Semen Plasma Research Articles

Cannabis consumption might exert deleterious effects on sperm nuclear quality in infertile men.

Can cannabis consumption alter sperm nuclear integrity in infertile men? A retrospective cross-sectional study conducted between July 2003 and December 2013, which included 54 men who consulted for male-factor infertility. Twenty-seven infertile men who were regular cannabis users were matched to 27 infertile men who were cannabis non-users. To complement the conventional semen parameter and plasma hormone level assessments, sperm nuclear alterations were explored using fluorescence in-situ hybridization to assess numerical chromosomal abnormalities, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling to investigate DNA fragmentation, aniline blue staining to examine chromatin condensation and a motile sperm organelle morphology examination to detect vacuoles in sperm heads. The rates of sperm aneuploidy (P=0.0044), diploidy (P=0.037), total chromosome abnormalities (P = 0.0027) and DNA fragmentation (P = 0.027) were significantly higher in cannabis users than in non-cannabis users. Cannabis consumption might have deleterious effects on sperm nuclear quality in infertile men by increasing numerical chromosome abnormalities and DNA fragmentation. Cannabis consumption induces these detrimental effects on the progression of spermatogenesis from meiotic stages to spermiogenesis and potentially on post-testicular sperm maturation in infertile men. Any potential findings, however, need to be validated with larger sample size, and our data are only exploratory findings.

Silk protein sericin pretreatment enhances osmotic tolerance and post-thaw sperm quality but reduces the ability of sperm cells to undergo in vitro induced acrosome reaction in rabbit

Effect of sericin pretreatment of sperm cells on the osmotic tolerance, ability to undergo acrosome reaction induced by calcium ionophore (CI), heparin (H) or lysophosphatidylcholine (LPC), post-thaw sperm quality and in vivo fertility was evaluated in four successive experiments in rabbit. In experiment 1, fresh semen was pretreated with sericin (0, 0.1% or 0.5% w/v) before exposing to fructose solutions adjusted to either 50, 100, 290, 500 or 1000 mOsm/L. Sericin pretreatment increased sperm livability in addition to live-membrane intact and total membrane intact sperm rates (P < 0.05) in 50 and 290 mOsm/L groups. In experiment 2, sperm samples were pretreated by either 0.1 or 0.5% sericin after removal of the semen plasma. CI, H or LPC were used to induce acrosome reaction in pretreated sperm samples. Sericin pretreatment, reduced the ability of sperm cells to undergo acrosome reaction (P < 0.05) in vitro. In experiment 3, ejaculates were frozen with or without sericin pretreatment in DMSO-sucrose extender. In post-thaw samples sericin pretreatment improved total and progressive motility, livability, membrane and acrosome integrity in a dose dependent manner (P < 0.05). In vivo fertility trials by artificial inseminations revealed contradictory results in experiment 4. Although 0.5% sericin pretreatment totally inhibited fertility, 0.1% sericin provided high pregnancy rates. In conclusion; sericin pretreatment enhances osmotic tolerance and post-thaw sperm quality, but reduces the ability of rabbit sperm cells to undergo in vitro induced acrosome reaction, but this effect is restored in vivo by dose dependent manner.

Seasonal effect on seminal constituents of swamp buffalo (Bubalus carabanesis) in the foot hill of Eastern Himalaya

ABSTRACT A total of 150 ejaculates, 10 from each of 15 swamp buffalo bulls comprising each group 5 animals were used in the study to observe the seasonal effect in sperm biochemical and semen quality. Methods: Clinical and andrological examinations, and monitoring of semen production and quality were carried out on fifteen mature, healthy swamp buffalo bulls in North East India. Seminal characteristics were compared between three seasons of the year (Summer, March-June; Monsoon, i.e. July–September; Autumn, i.e. October–November and winter, i.e. November–February) with distinct ambient temperature and humidity. Results: All bulls were diagnosed as clinically healthy and with good libido throughout the period of study. Ejaculate volume, mass activity, HOST and acrosomal integrity differ significantly (P < 0.05–0.001) between all seasons, whereas initial sperm motility, sperm concentration and abnormalities did not differ significantly. The total protein, total cholesterol and total lipid of seminal plasma of swamp buffalo semen differ significantly between all the seasons. In conclusion, climatic changes did not seem to largely affect semen sperm output or viability, although the proportions of some semen characteristics and biochemical constituents altered season wise. Conclusion: Seasonal changes moderately appear to cause deleterious changes in sperm quality in swamp buffalo.

Electrophoretic profile of seminal proteins and their correlation with in vitro sperm characters in Black Bengal buck semen.

Aim:This study aimed to study the electrophoretic properties of seminal plasma and sperm proteins of Black Bengal buck semen and their correlation with in vitro sperm characters and freezability.Materials and Methods:Semen ejaculates from nine Black Bengal bucks were collected by artificial vagina (n=20/buck). Ejaculates were evaluated for in vitro sperm characters and electrophoretic profile of seminal protein. In vitro sperm characters were evaluated immediately after collection, after completion of equilibration period, and after freeze-thawing. For seminal protein studies, seminal plasma proteins were precipitated by ice-cold ethanol method, and sperm proteins were extracted by Triton X detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to assess the molecular weight of seminal proteins. Correlation between in vitro sperm characters and protein bands was determined by Pearson’s correlation coefficient, and two-way ANOVA was applied to find the individual buck differences.Results:Significant difference (p<0.01) among the bucks was noticed in the in vitro sperm characters evaluated at all the three stages of semen evaluation such as immediately after collection, after completion of equilibration period, and post-freeze thawing. Progressive loss of sperm motility, membrane integrity, and other in vitro sperm characters were noticed during cryopreservation. A total of ten protein bands in the molecular weight ranging from 17 to 180 kDa were found in the SDS-PAGE of seminal plasma proteins, while nine bands of 17-134 kDa were observed in sperm proteins. Seminal plasma proteins of molecular weight 75, 62-49, 20, and 17 kDa and sperm proteins of 75, 20, and 17 kDa were present in all the nine bucks (100%) screened, and variation among the bucks was noticed for the presence of other proteins. Seminal plasma protein of 180-134 kDa showed a negative correlation with individual motility (−0.716) and functional membrane integrity of sperm cells (−0.724) in post-freeze–thaw analysis and 48 kDa protein had a positive correlation with individual motility (0.649) and functional membrane integrity of sperm cells (0.664) in post-thaw analysis. Sperm proteins of 63 kDa had a negative correlation (−0.616) with sperm concentration in neat semen.Conclusion:Variation among the bucks was noticed in the in vitro sperm characters and semen freezability. Correlation between seminal proteins and in vitro sperm characters and semen freezability had been found which might be useful as a tool to select breeding bucks.

Role of Oxidative Stress and Antioxidants in Male Infertility: An Interventional Study.

Aims:The study aims, in infertile men, (i) to assess oxidative stress parameters in semen plasma and (ii) to study the effect of antioxidants in those with abnormal semen parameters.Settings and Design:This was an interventional study.Population:Ninety men attending and infertility clinic in a tertiary center were enrolled in the study.Materials and Methods:The present study was conducted in the departments of obstetrics and gynecology and biochemistry in a tertiary center. Ethical approval was obtained from the institute ethics subcommittee, and the study was conducted between July 2014 and July 2016. The study was conducted on two groups of 45 men with normal semen parameters in group 1 and 45 men with abnormal semen parameters in group 2.Results:Malondialdehyde (MDA) value was higher in men with abnormal semen parameters, which was statistically significant. The total antioxidant assay was higher in men with abnormal semen parameters, which was not statistically significant. Oxidative stress index (OSI) value was higher in men with normal semen parameters, which was not statistically significant. After 90 days of antioxidants therapy to men with abnormal semen parameters, MDA value decreased, total antioxidant assay increased, and OSI value decreased, which were statistically significant. Semen parameters such as sperm concentration, motility, and normal morphology improved after 90 days of antioxidant therapy, which were statistically significant.Conclusions:Oxidative stress is increased in men with abnormal semen parameters. Antioxidant therapy improves sperm concentration and motility and decreases oxidative stress in the semen plasma.

EFFECTS OF PARSLEY SEEDS AT DIFFERENT LEVELS AS NATURAL ADDITIVES ON SEMEN QUALITY IN RABBIT MALES

This study is designed to evaluate the effect of using parsley seeds (PAS) at different levels as a natural on semen quality and reduce the effect of lipidperoxidation of bucks. Forty New Zealand rabbit bucks 8 month old were divided into four groups (each 10 rabbit bucks). The groups were divided as follows: Control diet (free from PAS), T1: Control diet + 5g/kg PAS, T2: Control diet + 10g/kg PAS; T3: Control diet + 15g/kg PAS. Semen sample were collected twice a week; from each rabbit buck by artificial vagina. The first semen collection was used to evaluate semen quality parameters. However, the second semen collection was used for analysis semen plasma. Results revealed that feeding diets containing different levels of PAS significantly (P&lt;0.05) increased semen volume, mass activity, individual motility, PSV, sperm concentration, total sperm and seminal plasma activity of ALP enzyme and significant (P&lt;0.05) decrease in dead sperm and seminal plasma concentrations of total lipids, triglycerides, cholesterol and MDA and seminal plasma activities of AST and ALT enzymes and TAC as compared with the control group. In conclusion, dietary supplementation with different levels of PAS, especially at the level of 15 g/kg (T3) resulted in significant improvement in semen quality, antioxidant status and reproductive traits of rabbit bucks.

Impact of selenium nano-particles in semen extender on bull sperm quality after cryopreservation

The aim of this study was to evaluate the effect of adding different concentrations of selenium nano-particles (Se-NPs) in semen extender on bull sperm cryopreservation. Five healthy, fertile Friesian bulls were used, and the ejaculates were obtained using an artificial vagina. Semen of all bulls were pooled and diluted in a tris-yolk fructose (TYF) extender supplemented with Se-NPs at concentrations of 0 (T1, control), 0.5 (T2), 1.0 (T3) and 1.5 (T4) μg/ml for a final sperm concentration of 80 × 106 sperm cells/ml. Diluted semen was packed in straws (0.25 ml) and stored in liquid nitrogen (−196 °C) for one month. After thawing, semen of each treatment was evaluated for sperm quality parameters, including sperm progressive motility, livability, morphological abnormalities, plasma membrane integrity and chromatin integrity. Apoptosis and sperm ultrastructure were also examined. Total antioxidant capacity and lipid peroxidation markers were determined in seminal plasma of semen in each treatment. Finally, the effect of Se-NPs on fertilization capacity was checked in vivo using n = 81cows. Results showed that T2 and T3 had a positive effect (P < 0.05) on post-thawing sperm progressive motility, livability and membrane integrity as compared with the control. Percentage of viable sperm increased (P < 0.05), while percentages of early apoptotic, apoptotic and necrotic sperm cells decreased (P < 0.05) in T3 as compared to T1. Total antioxidants capacity (TAC) in seminal plasma increased (P < 0.05) and malondialdhyde (MDA) concentration decreased (P < 0.01) in T3 as compared to T1, but did not differ in T4 from that in T1. In vivo fertility rate was higher in T3 (90%) than in T1 (59%).In conclusion, enrichment of semen extender with Se-NPs at a concentration of 1.0 μg/ml improved post-thaw sperm quality of Holstein bulls, and consequently in vivo fertility rate by reducing apoptosis, lipid peroxidation and sperm damage occurring by cryopreservation.

Cryopreservation and Quality Assessment of Buffalo Bull (Bubalus bubalis) Semen Using New Moringa Extender and Antioxidant Co-q10

Low fertility in buffaloes with frozen-thawed semen is attributed to sperm damage that is related to formation of ice crystals during freeze/thaw process. This study was conducted to evaluate the freezability of buffalo bull semen extended with four types of extenders, including citrate egg-yolk (CEY, control), CEY with ethanolic Moringa Olifera Leaf Extract (eMOLE) as replacement of antibiotics (CEYm), eMOLE and eMOLE with conventional antibiotic (eMOLEa) with or without Coenzyme Q10 as antioxidant. Semen was subjected to calculate progressive motility, livability, abnormality, acrosome status and integrity of sperm membrane in semen after extending, equilibrating and thawing processes. Four sexually mature buffalo bulls (4-5 y and 400-450 kg LBW) were used for semen collection, and semen was collected twice weekly for 4 successive weeks. Semen ejaculates of ≥70% as mass motility percentage were pooled and extended at 37°C by the four types of extenders without or with 30 µM Co-Q10). Results cleared that progressive motility, livability, abnormality, acrosomal damage and plasma membrane integrity percentages improved (P<0.05) in buffalo semen after dilution, equilibration and thawing, while activity of AST, ALT and LDH in seminal plasma of post-thawed semen reduced (P<0.05) by extension of semen with eMOLE as a new extender. All sperm parameters and enzyme activity were maintained by replacing antibiotics in CEY extender with 4 ml eMOLE. All sperm parameters were improved (P<0.05) and enzyme activity was maintained by adding 30 μM of coQ10 as compared to free extenders. Fertility rate of buffalo cows inseminated with semen cryopreserved in extender (eMOLE+coQ10) was the highest (91.6%), followed by eMOLE without coQ10 (83.3%) and the lowest for control CEY (66.6%). In conclusion, eMOLE as a new extender, is promising for the extension and preservation of buffalo semen. Also, adding 4 ml of eMOLE could be a good substitute for the antibiotic component of CEY as a conventional semen extender. Moreover, supplementing semen extenders with antioxidant addition (30 μM CoQ10) is important to improve function of spermatozoa and sperm fertility of cryopreserved semen of buffaloes.

Association between leukocytospermia and semen interleukin-6 and tumor necrosis factor-alpha in infertile men.

Seminal cytokines were previously reported to adversely affect process of spermatogenesis and ultimately induce poor semen quality. However, association between both IL-6 and TNF-α and leukocytospermia was not yet settled. The aim of this study was to evaluate the association between leukocytospermia and levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in semen of infertile men. This cross-sectional study included 100 age-matched (≥18-45years.) men. Participants were divided into four groups with 25 patients in each group: Group (A) - infertile patients with leukocytospermia and normal other semen parameters; Group (B) - infertile patients with leukocytospermia and abnormal semen parameters; Group (C) - infertile patients with oligospermia and/or asthenospermia and/or teratospermia but with no leukocytospermia; Group (D) - fertile patients with normal semen parameters and without leukocytospermia. All patients were assessed by detailed medical, sexual, fertility history, and complete physical examination. Laboratory assessment included hormonal and semen analysis and assessment of IL-6 and TNF-α in semen plasma. There were significant differences among the study groups regarding total sperm count, sperm concentration, and progressive motility (p<0.05 for each). There was significant increase in semen WBC counts in groups A and B vs. groups C and D (p=0.003). There were significant associations between increase levels of WBCs≥5/HPF and decrease levels of total sperm count (p=0.023), sperm concentration (p=0.001), and sperm progressive motility (p=0.02). There were significant upregulations in mean level of IL-6 (p=0.001) and mean level of TNF-α (p=0.003) in groups A and B vs. groups C and D. Overall, leukocytospermia is associated with reduction in sperm count, progressive motility and further upregulation of seminal IL-6 and TNF-α. The effect of treatment of leukocytospermia on the level of seminal cytokines is important point of future research.

The Effect of Oral Administration of Zinc Oxide Nanoparticles on Quantitative and Qualitative Properties of Arabic Ram Sperm and Some Antioxidant Parameters of Seminal Plasma in the Non-Breeding Season.

Zinc is an essential mineral and accepted as a trace element in the animal nutrition and for its role in biological enzymatic pathways. This study aimed to investigate the effect of oral administration of zinc oxide nanoparticles (ZnONPs) on some of the antioxidant parameters of semen plasma, quantitative and qualitative properties of Arabic ram sperm in the non-breeding season. Twelve adult Arabic rams (about 3-5 years old, 70 &plusmn;2.1 kg) were randomly assigned to receive one of the three levels of dietary ZnONPs (control; 0, group 1; 40 ppm and group 2; 80 ppm). Results showed that using different levels of ZnONPs increased the activity of superoxide dismutase enzyme (SOD) and total antioxidant capacity (TAC) of semen plasma significantly compared with the control group (P&lt;0.05). Motility (74.83%), viability (76.90%), semen volume (1.76 ml) and sperm concentration (1418&times;106/ml) were significantly (P&lt;0.05) higher in ZnONPs supplemented groups compared with the control group. The sperm morphological abnormalities reduced significantly in treated groups (10.46 and 9.07%) compared with the control group (12.66%; P &lt;0.05). Also, the results suggested that 80 ppm level of ZnONPs increased the functionality of sperm membrane (44.38%) compared with other groups (37 and 35.66%, respectively for groups 1 and control) (P&lt;0.05). Based on the results, using 80 ppm level of ZnONPs lead to an improvement in the activity of superoxide dismutase enzyme (48.62 ml) and total antioxidant capacity of semen plasma (111.88 &micro;g/ml) compared with other groups (P&lt;0.05). In conclusion using 80 ppm level of ZnONPs had a positive effect on the quantitative and qualitative properties of sperm and lead to a significant betterment in the activity of some antioxidant parameters of Arabic ram semen in the non-breeding season.

Influences of intrauterine semen administration on regulatory T lymphocytes in the oestrous mare (Equus caballus)

In the mare, early pregnancy loss is common, but involvement of the maternal immune system in the pathogenesis of this condition has not been investigated in detail so far. In the present study, we assessed effects of exposure of the endometrium to semen or seminal plasma in oestrous mares on the response of regulatory T lymphocytes (Tregs) in the peripheral circulation as well as in the endometrium. Raw semen, seminal plasma or PBS (control) were introduced into the uterus of oestrous mares (n = 12). Blood was collected immediately before insemination or PBS infusion (time 0), and 12, 24 and 48 h thereafter. Endometrial biopsies were collected at 24 h. In peripheral blood, Treg (CD4+Foxp3+) cells were determined by flow cytometry. In endometrial biopsies, Tregs were assessed as cells staining positive for Foxp3 by immunohistochemistry. The percentage of Tregs in blood decreased (p < 0.05) at 12 h after exposure to seminal plasma, tended to decrease in response to raw semen (p = 0.095) but not to PBS. Leukocyte and PMN counts were not affected. In the endometrium, numbers of Foxp3 positive cells at 24 h after insemination or PBS infusion were not changed by treatment. Results of the present study provide only little evidence that maternal tolerance of pregnancy in the horse is modulated already by exposure of the oestrous endometrium to seminal plasma at mating.

Effect of Propolis Ethanolic Extract Supplementation to Ram Semen Extenders on Sperm Characteristics, Lipid Peroxidation and some Enzymatic Activities in Seminal Plasma in Chilled Semen

This study designed to examine effects of adding different concentrations of propolis ethanolic extract (PEE) as antioxidant to ram semen extenders; tris egg yolk (TEY) or tris-soybean lecithin (TSBL) on sperm characteristics, lipid peroxidation and enzymatic activities in seminal plasma of ram semen preserved at 5°C for 48 hours. Semen was collected from five adult Rahmani rams using an artificial vagina twice/week for 5 weeks. Pooled semen was extended with TEY or TSBL supplemented with different concentrations from PEE (0, 0.1, 0.5 and 1.0 mg/ml). Polyphenols and flavonoid compounds in propolis ethanolic extract were identified. In each treatment, percentage of progressive motility, livability, and abnormality were determined. Also, percentages of sperm membrane integrity or chromatin damage were determined. Concentrations of total antioxidant (TAO) and malondialdehyde (MDA), and activity of acid phosphatase (ACP), lactate dehydrogenase (LDH), and asprtate (AST) and alanine (ALT) transaminase, were determined. Results showed that the most effective polyphenolic compounds in the prolopolis ethanolic extract was the chlorogenic acids, catechein, protocatchuic and pyrogallol (120.623, 53.006, 23.907 and 11.048 mg/100g extract, respectively). Luteo.6-arbinose 8-glucose was the predominant identified flavonoid component in propolis extract (5268.78ppm), followed by acacetin, hisperidin, apig.6-rhamnose8- glucose, apig.6-glucose8-rhamnose, kamp.3-(2-p-comaroyl) glucose and luteo.6-glucose 8-arbinose with concentrations )647.53, 561.09, 366.47, 150.46, 144.51 and 109.42 ppm, respectively. The effect of propolis ethanolic extract supplementation to ram semen extenders was significant (P<0.05) only on the overall percentages of sperm progressive motility and sperm chromatin damage while, sperm livability, abnormality and membrane integrity were not affected significantly (P<0.05). Concentration of TAC was numerically (P= 0.82) higher in TEY extender with different concentrations of PEE than TSBL. However, MDA was lower (P<0.05) in free TEY, followed by TSBL with PEE. Activity of LDH, AST and ALT tended to be lower in seminal plasma of semen extended with TSBL than TEY. These results showed beneficial effects of soybean lecithin and propolis ethanolic extract in tris based extender on progressive motility and chromatin integrity of spermatozoa up to 48 h of preservation at 5°C. Also, lipid peroxidation and enzyme activity were lower in TSBL than TEY. In addition, soybean lecithin can effectively replace egg yolk as a protective additive with propolis ethanolic extract for preservation extender without any detrimental effects on post-chilling semen quality in rams.

Effect of vibration emissions during shipping of artificial insemination doses on boar semen quality

The shipping of semen doses to sow farms can impair boar semen quality. Unfortunately, there is currently no practice-oriented information available regarding general shipping conditions of boar semen. For this reason, a special mobile sensing app (TransportLog 1.0), utilizing the built-in sensors of smartphones, has been programmed to capture vibration emissions during shipping of semen doses (QuickTip Flexitubes®, Minitüb). Data were analyzed, transformed and used as standards for simulating vibration emissions from an orbital shaker IKA MTS 4 (Laborgeräte München) in a spermatological reference laboratory. Twenty ejaculates were collected randomly and diluted using a one-step isothermal process in a split-sample procedure in Beltsville Thawing Solution (BTS, Minitüb). The sperm concentration was adjusted to 24 × 106 sperm/mL. The dose filling volume was 85 ± 1 mL. Samples were stored for seven days at 17 °C. The results showed that circular horizontal vibration emissions with frequencies of 300 rpm for a duration of 6 h led to a significant alkalization of the BTS-extended semen. Semen motility, mitochondrial activity, acrosome and plasma membrane integrity as well as thermo-resistance all demonstrated a frequency-dependent negative response to vibration emissions during long-term storage. This study leads to new insights and recommendations for the shipping of boar semen in the artificial insemination industry. Furthermore, a new monitoring tool for boar semen shipping was established using mobile sensing.

Human Semen or Seminal Plasma Does Not Enhance HIV-1BaL Ex Vivo Infection of Human Colonic Explants.

To determine whether human whole semen (WS) and seminal plasma (SP) either previously frozen or freshly acquired altered ex vivo infectibility of human colonic explants or was associated with histology or toxicity changes, which may influence mucosal HIV-1 transmission in vivo. Pooled human semen samples were freshly obtained from study volunteers (never frozen) and from commercial sources (frozen/thawed). Endoscopically acquired rectal biopsies were evaluated for toxicity following titered ex vivo WS/SP exposure by histological grading and by MTT assay. The ex vivo HIV-1 biopsy challenge model was used to evaluate effects of exposure to either previously frozen or freshly acquired WS/SP on HIVBaL infectibility at a range of viral inocula (104-100 TCID50). To evaluate the effects at lower viral inocula of HIV-1 (10-2-102), experiments in the presence or absence of WS/SP were also performed utilizing TZM-bl cells. MTT assays and histological scoring demonstrated no tissue degradation of biopsies when exposed for 2 h to concentrations of 10% or 100% of either fresh or previously frozen WS/SP. Ex vivo biopsy HIV-1 challenge experiments showed no differences in the presence of freshly acquired or previously frozen/thawed WS/SP compared with control; no differences were seen with lower infectious titers on TZM-bl cells. Within the limits of assay sensitivity and variability, these data show no toxicity or significant enhancement of HIV-1 infectibility of human rectal mucosa using the colorectal explant model with either pooled fresh or frozen/thawed nonautologous human semen.

Effect of age on lipid peroxidation of fresh and frozen-thawed semen of Nili-Ravi buffalo bulls

Buffalo bull spermatozoa are rich in polyunsaturated fatty acids and are prone to lipid peroxidation. We hypothesised that lipid peroxidation in buffalo bull semen will increase with age and will affect the semen quality. The objective was to compare malondialdehyde (MDA) concentration and the quality of fresh and frozen-thawed semen between aged (13.6 ± 1.0 years; n = 3) and young (3.4 ± 0.3 years; n = 3) Nili-Ravi bulls. The concentration of MDA did not differ (p >.05) between aged vs. young bulls in fresh (2.3 ± 0.2 vs. 2.9 ± 0.7 nmol mL−1), frozen-thawed (53.1 ± 2.8 vs. 48.4 ± 2.6 nmol mL−1) semen and seminal plasma (5.71 ± 0.97 vs. 5.19 ± 1.36 nmol mL−1), respectively. In fresh semen, sperm motility and total concentration did not differ (p > .05) between aged and young bulls. The volume of fresh semen increased (p > .05) while sperm viability and DNA integrity decreased (p < .05) in aged vs. young bulls. In frozen-thawed semen, sperm motility, viability and DNA integrity decreased (p < .05) in aged vs. young bulls. In frozen-thawed vs. fresh semen, MDA level increased within young (48.4 ± 2.6 vs. 2.3 ± 0.2 nmol mL−1) and aged bulls (53.1 ± 2.8 vs. 2.9 ± 0.7 nmol mL−1). Conversely, sperm motility and viability decreased (p < .05) within the age groups between fresh and frozen-thawed semen. In conclusion, (1) lipid peroxidation (MDA) does not increase due to age; however, it is negatively associated with semen quality and (2) cryopreservation steps up lipid peroxidation irrespective of age and deteriorates semen quality of Nili-Ravi bulls.

Corrected cholesterol, a novel marker for predicting semen post-thaw quality: a pilot study

Semen cryopreservation is the recommended method for protecting the fertility of men before they undergo iatrogenic treatments. However, post-thaw sperm quality is highly variable between and within individuals. Thus, prediction of post-thaw quality has not been achieved with great certainty. Men (n = 101) attending our andrology clinic each produced a single ejaculate for semen analysis and semen cryopreservation. Post-thaw semen quality was examined for associations with traditional and novel semen and seminal plasma parameters. This current research demonstrates that the cholesterol concentration of human sperm is associated with ejaculate tolerance to cryopreservation. Corrected cholesterol, calculated as the difference between the cholesterol of semen and the cholesterol of cell free seminal plasma, is predictive of post-thaw motility. This may have potential as a method to guide the number of ejaculates stored for individual patients before iatrogenic treatment and/or for the appropriate selection of ART with frozen/thawed semen.

Effect of Seminal Plasma Components on the Quality of Fresh and Cryopreserved Stallion Semen

The importance of seminal plasma (SP) components for stallion semen quality and freezability is little known. This study aimed to evaluate the relationship between SP components and fresh/cryopreserved stallion semen quality. Semen of 30 stallions was collected, and then, SP was recovered and lyophilized. Total protein (TP), vitamin C (CVIT), vitamin E (EVIT), vitamin A (AVIT), iron (Fe), copper (Cu), magnesium, and zinc (Zn) in SP were assessed. Sperm was frozen in an extender supplemented with lyophilized SP. In fresh semen motility, abnormal morphology (AM), sperm vitality (SV), and plasma membrane integrity (PMI) were evaluated. In post-thaw semen, additionally, total motility (TM), progressive motility (PM), straight line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH), and beat cross-frequency (BCF) were assessed. Levels of component of SP were established by a distribution analysis. Generalized linear models were fitted. Comparisons of means were done with Tukey's test. Correlation and regression analyses were performed. Vitamins and ions were found to be related to fresh semen quality. For post-thaw sperm, medium TP showed higher semen quality. Negative regression and correlation coefficients between CVIT and all post-thaw semen parameters were found. Low EVIT yielded the lowest PM, VSL, and VAP values, while a high level of AVIT yielded the best results for sperm quality. A high level of Cu yielded higher results for TM, PM, VCL, and ALH. Moreover, a negative correlation was found between Zn, SV, and PMI. In conclusion, SP composition influences fresh and post-thaw stallion semen quality.

Effect of some Semen Extenders as a Natural Source of Antioxidants on Quality of Frozen Friesian Semen

This study was designed to evaluate the possibility of using the optimal level of Moringa oleifera leaf aqueous extract (10, 15 or 20%) with 90, 85 or 80% saline (MOLE) as a semen extender of bull semen in comparing with three extenders, including Tris-egg yolk (TEY) without or with vitamin C at a level of 0.9 mg/ml (TEY+VC), and camel skim milk (CSM). Semen was collected from five Friesian bulls (350-450 kg LBW and 3-4 years old), twice a week for 5 weeks using artificial vagina. After ejaculation, mass motility was evaluated and only those of ≥70% were used in this study. Ejaculates were pooled, divided into 4 replicates and prepared for freezing. Semen was extended in TEY (control), TEY+VC, (CSM) and 15% MOE+85% saline (based on the best results of MOLE level). Semen was diluted, equilibrated at 5oC (4 h) and placed in liquid nitrogen (LN) at -196oC. Frozen semen stored for one month at least was warmed at 37oC for 30 sec. Semen evaluation for percentages of individual motility (IM), live sperm (LS), sperm abnormality (SA), acrosomal damage (AD) and hypo-osmotic swelling test (sperm cells with curled tail, CT) was estimated in diluted, equilibrated and thawed semen, while percentage of head to head agglutination (HHA) was determined in post-thawed semen only. Activity of AST, ALT, LDH and total antioxidant activity were determined in seminal plasma of post-thawed semen. Conception rate (CR) was recorded for Friesian cows inseminated with semen frozen with the four types of extenders. Results showed that percentages of IM, LS, SA, AD and CT in post-diluted and post-equilibrated semen, beside all previous characteristics and percentage of HHA in post-thawed semen showed the best results (P<0.05) in CSM, moderate in 15% MOE and TEY+VC, and the poorest in TEY. Enzyme activities of AST, ALT and LDH in post-thawed seminal plasma were the lowest (P<0.05) in CSM, followed by 15% MOE, while total antioxidants activity was higher (P<0.05) for TEY+VC, CSM and 15% MOE than for TEY, being the highest for 15% MOE. Artificial insemination of estrous synchronized cows with semen extended by different types of extenders indicated that conception rate of cows was significantly (P<0.05) the highest for Friesian cows inseminated with post-thawed semen extended with CSM (87.5%), moderate with both TEY+VC and 15% MOE (75%) and the lowest with TEY(62.5%). Extender containing 15% MOE showed the cheapest cost (4.80 L.E./100 ml) in comparing with CSM (6.45 L.E./100 ml), TEY+VC (13.95 L.E./100 ml), and even the conventional TEY extender (13.85 L.E./100 ml). In conclusion, extender containing 15% Moringa oleifera leaf extract and 85% saline solution could be successfully used, as a source of antibiotics and antioxidants, in extenders of frozen bull semen, being with the cheapest cost as compared to tris-egg yolk or camel skim milk extenders.

Impact of early cART on HIV blood and semen compartments at the time of primary infection

BackgroundHIV-infected cells in semen facilitate viral transmission. We studied the establishment of HIV reservoirs in semen and blood during PHI, along with systemic immune activation and the impact of early cART.MethodsPatients in the ANRS-147-OPTIPRIM trial received two years of early cART. Nineteen patients of the trial were analyzed, out of which 8 had acute PHI (WB ≤1 Ab). We quantified total cell-associated (ca) HIV-DNA in blood and semen and HIV-RNA in blood and semen plasma samples, collected during PHI and at 24 months of treatment.ResultsAt enrollment, HIV-RNA load was higher in blood than in semen (median 5.66 vs 4.22 log10 cp/mL, p<0.0001). Semen HIV-RNA load correlated strongly with blood HIV-RNA load (r = 0.81, p = 0.02, the CD4 cell count (r = -0.98, p<0.0001), and the CD4/CD8 ratio (r = -0.85, p<0.01) in acute infection but not in later stages of PHI. Median blood and seminal cellular HIV-DNA levels were 3.59 and 0.31 log10cp/106 cells, respectively. HIV-DNA load peaked in semen later than in blood and then correlated with blood IP10 level (r = 0.62, p = 0.04). HIV-RNA was undetectable in blood and semen after two years of effective cART. Semen HIV-DNA load declined similarly, except in one patient who had persistently high IP-10 and IL-6 levels and used recreational drugs.ConclusionsHIV reservoir cells are found in semen during PHI, with gradual compartmentalization. Its size was linked to the plasma IP-10 level. Early treatment purges both the virus and infected cells, reducing the high risk of transmission during PHI.Clinical trials registrationNCT01033760

Performance of microbiological, serological, molecular, and modified seminal plasma methods in the diagnosis of Brucella abortus in semen and serum of bovine bulls

Brucellosis remains as a major infectious disease of domestic animals and is considered a re-emerging zoonosis in several countries. B. abortus infections in bulls are related to reproductive tract infections, although infected animals show transient serological titers or nonreactor status. Thus, diagnosis of bovine brucellosis based exclusively on serological tests probably underestimates B. abortus infections in bulls. In this scenario, three hundred thirty-five serum samples from reproductively mature bovine bulls were subjected simultaneously to standard serodiagnosis using the rose Bengal test (RBT), 2-mercaptoethanol (2-ME), complement fixation (CFT), and fluorescence polarization assay (FPA). Furthermore, conventional semen plasma agglutination (SPA) and modified 2-ME, FC and, FPA were carried out in all bulls replaing serum by seminal plasma. Semen from all bulls was also analyzed for sperm viability, microbiological culture in Farrell media, and polymerase chain reaction (PCR). Only eight (2.38%) semen samples were considered improper for reproduction services (necrospermia and azoospermia), although none of these animals was positive in any of the diagnosis methods used. Five bulls (1.49%) were simultaneously positive in conventional RBT, 2-ME, SPA, modified 2-ME, microbiological culture in Farrell media, and in PCR for B. abortus strain 19. Two (1.67%) bulls were positive in PCR for B. abortus field strains and negative in all other tests, although semen was considered viable to reproduction service. The identification of B. abortus B19 strain in serum and semen of bulls occurred probably due to improper vaccination of males or infection by B19 strain shedding by vaccinated females that could to contaminated environment of farms. In addition, detection of B. abortus field strains only using PCR in bulls without sperm viability abnormalities indicate the need for including molecular methods to improve diagnosis of the disease in bovine bulls.