Can Ganoderma lucidum Be An Alternative Nutritional Supplement For Enhancing Sperm Motility Rate?

AbstractThis study aims developing and evaluate a protocol of semen cryopreservation of the lane snapper Lutjanus synagris. Firstly, sperm motility rate, motility time, density and spermatocrit were appraised to characterize the sperm quality of the lane snapper. The effect of three extenders with distinct ionic compositions and pH values combined with seven concentrations of cryoprotector dimethylsulfoxide (0; 2.5; 5.0; 7.5; 10.0; 12.5 e 15.0%), five cooling rates (110, 90, 60, 45 e 30°C –min), nine equilibration time (1; 2,5; 5; 10; 15; 20; 25; 30 e 60 minutes) e five dilutions ratio (1:1; 1:3; 1:6; 1:10 e 1:20) on the sperm motility rate and motility time were analyzed. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The higher sperm motility rate and motility time (P<0.05) was achieved by combining extender with pH 8.2 with 10% concentration of dimethylsulfoxide and cooling rate 60°C –min, 1 minute of equilibration time and 1:3 (v/v) dilution ratio. The use of cryopreserved sperm presented fertilization rates >60% validating the present protocol for lane snapper. The cryoconserved sperm of lane snapper is a viable alternative, being possible to maintain appropriate sperm viability.

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Randomized Clinical Trial: Effect of Walnuts on Semen Parameters and Male Fertility (P18-042-19)

Current Developments in Nutrition ◽

10.1093/cdn/nzz039.p18-042-19 ◽

2019 ◽

Vol 3 (Supplement_1) ◽

Author(s):

Wendie Robbins ◽

Howard Kim ◽

Justin Houman ◽

Geng-Wei Lee

Keyword(s):

Clinical Trial ◽

Los Angeles ◽

Randomized Clinical Trial ◽

Sperm Motility ◽

Clinical Care ◽

Sperm Concentration ◽

Nutritional Supplement ◽

Semen Parameters ◽

Male Factor Infertility ◽

Male Factor

Abstract Objectives Infertility affects ∼8–12% of couples worldwide with ∼40% attributed to male factors. Recent studies suggest a role for paternal diet in fertility. Walnuts contain a variety of nutrients essential in the development of spermatozoa. We conducted a randomized clinical trial (RCT) to determine if consumption of walnuts improves semen parameters and fertility in men seeking clinical care for male factor infertility. Methods This was a two arm, single blind, RCT. The comparison groups both received usual care for male factor infertility. One group added 42 gm/d walnuts to their diet, and the other group added a daily nutritional supplement recommended for male reproductive health. Participants (n = 75) were enrolled at an infertility clinic located in a large metropolitan medical center. Eligibility was determined by history, physical exam, and lab tests collected as part of clinical care. Research measures included semen analysis and blood sample at baseline and 3 months; ASA24 dietary recall at baseline, 2 and 3 months; and fertility report at 3 months and 1 year. Results Age range was 27 to 61 years (39.7 ± 7.0); BMI range 19.6 to 46.9 (26.8 ± 4.5); participant race was Asian 26.2%, White 44.3%, Hispanic White 16.4%, Black 8.2%, other 4.9%. Baseline sperm concentration was 39.4 ± 30 million per ml; sperm motility 31.1 ± 23.4%; and progressive motility 21.2 ± 15.8%. At 3 months, the walnut group demonstrated increased sperm motility and concentration, P = .04 and P = .07, respectively, whereas no significant changes from baseline were found in the nutritional supplement group. Both groups showed improved sperm morphology, P < .03. Preliminary data from the subset of men with 1-year follow-up data shows higher frequency of pregnancy in the walnut group compared to nutritional supplement, although not statistically significant, P = .09. We continue to follow the remaining participants until their 1-year fertility report. Conclusions This RCT demonstrated a beneficial effect of adding walnuts to the diet on sperm motility and morphology in men seeking care for infertility. Preliminary fertility data suggests walnuts may enhance the probability of pregnancy for men with male factor infertility. Funding Sources Center for Occupational and Environmental Health, University of California, Los Angeles; California Walnut Commission.

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76 THE EFFECT OF GLYCEROL CONCENTRATIONS IN EGG-YOLK CITRATE EXTENDER ON THE QUALITY OF CRYOPRESERVED NGUNI BULL SEMEN

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab76 ◽

2013 ◽

Vol 25 (1) ◽

pp. 185

Author(s):

M. M. Seshoka ◽

M. L. Mphaphathi ◽

F. V. Ramukhithi ◽

T. R. Netshirovha ◽

C. Hlungwani ◽

...

Keyword(s):

Sperm Motility ◽

Rural Areas ◽

Egg Yolk ◽

Positive Control ◽

Negative Control ◽

Glycerol Concentration ◽

Bull Semen ◽

Frozen Semen ◽

Thawing Process ◽

Motility Rate

There are bull shortages in South African poor rural areas. Artificial-insemination technology could play a significant role on breeding emerging farmer’s cattle. The objective of this study was to compare glycerol concentrations (0, 4, 8, or 12%) during freezing of Nguni bull semen to conduct AI in different villages. Semen was collected by electro ejaculator from 2 Nguni bulls of known and proven fertility. Collected semen samples were kept in a thermo flask (37°C) and transported to the laboratories within 10 min after collections. Semen samples were pooled and evaluated by Sperm Class Analyser® and allocated randomly per treatment. Semen was then diluted (1 : 2 v:v) with egg-yolk citrate extender supplemented with either 0% (negative control), 4, 8, and 12% of glycerol concentration or AndroMed® (positive control). Semen samples were equilibrated for 4 h at 5°C. After equilibration period, samples were loaded into 0.5-mL straws and placed horizontally into the controlled rate (–5, –8, –10, –12, –15, –25, –35°C min–1) from 5°C until target temperature of –80°C is reached. The frozen semen straws were stored in a liquid nitrogen tank (–196°C) until thawing. Treatment means were separated using Fisher’s protected t-test least, and data are presented as mean ± SD. There was a significant differences (P < 0.05) between raw total sperm motility (83.3 ± 19.3) and frozen–thawed sperm with either 0% (0.0 ± 0.0), 4% (30.2 ± 13.4), 8% (47.9 ± 12.5), or 12% (61.5 ± 4.7) of glycerol and on AndroMed® (27.7 ± 17.8) group. Regardless of the glycerol concentrations used, the freezing-thawing process reduced (P < 0.05) the Nguni total sperm motility rate compared to uncryopreserved sperm (83.3 ± 19.3). In conclusion, egg-yolk citrate extender supplemented with 12% glycerol yielded a better (P < 0.05) total sperm motility rate (61.5 ± 4.7) as compared with the 0% (0.0 ± 0.0), 4% (30.2 ± 13.4), 8% (47.9 ± 12.5), and AndroMed® (27.7 ± 17.8) group. Further studies are required to test other levels of glycerol concentrations (>12%) on freezing Nguni semen and conducting AI.

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163 EVALUATION OF VIABILITY OF BULL SEMEN COLLECTED BY ELECTRO-EJACULATION USING COMMERCIAL SEMEN EXTENDER AND 2 CULTURE MEDIA AT CONTROLLED ROOM TEMPERATURE

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab163 ◽

2017 ◽

Vol 29 (1) ◽

pp. 190

Author(s):

A. M. Raseona ◽

O. A. Ajao ◽

L. D. Nethengwe ◽

L. R. Madzhie ◽

T. L. Nedambale ◽

...

Keyword(s):

Sperm Motility ◽

Assisted Reproductive Technologies ◽

Culture Media ◽

Reproductive Technologies ◽

Sperm Viability ◽

Progressive Motility ◽

Bull Semen ◽

Significant Difference ◽

Semen Extender ◽

Motility Rate

Preservation of semen is an important process to ensure that semen quality is sufficient for assisted reproductive technologies. The aim of this study was to evaluate the viability of bull semen collected by electro-ejaculation using commercial semen extender and 2 modified culture media stored at controlled RT (24°C) for 72 h. Two Nguni bulls were used for semen collection; after collection, the semen was evaluated macroscopically for volume, pH, and colour, and microscopically for sperm motility, viability, and morphology. Uncontaminated semen samples with progressive motility >70% and morphological defects <20% were pooled after collection before being aliquoted into 3 extenders, namely Triladyl, modified Ham’s F10, and TCM-199 culture media, at a dilution ratio of 1:4 and then stored at controlled RT (24°C). Sperm motility rate was analysed using the computer-aided sperm analyser after 0, 24, 48, and 72 h of storage. Sperm morphology and viability was performed after staining the sperm cells with spermac and nigrosine-eosin stain, respectively. The study was replicated 4 times and data were analysed using ANOVA. Triladyl had a higher sperm viability rate (41.3%) and total motility rate (96.3%) for 72 h (P < 0.01) compared with the 2 modified culture media, Ham’s F10 (26.5 and 86.8%) and TCM-199 (25.0 and 86.7%), respectively. However, Ham’s F10 had higher progressive motility rate (37.8%) as compared with the other extenders, TCM-199 (31.7%) and Triladyl (23.4). There was no significant difference (P > 0.05), in viability rate between Ham’s F10 (26.5%) and TCM-199 (25.0%). No significant difference (P > 0.05) in straight line velocity was observed for the three extenders. Furthermore, no significant difference was observed in total sperm abnormalities, except for reacted acrosomes and absent tails (P > 0.05), between the 2 Nguni bulls. Nguni semen can be preserved in Triladyl or modified Ham’s F10 and TCM-199 culture media stored at 24°C and stay viable for 72 h. Triladyl proved to be the best suitable extender of the 3 extenders, showing higher sperm viability and total motility rate as compared with Ham’s F10 and TCM-199 modified culture media.

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An Edible, High Molecular Weight Glycoprotein Fraction from Ganoderma Lucidum Inhibits the Akt Pathway in Resistant Myeloma Cell Lines Decreasing Mcl-1 Expression, Up-Regulating FKHR and Inducing Apoptosis.

Blood ◽

10.1182/blood.v104.11.4895.4895 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4895-4895

Author(s):

Nazneen Sultana ◽

Larissa Verda ◽

Richard K. Burt ◽

Ann E. Traynor

Keyword(s):

Molecular Weight ◽

Ganoderma Lucidum ◽

Active Material ◽

Nutritional Supplement ◽

Myeloma Cell ◽

Akt Pathway ◽

Remission Induction ◽

Scid Mouse Model ◽

Induced Apoptosis ◽

Molecular Weight Fractions

Abstract We observed apparent synergy between dexamethsone and a nutritional supplement taken by a patient with poor prognosis plasma cell leukima. The rapidity of the clinical remission induction and the duration of remission, which was sustained without consolidation, suggested possible inhibition of the Akt pathway. A review of the supplements ingested indicated two mycelial derivatives, one of which was Ganoderma Lucidum, (Ling-Zhi, Reishi) a traditional Chinese medicine used to prevent kidney and liver disease and to promote anti-tumor immunity. We obtained a known quantity of certified shell broken spore of G Lucidum from China. Dissolved in 70 degree C water and then diluted as a 2.5% v/v solution in RPMI1640, it induced apoptosis of 80% of OPM6 cells by 72 hours, which was only marginally improved by the addition of1uM dexamethasone. It induced 70% cell death in MM.R cells by 72 hours. This was associated with a 90% reduction in measurable Mcl-1 by Western blot after two hours of incubation. In additon phosphorylation of FKHR was reduced 70% at 2 hours and was undetectable after 72 hours. Phosphorylation of Akt at threonine 308 was reduced 75%. To further purify this activity we separated out molecular weight fractions by Centricon centrifugation. We were able to 10 fold concentrate the active material in the >100,000 MW fraction by centrifugation. Used in the same relative dilution of 2.5% v/v, this concentrate killed between 90% and 96% of glucocorticoid resistant cells at 72 houyrs. Of interest the concentrate induced a lesser degree of apoptosis in corticosteroid sensitive cells, but enhanced the activity of dexamethasone. Spectrophotometry followed by oxidation with sodium meta-periodate and measurement absorption at 550 nm suggested a glycoprotein enrichment in this concentration. Further isolation of the active component of this fraction is underway in order to determine if it is the Lin-Zhi 8 protein sequenced in 1991 or a distinct glycoprotein from the Ganoderma Lucidum. It appears to be an orally tolerable inhibitor of the Akt pathway. It also shows activty in rheumatoid arthritis synovium RAS cells and may have usefulness in T cell supression and graft tolerance.Pre-clinical testing in a SCID mouse model has been begun.

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24 RELATIONSHIP BETWEEN BOAR SPERM TRAITS AND FERTILITY RATE FOLLOWING ARTIFICIAL INSEMINATION UNDER SMALLHOLDER PRODUCTION SYSTEMS

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab24 ◽

2017 ◽

Vol 29 (1) ◽

pp. 119

Author(s):

M. B. Matabane ◽

P. Nethenzheni ◽

R. Thomas ◽

D. Norris ◽

K. Nephawe ◽

...

Keyword(s):

Sperm Motility ◽

Production Systems ◽

Sperm Quality ◽

Conception Rate ◽

Mean Values ◽

The Mean ◽

The Relationship ◽

Fertility Traits ◽

Sperm Traits ◽

Motility Rate

The prediction of sperm fertility has a great economic importance to the pig breeding industry. The objective of the study was to determine the relationship between boar sperm quality and fertility following artificial insemination (AI) under smallholder production systems. A total of 18 ejaculates were collected from 3 breeding boars using a hand-gloved technique. Aliquots of diluted semen were assessed for sperm motility using a computer aided sperm analysis before AI. Sperm viability was evaluated using Synthetic Binding CD-14 (SYBR-14+)/propidium iodide (PI–), whereas sperm morphology was evaluated using Eosin Nigrosin staining. Fluorescent microscope was used at 100× magnification to count 200 sperm per slide. The semen was extended with Beltsville Thawing Solution and contained 3 × 109 sperm/dose. A total of 73 multiparous sows were inseminated twice. Fertility was measured by conception rate, farrowing rate, litter size and number of piglets born alive following AI. Sperm quality and fertility data were analysed using one-way ANOVA. Spearman’s rank correlation was used to determine the relationship between sperm quality and fertility traits. The mean values for total sperm motility ranged from 93.5 to 96.8%. Progressive and rapid sperm motility differed significantly (P < 0.05) among the boars. However, no significant differences were found for sperm velocity traits. The mean values for morphologically normal sperm ranged from 47.8 to 60.9% and live sperm ranged from 71.8 to 77.2%, but did not differ significantly among the boars (P > 0.05). Conception rate from different boars varied (P < 0.05) from 63.6 to 93.3%. Of all fertility traits studied, conception rate was significantly related to total sperm motility rate (r = 0.34, P < 0.0029), progressive motility (r = 0.29, P < 0.0141) and rapid motility (r = 0.34, P < 0.0032), although relatively low. There was a low positive relationship between morphologically normal sperm and fertility traits (P > 0.05). In conclusion, total, progressive, and rapid sperm motility rate were the only sperm traits significantly related to conception rate. Conversely, litter size and number born alive were not correlated with sperm motility, viability, or morphology traits.

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Short-term cold storage of sperm from six neotropical characiformes fishes

Brazilian Archives of Biology and Technology ◽

10.1590/s1516-89132004000500016 ◽

2004 ◽

Vol 47 (5) ◽

pp. 799-804 ◽

Cited By ~ 19

Author(s):

Simone Marques ◽

Hugo Pereira Godinho

Keyword(s):

Sperm Motility ◽

Cold Storage ◽

Storage Duration ◽

Plastic Tube ◽

Prochilodus Lineatus ◽

Practical Applications ◽

Reproductive Management ◽

Sperm Sample ◽

Plastic Bag ◽

Motility Rate

Sperm of the following Neotropical Characiformes fish species were tested for cold storage: Brycon lundii, Piaractus mesopotamicus, Leporinus elongatus, Leporinus friderici, Prochilodus lineatus and Prochilodus marggravii. Each sperm sample was split into two aliquots. The first was placed into a plastic bag with air or oxygen and the second, in a plastic tube with air. The samples were maintained at temperatures between 1.7-4.9 ºC. The rate of sperm motility was estimated using a 50 mM NaCl solution as the activating solution. The shortest sperm storage duration (7 h) was recorded for L. friderici, when the sperm motility rate reached ~ 30%, whereas the longest duration (20 h) was obtained with the sperm of P. lineatus. A fertilisation test using Prochilodus marggravii sperm refrigerated for 8 h yielded 88-90% of viable embryos. The refrigerated storage method could be of practical applications, especially in fish reproductive management at hatchery stations.

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Tenability of ram semen

Acta Agraria Debreceniensis ◽

10.34101/actaagrar/31/3008 ◽

2008 ◽

pp. 63-66

Author(s):

János Oláh ◽

Tamás Pécsi ◽

András Kovács ◽

Anna Pécsi ◽

András Jávor

Keyword(s):

Individual Differences ◽

Heat Resistance ◽

Breeding Season ◽

Sperm Motility ◽

Resistance Test ◽

Linear Decrease ◽

Different Temperatures ◽

Motility Rate

It could be stated that the diluted semen of Awassi rams taken in the breeding season preserved its fertilizing abilityat different temperatures for different periods of time. The motility of spermatozoa kept at 23 vs. 8oC was checked daily. The largest spread of data was observed 24 hours after taking the semen, then the motility rate of cells showed a linear decrease. Motility results of fresh and frozen-thawed samples were compared also after heat resistance test and significant differences were found between these groups. Significant individual differences were observedin the sperm motility after heat resistance test.

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53 COMPARISON OF 4 DIFFERENT CRYOPROTECTANTS ON FREEZING NGUNI BULL SPERMATOZOA EVALUATED BY COMPUTER-AIDED SPERM ANALYSIS

Reproduction Fertility and Development ◽

10.1071/rdv26n1ab53 ◽

2014 ◽

Vol 26 (1) ◽

pp. 140

Author(s):

M. M. Seshoka ◽

M. L. Mphaphathi ◽

T. L. Nedambale

Keyword(s):

Sperm Motility ◽

Artificial Insemination ◽

Egg Yolk ◽

Cleavage Rate ◽

Sperm Analysis ◽

Bull Semen ◽

Significant Difference ◽

Computer Aided ◽

Freezing Curve ◽

Motility Rate

Cryopreservation has been reported to damage approximately 40 to 50% of viable spermatozoa in bulls. It is critical to evaluate frozen-thawed spermatozoa with computer-aided sperm analysis (CASA) and find a suitable cryoprotectant for Nguni semen. The study was conducted to compare cryo-effectiveness of glycerol (GLY), dimethyl sulfoxide (DMSO), propanediol (PND), and ethylene glycol (EG) cryoprotectants at 12% concentrations during freezing of Nguni bull semen. Semen was collected from 18 stud Nguni bulls of proven fertility with the use of an electro-ejaculator. Collected semen samples were kept in a thermo flask (37°C) and transported to the laboratories. Semen samples were pooled and sperm motility rate was evaluated using CASA. Semen was then diluted (1 : 2; vol : vol) with egg-yolk citrate extender supplemented with either 12% GLY, DMSO, PND, or EG. Semen samples were equilibrated for 4 h at 5°C. After equilibration, samples were loaded into 0.25-mL straws and placed into the controlled rate programmable freezer using a customized freezing curve (from 5 to –5°C at 0.008°C min–1 and from –3 to – 130°C at 6°C min–1). Following thawing of semen, artificial insemination was conducted on 104 oestrus-synchronised Nguni cows. The IVF was also conducted on 120 oocytes to check the cleavage rate. Data were analysed using ANOVA. There was a significant difference (P < 0.05) between raw total sperm motility (94.70 ± 2.63) and frozen-thawed sperm total motility with GLY (77.80 ± 11.03), EG (20.35 ± 11.86), DMSO (15.68 ± 10.14), and PND (11.19 ± 11.27) groups. The pregnancy rate following artificial insemination was 75.9% and a total of 86.6% oocytes had cleaved after fertilization with frozen (12% GLY)/thawed semen. In conclusion, cryopreservation process reduced sperm motility and velocity rates, regardless of cryoprotectant. Egg-yolk citrate extender supplemented with 12% glycerol had recorded the highest post-thaw sperm motility rate.

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41 THE EFFECT OF AMBIENT TEMPERATURE ON SPERM MOTILITY DURING LIQUID STORAGE OF VENDA COCK SEMEN AND INDIVIDUAL DIFFERENCES IN SPERM CRYOTOLERANCE

Reproduction Fertility and Development ◽

10.1071/rdv24n1ab41 ◽

2012 ◽

Vol 24 (1) ◽

pp. 133

Author(s):

M. L. Mphaphathi ◽

D. Luseba ◽

M. B. Masenya ◽

B. Sutherland ◽

T. L. Nedambale

Keyword(s):

Cell Motility ◽

Sperm Motility ◽

Sperm Cell ◽

Agricultural Research ◽

Germplasm Conservation ◽

Storage Period ◽

Liquid Storage ◽

Frozen Semen ◽

Significant Difference ◽

Motility Rate

Improving techniques for liquid storage of cock semen can increase the efficiency of AI programs in the poultry industry. The aims of the present study were (1) to compare storage of cock sperm for 24 h at 5 and 25°C and (2) to test the cryotolerance of sperm cell motility in individual Venda cocks. Semen was collected with the abdominal massage method, from 6 indigenous Venda cocks. Cocks were 26 weeks of age and were kept under the same conditions. After macroscopic analysis, semen was pooled and diluted (1:2) with Kobidil+ extender and divided into 3 equal parts. Part 1 was evaluated immediately (0 h), part 2 was stored at 5°C and part 3 was stored at 25°C and evaluated for sperm motility and velocity parameters at 4, 8, 12 and 24 h of storage. For cryopreservation, semen was diluted (1:2) with modified Kobidil+ extender supplemented with 8% of dimethyl sulfoxide. Individual ejaculates were equilibrated at 5°C for 4 h and then loaded into the programmable freezer. Then, semen straws were thawed at 5°C. Sperm motility and velocity parameters were evaluated using the Sperm Class Analyzer® system. Six replicates were done per trial. Data were analysed using the statistical programme GenStat®. Treatment means were separated using Fisher's protected t-test least significant difference (P < 0.05). Total sperm cell motility rate was 87.5% and decreased significantly during in vitro storage and was <31% after 24 h at 25°C. Semen samples stored at 5°C showed a total sperm cell motility rate of above 50% after 24 h. There was a slight linear decrease in the percentage of sperm with progressive motility and rapid velocity as the storage period increased, irrespective of the storage temperature. The rapid and medium motility percentages were higher in fresh semen and significantly decreased (P < 0.05) during the incubation period. There was variation in the total sperm cell motility of fresh and frozen semen among cocks. There was no significant difference in variation in non-progressive and medium percentage (P > 0.05) motility in diluted fresh or frozen sperm cells or in the percentage of sperm with rapid motility in thawed semen. There was variation in <25% of the cocks in total sperm motility rate. In summary, cryopreservation reduced sperm cell motility and velocity rates in all the cock semen donors. We found that cryotolerance of cock sperm does vary among males. Furthermore, the lower temperature 5°C was suitable for semen storage of Venda cocks. This temperature (5°C) could potentially improve methods of semen equilibration before cryopreservation. The study was supported by an Agricultural Research Council Parliamentary Grant, Department of Agriculture Forestry and Fisheries and National Research Foundation-GUN No RT21 and 24000 (NRF). The Germplasm Conservation & Reproduction Biotechnologies (GCRB) group is thanked for their support.

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