Extenders with vitamins C and E applied to Rhamdia quelen sperm cryopreservation / Extensores com vitaminas C e E aplicados à criopreservação de esperma de Rhamdia quelen

We performed this experiment to evaluate the effects of adding vitamins C and E on extenders for sperm cryopreservation of Rhamdia quelen over spermatic mobility after thawing. At cryopreservation, sperm samples were diluted in a proportion of 1:3 (v/v), following pre-freezing in nitrogen steam and subsequent immersion in liquid nitrogen. The diluents were composed by 5% milk powder, 5% glucose, 10% methanol and different levels of vitamin. Three sperm cryopreservation tests were carried out with (1) diluent containing 0.0; 4.0; 6.5; 9.0 and 11.5 mg of vitamin C mL-1, (2) diluent containing 0.0; 2.0; 4.0; 6.0 and 8.0 mg of vitamin E mL-1; (3) diluent containing 0.0; 4.0 + 2.0; 6.5 + 4.0; 9.0 + 6.0 and 11.5 + 8.0 mg of vitamin C mL-1 plus vitamin E mL-1, respectively. The spermatic motility rate, spermatic curvilinear velocity, average path and straight line velocities were measured in thawed semen by CASA. Data were submitted to ANOVA and Duncan´s test at 5% of significance. After thawing the effect (P0.05) of vitamin C was observed only for sperm motility, with higher values (38.2±20.7%) on solution containing 4.0 mg of vitamin C mL-1. The concomitant addition of both vitamins influenced (P0.05) only the curvilinear velocity, reducing the velocity at any concentration. In conclusion, diluents with 4.0 mg vitamin C mL-1 to cryopreservation of the silver catfish semen improve the sperm quality after thawing, and the use of diluents with vitamin E or both vitamins are not recommended because do not ensure the cells protection.

Seminal features and reproductive index of Thai tilapia (Oreochromis sp.) on different reproductive periods

Learning about the biology of the species is essential to the success of intensive farming. This study aimed to evaluate the semen of Thai tilapia during the four seasons of the year and thereby analyze their reproductive indices. Thus, 60 breeding males of Tilapia were used and were randomly divided into four water tanks and fed with isoproteic and isocaloric feed. The experiment lasted 12 months, starting from October 2014 and ending on September 2015. Thus, it was possible to collect sperm material of animals during the four seasons, twice a month, as well as to evaluate the water quality parameters in the tanks (temperature, pH and dissolved oxygen). The semen was evaluated from a light microscope in an increase of 100 x, then was activated with water. Motility was measured subjectively in the light microscope, as well as the percentage of sperm showing progressive motility. The duration was evaluated with the addition of a timer. For analysis of the morphology of the semen, the test consisted of morphopathology observation of 100 sperm focused in various fields throughout the slide in the light microscope. Once obtained, these data were analyzed through ANOVA and Tukey test as post-hoc analysis, with the help of the software R Statistics. Water quality factors (temperature, pH and2 dissolved) were acceptable and during the 12 month period the sperm of tilapia (Oreochromis sp.) had a good ability for fertilization, seen that it performed below the average of the percentage of critical abnormalities, and quality was perceived by the parameters that also influence fertilization (motility rate, duration of motility and vigor).

Effects of Inositol Supplementation in Sperm Extender on the Quality of Cryopreserved Mesopotamian Catfish (Silurus triostegus, H. 1843) Sperm

This study was based on the determination of post-thaw semen quality, oxidant–antioxidant status and DNA-damage status of frozen Mesopotamian catfish semen. To this end, the sperm was frozen in diluent containing different inositol concentrations (5, 10, 20 and 40 mg). Increasing levels of inositol linearly increased the spermatozoa motility rate and duration significantly (p < 0.05). High doses of inositol increased antioxidants and decreased oxidants. In addition, lower intracellular DNA damage and percentage of apoptotic spermatozoa occurred in high doses of inositol (p < 0.05).Abstract: In this study, the effects of supplemented inositol on sperm extenders were examined on the spermatozoa motility rate and duration, total antioxidant and oxidant status, apoptotic spermatozoa and DNA damage, during the sperm post-thaw process of Mesopotamian Catfish (Silurus triostegus, H. 1843). The semen was frozen in diluents containing different inositol concentrations (5, 10, 20 and 40 mg). Increasing levels of inositol linearly improved the spermatozoa motility rate and duration significantly (p < 0.05). MDA and TOS were linearly decreased, however, TAS and GSH linearly increased (p < 0.05). The increasing inositol levels resulted in a linear and quadratic decrease in DNA damage in the comet assay, 8-hydroxydeoxyguanosine and the determined percentage of apoptotic spermatozoa (p < 0.05). These results suggest that there are many positive effects of the use of supplemental inositol on enhancing sperm cryopreservation efficiency in Silurus triostegus.

Using Gum Arabic in Place of Egg Yolk as Cryoprotectant for Cryopreservation of the Buck Semen

Background: Egg yolk (EY) is well known to be toxic for buck spermatozoa, which creates restrictions of its use in cryopreservation. Therefore, this study is to compare the effect of different levels of gum Arabic (GA) in an extender on quality and fertility of cryopreserved buck sperm. Methods: Each ejaculate of six bucks was frozen in Tris with one of concentrations of GA which contained 3, 6, 9 and 12 gm/ 100 ml in place of the EY. Control was Tris extender containing 2.5% of EY. Result: A percentages of total motile sperm (54.92%; P less than 0.05) and progressively motile sperm (26.22%; P less than 0.05) of semen was frozen in Tris containing 9% of GA. Similar to control group, the pregnancy rate of does inseminated with extender containing 9% (50.0%) were significantly higher than those of does inseminated with extender containing 6% (8.33%), 3% (0.0%) and 12% (0.0%). Semen evaluation and fertility rate were similar when replacing the EY with GA in the Tris cryodiluent, after cryopreservation of buck semen. The present study shows that high motility rate of frozen semen and acceptable pregnancy rate can be obtained when using GA in place of EY for cryopreserving the buck sperm.

Echinacoside Protects Against Dysfunction of Spermatogenesis Through the MAPK Signaling Pathway

Dysfunction at various levels of spermatogenesis (SD) is one of the important causes of infertility in men of reproductive age and requires advanced treatment strategies. Increasing evidence suggests that the therapeutic effects of echinacoside (ECH) mainly depend on their capacity to inhibit cell death. This study aimed to explore the therapeutic potential of ECH in SD rat models. Treatment with ECH reverted the morphological changes observed in testes with spermatogenesis dysfunction. It improved total sperm number, decreased the sperm deformity rate, and increased the sperm forward motility rate. The level of glutathione (GSH) was significantly higher in ECH-treated mice, whereas the lactate dehydrogenase (LDH) and SOD activities were improved compared with those in the spermatogenesis dysfunction model. Moreover, the increased expression of p38 and JNK was partially reversed by ECH. The number of normal TM3 cells increased gradually in an Echinacea dosage-dependent manner, suggesting that ECH promoted the proliferation of TM3 cells. In addition, treatment with ECH partially reversed the increased expression of p38 and JNK in TM3 cells. ECH protects against oxidative stress damage by activating antioxidant enzymes and MAPK signaling-related factors (p38 and JNK). It suggested that treatment with ECH alleviated spermatogenetic dysfunction of testes in male mice and it could be a promising strategy for patients suffering severe SD.

Effect of Semen Freezing and Thawing on Sperm Survival And Motility Rate : A Comparative Analysis

Objective : To compare the rate of sperm survival and motility through semen freezing and thawing during infertility treatment. Methodology: In this bidirectional observational study, we enrolled 31 patients who underwent semen analysis for infertility treatment at the Institute of Reproduction and Child Cares & IRCC IVF Centre, Panchkula, Haryana, from June 2020 to December 2020. Out of these patients, 21 (67.74 %) were considered for semen freezing and thawing. For the rest of the ten patients (32.25 %), sperm count and motility were not good, and we excluded them from this study. Semen freezing based upon sperm count and motility were done. We did semen thawing after two weeks of semen freezing and recorded the sperm survival and motility. Results: Post thaw sperm survival rate and motility was 37.66% compare to pre-cryopreservation (61.82%). The observed rate of sperm motility declining was 24.16 % after cryopreservation/freezing. Conclusion: The present study results concluded that sperm's cryopreservation results in a decrease in sperm motility. There is a need of finding more accurate and reliable methods to freeze and thaw semen.

Effect of Bioxcell® and Triladyl® Extenders and Removal of Seminal Plasma on Equilibrated and Cryopreserved Semen from South African Unimproved Indigenous Bucks

The objectives of the study were to evaluate the effect of two extenders (Triladyl® and Bioxcell®) and the removal of seminal plasma on indigenous buck’s semen. Semen was collected from six indigenous bucks using an electro-ejaculator. Raw semen was pooled and randomly allocated into six groups as follows: (i) Raw non-washed, (ii) Raw washed, (iii) Triladyl®-washed, (iv) Triladyl®-non-washed, (v) Bioxcell®-washed and (vi) Bioxcell®-non-washed. Both the Triladyl® and Bioxcell® washed semen samples groups were diluted (1:4 v/v) with Phosphate Buffered Saline (PBS) then centrifuged at 1500x g for ten min and seminal plasma was removed. The groups were analysed for spermatozoa motility rates using Computer-Aided Sperm Analysis (CASA). The spermatozoa viability was assessed using Eosin-Nigrosin, acrosome integrity using Spermac, chromatin structure using Acridine Orange, and mitochondria using JC-1 staining solutions. Semen samples were diluted (1:4 v/v) as follows: Triladyl® (washed and non-washed) or Bioxcell® (washed and non-washed) and then equilibrated at 5°C for 2 hours. Equilibrated semen samples in 0.25 mL French straws were placed 5 cm above a Liquid Nitrogen (LN2) vapour for 10 min, and stored for one month. Frozen semen straws per treatment group were thawed at 37°C for 30 seconds. Significant differences among the mean values of semen parameters were determined by Tukey’s test using ANOVA, GLM procedure of SAS version 12.1 of 2010. The spermatozoa progressive motility rate in non-washed semen extended with Bioxcell® was significantly higher (89.6±7.5a) compared with that of non-washed Triladyl®, washed Bioxcell® and Triladyl® (P<0.05). Live spermatozoa percentage in washed semen extended with Triladyl® extender was reduced (27.7±17.1) significantly compared with the other groups (P<0.05). There was a lower percentage of spermatozoa with high mitochondrial membrane potential in non-washed and washed semen extended with Bioxcell® (39.5±23.2 and 37.9±28.6, respectively) compared with that of non-washed and washed semen extended with Triladyl® (P>0.05). The spermatozoa progressive motility rate in non-washed semen extended with Bioxcell® (58.5±10.0) extender was significantly higher compared with that of the other groups (P<0.05). There was a higher live and normal spermatozoa percentage in non-washed semen extended with Bioxcell® (45.7±21.2) compared with that of the other groups (P<0.05). In conclusion, Washing of seminal plasma in semen extended with Triladyl® was not essential, as it lowered viability, progressive motility and chromatin membrane integrity prior and post-cryopreservation. However, Bioxcell® extender was found to be more suitable for preserving spermatozoa during equilibration and freezing/thawing process of non-washed and washed buck semen.

Mild Effects of Sunscreen Agents on a Marine Flatfish: Oxidative Stress, Energetic Profiles, Neurotoxicity and Behaviour in Response to Titanium Dioxide Nanoparticles and Oxybenzone

UV filters are potentially harmful to marine organisms. Given their worldwide dissemination and the scarcity of studies on marine fish, we evaluated the toxicity of an organic (oxybenzone) and an inorganic (titanium dioxide nanoparticles) UV filter, individually and in a binary mixture, in the turbot (Scophthalmus maximus). Fish were intraperitoneally injected and a multi-level assessment was carried out 3 and 7 days later. Oxybenzone and titanium dioxide nanoparticles induced mild effects on turbot, both isolated and in mixture. Neither oxidative stress (intestine, liver and kidney) nor neurotoxicity (brain) was found. However, liver metabolic function was altered after 7 days, suggesting the impairment of the aerobic metabolism. An increased motility rate in oxybenzone treatment was the only behavioural alteration (day 7). The intestine and liver were preferentially targeted, while kidney and brain were unaffected. Both infra- and supra-additive interactions were perceived, with a toxicodynamic nature, resulting either in favourable or unfavourable toxicological outcomes, which were markedly dependent on the organ, parameter and post-injection time. The combined exposure to the UV filters did not show a consistent increment in toxicity in comparison with the isolated exposures, which is an ecologically relevant finding providing key information towards the formulation of environmentally safe sunscreen products.

Establishment of a Mouse Asthenospermia Model through Triggering DGalactose Mediated Oxidative Stress Injury

Background: Asthenospermia is defined as forward motility of sperm less than 32%. Aim/Objective: This study aimed to establish mouse model of asthenospermia through triggering D-galactose mediated oxidative stress. Materials and methods: Total of 40 Kunming male mice were randomly divided into control group, low-dose group (administrating D-galactose at 60 mg/kg), high-dose group (administrating D-galactose at 120 mg/kg) and high-dose+feed addition group (administrating D-galactose at 120 mg/kg together with oral D-galactose). The testicular weight, testicular organ coefficient, sperm viability, sperm concentration and survival rate of tail of epididymis were measured. Oxidative damage of D-galactose to reproductive system of mice was evaluated by measuring superoxide dismutase (SOD) and malondialdehyde (MDA) in testicular homogenate of mice. Findings: The sperm motility, motility rate, concentration and survival rate of low-dose, high-dose and high-dose+feed addition group were decreased, compared to that in control group. However, there were significant difference between highdose group/high dose+feed group and control group (p<0.05). The forward motile sperm motility rate and total motility rate accorded with critical criteria of asthenospermia. Comparing with the control group, activity of SOD of model group mice significantly decreased, and MDA concentration significantly increased (p<0.05), excepting for low-dose versus control group for SOD activity. This suggests that testicular tissues suffered from oxidative damage. Conclusions: This study successfully established a mouse asthenospermia model through D-galactose mediated oxidative stress injury. The establishment of asthenospermia model in this study would provide a new promising insight and act as a potential approach for studying asthenospermia in vivo levels.