Sperm Activation Research Articles

Do egg cell‐secreted aspartic proteases promote gamete attachment?

During the double fertilization process of flowering plants (angiosperms), pollen tubes transport a pair of immobile sperm cells towards and inside the embryo sac for fusion with egg and central cell, respectively. Male-female gamete interactions during fertilization are critical for reproductive success and seed development, and thus have interested biologists since decades. Compared to animals, little is known about gamete interaction mechanisms in angiosperms. This article is protected by copyright. All rights reserved.

The Impact of Adding Melatonin and Other Antioxidants on Post-Thaw Human Sperm Quality During Cryopreservation

Sperm cryopreservation is the process of storing sperm for an extended period of time in order to maintain male fertility. Cryopreservation involves exposing germ cells to substances that prevent freezing, cooling them to below-freezing temperatures, storing, melting, and then removing the anti-freezing material when it has been used. It is helpful in cancer patients before chemotherapy and radiotherapy. Spermatozoa can be adversely impacted by reactive oxygen species by its detrimental effects on sperm membrane lipids, which cause ice crystal formation and the induction of oxidative stress (OS) during cryopreservation. Owing to the tight relationship between OS induction and cryopreservation, several recent researches have concentrated on the function of antioxidants in preserving male fertility. A variety of antioxidants have been developed for in vitro supplementation in an attempt to prevent the cellular harm brought on by cryopreservation. Examples of antioxidants include melatonin, catalase, superoxide dismutase, tocopherol, ascorbic acid, and carotenoids. When added to sperm extenders, melatonin, a natural hormone that plays a role in a number of sperm physiological processes, has frequently increased sperm viability and fertility. In order to determine whether it can protect human sperm from the damaging effects of cryopreservation, it was added to the sperm cryopreservation solution. Melatonin was added to freezing extenders in recent studies on mammals, and this increased the post-thaw activities of human sperm. Therefore, this study was aimed to review the background documents on the state-of-the-art scientific literature in this area of work. Also, this study reviewed the feasibility of employed melatonin in cryopreservation because it has antioxidant ability.

Improving Capacities of Chilled- and Cryo-Preserved Buck Spermatozoa by Enriching Preservation Medium with Moringa oleifera leaves extract

In the present work, two experiments were conducted to investigate the efficiency of supplementing buck sperm medium with Moringa oleifera leaves extract on liquid-chilled storage and cryopreservation capacities of spermatozoa. Twenty ejaculates were collected by an artificial vagina from 5 Damascus bucks, 4 ejaculates each, during September-October. The pooled specimens of each collection session were diluted (1:10) with Tris-citric egg yolk extender, and were split into 6 equal aliquots. The first aliquot served as control (moringa-free), whereas the other five aliquots were supplemented with 0.05, 0.1, 0.2, 0.3 or 0.4 mL/ml (v/v) moringa leaves extract, respectively. The samples were then stored and maintained at 4 oC for the subsequent 48 h during which sperm traits were assessed alongside malondialdehyde (MDA) production and glutathione peroxidase (GPX) activity. The optimum concentration of moringa leaves extract that showed potential for maintaining sperm traits throughout the chilled storage period was further investigated for preserving the lifespan of spermatozoa after exposure to the cryopreservation stress against control. The results showed that moringa leaves extract supplementation reduced (P<0.05) the oxidative stress on spermatozoa and recorded a positive correlation (P<0.01) with sperm motility (%), viability (%), normal sperm (%), acrosome integrity and GPX activity (r= 0.36, 0.42, 0.61, 0.79 and 0.35, respectively) while recording a negative correlation (P<0.01) with production of MDA (r= - 0.56). Furthermore, supplementing buck sperm medium with 0.4 mL/ml moringa leaves extract efficiently improved (P<0.05) all sperm characteristics and decreased (P<0.05) DNA fragmentation index (DFI) post thawing compared to those of control. These results imply the powerful potential of moringa leaves extract as an exogenous antioxidant supplement in buck sperm medium particularly when spermatozoa are used for AI and IVFM applications.

Resfriamento de sêmen suíno em um diluente suplementado com isoespintanol

ABSTRACT: Spermatozoa experience oxidative, osmotic, chemical, and thermal stresses when cooled, which degrade the quality and fertilizing capacity of the cells. Adding antioxidants to the sperm extender mitigates these alterations. This study evaluated the effect of isoespintanol (ISO) on boar semen subjected to cooling. Fifteen ejaculates from five boars (Susscrofadomestica) were extended in Beltsville thawing solution (BTS) supplemented with 0 µM (control), 5 µM (ISO5), 10 µM (ISO10), 15 µM (ISO15), 20 µM (ISO20), 25 µM (ISO25), and 30 µM (ISO30) of ISO, which were then cooled for five days at 16 °C. Sperm kinetics, total motility (TM), and progressive motility (PM) were evaluated every 24 h using an IVOS computer-assisted sperm analysis (CASA) system. On day 1 and day 5 of cooling, a hypoosmotic test, spectrofluorometry, and flow cytometry were performed to evaluate the following: membrane functionality, measured as a function of hypoosmotic swelling (HOS); total antioxidant capacity (TAC); reactive oxygen species (ROS); and mitochondrial membrane potential (Δ¥M). Regression analysis and comparison of means using the Duncan test were performed. The ISO added had a slight impact on sperm motility, as evidenced by a reduction in TM at 24 h of cooling (but not prior) with the addition of 20 µM of ISO. Similarly, no effect of the ISO on the kinetics and functional integrity of the sperm membrane was observed at 96 h of cooling; however, the regression coefficients indicated that the ISO lowered the rate of decrease in sperm motility and the proportion of rapid spermatozoa relative to the concentration of ISO used. The ISO did not affect the TAC of the cooled semen; however, different concentrations of ISO lowered ROS production in the semen after 96 h of cooling. ISO also impacted the Δ¥M of the spermatozoa at 0 h of cooling, increasing the proportion of low Δ¥M cells and decreasing the proportion of high Δ¥M cells. In conclusion, ISO can reduce the loss of quality and oxidative stress occurring in boar semen during cooling and can modulate the mitochondrial activity of sperm.

Quality of ram semen in relation to scrotal size

Aim of this study was to select superior rams by assessing their scrotal size and quality of semen. In ten indigenous Bangladeshi rams, the body weight, scrotal circumference, and scrotal volume of rams increased with age (P<0.05). Body weight at 381-410 days (13.6 ± 1.5 kg) was higher than at 169-200 days (10.2 ± 0.1 kg) and 201-230 days (10.4 ± 1.2 kg) (p<0.05). Scrotal circumference at 381 - 410 days (19.1 ± 0.3 cm) was higher (p<0.05) than at 169 - 200 days (17.2 ± 0.1 cm), 201 - 230 days (17.2 ± 0.6 cm), and 231 - 260 days (17.2 ± 0.5 cm). At 381 - 410 days, scrotal volume (87.4 ± 5.5 cm3) was higher than in the five youngest groups (p<0.05). Both scrotal circumference and volume were positively correlated (R2=0.519) with the increased body weight. Semen volume increased with age, which varied from 0.4 ± 0.0 - 1.2 ± 0.3 ml. The semen volume at 381 - 410 days (1.2 ± 0.3 ml) was higher than other age groups (P<0.05) except for 351 - 380 days. The mass activity of sperms of different age groups varied from 2.8 ± 0.4 to 4.3 ± 0.3. Sperm concentration, progressive motility (%), and normal spermatozoa (%) improved progressively with age, but significant changes in these parameters were seen when the animals reached 291 days or more (P<0.05). However, normal spermatozoa (%) improved from the age of 261 days. The mass activity was positively correlated with sperm concentration (R2 = 0.568) and with progressive motility (%) (R2 = 0.616). Measurements of scrotum could provide a reliable guide in estimating sperm production capacity, which are important for breeding soundness evaluation, and genetic improvement of indigenous rams.
 Bangl. vet. 2021. Vol. 38, No. 1-2, 1–9

Ovarian lavage method as an alternative route for hormonal administration and short-term sperm storage in Clarias gariepinus

Ovarian lavage is a term used to describe the injection of fish with a catheter through the oviduct into the ovary. In this study, the efficacy of this technique was evaluated as a route for hormone administration and sperm preservation in the African catfish Clarias gariepinus. Firstly, the effects of hormone injection routes (namely, intramuscular, intraperitoneal, and ovarian lavage) were evaluated on breeding and haematological parameters. In the second study, the fish's spermatozoa were stored in the ovaries for 1, 2, 3, and 4 days before stripping, sperm activation with freshwater, and fertilization. The breeding performance was then compared with eggs fertilized using spermatozoa refrigerated for the same duration. The study showed that the administration of synthetic hormone (ovaprim®) through the ovaries was comparable to the intramuscular route, while those injected intraperitoneally had the least values (P < 0.05) for breeding performance. The trend of the haematological parameters also suggests the intraperitoneal administration procedure elicited more stress compared to intramuscular or ovarian lavage routes (P < 0.05). In the second experiment, the use of sperm stored in the ovaries or refrigerated for 24hr didn't affect (P > 0.05) the fertilization (92–93%) and hatching (81–83%) of the eggs when compared to the control (91% and 82%). Beyond this 24hr threshold, breeding performances were significantly reduced in the ovarian lavage treatments compared to those fertilized with refrigerated sperm (P < 0.05). The loss of fertilizing capacity observed herein was justified by the reduction in sperm quality over time (P < 0.05). It was concluded that ovarian lavage is a viable route for hormonal administration and 24hr sperm storage in C. gariepinus.

Comparative assessment of the effect of titanium dioxide – based nanoparticles on boar germ cells in vitro

Modern titanium dioxide nanomaterials, in particular modified with nanosilver, have antiviral, antibacterial, antifungal activity, cytotoxic effects in vitro experiments and can affect germ cells of warm-blooded laboratory animals. The research aims to study the effect of different types of titanium dioxide nanoparticles on the physiological and biochemical characteristics of wild boar germ cells in vitro. When applying the maximum dose (LD50) of the studied nanopowders in all samples, sperm survival was probably lower than in the control: by 41.9% (р&lt;0.001) ‒ under the action of TiO2 nanoparticles (hereinafter NPs) (Frantsevich Institute for Problems of Materials Science of NAS of Ukraine, hereinafter IPM), by 28.0% (р&lt;0.05) ‒ under the action of TiO2 NPs (Acros Organics), by 53.5% (р&lt;0.001) ‒ under the action of 4% Ag-TiO2 NPs (IPM) and by 55.9% (р&lt;0.001) ‒ are the action of NPs 8% Ag-TiO2 (IPM). Nanopowders of different based on titanium dioxide manufacturers, characterized by similar morphological characteristics and varying degrees of nanosilver addition (Ag content from 0 to 8%), starting from a dose of 1/10 LD50, can have a damaging effect on the germ cells of wild boars, which is manifested by a decrease in the respiratory activity of sperm and cytochrome oxidase activity, by increasing succinate dehydrogenase activity. The most sensitive marker was the sperm survival rate, the value of which significantly decreased under the action of 8% Ag-TiO2 nanoparticles (IPM) at a dose of 1/100 LD50, which is a consequence of disruption of the activity of mitochondrial enzymes and, accordingly, resynthesis of ATP. Our results confirm and extend the data on the nature of the damaging effect of titanium dioxide-based nanoparticles on germ cells of warm-blooded animals due to oxidative stress. The obtained experimental data will be taken into account in the hygienic regulation of the investigated nanopowders.

Protective effects of different doses of MitoQ separately and combined with trehalose on oxidative stress and sperm function of cryopreserved Markhoz goat semen

The mitochondria-targeted antioxidant MitoQ has been regarded as an effective antioxidant agent against cryo-induced oxidative cellular damage. This study aimed to evaluate the use of different doses of MitoQ combined with trehalose to minimize mitochondrial impairment and oxidative stress during sperm cryopreservation of Markhoz goat. For this, semen samples (n = 50) were collected by electroejaculation every 5 days from 5 bucks in 10 replicates. On each collection day, 5 ejaculates (one ejaculate for each buck) were pooled and then diluted in eight different Tris-based extenders as follows: no additives (control), 20, 200, 2000 nM of MitoQ (MT20, MT200, MT 2000, respectively), 150 mM of trehalose (Tr), MT20+Tr, MT200+Tr, MT2000+Tr. The semen samples were frozen using a standard protocol, and sperm function and oxidative stress were evaluated after thawing. The semen extender supplemented with MT200+Tr had higher (P < 0.05) total and progressive motility, acrosome and membrane integrity, superoxide dismutase, glutathione peroxidase, total antioxidant capacity, and lower (P < 0.05) DNA fragmentation, malondialdehyde and intracellular hydrogen peroxide levels than the all other groups except MT200; meanwhile, MT200 was also improved (P < 0.05) in these parameters than in the control group. Furthermore, MT200 and MT200+Tr showed higher (P < 0.05) percentages of live cryopreserved sperm with high mitochondrial activity than other groups. However, abnormality percentage and catalase activity of frozen-thawed sperm were not affected by treatments (P > 0.05). To conclude, we have found that supplementation of 200 nM MitoQ alone or in combination with 150 mM trehalose to semen extender improved the quality of cryopreserved sperm in goats, which is associated with enhanced antioxidant enzymatic defense and mitochondrial activity and reduced DNA fragmentation.

THE REPRODUCTIVE ABILITY OF MALE RABBITS DEPENDING ON THE INFLUENCE OF PARATYPICAL AND GENOTYPICAL FACTORS

THE REPRODUCTIVE ABILITY OF MALE RABBITS DEPENDING ON THE INFLUENCE OF PARATYPICAL AND GENOTYPICAL FACTORS

Effect of Silybum marianum (L.) Geartn extract on in vitro fertilization in mice

Composition of Silybum marianum (L.) Geartn extract rich with flavonoid sylimarin which have antioxidant properties. Aim of this project study the effects of sylimarin on In vitro fertilization (IVF) and cleavage stages of embryos in mice. Fifteen adult males and females mice were divided into three groups , each group containing 5 mice , first and second groups were treated with (25 and 50 mg/kg body weight) of Silybum marianum (L.) Geartn extract (IP) while the third group treated with normal saline as control group . After 35 days of treatment the animals (male and female) were sacrificed and sperms isolated from tail of epidydymis were used for the following tests: Sperm motility, dead / live sperm, sperm abnormalities and serum was collected to assay testosterone. Isolation of oocyte from oviduct and serum blood to hormonal assay (FSH and LH) . Sperm and matured ova were incubated in TCM-199 media for insemination. The results showed increase in body weight and testes weight in groups treated with Silybum marianum (L.) Geartn. Significant increase in sperm activity and decrease in percentage of dead sperm and abnormalities in groups treated with Silybum marianum (L.) Geartn. The male testosterone and female FSH, LH increased significantly in groups treated with Silybum marianum (L.) Geartn. IVF study showed significant increase in percentage of maturation, fertilization and cleavage (2-4 cells). In conclusion sylimarin have benefits effect on IVF and cleavage stages of embryo development when injected before IVF by improve fertilization of males and females.

CX3CR1 deficiency leads to impairment of immune surveillance in the epididymis.

Mononuclear phagocytes (MPs) play an active role in the immunological homeostasis of the urogenital tract. In the epididymis, a finely tuned balance between tolerance to antigenic sperm and immune activation is required to maintain epididymal function while protecting sperm against pathogens and stressors. We previously characterized a subset of resident MPs that express the CX3CR1 receptor, emphasizing their role in antigen sampling and processing during sperm maturation and storage in the murine epididymis. Bacteria-associated epididymitis is the most common cause of intrascrotal inflammation and frequently leads to reproductive complications. Here, we examined whether the lack of functional CX3CR1 in homozygous mice (CX3CR1EGFP/EGFP, KO) alters the ability of MPs to initiate immune responses during epididymitis induced by LPS intravasal-epididymal injection. Confocal microscopy revealed that CX3CR1-deficient MPs located in the initial segments of the epididymis displayed fewer luminal-reaching membrane projections and impaired antigen capture activity. Moreover, flow cytometry showed a reduction of epididymal KO MPs with a monocytic phenotype under physiological conditions. In contrast, flow cytometry revealed an increase in the abundance of MPs with a monocytic signature in the distal epididymal segments after an LPS challenge. This was accompanied by the accumulation of CD103+ cells in the interstitium, and the prevention or attenuation of epithelial damage in the KO epididymis during epididymitis. Additionally, CX3CR1 deletion induced downregulation of Gja1 (connexin 43) expression in KO MPs. Together, our study provides evidence that MPs are gatekeepers of the immunological blood-epididymis barrier and reveal the role of the CX3CR1 receptor in epididymal mucosal homeostasis by inducing MP luminal protrusions and by regulating the monocyte population in the epididymis at steady state as well as upon infection. We also uncover the interaction between MPs and CD103+ dendritic cells, presumably through connexin 43, that enhance immune responses during epididymitis. Our study may lead to new diagnostics and therapies for male infertility and epididymitis by identifying immune mechanisms in the epididymis.

Mitochondrial succinate dehydrogenase function is essential for sperm motility and male fertility.

Mitochondrial health is crucial to sperm quality and male fertility, but the precise role of mitochondria in sperm function remains unclear. SDHA is a component of the succinate dehydrogenase (SDH) complex and plays a critical role in mitochondria. In humans, SDH activity is positively correlated with sperm quality, and mutations in SDHA are associated with Leigh Syndrome. Here we report that the C.elegans SDHA orthologue SDHA-2 is essential for male fertility: sdha-2 mutants produce dramatically fewer offspring due to defective sperm activation and motility, have hyperfused sperm mitochondria, and disrupted redox balance. Similar sperm motility defects in sdha-1 and icl-1 mutant animals suggest an imbalance in metabolites may underlie the fertility defect. Our results demonstrate a role for SDHA-2 in sperm motility and male reproductive health and establish an animal model of SDH deficiency-associated infertility.

Opioid replacement therapy with methadone or buprenorphine effects on male mice reproduction.

Opioid use disorders are commonly treated by long-acting agonist opioids including methadone and buprenorphine which could affect various aspects of male reproduction especially spermatogenesis. We aimed to determine whether detoxification with methadone or buprenorphine was associated with reproductive disorders in male mice. We orally induced morphine dependence in NMRI male mice, and then performed detoxification programs using either methadone or buprenorphine. Testis architecture and sperm parameters including sperm nuclear DNA integrity, mitochondrial activity, oxidative stress in seminal plasma, and routine sperm parameters were assessed to find the involved mechanisms. The number of Leydig cells and the thickness of germinal epithelium reduced following morphine use and increased differently after detoxification with methadone or buprenorphine. Morphine dependence and detoxification with methadone and buprenorphine had different effects on sperm parameters. Morphine altered chromatin integrity, mitochondrial activity, and oxidative stress in sperm. Detoxification with methadone improved mitochondrial activity but worsened chromatin integrity, whereas detoxification with buprenorphine improved neither chromatin integrity nor mitochondrial activity. Seminal plasma oxidative stress was higher in the treated groups compared to control groups but was comparable among treatment groups. Our study revealed that long-term morphine use followed by detoxification with methadone or buprenorphine impairs testis structure and sperm parameters. Detoxification from morphine use with methadone and buprenorphine led to different preclinical outcomes in semen quality parameters, including chromatin integrity. Therefore, clinical detoxification protocols should be performed more cautiously, considering the desire of the individuals to reproduce.

Sperm productivity of Texel breed rams depending on the period of seasonal activity and mode of use

The results of laboratory studies of freshly obtained and deep-frozen semen of Texel breeding rams in the non-mating and mating periods under different modes of using 4, 6, and 8 ejaculates per week are given. It has been proven that during the non-mating period, when receiving four ejaculates per week compared to 6 and 8 ejaculates, the ejaculate volume was 11.3 and 22.6 % higher, respectively. The sperm concentration in it was 5.1 and 12.2 %, sperm activity was higher by 1.7 and 3.2 %, and their resistance by 7.1 and 17.3 percent. In the mating period under the above regimen, the ejaculate volume was 9.9 and 19.7 % higher; sperm concentration was 4.1 and 5.9 % higher; sperm activity was higher by 5.7 and 8.3 %, resistance was higher by 7 .7 and 19.9 percent. Higher quality indicators were found in the mode of obtaining four ejaculates after sperm cryopreservation in both research periods, on average, according to the activity of deconserved sperm by 3.2 and 8.0 and 5.8 and 11.2 %, freezing resistance by 0.7 and 2,0 and 2.6 and 4.6 %, survival by 7.6 and 15.5 and 2.2 and 10.6%, absolute survival rate by 3.4 and 8.1 and 4.7 and 8.5 percent Based on the analysis of the investigated indicators of freshly obtained and de-preserved sperm, it was experimentally confirmed that in the mating period compared to the non-mating period, they were significantly higher on average in freshly obtained sperm from 7.7 to 40.1 %, in cryopreserved sperm – from 2.9 to 16.7 percent. The preservation of acrosomes of sperm in the non-mating and mating period, when receiving four ejaculates per week compared to receiving 6 and 8, was higher by periods by 3.5 and 6.0 %, and 2.5 and 5 %, respectively. The studies established significantly higher quantitative and qualitative indicators of freshly collected and deconserved sperm in both seasonal activity periods under the regime of obtaining four ejaculates per week from breeding rams. Under this regime, the sperm is biologically complete, and its quality indicators meet the requirements of the instructions for the artificial insemination of sheep.

EPICATECHIN IMPROVES FROZEN SPERM VITALITY BY ITS ANTIOXIDANT AND CRYOPROTECTIVE ACTIONS

This study served to elucidate the effects of three selected doses of epicatechin (EPI; 25, 50 or 100 μmol/L) on the motion and oxidative profile of bovine spermatozoa subjected to the process of cryopreservation. Furthermore, we focused on the impact of EPI on the expression patterns of heat shock proteins 90 and 70 as well as the BAX and Bcl-2 proteins involved in the process of apoptosis. The experimental samples were compared with a native control as well as specimens cryopreserved in the absence of any supplement. Our dada indicates that particularly 50 and 100 μmol/L EPI assured a higher preservation of the sperm motility (p&lt;0.001) in comparison with the cryopreserved control. The best protection of the sperm DNA (p&lt;0.05), lipids (p&lt;0.05) and proteins (p&lt;0.01) were observed in the samples exposed to 100 μmol/L EPI in comparison to untreated frozen samples. The presence of higher EPI concentrations in the cryopreservation medium led to significant stabilization of the heat shock protein 90 (p&lt;0.01 in case of 50 μmol/L EPI; p&lt;0.001 with respect to 100 μmol/L EPI) as well as a higher maintenance of the BAX:Bcl-2 ratio (p&lt;0.001) when compared to the untreated frozen control. In summary, we may assume that EPI exhibits antioxidant properties which enable the molecule to prevent excessive damage to biomolecules essential for the cell survival and offer an additional layer of protection to spermatozoa exposed to low temperatures, all of which may be translated into a higher post-thaw sperm motility and activity.

Ameliorative effect of silymarin on the quality of frozen-thawed boar spermatozoa.

Although Silymarin (SMN) has powerful antioxidant properties, little is known about its effects on the quality of frozen-thawed boar sperm. The present study aimed to evaluate the influences of SMN added to the thawing extender on boar sperm parameters essential for fertilization. The frozen-thawed semen was diluted in a Modena thawing extender supplemented with different concentrations of SMN (0, 5, 10, 20 and 50 μM respectively), and then the changes in quality parameters, antioxidant capacity, mitochondrial function and in vitro fertilization (IVF) capability of frozen-thawed sperm were assessed. Here we demonstrated that the motility, plasma membrane integrity and acrosomal integrity of frozen-thawed sperm improved efficiently by SMN (p < .05). In antioxidant parameters evaluation, the tROS level and MDA content of frozen-thawed spermatozoa were reduced in the 20 μM SMN group, while the T-AOC activity significantly increased (p < .05), indicating that the supplementation with SMN can promote the antioxidant capacity of frozen-thawed boar sperm. Besides, we also discovered that the addition of SMN significantly upregulated ATP content and enhanced the mitochondrial activity of sperm. More interestingly, SMN promoted the activities of mitochondrial respiratory chain complexes (MRCC) I, II, III and IV in frozen-thawed sperm significantly. Functionally, the higher penetration rate and increased total efficiency of fertilization were observed in the 20 μM SMN group. In summary, supplementation with SMN in the thawing medium ameliorates the quality of frozen-thawed boar sperm by enhancing mitochondrial respiratory capacity, producing large amounts of ATP and regulating ROS formation.

Effect of Trigonella Foenum-Graecum Foenugreek oil extract on some parameters of sperms and in vitro fertilization in mic

This project, aims to study the effect of trigonella seeds oil extract on body weight , testes weight and some parameter of reproductive system in male mice , and on In vitro fertilization (IVF) , cleavage stages of embryos in females. Fifteen adult (8-9 weeks) males and females mice were divided into three groups , each group containing 5 mice , first and second groups were treated with (2 and 4 mg/kg body weight) of Trigonella oil extract (intraperitonally) while the third group treated with normal saline as control group . After 35 days of treatment with Trigonella the animals (male and female) were sacrificed by dislocation of cervical vertebrate. Isolated sperms from tail of epidydymis were used for the following tests. Sperm motility, dead / live sperm, sperm abnormalities and serum ws collected to assay testosterone. Isolatation of ova from oviduct after micing in 500μl of TCM-199 media serum blood to assay FSH and LH . Sperm and matured ova were incubated in TCM-199 media for insemination. The results showed increase in body weight and testes weight in groups treated with trigonella. significant increase in sperm activity and decrease in percentage of dead sperm and abnormalities in groups treated with trigonella oil. The male testosterone and female FSH, LH increased significantly in groups treated with trigonella. IVF study showed significant increase in percentage of maturation, fertilization and cleavage (2-4 cells).

Analysis of reproductive damage in earthworms (Amynthas corticis) exposed to cypermethrin.

Cypermethrin contamination was a potential threat to soil organisms. In the present work, reproductive damage in earthworms (Amynthas corticis) exposed to cypermethrin was investigated. It was found that earthworms could absorb and accumulate residual cypermethrin in soil, and also earthworm activities helped accelerate the degradation of cypermethrin in soil. The accumulation of cypermethrin in earthworms induced sperm damage, and cypermethrin not only caused the imbalance of calcium homeostasis in earthworm sperm cells by inhibiting earthworm sperm Ca2+-ATP and Ca2+-Mg2+-ATP enzyme activities but also caused barriers in acrosome reaction. It also affected sperm energy supply of earthworms by inhibiting the activity of Na+-K+-ATPase and Mg2+-ATPase of earthworm sperm. Meanwhile, the inhibition of acrosome enzyme activity of earthworm sperm by cypermethrin led to hinder fertilization and reduced cocoon production of earthworms, and the damage of cypermethrin to sperm of earthworm was a significant cause of its reproductive toxicity. The results of the evaluation of IBR index showed that reproductive toxicity of cypermethrin to earthworms reduced with the increasing time. The decreased reproductive toxicity of cypermethrin to earthworms at the later stage of exposure (42-56 d) might be due to a combination of reduced absorption of cypermethrin in soil by earthworms, decreased accumulation of cypermethrin in the body, and improved sperm capacitation.

INFLUENCE OF THE STRUCTURE OF DIETS ON THE PRODUCTIVITY OF SERVICING BULLS

Complete feeding of young replacement and adult breeding animals makes it possible to provide the diets of animals with the necessary nutrients, which have a positive effect on the semen quality and, consequently, increase the period of economic use. Therefore, the purpose of the study was to optimize the structure of diets for servicing bulls. It was found that feeding the experimental diets of winter and summer periods with the same type of feeding increased the ejaculate volume by 4.0-4.2%, sperm activity - by 3.1-6.4%, sperm concentration in ejaculate - by 13.8-15.3%.

Electromagnetic radiations on the functional potential of spermatozoa

The growing exposure to digital communication system and tools leads to radiation toxicity to the users. Unaware of the safety measures, even a kid at the age of one starts operating digital gadgets emitting radio frequency-electromagnetic radiations. Radiations from mobile phones, laptops, note pad, Wi-Fi or other devices are reported to be harmful beyond the permissible limit. So there is a growing concern for the overall health, reproductive and hormonal functions. Experimental studies were conducted by using a Wi-Fi network active laptop and live spermatozoa. The exposure of sperms to the source of EMF showed that the activity of the live sperms got reduced and mortality was observed depending on the exposure duration and the distance from EMF sources. After 5h of exposure to RF-EMF source, the semen quality changed when compared with control. The vitality of sperm in the control was 95±1.0 after 2 hr and it was reduced to 60±1.5 percent at a distance of 1cm and 55 ±2.2 percent at a distance of 10 cm. After 5 hour exposure, the vitality was reduced to 63±1.86 at 1 cm distance and 70± 1.42 at 10 cm distance. The reduction in vitality of sperm after exposure to RFEMF source for 2 hour was 35% at 1cm and 40% at 10cm distance.