Decrease In Motility Of Spermatozoa Research Articles

Detection of Melanocortin Receptor Type 4 (MC4R) Gene in Semen of Etawah Crossbreed and Senduro Goats

Etawah crossbreed and Senduro goats as superior local goats and biological genetic resources that must be preserved in Indonesia. Melanocortin receptor type 4 (MC4R) is the primary gene that regulates food intake and energy balance which can be used as a candidate marker for livestock selection genes. This study aimed to analyze the comparison of cement quality and detect the profile of the MC4R gene in Etawah crossbreed and Senduro goat semen. The samples of this study were Etawah crossbreed and Senduro goat semen. Macroscopic examination of cement includes the evaluation of volume, consistency, odor, color, and pH. Microscopic examination of semen includes motility, abnormalities, and spermatozoa concentration. Examination of motility was performed in the condition of fresh semen, after the addition of diluents, and post-thawing of frozen straw. The polymerase chain reaction (PCR) method was performed to amplify specific fragments of the MC4R gene. There was a decrease in spermatozoa motility before freezing and post-thawing frozen semen compared to fresh semen. The quality of frozen straw post-thawing Etawah crossbreed and Senduro goat was still worth using for artificial insemination and in vitro embryo production. The MC4R gene can be detected in liquid and frozen semen from Etawah crossbreed and Senduro goats using a PCR test in the form of a single band along 642 bp.

Effect of Vitamin C and E Supplementation and Combination of Both in Egg Yolk Tris Diluter on the Quality of Sapera Goat Spermatozoa in the 5 °C Cooling Process

This study was aimed to determine the effect of vitamin C, E supplementation and both combination in the egg yolk tris diluent on the quality of Sapera goat spermatozoa in 5˚C cooling process. The samples were fresh semen collected from Sapera goat. It was divided into four different treatment groups. The control treatment or P0 contained semen sample + egg yolk tris diluent, P1 contained semen sample + egg yolk tris diluent + vitamin C 0,9 mg/ml, P2 contained semen sample + egg yolk tris diluent + vitamin E 1 mg/ml, P3 contained semen sample + egg yolk tris diluent + vitamin C 0,9 mg/ml + vitamin E 1 mg/ ml. The least decrease of spermatozoa motility, viability and membrane integrity was found in P3, which was significantly different to P0 (p<0,05). However, P1 and P2 did not show significant difference in the results of motility and viability evaluation (p>0,05). It can be concluded that the combination of vitamin C and E supplementation can be an optimal antioxidants supplement in egg yolk tris diluent on the quality of Sapera goat spermatozoa in 5˚C cooling process.

Effects of Selected Metal Nanoparticles (Ag, ZnO, TiO2) on the Structure and Function of Reproductive Organs.

Various studies have shown that the reproductive organs are highly sensitive to toxic elements found in the environment. Due to technological progress, the use of nanoparticles has become more common nowadays. Nanoparticles are used for drug delivery because their dimensions allow them to circulate throughout the body and enter directly into the cell. Antimicrobial properties are increasingly used in the manufacture of medical devices, textiles, food packaging, cosmetics, and other consumer products. Nanoparticles provide several benefits, but aspects related to their effects on living organisms and the environment are not well known. This review summarizes current in vivo, and in vitro animal studies focused on the evaluation of toxicity of selected metal nanoparticles (Ag, ZnO, TiO2) on male and female reproductive health. It can be concluded that higher concentrations of metal nanoparticles in the male reproductive system can cause a decrease in spermatozoa motility, viability and disruption of membrane integrity. Histopathological changes of the testicular epithelium, infiltration of inflammatory cells in the epididymis, and prostatic hyperplasia have been observed. Nanoparticles in the female reproductive system caused their accumulation in the ovaries and uterus. Metal nanoparticles most likely induce polycystic ovary syndrome and follicular atresia, inflammation, apoptosis, and necrosis also occurred.

New Insight on the In Vitro Effects of Melatonin in Preserving Human Sperm Quality.

Spermatozoa (SPZ) are sensitive to stressful conditions, particularly oxidative stress, which alters their quality; thus, the use of protective molecules as an antioxidant is encouraged. Herein, we used melatonin (MLT) to investigate its in vitro effects on human sperm parameters under conditions of oxidative stress induced by cadmium (Cd). Fifteen human semen samples were divided into control, Cd-treated, MLT-treated, and Cd+MLT-treated groups and analyzed after 30 min, 6 h, and 24 h of exposure. Results showed a time-dependent decrease in SPZ motility, DNA integrity, and increased apoptosis induced by oxidative stress, and these effects were counteracted by MLT co-treatment. Based on these data, we further explored additional parameters just at 24 h. The induced oxidative stress, highlighted by the increased lipid peroxidation, reduced the percentage of SPZ able to undertake acrosome reaction and altered the levels and localization of some protein markers of motility (PREP, RSPH6A), morphology (DAAM1), and acrosome membrane (PTMA, IAM38); all these effects were counteracted by MLT co-treatment. Interestingly, MLT alone was able to ameliorate motility at 30 min of incubation compared to the control, while at 24 h, it prevented the physiological alteration in terms of motility, DNA integrity, and apoptosis. Collectively, the data encourage MLT use as an integrative molecule to ameliorate human gamete quality when compromised by stressful conditions.

Comparative Effects of Methanol and Aqueous Extracts of Corchorus olitorius Plant on Haematology and Some Reproductive Indices of Male Albino Rats (Wistar Strain)

Corchorus olitorius is a vegetable plant/shrub and the leaves are very nutritious and rich in vitamins, minerals and dietary fibers. The study was carried out to identify the effects two extracts of this plant on the reproductive indices of male Wistar rats&#x0D; Forty-five adult male rats of about 12 weeks old, each weighing about 120g were used for this study, kept in a cage and fed with commercial rat pellets and water was given at ad-libitum. They were randomly divided into groups A, B and C of 15 rats per group. 250mg/kg of the leaf extract was administered orogastrically using once daily for 21 days. Group A and B were treated with methanol and aqueous leaves of Corchorus extract respectively and Group C was given distilled water and served as the control. The weekly sacrifice of five rats per group was done following&#x0D; treatments. In this study, it was observed at the first-week post -treatment that there was significant (p≤0.05) decrease in the PCV and haemoglobin values of group A rats compared to the control group but the values later increased at the second week and third-week post-treatments. There was also a significant decrease in spermatozoa motility in the treated groups A and B compared to the control group C at the first, second- and third-week post treatments. Also, with the results of the serum testosterone level of group A and B compared to group C. The value was higher in group A followed by group B whereas, at the second week and third week, there was no significant difference in the values of the serum testosterone levels compared to the control groups.&#x0D; It is therefore concluded that the methanol and aqueous extract of Corchorus olitorius leaves significantly decreased sperm motility in male albino rats hence could decrease male fertility.

Comparative Effects of Methanol and Aqueous Extracts of Corchorus olitorius Plant on Haematology and Some Reproductive Indices of Male Wistar Rats.

Corchorus olitorius is a vegetable plant/shrub and the leaves are very nutritious and rich in vitamins, minerals and dietary fibers. The study was carried out to identify the effects two extracts of this plant on the reproductive indices of male Wistar rats Forty-five adult male rats of about 12 weeks old, each weighing about 120g were used for this study, kept in a cage and fed with commercial rat pellets and water was given at ad-libitum. They were randomly divided into groups A, B and C of 15 rats per group. 250mg/kg of the leaf extract was administered orogastrically using once daily for 21 days. Group A and B were treated with methanol and aqueous leaves of Corchorus extract respectively and Group C was given distilled water and served as the control. The weekly sacrifice of five rats per group was done following treatments. In this study, it was observed at the first-week post -treatment that there was significant (p≤0.05) decrease in the PCV and haemoglobin values of group A rats compared to the control group but the values later increased at the second week and third-week post-treatments. There was also a significant decrease in spermatozoa motility in the treated groups A and B compared to the control group C at the first, second- and third-week post treatments. Also, with the results of the serum testosterone level of group A and B compared to group C. The value was higher in group A followed by group B whereas, at the second week and third week, there was no significant difference in the values of the serum testosterone levels compared to the control groups. It is therefore concluded that the methanol and aqueous extract of Corchorus olitorius leaves significantly decreased sperm motility in male albino rats hence could decrease male fertility.

Drosophila Lysophospholipase Gene swiss cheese Is Required for Survival and Reproduction.

Simple SummaryBiological evolution implies fitness of newly evolved organisms that have inherent adaptive traits because of mutations in genes. However, most mutations are detrimental, and they spoil the organism’s life, its survival and its ability to leave progeny. Some genes are extremely vital for an organism, and therefore, they tend to save their structure and do not mutate or do it very composedly. That is the case of the gene encoding PNPLA6 lysophospholipase domain that evolved in bacteria, and evolution obliged it to save its function in higher animals. In mammals, complete dysfunction of such a gene is lethal because of its high importance in placenta for early embryo development. Why is it conserved in other species, for instance insects, that have no placenta? Here we studied the role of the PNPLA6-encoding gene named swiss cheese in Drosophila melanogaster fitness. We have found that its dysfunction results in premature death of specimens and their inability to leave enough progeny. Thus, we provide the first evidence for significance of the gene that encodes the lysophospholipase enzyme in fitness of insects.Drosophila melanogaster is one of the most famous insects in biological research. It is widely used to analyse functions of different genes. The phosphatidylcholine lysophospholipase gene swiss cheese was initially shown to be important in the fruit fly nervous system. However, the role of this gene in non-nervous cell types has not been elucidated yet, and the evolutional explanation for the conservation of its function remains elusive. In this study, we analyse expression pattern and some aspects of the role of the swiss cheese gene in the fitness of Drosophila melanogaster. We describe the spatiotemporal expression of swiss cheese throughout the fly development and analyse the survival and productivity of swiss cheese mutants. We found swiss cheese to be expressed in salivary glands, midgut, Malpighian tubes, adipocytes, and male reproductive system. Dysfunction of swiss cheese results in severe pupae and imago lethality and decline of fertility, which is impressive in males. The latter is accompanied with abnormalities of male locomotor activity and courtship behaviour, accumulation of lipid droplets in testis cyst cells and decrease in spermatozoa motility. These results suggest that normal swiss cheese is important for Drosophila melanogaster fitness due to its necessity for both specimen survival and their reproductive success.

Pubertal exposure to low doses of zearalenone disrupting spermatogenesis through ERα related genetic and epigenetic pathways

Endocrine disruptor zearalenone (ZEA) has been found to damage the reproductive system especially spermatogenesis. In our previous report, we have found that low dose (lower than No-Observed Effect Level, NOEL) ZEA exposure disturbed mouse spermatogenesis and diminished mouse semen quality. The purpose of current investigation was to explore the underlying mechanisms of pubertal low dose ZEA exposure upsetting spermatogenesis. And it was demonstrated that pubertal low dose ZEA exposure disrupted the meiosis process and the important genetic pathways to inhibit the spermatogenesis and even to diminish the semen quality with the decrease in spermatozoa motility and concentration. The DNA methylation markers 5mC and 5hmC were decreased, the histone methylation marker H3K27 was increased, at the same time estrogen receptor alpha was diminished in mouse testis after pubertal low dose ZEA exposure. The data indicate that the disruption in spermatogenesis by pubertal low dose ZEA exposure may be through the alterations in genetic and epigenetic pathways, and the interactions with estrogen receptor signaling pathway. Therefore, we should pay great attention on ZEA exposure to reduce its adverse impacts on male reproductive health.

Gestational exposure to low-dose zearalenone disrupting offspring spermatogenesis might be through epigenetic modifications.

Zearalenone (ZEA), a F-2 mycotoxin produced by Fusarium, has been found to be an endocrine disruptor through oestrogen receptor signalling pathway to impair spermatogenesis. The disruption on reproductive systems by ZEA exposure might be transgenerational. In our previous report, we have found that low dose (lower than no-observed effect level, NOEL) of ZEA impaired mouse spermatogenesis and decreased mouse semen quality. The purpose of the current investigation was to explore the impacts of low-dose ZEA on spermatogenesis in the offspring after prenatal exposure and the underlying mechanisms. And it demonstrated that prenatal low-dose ZEA exposure disrupted the meiosis process to inhibit the spermatogenesis in offspring and even to diminish the semen quality by the decrease in spermatozoa motility and concentration. The DNA methylation marker 5hmC was decreased, the histone methylation markers H3K9 and H3K27 were elevated, and oestrogen receptor alpha was reduced in the offspring testis after prenatal low-dose ZEA exposure. The data suggest that the disruption in spermatogenesis by prenatal low-dose ZEA exposure may be through the modifications on epigenetic pathways (DNA methylation and histone methylation) and the interactions with oestrogen receptor signalling pathway. Moreover, in the current study, the male offspring were indirectly exposed to low-dose ZEA through placenta and the spermatogenesis in offspring was disrupted which suggested that the toxicity of ZEA on reproductive systems was very severe. Therefore, we strongly recommend that greater attention should be paid to this mycotoxin to minimize its adverse impact on human spermatogenesis.

Time-dependent effect of ground water fluoride on motility, abnormality and antioxidant status of spermatozoa: An in vitro study.

The present study was undertaken to investigate the toxic effect of ground water fluoride (F) on motility, abnormality, and antioxidant status of spermatozoa. Treatment of ground water F with epididymal sperm suspension caused complete loss of sperm motility and decrease in the activities of superoxide dismutase (SOD) and catalase (CAT) and increase in the concentration of malondialdehyde (MDA) and abnormality of spermatozoa at 15 and 30 min time intervals. Further, incubation of spermatozoa with ground water F for 5, 10, and 15 min time intervals significantly reduced the sperm motility and activities of SOD and CAT and increased the concentration of MDA and abnormality of spermatozoa. The study revealed that F-induced effect on sperm motility and antioxidant status is time dependent. Increase in oxidative stress and concomitant decrease in motility of spermatozoa in ground water F clearly indicates that F-induced oxidative stress affected the sperm motility. The present study for the first time demonstrated the toxic effect of ground water F on spermatozoa at shorter duration of exposure, which affects the capability of spermatozoa in fertilization.

Low Concentration of Ethylene Glycol Improved Recovery Rate of Human Spermatozoa After Vitrification (ETILEN GLIKOL KONSENTRASI RENDAH MENINGKATKAN RECOVERY RATE SPERMATOZOA MANUSIA PASCAVITRIFIKASI)

The use of cryoprotectants for the cryopreservation of human spermatozoa, oocytes, zygote, earlycleavage stage of embryos and blastocyst is an integral part of almost every human In Vitro Fertilizationprogram. Moreover, the cryopreservation of these types of cells by direct plunging into liquid nitrogen (-196°C) usually requires a high concentration of cryoprotectant with a consequent of cytotoxic effect. Theaim of this study was to observe the effect of ethylene glycol concentration on the spermatozoa recoveryrate following vitrification process. Earle’s balanced salt solution + 0.25 M sukrosa + 1 % human albumineserum as basic solution supplemented with some different concentrations of etylene glycol (ie: 36.25%;18.25%; 9.12%; 4.56%; 1.14% and 0.57%) were used to evaluate the motility and viability of spermatozoafollowing vitrification. Human’s spermatozoa from ejaculates with progressive motility and viability above50% were used as samples. Samples were mixed with vitrification solution and then loaded into 0.25 mLstraws, equilibrated for 10 minutes at room temperature before plunged into liquid nitrogen. Spermatozoathawing was done at 24 hours after the vitrification. The results showed that, the decrease of spermatozoamotility and viability were observed at the highest (100%, 96.70%, respectively) in the samples that wereadded with vitrification medium contained 36.25% of ethylene glycol. On the other hand, the decrease ofthe spermatozoa motility and viability were found at the lowest (14.11%, 43.81 %, respectively) in thesamples without ethylene glycol supplementation. It can be concluded that the highest spermatozoa recoveryrate was obtained from the vitrification using a low concentration of ethylene glycol.

Lead induced alterations in rabbit spermatozoa motility and morphology in vitro

The aim of this in vitro study was to determine the effect of lead chloride (PbCl2) on rabbit spermatozoa motility and morphology. Lead concentrations in the medium ranged between 0.45 and 11.17 μg/ml; incubation time was 240 min (analyzed immediately after Pb addition followed by 30, 60, 120, 180, and 240 min), and temperatures of the culture environment were 22°C (laboratory), 4°C (refrigerator), and 37°C (incubator). Results were compared with a control group without Pb addition. After 30 min of culture at 22°C, a negative effect of Pb was noted as spermatozoa motility significantly decreased in groups with higher concentrations. After 120 and 240 min, a dose-dependent effect on spermatozoa motility was noted. At 4°C, spermatozoa motility analysis detected no significant differences between any of the experimental groups and control. At 37°C, a negative effect of Pb incubation on motility was detected at Times 30, 60, 120, 180, and 240 in groups with high concentrations. At Times 120, 180, and 240 a significant decrease in spermatozoa motility was also noted in all experimental groups in comparison to control. The analysis of pathological spermatozoa at Time 240 revealed an increasing trend of morphological abnormalities after incubation with Pb. Across three temperature regimes an increase of morphological changes was noted, particularly in the group with the highest Pb concentration. The predominant morphological abnormalities were knob twisted flagellum, flagellum ball, separated flagellum, and broken flagellum. Knob twisted flagella represented the most frequent pathological changes in the experimental group with the highest Pb concentration. Results suggest that the inhibitory effect of Pb on spermatozoa motility parameters depends on the concentration, incubation time, as well as environmental temperature during incubation. Furthermore, a negative effect of Pb in vitro on spermatozoa morphology indicates possible reproductive problems under in vivo conditions, too.

PENENTUAN KONSENTRASI SODIUM DODECYL SULFATE DALAM PENGENCER RINGER LAKTAT-KUNING TELUR UNTUK PRESERVASI SEMEN AYAM PELUNG (Determination of Sodium Dodecyl Sulfate Concentration in Ringer Lactate-Egg Yolk Extender for Pelung Rooster Semen Preservation)

The objective of this experiment was to determine the addition of sodium dodecyl sulfate (SDS) to Ringer Lactate-Egg Yolk (RL-EY) extender on pelung chicken semen preservation. Semen was collected three times a week from three pelung chickens. Collected semen was evaluated macroscopically and microscopically. Only ejaculates of at least 70% sperm motility was then divided into three equal microtubes. Each of them diluted with RL-EY with ratio 90%:10%, then added with 0.00, 0.025, and 0.05% SDS respectively. The liquid semen then stored at 5 °C for 72 hours. Sperm motility and viability were observed every 12 hours. The addition of 0.025% SDS showed higher spermatozoa motility and viability (72.08±1.44% and 80.82±1.30%) which were significantly higher (P&lt;0.05) than 0.00 and 0.05% SDS addition at 24 hours of storage. There was no differences on the spermatozoa motility and viability between 0.00 and 0.05% SDS addition. The decrease of spermatozoa motility and viability was observed in 0.025% SDS addition (4.17±0.56% and 4.65±0.59%, respectively) that significantly lower compared to 0% and 0.05% SDS addition at 24 hours of storage. In conclusion, 0.025% SDS addition in a Ringer Lactate -Egg Yolk extender maintained pelung spermatozoa motility and viability better than 0.00 and 5% SDS addition.

Effect of Nickel on Male Reproduction

In this study distribution of nickel as a risk factor of environment in testis and its effects on the testicular structure in experimental animals and effect on spermatozoa was analyzed. In this study the effect of Ni on the testicular structure after an experimental intraperitoneal (i.p.) administration, concentration of nickel in semen of different animal species and the effect of in vitro spermatozoa incubation with nickel on the spermatozoa motility and membrane changes is reported. Our findings clearly suggest a negative effect of nickel on the structure as well as on the function of seminiferous epithelium. In experimental groups with nickel a significant (p 250 μM Ni/ml) cause decrease of spermatozoa motility in vitro.

Oxidative Stress Negatively Affects Human Sperm Mitochondrial Respiration

To correlate the level of oxidative stress in serum and seminal fluid and the level of sperm deoxyribonucleic acid (DNA) fragmentation with sperm mitochondrial respiratory efficiency. Sperm mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically treated sperm cells. A possible relationship between sperm mitochondrial respiratory efficiency, the level of oxidative stress, and the level of sperm DNA fragmentation was investigated. Sperm motility was positively correlated with mitochondrial respiration but negatively correlated with oxidative stress and DNA fragmentation. Interestingly, sperm mitochondrial respiratory activity was negatively affected by oxidative stress and DNA fragmentation. Our data indicate that sperm mitochondrial respiration is decreased in patients with high levels of reactive oxygen species by an uncoupling between electron transport and adenosine triphosphate synthesis. This reduction in mitochondrial functionality might be 1 of the reasons responsible for the decrease in spermatozoa motility.

Studies on the toxicological effect of nevirapine, an antiretroviral drug, on the liver, kidney and testis of male Wistar rats

We investigated the toxic effect of nevirapine (NVP; Viramune(®)), an antiretroviral drug, on the liver, kidney and testis of Wistar rats. Twenty-one rats were assigned into 3 groups of 7 animals each. The first group served as control, and the second and third groups received NVP at 18 and 36mg/kg body weight, respectively. Clinical signs of toxicity were not observed in the animals. NVP at both doses did not significantly (p>0.05) alter the body weight gain, relative weights of kidney and testis, serum protein, urea, creatinine and alkaline phosphatase levels of the animals. However, NVP2 significantly (p<0.05) increased the relative weight of liver, level of serum total bilirubin and activities of γ-glutamyl transferase, alanine and aspartate aminotransferases. NVP administration caused a dose-dependent, significant (p<0.05) elevation of lipid peroxidation measured as malondialdehyde (MDA) content in the liver, kidney and testis of the rats. Hepatic, renal and testicular MDA were increased by 107%, 80% and 163%, respectively, in NVP2-treated rats. Elevation in MDA was accompanied by a significant (p < 0.05) decrease in the activities of hepatic, renal and testicular superoxide dismutase and catalase. NVP2 caused 43% and 32% decrease in spermatozoa motility and live/dead sperm count, respectively, and 94% increase in total sperm abnormalities. Histopathological findings showed that NVP2 caused degeneration of seminiferous tubules in testis, and severe necrosis in liver slides. NVP induced oxidative stress with corresponding decrease in antioxidant status of the rats. The changes in sperm parameters and, elevation of liver marker enzymes suggest an interference of NVP2 with these organs.

Alterations in mitochondrial function and spermatozoal motility in goat spermatozoa following incubation with a human lysozyme plasmid

Spermatozoa mediated gene transfer has become a promising technology to generate transgenic animals with disease resistance. However, exogenous DNA invasion may cause changes in spermatozoon natural defense system which result in spermatozoon dysfunction. The objective of this study was to investigate the changes of mitochondrial function and motility in goat spermatozoa after pre-incubation and incubation with and without the human lysozyme plasmid pFLAG-hLY. The results demonstrated that human lysozme plasmid pFLAG-hLY could bind to the surface of the spermatozoon membrane at 186,000 copies/spermatozoa and incorporate to spermatozoon nucleus at 78 copies/spermatozoa after incubation. However, the treated spermatozoon samples showed a significant lower motility (29.7 ± 2.2% vs. pre-incubation control 48.0 ± 1.4% and incubation control 54.5 ± 1.5%, P < 0.05), and percentage of rapid progressive motile spermatozoa (16.4 ± 2.4% vs. pre-incubation control 31.4 ± 0.6% and incubation control 37.0 ± 0.5%, P < 0.01). Meanwhile, the incubation with plasmids caused significant reduction of mitochondrial membrane potential (31.44 ± 2.17% vs. pre-incubation control 51.79 ± 2.08% and incubation control 58.81 ± 1.76%, P < 0.05). In addition, dichlorofluorescin relative fluorescence intensity (32.81 ± 2.41%) and malondialdehyde levels (2.18 ± 0.21 nM/10 8 spermatozoa), which represents mitochondrial function, showed a significant increase after incubation ( P < 0.05). The cytochrome c release from the mitochondrial inner membrane space, and enzymatic activities of caspase-3 (0.086 ± 0.024) and caspase-9 (0.083 ± 0.019) ( P < 0.05) also increased, in which resulted in spermatozoon dysfunction. In conclusion, this study confirmed that goat spermatozoa could capture human lysozyme plasmid pFLAG-hLY, but the incubation with the plasmids resulted in a decrease of spermatozoa motility and partial rupture of mitochondrial membrane, and further prompted the expression of cytochrome c, and generation of oxidative stress in vitro and finally led to spermatozoon dysfunction.

Curcumin and Kolaviron Ameliorate Di‐n‐Butylphthalate‐Induced Testicular Damage in Rats

The present study was carried out to evaluate the ameliorative effects of kolaviron (a biflavonoid from the seeds of Garcinia kola) and curcumin (from the rhizome, Curcuma longa L.) on the di-n-butylphthalate (DBP)-induced testicular damage in rats. Administration of DBP to rats at a dose of 2 g/kg for 9 days significantly decreased the relative testicular weights compared to the controls, while the weights of other organs remained unaffected. Curcumin or kolaviron did not affect all the organ weights of the animals. While only DBP treatment significantly increased the testicular malondialdehyde level and gamma-glutamyl transferase activity (gamma-GT), it markedly decreased glutathione level, the testicular catalase, glucose-6-phosphate dehydrogenase, superoxide dismutase, sperm gamma-GT activities and serum testosterone level compared to the control group. Data on cauda epididymal sperm count and live/dead ratio were not significantly affected in the DBP-treated rats. Alone, DBP treatment resulted in a 66% decrease in spermatozoa motility and a 77% increase in abnormal spermatozoa in comparison to control. DBP-treated rats showed marked degeneration of the seminiferous tubules with necrosis and defoliation of spermatocytes. The DBP-induced injuries in biochemical, spermatological parameters and histological structure of testis were recovered by treatment with kolaviron or curcumin. The pattern in the behaviour of these compounds might be correlated with their structural variations. Our results indicate that kolaviron and curcumin protect against testicular oxidative damage induced by DBP. The chemoprotective effects of these compounds may be due to their intrinsic antioxidant properties and as such may prove useful in combating phthalate-induced reproductive toxicity.

Chilled storage of semen from Atlantic halibut, Hippoglossus hippoglossus L. II: Effect of spermiation advancement, catheterization of semen, and production-scale application

A method for in vitro storage of Atlantic halibut Hippoglossus hippoglossus L. semen can facilitate seed production. The study aimed at determining the effect of male spermiation advancement on viability of chilled stored spermatozoa. The use of catheterization of semen from the sperm duct was examined. Also, large volumes of semen were stored under sub-optimal production-like conditions in order to determine the suitability of the method into practical use. Semen was collected from two broodstocks: natural photoperiod males, being at the first phase of the reproductive season and 3-month advanced photoperiod males, being at the end of the reproductive season. Semen samples were diluted 1:5 (v/v) with modified Hanks’ Balanced Salt Solution supplemented with antibiotics, and stored in Ziploc bags filled with air. Sperm motility parameters, assessed by computer-assisted sperm analysis (CASA), were assessed weekly. Experimental and production-scale fertilization trials were performed. Sperm samples from natural photoperiod males showed significantly longer viability under in vitro storage conditions than sperm from advanced photoperiod males. In the natural photoperiod group, the decrease in spermatozoa motility, curvilinear velocity and straight-line velocity occurred on day 50, 14, and 28 of storage, respectively. Spermatozoa from one of five males were still motile on day 80 of storage, and fertilization rates and embryo survival rates obtained using semen stored for 70 days did not differ from control values and they were significantly higher than values obtained with the use of fresh semen of the same male, but being at the end of reproductive season. Catheterization of semen showed no advantage to stripping the semen without a catheter, even for samples stored undiluted for 1 day of collection, before dilution. Under sub-optimal conditions, spermatozoa stored in large volumes (10–100 mL of diluted semen) without any special treatments except for weekly swirling, remained viable for more than 1 month. Production-scale fertilizations with samples stored for 5–21 days resulted in high survivals of embryos and hatchlings. Because of its simplicity and efficiency, the method shows a high potential for use in commercial Atlantic halibut farming. It has already been applied to a halibut breeding programme for the next reproductive season at our research station.

MENINGKATNYA HIDROGEN PEROKSIDA PADA VARIAN T16189C mtDNA SEMEN MANUSIA

Mitochondria are a site of cellular respiration through oxidative phosphorylation enzymatic reaction (OXPHOS), which is producing energy in the form of ATP (Adenosine Triphosphate). If abnormalities occur along cellular respiratory chain, ATP will decrease and Reactive Oxygen Species (ROS), will increase, one of which is hydrogen peroxide. ROS is an oxidation whose targets are lipid, protein and DNA, all of which may result in the decrease of spermatozoa motility. The detection of hydrogen peroxide was conducted by means of chemiluminescence using luminol, while the detection T16189C mtDNA variant was done using PCR-RFLP with restriction enzyme MnLI. In normozoospermia, hydrogen peroxide in 16189T was 4.4 1.8 CPM/106 sp and in 16189C was 6.4 1.8 CPM/106 sp. In asthenozoospermia, hydrogen peroxide in 16189T was 20.3 8.3 CPM/106 sp while in 16189C was 62.5 9.0 CPM/106 sp. Hydrogen peroxide in normozoospermia and asthenozoospermia 16189T and 16189C showed significant difference (p less than 0.00; p less than 0.01). In normozoospermia and asthenozoospermia, 16189T and 16189C has correlation with the decrease of motile spermatozoa motility (normozoospermia, p = 0.02; p less than 0.05; asthenozoospermia p = 0.03; p less than 0.05).