Effects of intratesticular injection of hypertonic mannitol and saline on the quality of donkey sperm, indicators of oxidative stress and testicular tissue pathology.

OBJECTIVES: The aim of the present study was to examine donkey sperm quality after intratesticular injection of hypertonic mannitol (HM) and saline (HS). METHODS: Randomly assigned to five treatment groups were 15 adult male donkeys: (1) Control group (no treatment), (2) Surgery group (surgical castration for testosterone control), (3) NS group (normal saline intratesticular injection), (4) HS group (hypertonic saline), and (5) HM group. We injected 20 mL per testicle. We took 5 mL blood from all donkeys before injection. Castration was performed under general anesthesia 60 days later. Samples included blood and testicular tissue. Total motility (TM), progressive motility (PM), movementy features, DNA damage, morphology, viability, and plasma membrane functionality were evaluated. Hormone analyses, histomorphometric studies and oxidative stress indices including total antioxidant capacity (TAC), glutathione peroxidase (GPx), glutathione (GSH), superoxide dismutase (SOD), malondialdehyde (MDA), and NADP+/NADPH were evaluated. Apoptosis, pyroptosis-related Bax, Caspase-1, GSDMD, and Bcl-2 expression were also assessed. RESULTS: In HS and HM groups, testosterone, epididymal sperm count, motility, viability, and plasma membrane functionality dropped while sperm DNA damage increased. HS and HM groups had significantly lower histomorphometric parameters, TAC, GPx, SOD, GSH, and Bcl-2 gene expression. MDA, NADP+/NADPH, Bax, Caspase-1, and GSDMD gene expression were substantially higher in the HS and HM groups than in the control group. CONCLUSIONS: Toxic effects of hypertonic saline and mannitol on reproductive parameters were seen following, hence, they might be considered as a good chemical sterilizing treatment in donkeys.

Comparison of sperm characteristics and antioxidant and oxidant levels in bull semen frozen with four widely used extenders.

Sperm survives for a very short time in fresh semen, and slow cooling to 5.00 ˚C kills a large number of sperms. This study was aimed to compare the semen quality parameters and anti-oxidant levels in four extenders (manual, Triladyl, Steridyl and AndroMed). Semen samples were obtained from a total number of 12 dual-purpose Simmental bulls kept in the Simmental Cattle Breeding Center for a period of 3 months using an artificial vagina. Sperm viability, motility, abnormal morphology, plasma membrane integrity, DNA damage, chromatin quality, total antioxidant capacity (TAC) and lipid peroxidation were evaluated. The highest progressive motility, viability, plasma membrane integrity, and TAC and the lowest levels of malondialdehyde in the frozen-thawed semen belonged to the semen group frozen with Triladyl. Parameters of motility were higher in the frozen-thawed semen with Triladyl than in other groups, indicating a significant difference from the manual extender. Among the extenders studied, Triladyl was the most suitable for semen freezing in Simmental bulls.

The influence of L-proline and fulvic acid on oxidative stress and semen quality of buffalo bull semen following cryopreservation.

BACKGROUND: This study investigates the effects of cryopreservation and supplementation of Azeri water buffalo's semen with proline (Lp) and fulvic acid (FA). OBJECTIVES: Therefore, this study aimed to assess motility parameters, sperm viability, oxidative stress parameters, and DNA damage to detect the optimum concentrations of Lp and FA for buffalo semen cryopreservation. METHODS: Thirty semen samples of three buffalo bulls were diluted in Tris-egg yolk extender and divided into 12 equal groups including control (C), Lp-10, Lp-20, Lp-40, Lp-60, Lp-80 (containing 10, 20, 40, 60, 80 mM L-proline, respectively), FA-0.2, FA-0.5, FA-0.8, FA-1.1, FA-1.4 and FA-1.7 (containing 0.2%, 0.5%, 0.8%, 1.1%, 1.4% and 1.7% fulvic acid, respectively). RESULTS: The velocity parameters, TM and PM were improved by FA-1.7, FA-1.4, Lp-40 and Lp-60 groups compared to the C group but no significant difference was found regarding the amplitude of lateral head displacement and straightness compared to the control groups. The percentage of sperm viability and PMF were increased by FA-1.7, FA-1.4, FA-1.1, Lp-40 and Lp-60 groups compared to C group, while in terms of sperm DNA damage FA-1.7, FA-1.4, FA-1.1, Lp-10, Lp-20, Lp-40 and Lp-60 groups showed better results compared to C group. The results also showed that FA-1.7, FA-1.4, FA-1.1, Lp-20, Lp-40 and Lp-60 groups could improve TAC, SOD, GSH and decrease MDA levels. Also, FA-1.7, FA-1.4, Lp-20 and Lp-40 groups could improve GPx levels but just FA-1.7, and Lp-40 groups could improve CAT levels compared to C group. CONCLUSIONS: Thus, it can be concluded that L-proline and fulvic acid supplementations can improve the quality parameters of post-thawed buffalo bull semen.

Tribulus terrestris aqueous extract supplementation effects on sperm characteristics and anti-oxidant status during chilled storage of canine semen.

The reduction of spermatozoa survival time is a major problem of canine chilled sperm for artificial insemination. The current study looks at the possible advantages of chilling canine sperm to 4.00 ˚C for three days using Tribulus terrestris aqueous extract (TTAE). Three mixed-breed dogs were utilized to extract 24 ejaculates, which were then diluted in a Tris-based extender. The ejaculates were then divided into five groups including 20.00, 40.00 and 50.00 µg mL-1 of TTAE, sham (distilled water devoid of TTAE) and control (without TTAE) groups. During the three days of experiment, several parameters were measured every 24 hr. It was noticed that after 48 and 72 hr of liquid storage, total and progressive motilities were greater in the group with the 40.00 µg mL-1 TTAE concentration than the control group. Compared to the control group, the group with the 40.00 µg mL-1 TTAE concentration exhibited superior motility and viability. The percentages obtained from the hypo-osmotic swelling test were much greater. In contrast to the control group, DNA integrity was poorer in the 40.00 µg mL-1 TTAE concentration. After 72 hr of storage, the group with 40.00 µg mL-1 TTAE concentration had lower malondialdehyde levels but considerably greater total anti-oxidant capacity, superoxide dismutase, glutathione peroxidase and catalase levels than the control groups. The current study found that supplementing the semen extender with 40.00 µg mL-1 TTAE improves semen parameters after 72 hr of storage at 4.00 ˚C, and therefore can improve fertilization efficiency.

Natural Products/Bioactive Compounds as a Source of Anticancer Drugs.

Cancer is one of the major deadly diseases globally. The alarming rise in the mortality rate due to this disease attracks attention towards discovering potent anticancer agents to overcome its mortality rate. The discovery of novel and effective anticancer agents from natural sources has been the main point of interest in pharmaceutical research because of attractive natural therapeutic agents with an immense chemical diversity in species of animals, plants, and microorganisms. More than 60% of contemporary anticancer drugs, in one form or another, have originated from natural sources. Plants and microbial species are chosen based on their composition, ecology, phytochemical, and ethnopharmacological properties. Plants and their derivatives have played a significant role in producing effective anticancer agents. Some plant derivatives include vincristine, vinblastine, irinotecan, topotecan, etoposide, podophyllotoxin, and paclitaxel. Based on their particular activity, a number of other plant-derived bioactive compounds are in the clinical development phase against cancer, such as gimatecan, elomotecan, etc. Additionally, the conjugation of natural compounds with anti-cancerous drugs, or some polymeric carriers particularly targeted to epitopes on the site of interest to tumors, can generate effective targeted treatment therapies. Cognizance from such pharmaceutical research studies would yield alternative drug development strategies through natural sources which could be economical, more reliable, and safe to use.

Evaluation of dietary betaine on post-thawed semen quality in mature bulls during summer heat stress.

Heat stress (HS) has caused relative hypoxia, oxidative stress and high level of homocysteine, which contributes significantly to fertility failures in bulls. The aim of present study was to evaluate the role of dietary betaine (BET) in improving dual purpose Simmental (Fleckvieh) post-thawed semen quality especially during the hottest summer days. A total number of 16 mature bulls were randomly assigned to three equal groups including: 1) Control condition (without betaine), 2) BET1: 57.00 mg of betaine kg-1 per day and 3) BET2: 114 mg of betaine kg-1 per day, through daily intakes for 90 days in summer. Plasma levels of homocysteine, seminal plasma antioxidants levels and sperm parameters such as DNA fragmentation, chromatin integrity, motility, viability, morphology and membrane integrity were evaluated. Under maximal HS, serum homocysteine concentrations were reached 16.67 ± 0.09 µmol L-1. Dietary betaine supplementation influenced DNA fragmentation of sperm and was higher in the control group compared to BET2 group. There were significant decreases in seminal plasma superoxide dismutase (SOD), glutathione peroxidase (GPx) activity and sperm viability and motility in bulls treated with betaine. The activity of GPx and SOD in the control group was increased up to 0.08 ± 0.00 U mg-1 protein and 0.52 ± 0.01 U mg-1 protein in seminal plasma. There were no significant differences between groups in the percentage of swollen spermatozoa, membrane integrity, sperm morphology, abnormal head morphology and percentage of spermatozoa stained with aniline blue. In conclusion, BET supplements improved semen parameters in sperm motility, sperm viability and influenced DNA fragmentation during HS with reduction in serum homocysteine concentrations.

The effects of varying concentrations of glutathione and trehalose in improving microscopic and oxidative stress parameters in Turkey semen during liquid storage at 5 °C.

Since turkey reproduction is mainly through artificial insemination, short-term preservation of turkey semen is one of the most important issues in turkey reproduction management. The present study investigates the effects of glutathione (GSH) and trehalose on lipid peroxidation degree and turkey semen quality while being stored at 5 °C for 72 h. To this end, semen samples were collected from 20 turkeys with a weekly frequency for 12 weeks. A glucose-based extender was used to dilute the pooled semen. It was divided into seven equal parts with varying levels of glutathione [0.5, 1 and 2 mM), trehalose [50, 75 and 100] and control [extender without antioxidant]. Subsequently, the divided semen samples were stored at 5 °C for 72 h. Several sperm parameters such as motility and motion parameters, plasma membrane integrity (PMI), plasma membrane functionality, DNA integrity, and oxidative parameters were assessed following storage for 0, 24, 48, and 72 h. The obtained results indicated an improvement in the plasma membrane functionality and DNA integrity, along with the percentages of PMI in GSH-2 mM group in comparison to the control group following storage at 5 °C for 72 h (P ≤ 0.05). It is also notable that the 2 and 1 mM concentrations of GSH increased the spermatozoa motility and motion parameters in comparison to the control group, respectively (P ≤ 0.05). The study results indicated that GSH-2, 1 mM and trehalose- 100 mM concentrations reduced lipid peroxidase levels and increased total antioxidant activity, catalase, superoxide dismutase, and glutathione peroxidase in comparison to the control group (P ≤ 0.05). Our study's data show that improvement of semen parameters and oxidative stress parameters of turkey semen can be improved by glutathione at 2 and 1 mM and trehalose at 75 mM while storing it 5 °C.

Antioxidant effects of supplementation of 3,4-dihydroxyphenyl glycol on sperm parameters and oxidative markers following cryopreservation in canine semen.

Supplementing the extender with antioxidants with low molecular weight can enhance the quality of the post-thaw sperm during the freezing process. This study was aimed at determining the impacts of 3,4-dihydroxyphenyl glycol (DHPG) on the spermatozoa of the canine undergoing freeze-thawing process. In this study, 24 ejaculates were obtained from three mixed-breed dogs and were diluted in a Tris-based extender. The diluted semen was divided into aliquots for supplementation of 10, 30, 50 and 70 µg/ml of DHPG, control (without antioxidant) and control sham (DMSO). After being extended, the semen was equilibrated at a temperature of 4°C and then transferred to the straws and kept 4 cm above the liquid nitrogen for 20 min and was finally immersed in the liquid nitrogen. They were cryopreserved for seven days; then, sperm parameters including sperm motility evaluation, motility characteristics, viability, DNA and plasma membrane integrity, total antioxidant capacity (TAC), reduced glutathione content (GSH), antioxidant enzymes (superoxide dismutase [SOD], catalase [CAT] and glutathione peroxidase [GPx]) activity malondialdehyde (MDA) levels were evaluated. This study showed that spermatozoa cryopreservation with 50, 30 and 70 µg/ml of DHPG concentrations had better progressive motility, Curvilinear Velocity, Linearity, viability, intact plasma membrane and the levels of TAC, GPx and GSH were higher than the control group. The 50, 30 and 70 µg/ml of DHPG concentrations led to the significant decrease of DNA damage compared to the control group. Total motility, average path velocity, straight-line velocity and CAT activity were significantly improved in 30 and 50 µg/ml of DHPG concentrations, compared to the control group. Also, the 50 and 30 µg/ml of DHPG concentrations, decreased MDA levels compared to the other groups, significantly. In conclusion, our study showed that the addition 50 µg/ml of DHPG to the canine semen extender improved the semen characteristics and oxidative markers in the cryopreservation process.

Bisphenol-A analogue (bisphenol-S) exposure alters female reproductive tract and apoptosis/oxidative gene expression in blastocyst-derived cells.

OBJECTIVES: One of the major endocrine-disrupting chemicals, bisphenol-S (BPS) has replaced bisphenol-A due to public health anxiety. The present study evaluated low dosage BPS effect on female reproductive potential, hormonal disruption, and gene expression pathways of blastocyst-derived cells. MATERIALS AND METHODS: NMRI female mice (5-6 weeks) in the estrous stage were chosen following vaginal smear examination for estrus cycle detection and BPS (0, 1, 5, 10, 50 and 100 µg/kg) was administrated subcutaneously for twenty-one consecutive days. After the last administration, blood, ovary tissue and oocytes were collected for further examination. RESULTS: BPS induced oxidative stress in ovarian tissue and reduced hormonal status, LH and FSH, even at low concentration. Furthermore, apoptosis was induced in blastocyst derived cells in BPS administrated mice groups even at low BPS concertation, however, P53 and E2f1 expression were downregulated in doses more than 50 µg/kg, which might indicate apoptosis pathway exchange from P53 dependent to p53 independent pathways. IVF outcome was negatively associated with blastocyst apoptosis gene expression, estrogen receptor beta (ERß) as well as oxidative status in ovaries. Finally, Stepwise regression indicated that E2f1, Nrf2, catalase (CAT), and malondialdehyde (MDA) could be chosen as predictor values for hatch percentage in IVF outcome. CONCLUSION: In summary, this study revealed BPS might have detrimental potential in the female reproductive system by oxidation induction and hormonal alteration as well as next generation blastocyst derived cells apoptosis induction. Further studies are recommended for public health assurance of BPS safety especially for female consumed products.

The evaluation of lycopene and cysteamine supplementation effects on sperm and oxidative stress parameters during chilled storage of canine semen.

The aim of this study was to evaluate the effects of different concentrations of lycopene and cysteamine on characteristics of sperm, liquid peroxidation and enzymatic activities in seminal plasma of canine semen preserved at 5°C for 72 hr. The semen samples were divided into eight aliquots: control, control sham (dimethyl sulfoxide 5%), lycopene groups (250, 500 and 750 µg/ml) and cysteamine groups (2.5, 5 and 10 mM). Motility, viability, membrane integrity, DNA integrity, total antioxidant capacity (TAC) and malondialdehyde (MDA) levels were evaluated. Progressive motility and total motility were higher with the 500 and 750 µg/ml lycopene concentrations, respectively, compared to the control group and the cysteamine groups following 72 hr of storage in the liquid storage. Motility characteristics, viability and hypo-osmotic swelling test (HOST) percentages were significantly improved in 500 µg/ml lycopene compared to other groups. The 500 and 750 µg/ml lycopene concentrations, respectively, showed significantly reduced percentages of spermatozoa with DNA integrity compared to the control group. The 500 and 750 µg/ml lycopene concentrations, respectively, led to the significant decrease of MDA levels. The 500 µg/ml lycopene enhanced TAC levels after 48 and 72 hr that was not observed in other groups. In conclusion, the findings of this study showed that lycopene supplementation in canine semen extenders improved canine semen parameters and TAC levels and decreased MDA levels in the chilling process.

Caffeic acid improves microscopic sperm parameters and antioxidant status of buffalo (Bubalus bubalis) bull semen following freeze-thawing process.

This study investigates the effects of cryopreservation and supplementation of buffalo's semen with Caffeic acid. It studies the effects of different Caffeic acid concentrations on cryopreservation capacity of the buffalo and evaluates their influence on various sperm parameters like motility, viability, progressive motility, sperm plasma membrane integrity, and antioxidant status. Twenty-four semen samples were collected with an artificial vaginal from three adult water buffalos. The semen samples were evaluated and the qualified ejaculates were separated and were diluted in a Tris-based extender. The resulting samples were classified into 5 groups: No antioxidant (control), Control sham (NaOH), Caffeic acid 50 µM, Caffeic acid 100 µM, and Caffeic acid 200 µM. The semen samples encountered cryodamage and the quality was deteriorating during the cryopreservation (P < 0.05). The semen evaluation after thawing showed that the groups of samples receiving 100 µM Caffeic acid had higher viability, total motility, and lower abnormal sperm and better linearity (LIN), curvilinear velocity (VCL), straight-line velocity (VSL) and path velocity and higher intact plasma membrane (P < 0.05) compared to other groups. It is notable that adding 100 µM Caffeic acid to freezing extenders enhances the CAT, GPx, SOD, and GSH and also ameliorates total antioxidant capacity of spermatozoa after thawing. It is notable that the addition of 100 µM Caffeic acid decreases the amount of Malondialdehyde. These reactions lead us to conclude that 100 µM Caffeic acid enhances the semen quality of water buffalo after thawing.

Carob (Ceratonia siliqua L.) fruit hydro-alcoholic extract alleviates reproductive toxicity of lead in male mice: Evidence on sperm parameters, sex hormones, oxidative stress biomarkers and expression of Nrf2 and iNOS.

OBJECTIVE: Carob (Ceratonia siliqua L.) is an evergreen tree with fruits that have potent antioxidant activity. The aim of this study was to investigate alleviative effects of carob fruit hydro-alcoholic extract (CFHAE) against reproductive toxicity induced by lead (Pb) in male mice. MATERIALS AND METHODS: Forty-two NMRI adult male mice were randomly categorized into 7 groups (N=6). Group I was the control group and received no treatment. Group II was the sham group and received 0.2 ml distilled water per day. Group III (Pb group) received Pb acetate 1000 ppm/kg/day. Groups IV and V received CFHAE 500 and 1000 mg/kg/day, respectively. Groups VI and VII received both Pb 1000 ppm/kg/day and CFHAE at doses of 500 and 1000 mg/kg/day, respectively at the same time. The groups were treated by gavage. After 35 days, sperm parameters (count, motility, morphology, viability, DNA damage, and teratozoospermia index), total antioxidant capacity (TAC), reduced glutathione content (GSH), antioxidant enzymes (SOD, CAT, and GPx) activity, MDA levels, and sex hormones (FSH, LH, and testosterone) concentrations in serum, testicular expression of Nrf2 and iNOS genes and histopathological alterations were evaluated. RESULTS: Our findings revealed that co-administration of CFHAE with Pb significantly (p<0.05 to p<0.01) improved sperm parameters, elevated sex hormones, TAC, GSH content, and antioxidant enzymes activity of serum, decreased serum MDA levels, and down-regulated testicular expression of Nrf2 and iNOS genes compared with Pb group. Also, CFHAE ameliorated histopathological alterations in testis tissue caused by Pb. CONCLUSION: CFHAE can alleviate reproductive toxicity following Pb exposure in male mice.

Effects of nano-selenium on mRNA expression of markers for spermatogonial stem cells in the testis of broiler breeder males.

Fertility is one of the most important parameters in breeder farms and cockerels play an outstanding role in the fertility of eggs in broiler breeder farms. Todays, supplementation of chicken diet with additives such as organic selenium is used to increase fertility. The aim of this study was to evaluate the effects of different levels of nano-selenium (Nano-Se) on the expression of molecular markers of spermatogonial stem cells (SSCs) in the testis of broiler breeder males. A total of 30 roosters of 40 weeks of age were randomly divided into five groups. Groups were as follows: 1) control (normal diet) group, 2) diet supplemented with 0.30 mg kg-1 sodium selenite, 3) diet supplemented with 0.15 mg kg-1 Nano-Se, 4) diet supplemented with 0.30 mg kg-1 Nano-Se, and 5) diet supplemented with 0.60 mg kg-1 Nano-Se. At the end of the experimental period (5th week), birds were autopsied and samples from testis of all birds were collected. The testis samples were used to examine the ß1-integrin (CD29), thy-1 (CD90) and NANOG mRNA expression by real-time PCR. The results showed that testis of the groups fed with the diets supplemented with 0.60 mg kg-1 and 0.15 mg kg-1 of Nano-Se had the highest and lowest mRNA expression of SSCs markers, respectively. In conclusion, the present study indicated that Nano-Se had advantages over sodium selenite. Diet supplemented with 0.60 mg kg-1 of Nano-Se may contribute to optimal fertility via increasing the mRNA expression of SSCs markers of roosters' testis and could be used to delay the reduction of fertility caused by aging in broiler breeder males.

Protective effects of hydro-alcoholic extract of Quercus brantii against lead-induced oxidative stress in the reproductive system of male mice.

OBJECTIVE: Exposure to heavy metals such as lead (Pb) results in oxidative stress induction in the male reproductive system. Herbal medicine can be utilized as antioxidant agents against oxidative stress. Quercus brantii (QB) has shown antioxidant activity in previous studies. The aim of the present study was to evaluate effects of QB hydro-alcoholic extract against Pb-induced oxidative stress in the male mice reproductive system. MATERIALS AND METHODS: Forty-two NMRI adult male mice were randomly divided into 7 groups of 6 animals each. Group I was the control group that received no treatment. Group II was the sham group and received 0.2 ml distilled water. Groups III and IV received QB hydro-alcoholic extract 500 and 1000 mg/kg bw, respectively. Group V received Pb 1000 ppm/kg bw. Group VI and VII received Pb 1000 ppm/kg bw and QB extract 500 and 1000 mg/kg bw, respectively. All groups received treatment via oral gavage. After 35 days, sperm parameters (i.e. sperm motility, count and morphology) were evaluated. Levels of sex hormones including LH, FSH, and testosterone, total antioxidant capacity (TAC), superoxide dismutase (SOD) and malondialdehyde (MDA) were measured in animals' serum. RESULTS: Exposure to Pb negatively affected sperm parameters (i.e. sperm motility, count and morphology), decreased serum concentrations of sex hormones (i.e. LH, FSH, and testosterone), TAC and SOD activity but increased MDA levels. However, co-administration of 500 and 1000 mg/kg bw QB hydro-alcoholic extract and Pb considerably improved sperm parameters (i.e. sperm motility, count and morphology), increased sex hormones (i.e.LH, FSH, and testosterone), TAC, and SOD activity while decreased MDA levels in animals' serum. CONCLUSION: Administration of QB extracts (Low dose and high dose) is able to protect the male reproductive system of mice against Pb-induced oxidative stress.

Ameliorative effect of Allium sativum extract on busulfan-induced oxidative stress in mice sperm.

Busulfan is known to cause several adverse effects including reproductive toxicity in humans. Garlic (Allium sativum), a widely distributed medicinal plant, is highly regarded for its medicinal activities including antioxidant property. This study was conducted to assess whether garlic extract could serve as protective agents against testicular toxicity during busulfan treatment in a mice model. Seventy-two adult male mice were randomly divided into nine groups. In groups 1,2 and 3, distilled water, busulfan, and dimethyl sulfoxide and in the treatment groups hydro-alcoholic extract of garlic was administered orally at different doses per day (groups 4, 5 and 6; 200, 400, 800 mg kg-1 respectively). Groups 7, 8 and 9 were treated with the extract (200, 400 and 800 mg kg-1, respectively) plus busulfan. Following euthanasia, blood samples and epididymal sperm were collected. The busulfan-treated group showed significant decreases in sperm quality parameters, and serum levels of testosterone, LH and FSH was observed in the busulfan-treated mice. In addition, the TAC levels and antioxidant enzymes activities were reduced and malondialdehyde (MDA) levels were increased in the busulfan-treated mice. Notably, garlic extract co-administration caused a considerable recovery in sperm quality parameters, TAC levels, antioxidant enzymes activities, hormonal changes and MDA level. Based on our results, garlic has antioxidant effects against busulfan-induced testicular damages in mice.

Comparative study of the protective effects of chicken embryo amniotic fluid, vitamin C and coenzyme Q10 on cyclophosphamide-induced oxidative stress in mice ovaries.

Cyclophosphamide is a chemotherapy drug for the treatment of cancer. Chicken embryo amniotic fluid, vitamin C and coenzyme Q10 have anti-oxidant properties. Total of 70 adult female mice were selected and divided into seven groups. The first group that received 2 ml kg-1 of inactivated amniotic fluid subcutaneously. The second group treated with 75 mg kg-1of cyclophosphamide by intraperitoneal injection. Third to fifth groups received 1, 2, and 4 ml kg-1 of chicken embryo amniotic fluid, respectively. The sixth group received vitamin C at a dose of 0.2 mg g-1 of body weight by oral gavages. Seventh group received 10 mg kg-1 coenzyme Q10 intraperitoneally. All cyclophosphamide treated groups (3-7) received 75 mg kg-1 of cyclophosphamide intraperitoneal on day 22. The mice were euthanized on day 29 and ovarian tissue antioxidant enzymes including glutathione peroxidase, superoxide dismutase and catalase activities and malondialdehyde (MDA) were evaluated. Activities of above mentioned enzymes in treatment groups (3-7) was significantly higher than patient control group (2). The results also revealed that MDA levels were higher in the control group in comparison to other treatment groups. Therefore, it is concluded that the chick embryo amniotic fluid and coenzyme Q10 can compete with compounds like vitamin C in increasing the anti-oxidant level in ovarian tissue.

Effects of bisphenol-S low concentrations on oxidative stress status and in vitro fertilization potential in mature female mice.

Bisphenol-S (BPS) is a new bisphenol-A substitute widely used in many plastic products. Bisphenol-A as a main member of bisphenol family has been known as an endocrine system disrupter chemical compound. Like other members of bisphenol family, there is public health concern about the toxic effects of BPS on reproductive system, thus, we examined BPS effects on in vitro fertilization (IVF) potential and oxidative stress status in a murine model. Adult female mice (n = 70) were randomly divided into control and BPS-treated groups. Bisphenol-S was administered at doses of 0, 1, 5, 10, 50 and 100 µg kg-1 body weight per day intraperitoneally for 21 consecutive days. Twenty-Four hr after the last treatment, five mice in each group were super-ovulated and the oocytes were harvested for IVF. All ovaries were collected and used for biochemical factors analyses. Bisphenol-S exposure at doses more than 10 µg kg-1 induced developmental arrest of pre-implantation embryos. Further, lipid peroxidation measurement in ovaries indicated that all doses of BPS cause oxidative stress in female mice. In conclusion, BPS administration even in low doses can result in female reproductive toxicities and oxidative stress in mice.

Effect of curcumin on rat sperm morphology after the freeze-thawing process.

Reactive oxygen species (ROS) generation, induced by the cryopreservation process, can be responsible for mammalian sperm damage. Curcumin is known as an effective antioxidant against oxidative stress. The aim of this study was to evaluate the effects of curcumin on sperm count, motility and viability, semen total antioxidant capacity and DNA integrity of rat spermatozoa during semen freeze-thawing process. Sperm collected from 10 adult rats were divided into two groups (n=10 for each group): control and a test group supplemented with 2.5 mM curcumin. After freezing-thawing, the number of spermatozoa, motility, viability, total antioxidant capacity (TAC), and DNA integrity of the sperm were analyzed. Motility, viability and DNA integrity of sperm were significantly preserved in treatment groups compared to the control (p < 0.05) after freezing-thawing. Following cryopreservation, TAC was significantly preserved in thawing semen supplemented with curcumin compared to the control group (p<0.05). Based on our results, it is concluded that curcumin addition during freezing resulted in positive effects on sperm parameters after thawing in adult rats.

Susceptibility of different life stages of Ornithodoros lahorensis to entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana.

The use of chemicals for the control of arthropod pests can be problematic due to the potential for both environmental contamination and resistance development. As a result, there is an increasing interest in nonchemical alternatives, such as biocontrol by entomopathogenic fungi. In the present study, three strains of Metarhizium anisopliae (V245, 3247, and 4456) and one strain of Beauveria bassiana (LM 3.2) were evaluated under laboratory conditions for their virulence towards three life stages of Ornithodoros lahorensis. Groups of eggs, larvae, and adult ticks were treated by immersion in two different suspensions (10(5) and 10(7) conidia/ml) of each fungal strain. All treatment and control groups were observed during a 3-week period, and the hatchability of eggs and mortality percentage of larvae and adult ticks were assessed. The effect of fungal strains on egg hatchability and larva and adult mortality was significant and dose dependent compared to the control groups (P < 0.05). The results also showed that the greatest biopesticidal effect was due to strain 4456 of M. anisopliae and LM 3.2 strain of B. bassiana at all tested concentrations, making these fungi potential biological control agents of O. lahorensis reducing the use of chemical acaricides.