concluded that increased temperature per se was the underlying pathophysiology of testicular impairment, more studies are required to better understand the mechanisms. Therefore, our objective was to investigate the impacts of mild acute heat stress on sperm and testes, and based on mRNA, elucidate involvement of StAR, Trp53 and Trp53-dependent intrinsic and extrinsic apoptotic pathways in pathophysiology of testicular heat stress. Forty-eight C57 BCL6 elite male mice were equally allocated into six groups, anesthetized and the distal third of their body immersed in a water-bath at 40 or 30 °C (heat treatment and control, respectively) for 20 min. Intervals from heat exposure (Day 0) to euthanasia were: 8 and 24 h and 7, 14 and 21 d (plus a control group at 14 d). The epididymides were excised, minced and placed in Tyrode albumin lactate pyruvate hepes (TALPH) at 37 °C for 15 min to recover sperm. Based on computer assisted sperm analysis (CASA), heat treatment reduced total and progressive motility ∼40% (P < 0.05) on Days 14 and 21. Furthermore, percentage morphologically normal sperm was significantly decreased on Day 7, with greater reductions on Days 14 and 21, mostly due to increased midpiece defects. Acrosome integrity (FITC PSA) was decreased ∼35% at 8 h (P < 0.05) and reached a nadir on Day 14. There were decreases (P < 0.05) in seminiferous tubule diameter and testicular weight (relative to body weight) on Day 14. Testicular RNA was extracted, reverse-transcribed and cDNA used for PCR. Expression of genes Hspa1b (Hsp70) and Gpx1 had 7- and 10-fold increases (P < 0.001 for each) at 8 and 24 h, respectively, with Hspa1b remaining upregulated at 24 h, whereas StAR peaked at Day 14 (15-fold, P < 0.0001) and had returned to baseline on Day 21. Both Trp53 and Casp8 were upregulated (P < 0.05) on Day 14, whereas Bcl-2 was decreased (P < 0.05) on Days 7 and 14. In conclusion, acute mild heat stress severely reduced sperm quality and based on mRNA, there was upregulation of chaperone and antioxidant systems and Trp53-dependent intrinsic and extrinsic apoptotic pathways, with deleterious effects on sperm, spermatocytes and spermatids. These findings provided insights into the pathophysiology of heat stress and should contribute to development of evidence-based approaches to mitigate effects of testicular heating.

First Evaluations and Cryopreservation of Semen Samples from Sunda Clouded Leopards (Neofelis diardi).

Simple SummaryThe Sunda clouded leopard is a medium-sized cat from Borneo and Sumatra. The captive population has less than 20 individuals on both islands. Conservation breeding of this species has been unsuccessful due to the small numbers of individuals and the difficulty of pairing them. The present study aimed at evaluating for the first time the reproductive organs, semen characteristics, and sperm freezing for two wild-caught males (8 and 11 year old) living singly. The morphometric and ultrasound examination showed some degree of abnormalities in both leopards. Semen evaluation obtained by electro-ejaculation revealed a high proportion of abnormal sperm cells (similar to mainland clouded leopards). While only the younger male produced samples that could be frozen, post-thaw evaluations showed that the few surviving spermatozoa could potentially be used for in vitro fertilization or sperm injection. Studies on more individuals are needed to validate those encouraging but preliminary results.A better understanding of semen characteristics and resilience to freezing temperatures is necessary before developing assisted reproductive techniques and systematic biobanking for the Sunda clouded leopard. The objective of this study was to evaluate for the first time the semen and sperm quality (in fresh and frozen samples) of two captive Sunda clouded leopards in Malaysia. A total of 17 examinations of the reproductive tract (using ultrasonography) and electro-ejaculations were performed on the two leopards over a 2-year period. Samples obtained from Leopard 1 (8 years old) varied in terms of volume (402 ± 92 µL), pH (7.9 ± 0.9), sperm motility (54.5 ± 24.2%), sperm concentration (122.4 ± 84.7 × 106 sperm/mL), normal morphology (23.9 ± 12.3%), and viability (55.2 ± 18.2%). Midpiece defects represented the most common structural abnormality followed by abnormal tail and head defects. Samples from Leopard 2 (11 year old with abnormal testicular tissue) were of lesser quality. Two frozen semen samples from Leopard 1 were thawed and examined for acrosome integrity. Post-thawed samples contained <10% of motile spermatozoa but almost 50% of abnormal acrosomes. The present results emphasized the high incidence of structurally-abnormal spermatozoa, similar to the mainland clouded leopard. Post-thaw evaluations showed that the few surviving spermatozoa could potentially be used for in vitro fertilization or sperm injection. However, more individuals must be studied to validate those first findings that are exciting but still preliminary.

Impact of sperm characteristics on time-lapse embryo morphokinetic parameters and clinical outcome of conventional in vitro fertilization.

Sperm abnormalities may negatively affect embryo development. To determine the influence of sperm abnormalities (morphology, motility, DNA fragmentation) on embryo morphokinetic variables and clinical outcome of conventional IVF. Participants were 86 couples undergoing in vitro fertilization (IVF). Sperm morphology was evaluated according to the strict criteria proposed by Kruger/Tygerberg. CASA system was applied for sperm motility assessment. Sperm DNA fragmentation was assessed by the chromatin structure assay (SCSA). Morphokinetic parameters were determined in 223 embryos obtained from conventional IVF only and cultured in a single-step medium using time-lapse imaging technology. Time-lapse variables from the initial embryo development, such as time of pronuclei fading (tPNf) and time for two cells (t2), were those more strongly related with abnormalities of sperm motility, morphology, and DNA fragmentation. Sperm morphological abnormalities rather than sperm motility were more closely associated with embryo morphokinetics. Sperm head defects were mainly correlated with the last stages of embryonic development (t9 to tHB), sperm midpiece defects with intermediate cleaving embryos (t5-t9), and sperm tail defects with the initial stages of embryonic development (tPNa-t4). Excess residual cytoplasm was positively correlated with all embryo morphokinetic parameters except t2 and tM. Absence of acrosomes, pinheads, coiled tails, and multiple sperm morphological defects correlated negatively with time-lapse embryo morphokinetic variables. A large number of sperm-related variables, including frequency of specific morphological defects, morphological indexes, DNA fragmentation and motility, and time-lapse embryo variables, such as time intervals based mainly of 15 time points were recorded. There were strong associations between specific sperm defects of the head, midpiece, and tail with certain stages of embryonic development from observation of pronuclei to the hatched blastocyst. Coiled tail, cumulative head defects, and multiple abnormalities index (MAI) were associated both with embryo morphokinetics and the implantation success.

25 A preliminary study of the effects of breed and nanowater as extender diluent on ram semen characteristics post-thawing

This study examined the effect of genotype and extender diluent on the characteristics of frozen-thawed ram semen. Twenty ejaculates collected from 10 rams aged 4-12 years (2 Polish Lowland (PON), 4 synthetic line BCP (Berrichon du Cher×Charolais×PON/Polish Merino), and 4 SCP (Suffolk×Charolais×PON/Polish Merino) in mid-breeding season were divided into six equal portions each and frozen in semen extender prepared by mixing commercial Triladyl extender (MiniTub GmbH) with deionized water (Aqua Purificata, Prolab) or nanowater (NW; deionized water declusterized in a cold plasma generator for 15, 30, 45, 60, or 90 minutes (15’, 30’, 45’, 60’, or 90’, respectively); Nanotechnology Systems) and Gallus domesticus egg yolk (1:3:1 vol/vol/vol). All semen samples were evaluated for progressive motility (Sperm Class Analyzer) and sperm morphology (Nikon Eclipse 80i microscope, Nikon Corp.). In addition, the ex situ survival time at 37°C and extender content of ALT (alanine transferase), ALP (alkaline phosphatase), and aspartate aminotransferase were measured, and the numbers of apoptotic, necrotic, and live spermatozoa were determined post-thawing with flow cytometry (BD Accuri™ C6 Plus, Becton Dickinson). Data were analysed by two-way analysis of variance and least significant difference test (SigmaPlot, Systat Software Inc.). The PON exceeded (P&amp;lt;0.05) SCP rams in the occurrence of mid-piece and double tail defects using NW30’ and NW45’ extenders. The proportion of spermatozoa with proximal droplets was greater (P&amp;lt;0.05) in PON than in SCP or BCP rams with NW15’ and NW30’ extenders, respectively, but the percentage of sperm with detached heads was greater (P&amp;lt;0.05) in BCP and SCP compared with PON using NW15’ extender. Overall, spermatozoa in PON rams’ semen had survival times greater by 25-50min (P&amp;lt;0.05) compared with those for semen from BCP and SCP synthetic lines. The ALT levels were significantly elevated in SCP compared with PON and BCP rams only in NW90’ extender. The numbers of necrotic spermatozoa were greater (P&amp;lt;0.05) in SCP and BCP compared with PON rams, and the numbers of apoptotic spermatozoa were greater (P&amp;lt;0.05) in BCP than in SCP rams with NW45’, NW60’, and NW90’ extenders. Live spermatozoa were more prevalent (P&amp;lt;0.05) in PON than in BCP rams in NW45’ extender post-thawing. To summarise, semen from PON rams had higher survival times and fewer spermatozoa with detached heads but more mid-piece defects and sperm with double tails post-thawing compared with BCP and SCP breeds. The number of necrotic cells was greater in semen from BCP and SCP rams compared with PON. It can be concluded that breed-related differences in post-thaw sperm morphology were apparent in extenders containing NW15-60’, whereas the variations in cell necrosis/apoptosis were primarily seen with the use of NW45-90’ extenders.

39 Nanowater enhances cryoprotective effects of glycerol during ram semen freezing

It was suggested that unique physicochemical properties of nanowater (NW, water declustered in the cold plasma generator), such as a low dielectric constant and high diffusivity, might ameliorate ram semen freezing by improving bioavailability of extender constituents. This study was undertaken to evaluate the effects of NW added to glycerol-containing semen extenders on the characteristics of frozen-thawed ram semen. A total of 18 ejaculates collected from 6 Olkuska breed rams during the breeding season were divided into six equal portions. The ejaculates were frozen in the modified fructose-skimmed milk-egg yolk Kareta extender containing 3 or 7% glycerol and diluted in deionized water (control extenders; Aqua Purificata, Prolab; C3% and C7%, respectively) or NW (declustered for 15min (NW15’) or 30min (NW30’); Nanotechnology Systems) to a final concentration of 800×106 spermatozoa/mL. They were placed in 0.25-mL plastic straws and frozen in liquid nitrogen. The two declusterization times were chosen on the basis of previous laboratory tests and fertility trials yielding the best results in terms of semen quality post-thawing and pregnancy rates after AI. All semen samples were evaluated for sperm concentration, progressive motility (Sperm Class Analyzer), and morphological defect rates (Nikon Eclipse 80i microscope, Nikon Corp.). In addition, the ex situ survival time at 37°C and extender content of alanine transferase, alkaline phosphatase, and aspartate aminotransferase were measured, along with the proportions of apoptotic, necrotic, and live spermatozoa determined post-thawing with flow cytometry (BD Accuri™ C6 Plus, Becton Dickinson) at 0 and 1h post-thawing. Data were analysed by one-way analysis of variance and Holm-Sidak method using SigmaPlot statistical software (Systat Software Inc.). The proportion of spermatozoa with mid-piece defects was significantly (P&amp;lt;0.05) reduced in NW15’-3% compared with C3% (1.3±0.3% vs. 4.0±06%; mean±s.e.m.). The proportion of necrotic spermatozoa 1h after thawing was greater (P&amp;lt;0.05) in C7% compared with NW30’-7% (20.7±0.4% vs. 17.6±1.0%), whereas the proportions of live cells detected immediately (0h) and 1h after thawing were greater (P&amp;lt;0.05) in NW30’-7% than in C30’ (0 h: 54.0±0.6% vs. 50.4±1.2%; 1 h: 54.4±1.2% vs. 59.2±0.4%, respectively). The mean survival time of spermatozoa was greater (P&amp;lt;0.05) in extenders dissolved in NW30’compared with their respective controls (NW30’-3% vs. C3%: 215±4min vs. 189±6 min; and NW30’-7% vs. C7%: 242±8min vs. 217±4min, respectively). Alkaline phosphatase concentrations in extenders prepared with NW30’ were lower (P&amp;lt;0.05) compared with the control groups (NW30’-3% vs. C3%: 3105±231IU vs. 4490±458 IU; and NW30’-7% vs. C7%: 2516±128IU vs. 3956±263IU, respectively). Our results indicate that NW significantly improves the effects of glycerol on ram sperm viability post-thawing, with the reduction in sperm necrosis/overall enhancement of sperm survivability being greater with 7% compared with 3% of glycerol in semen extender.

138 Associations of sperm head morphometrics with quality parameters of frozen-thawed ram semen

Current methods of mammalian semen evaluation focus on determining spermatozoa motility, concentration, mitochondrial status, and nucleus or chromatin structure integrity, quantifying their ability to bind to ova or measuring seminal plasma content of various biochemical markers. However, there is a paucity of studies that address relationships between sperm head morphometry (the external shape and dimensions of the sperm) and fertilising ability. Sperm head morphometrics are influenced by many molecular and biochemical factors such as genetics, DNA or protein condensation, and cell membrane permeability, all of which can affect semen viability. The objective of this experimental work was to determine quantitative correlations between sperm head dimensions and various indices of sperm quality in frozen-thawed ram semen. Ejaculates were collected from 16 clinically healthy rams (4 Polish Lowland (PON), 4 Olkuska, 5 synthetic line BCP (Berrichon du Cher×Charolais×PON/Polish Merino), and 3 synthetic line SCP (Suffolk×Charolais×PON/Polish Merino) aged 4-12 years) into an artificial vagina in the middle portion of the breeding season. Ejaculates from each ram were divided into two equal portions, diluted with a commercial semen extender prepared in deionised water or nanowater (water declusterised using cold plasma treatment) to a final concentration of 400×106 spermatozoamL−1, and frozen in 0.25-mL plastic straws. After 6 months of being cryogenically preserved, semen samples were thawed and used for the preparation of smears stained with eosin or SpermBlue. Images of the samples containing at least 100 spermatozoa were taken under 200× magnification and used for determination of sperm head morphology with the image analytical software Image Pro Plus (Media Cybernetics Inc.). Sperm progressive motility and survival time, as well as extender concentrations of alkaline phosphatase and aspartate aminotransferase, were measured. Finally, 128 BCP ewes were inseminated laparoscopically with the ram semen and fertility parameters were recorded. The present data were analysed using a multivariate analysis of variance in SAS (SAS Institute Inc.) and Spearman correlation tests. There were no significant effects or interactions of breed, staining method, or extender diluent on sperm head dimensions (head length, width, area, perimeter, and roundness). The mean head length was negatively correlated (P&amp;lt;0.05) with the percentages of spermatozoa with vacuolated, detached, or amorphous heads or small acrosomes; thick and thin midpiece defects, distal droplet, broken tail plus distal droplet, short tail plus distal droplet, and thick midpiece plus proximal droplet; and sperm progressive motility. In addition, sperm head roundness was negatively correlated with the proportion of spermatozoa with coiled tails. There were no correlations of sperm head dimensions with survival time, alkaline phosphatase and aspartate aminotransferase concentrations, or conception and pregnancy rates of artificially inseminated ewes. Sperm length and roundness (but no other measurements) were significantly correlated with segmental sperm defects and motility that may impinge the fertilising ability of frozen-thawed ram semen.

Use of testicular tone and echogenicity in identifying boars with low sperm quality

ABSTRACT This study aimed to evaluate the use of some testicular traits to identify boars with low sperm morphological quality. The consistency (scores from 1 to 5), tone with tonometry (mm), parenchyma echogenicity and heterogeneity with ultrasound (pixel) were assessed in 402 mature boars (18.5 months on average). Sperm abnormality thresholds (≤ 25% of total sperm abnormalities, ≤ 5% of heads, acrosome, neck or midpiece defects, ≤ 10% tail defects, and 15% cytoplasmic droplets) were used to classify boars as approved or disapproved. Three classes of testicular traits were formed (extremely low and high values, approximately 15% each, were kept in separated classes). When the traits were individually evaluated, fewer boars were approved if the echogenicity or heterogeneity were high, or if the tone was rigid. When evaluated in combination, the interaction between heterogeneity and tone, and between heterogeneity and echogenicity were significant. The high heterogeneity combined with soft tone or with hypo-echogenicity resulted in lower approval of boars. Tonometry and ultrasonography have a moderate potential to be included in breeding soundness examination of boars. When combined, they provide more reliable information about the impact of testicular parenchymal alterations on morphology of sperm cells.

Disruption of bovine sperm functions in the presence of aplastic midpiece defect.

Bulls are of great importance in the productive chain and for this reason they should have a good semen quality. There is no doubt that sperm morphology is very important to bull fertility, although little is known about how exactly the abnormal morphologies may affect sperm functions. To detail the morphological description of the aplastic midpiece defect (AMD), as well as to understand its consequences for male fertility based on membrane and acrosome status, mitochondrial membrane potential and DNA integrity parameters. The bulls were divided into two groups: control, consisting of satisfactory potential breeders (n=3); and AMD, consisting of unsatisfactory potential breeders with a high percentage of AMD (n=3). Bulls were evaluated by the breeding soundness evaluation; five ejaculates were collected from each animal and analyzed by flow cytometry. Spermatozoa from AMD group exhibited lower sperm motility and vigor (p<0.05). In addition, it also exhibited lower mitochondrial membrane potential (p<0.05), a higher percentage of spermatozoa with DNA fragmentation (p<0.05), lower acrosome and plasma membrane integrity (p<0.05), and higher lipid bilayer sperm membrane disorganization (p<0.05) in comparison with control bulls. These findings may be due to oxidative stress and a reduction of the energy production capacity in addition to an alteration in the structural composition of the sperm cell. Moreover, semen with a high percentage of AMD may also be undergoing apoptosis. Bulls with a high percentage of AMD in their semen are not suitable for reproduction. Furthermore, it suggests there is a putative genetic basis for this sperm defect.

Storage of boar semen at 16-18 °C in the long-term commercial extender prepared with deionized water or nanowater.

The aim of this study was to assess the usefulness of nanowater (NW; water declusterized using cold plasma treatment) as a diluent for a commercial boar semen extender during the 15-day storage (Days 1 to 15) at 16-18 °C. Ejaculates collected from 8 boars were subjected to the standard evaluation and then diluted in the extender prepared with deionized water (DW) or NW to a final concentration of 3×109 spermatozoa/ml. The proportion of defective spermatozoa increased (P<0.05) from Day 10 to Day 15 of storage (22.8±16.6% to 41.8±26.4% in DW group and 18.6±11.7% to 34.8±25.4% in NW group) and it was significantly greater in DW group compared with NW group on Days 5 and 10 due mainly to a greater (P<0.05) number of mid-piece defects in semen stored in the DW-containing extender. Sperm progressive motility decreased (P<0.05) in both groups between Days 2 and 6, Days 6 and 10, and Days 10 and 12, whereas the percentage of motile spermatozoa declined (P<0.05) to Day 14 only in NW group. Sperm motility was greater (P<0.05) in NW group compared with DW group from Day 5 to Day 13. A decline in sperm progressive motility below 40% in all semen samples occurred by Day 11 in DW group and by Day 12 in NW group. The mean survival time of sperm at 37 °C ex situ was greater in NW group than in DW group on Day 5 (314±87 min compared with 284±87 min) and Day 10 (223±34 min compared with 182±27 min; NW group compared with DW group, respectively). There were no differences (P>0.05) between the two groups in the concentrations of alkaline phosphatase and aspartate aminotransferase in semen extender. To summarize, the use of NW as an extender diluent exerts cytoprotective effects on boar spermatozoa and delays a decline in sperm progressive motility.

Daily administration of a GnRH analogue enhances sperm quality in bucks during the non-breeding season

The aim of this study was to determine if daily administration of a GnRH analogue (buserelin acetate) to bucks during the non-breeding season increases testosterone concentration and improves sperm quality. Five bucks received a daily dose of buserelin for 10 days, starting on Day 0 (first administration), and another five bucks remained as controls. Testosterone concentrations were greater in treated than in control bucks during the first hours after buserelin administration (P = 0.05), but greater in controls 10 h later (P < 0.01). Sperm mass motility and percentage of motile sperm were greater in treated (3.9 ± 0.6 and 70.1 ± 7.9%, respectively) than in control bucks (1.0 ± 0.6, P < 0.01; 45.0 ± 7.9%, P < 0.05 respectively) on Day 4. Percentage of sperm with normal morphology tended to be greater in treated than in control bucks (81.8 ± 6.2% compared with 63.5 ± 6.4% respectively, P = 0.08). The treatment decreased the percentage of sperm with mid piece defect and with bent tail (7.0 ± 1.5% compared with 12.0 ± 1.5%; 8.0 ± 1.7% compared with 13.5 ± 1.7%, treated and control bucks, respectively, P = 0.05 for both). The square root percentage of sperm with loose but heads with normal structures tended to be less in treated than control bucks (1.3 ± 0.3% compared with 0.4 ± 0.3% respectively, P = 0.06). It was concluded that daily administration of buserelin during the non-breeding season led to a rapid increase in testosterone concentration and improved sperm quality.

The Abnormality of Spermatozoa Goat after Freezing on -80°C Using Tris Diluent Added Combination Hatching Egg Yolk and Amnion Fluid

This study aims to determine the combination of the hatching egg yolk (HEY) and amniotic fluid (AF) in Tris diluent to the type of goat spermatozoa abnormalities after frozen at temperature -80°C. This study used four treatments and 6 replications. The semen is diluted using tris extender and added treatment, there are control group (T 0 ) (Tris egg yolk 20%), T 1 (15% HEY + 5% AF), T 2 (10% HEY + 10% AF), and T3 (5% HEY + 15% AF). After dilution, the semen was equilibrated for 2 hours, then stored using Mr. Frosty and frozen at temperature -80°C in the ultra-freezer for 24 hours. The type of spermatozoa abnormalities type was evaluated on fresh semen and after thawing. The data were analyzed using ANOVA (P<0.05). The results showed that there is no significant difference among treatments. The primary abnormalities in fresh semen, there were macro-cephalic, amorphous, and the secondary spermatozoa abnormalities such as detached head, mid-piece defect, coiled tail, loosehead, shoe-hook tail, and a broken tail. The secondary abnormalities in post-thawing were found both in the control group and treatment, which are detached head, loosehead, coiled tail, shoe-hook tail, and broken tail. The conclusion of this study was the percentage of secondary abnormalities after thawing was increasing both in the control and treatment groups. Keywords: Amniotic Fluid, Hatching Egg Yolk, Mr. Frosty , Type of Spermatozoa Abnormality.

Variables affecting semen quality and its relation to fertility in the dog: A retrospective study

Although semen analysis is routinely used to evaluate the male reproductive potential in the canine species, only a few authors have attempted to relate semen characteristics to donor traits, such as age or weight, and fertility. This study evaluated the effects of dogs' age (young: 0–24 months, adult: 25–84 months, old: >84 months), size (small: <15 kg, medium: 16–40 kg, large: >40 kg) and reason for semen collection (evaluation, artificial insemination, cryopreservation or research purposes) on the seminal characteristics and related the main seminal characteristics with fresh AI results. Overall, 251 semen examinations were performed from 140 dogs presented between 2000 and 2015 at the University of Pisa Veterinary Teaching Hospital. Conventional semen parameters (semen volume, sperm concentration and total number, subjective motility and sperm morphology) were evaluated. Overall, 227 (90.4%) samples contained spermatozoa. Azoospermic samples resulted from complete (n = 13, 9 dogs), or incomplete ejaculations (n = 11, 8 dogs), based on seminal alkaline phosphatase concentration and/or ancillary examinations. Reason for semen collection had a significant effect on semen volume, sperm total number and sperm concentration. The proportion of normal spermatozoa was lower in dogs presented for semen evaluation (P < 0.05). Dog size affected ejaculate volume and total sperm number, which was lower in small dogs compared to medium (P < 0.01 and P < 0.05, respectively) and large-sized dogs (P < 0.01). Age had no influence on volume, sperm number or motility. The proportion of normal spermatozoa was higher in young animals than in old ones (P < 0.05). There was a significantly higher proportion of midpiece defects in old dogs compared to young ones (P < 0.01). Total sperm number, motility and proportion of morphologically normal spermatozoa in the semen used in AIs resulting in a pregnancy were 627.6 × 106, 83.9% and 64.9%, respectively, all significantly higher than in the unsuccessful AIs, where they were 389.4 × 106, 66.5% and 42% (P < 0.05). In conclusion, age, size and reason for semen collection may affect main sperm parameters and must be taken into consideration during the interpretation of a routine semen evaluation. In turn, altered semen parameters may affect fertility after artificial insemination.

The impact of Corynebacterium glucuronolyticum on semen parameters: a prospective pre-post-treatment study.

Corynebacterium glucuronolyticum (C.glucuronolyticum) is a rare isolate that is only recently being acknowledged as a potential urogenital pathogen. The bibliographical references on this bacterial species are scarce, and its influence on all semen parameters was hitherto unknown - therefore, the aim of this study was to evaluate its effects on a range of sperm quality parameters. A prospective approach to compare semen parameters before and after treatment was used in this study. C.glucuronolyticum in semen specimens was identified using analytical profile index biotyping system (API Coryne) and additionally confirmed by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS), with the determination of antimicrobial susceptibility by Kirby-Bauer method. Semen analysis was performed according to the criteria from the World Health Organization (with the use of Tygerberg method of sperm morphology categorization). Very strict inclusion criteria for participants also included detailed medical history and urological evaluation. From a total of 2169 screened semen specimens, the inclusion rate for participants with C.glucuronolyticum that satisfied all the criteria was 1.01%. Antibiogram-guided treatment of the infection with ensuing microbiological clearance has shown that the resolution of the infection correlates with statistically significant improvement in the vitality of spermatozoa, but also with a lower number of neck and mid-piece defects. Parameters such as sperm count, motility and normal morphology were not affected. In addition, susceptibility testing revealed a trend towards ciprofloxacin resistance, which is something that should be considered when selecting an optimal treatment approach. Albeit it is rarely encountered as a monoisolate in significant quantities, C.glucuronolyticum may negatively influence certain sperm parameters; therefore, it has to be taken into account in the microbiological analysis of urogenital samples.

Exposure to Leather Tanning Factories Lowers Semen Quality in Mongolian Men

Objectives: To examine the difference in semen qualities of male workers in leather tanning factories and of those in other industries. Methods: Eighty reproductive aged male workers from six leather tanning factories and eighty-five from non-tanning workplaces participated in the study as the exposed subjects and non-exposed subjects, respectively. Each subject was given a physical examination, required to complete a questionnaire, and asked to supply a semen specimen. Semen analysis on fresh specimens followed the latest World Health Organization guideline. Semen parameters were compared between the exposed and non-exposed subjects using tabulation and chi-square test for categorical variables and t-test or rank-sum test for continuous variables. Multiple linear or logistic regression models were constructed to examine the independent effects of exposure. Results: After adjusting for confounding variables, exposed subjects had significantly higher percentages of grade C motility (p=0.008), mid-piece defect (p=0.009), and tail-defect (p=0.029) in their spermatozoa compared to non-exposed subjects. Liquefaction time was also longer (p=0.046). Conclusion: Leather tanning workers have significantly higher percentages of grade C motility, mid-piece defect, and tail-defect in their spermatozoa.

Hyperthermia and not hypoxia may reduce sperm motility and morphology following testicular hyperthermia

The mammalian testis typically operates on the brink of hypoxia; the long-standing dogma is that increased testicular temperature increases metabolism, but blood flow is unaffected and the resulting hypoxia reduces sperm motility and morphology. In rats and mice, oxygen (O&lt;sub&gt;2&lt;/sub&gt;) content of inspired air affected O&lt;sub&gt;2&lt;/sub&gt; content of testes, enabling the latter to range from approximately 50 to more than 200% of physiologic concentrations. A ram model was used to test the hypotheses that hypoxia would disrupt sperm motility and morphology and that hyperoxia would prevent hyperthermia-induced reductions in sperm motility and morphology. Eighteen Canadian Arcott rams (approximately 10 months old) were used in a 2 × 3 factorial, with factors being scrotal insulation (insulated or not insulated) and O&lt;sub&gt;2 &lt;/sub&gt;concentrations in inspired air (14, 21 or 85%). Six rams, three with and three without scrotal insulation, were placed in each of three enclosed areas for 30 h to expose them to their respective oxygen concentrations, with scrotal insulation removed at the end of the exposure. Semen was collected by electro-ejaculation twice before insulation, bi-weekly for four weeks starting one week after exposures, and then once weekly for two weeks. There were effects of insulation, time and an insulation × time interaction for motile sperm and sperm that had normal morphology or head or midpiece defects (P &amp;lt; 0.01 for each). Sperm motility and morphology exhibited alterations between approximately two and five weeks after insulation, with mean progressively motile and morphologically normal sperm decreasing from approximately 55 to 35% and from 80 to 30%, respectively, and with head and midpiece defects increasing from approximately 3 to 50% and from 10 to 20% (P &amp;lt; 0.05 for each). The hypotheses that hypoxia would disrupt sperm quality and production, whereas hyperoxia would prevent hyperthermia-induced reductions in sperm quality and production, were not supported. This is apparently the first report that heat-stress induced damage to sperm was due to increased temperature per se and not testicular hypoxia, calling into question a long-standing paradigm.

Study on the short-term effects of increased alcohol and cigarette consumption in healthy young men\u2019s seminal quality

Many studies have reported a negative impact of lifestyle factors on testicular function, spermatozoa parameters and pituitary-gonadal axis. However, conclusions are difficult to draw, since studies in the general population are rare. In this study we intended to address the early and late short-term impact of acute lifestyle alterations on young men’s reproductive function. Thirty-six healthy male students, who attended the Portuguese academic festivities, provided semen samples and answered questionnaires at three time-points. The consumption of alcohol and cigarette increased more than 8 and 2 times, respectively, during the academic festivities and resulted in deleterious effects on semen quality: one week after the festivities, a decrease on semen volume, spermatozoa motility and normal morphology was observed, in parallel with an increase on immotile spermatozoa, head and midpiece defects and spermatozoa oxidative stress. Additionally, three months after the academic festivities, besides the detrimental effect on volume, motility and morphology, a negative impact on spermatozoa concentration was observed, along with a decrease on epididymal, seminal vesicles and prostate function. This study contributed to understanding the pathophysiology underlying semen quality degradation induced by acute lifestyle alterations, suggesting that high alcohol and cigarette consumption are associated with decreased semen quality in healthy young men.

Effect of Lifestyle Factors on Semen Quality

Effect of Lifestyle Factors on Semen Quality

Physical and morphological characteristics of the first three jets of Pêga jackasses sperm-rich fraction

ABSTRACT The first three jets of the sperm-rich fraction of Pêga jackasses were collected and assessed separately. Five fertile Pêga jackasses were used as semen donors and underwent fractionated semen collection, using an open model artificial vagina. The first three jets of the semen were collected separately and assessed for volume, sperm motility, vigor, concentration/mL of semen, and sperm morphology. These characteristics were compared between first, second and third jets and between jackasses. It was observed that the jet volume differed (P&lt;0.05) between jackasses, although it was similar (P&gt;0.05) between first, second and third jets. Sperm motility did not differ (P&gt;0.05) between jets and jackasses. Vigor was similar (P&gt;0.05) between jets of the same jackass, and only the first jet differed (P&lt;0.05) between jackasses. The first, second and third jets of the sperm-rich fraction had decreased sperm concentrations (P&lt;0.05) of 955.56, 725.56 and 280.56x 106 sperm/mL of semen, respectively. Sperm morphology differed between the first three jets only for the incidence of mid-piece defect, higher in the third one (4.26%), compared to the first (3.36%) and second (3.38%) ones. When comparing the morphological characteristics of the sperm-rich fraction between five jackasses, regardless of the jet, there were differences in the percentage of normal sperm, proximal cytoplasmic droplet, mid-piece and head defects.