Assessment of the effect of kinetin against formic acid toxicity in chicken embryo model.

The current study was designed to investigate the in ovo injection of formic acid (FA) on hatchability rate (HR; Experiment 1) and the potential ameliorative role of kinetin concurrent with FA on biochemical parameters of hatched broilers (Experiment 2). In Experiment 1, live embryonated eggs (n = 280; Day 4 of incubation) were in ovo injected with 0.03, 0.06, 0.125, 0.25, 0.50, 1, 2, 4, 8, 16 and 32 m m FA. In Experiment 2, intra-yolk-sac administration of toxic doses of FA (2 m m) concurrent with kinetin at 50, 100 or 200 µ m were evaluated on hatched embryos. The amount of malondialdehyde (MDA), total antioxidant capacity (TAC), total nitrate-nitrite (TNN), total lipid hydroperoxide (TLHP) and superoxide dismutase (SOD) activity was measured in serum, liver, heart and brain tissues. The results revealed that injection of 2 mM FA significantly increased mortality compared to the control group (p < 0.05). Concurrent administration of 50 or 100 µ m kinetin + 2 m m FA increased HR to 10% and 20% compared to the FA-alone-treated group, respectively. Intra-yolk-sac-received FA group showed greater amounts of MDA, TLHP and TNN and lesser amounts of TAC and SOD activity in serum and tissue samples of liver, heart and brain compared to control groups (p < 0.001). In comparison to the FA-alone-treated group, all doses of kinetin were able to increase the TAC levels in serum and tissue samples when administered concurrently with FA. The doses of 50 and 100 µ m kinetin were efficacious to ameliorate the toxic role of FA injection on SOD activities (p < 0.001). Co-injection of 100 µ m kinetin plus FA significantly reduced the amounts of MDA, TNN and TLHP in measured samples compared to the FA-alone-injected group (p < 0.001). Our results indicated that kinetin (especially at 100 µ m doses) would ameliorate the toxic effects of FA on developing live chicken embryos.

ß-Cyfluthrin-Mediated Cytotoxicity of Cultured Rat Primary Hepatocytes Ameliorated by Cotreatment with Luteolin.

The current study was designed to evaluate the possible protective effects of luteolin against ß-cyfluthrin-mediated toxicity on the primary culture of rat hepatocytes (RHs). In the first step, the exposure of RHs to ß-cyfluthrin (10, 20, 40, and 80 µM) was assessed by MTT. Second, redox condition was evaluated in cotreatment of cells with luteolin (20, 40, and 60 µM) and ß-cyfluthrin (40 µM) at both medium and intra levels. In comparison to control, viability was lower in 40 and 80 µM ß-cyfluthrin-treated groups at 24 h and all ß-cyfluthrin-treated groups at 48 h (P < 0.05). Cotreatment with 20 or 40 µM luteolin + 40 µM ß-cyfluthrin resulted in a higher viability value compared to ß-cyfluthrin alone at 24 and 48 h of incubation (P < 0.05). Administration of 20 or 40 µM luteolin with ß-cyfluthrin led to the decrease of malondialdehyde and total nitrate/nitrite and the increase of total antioxidant capacity (TAC) values in both medium and intrahepatocyte levels compared to the ß-cyfluthrin-treated group at 48 h (P < 0.05). It seems that low and medium doses of luteolin possess the potential to reduce ß-cyfluthrin-mediated hepatotoxicity via attenuation of peroxidative/nitrosative reactions and augmentation of TAC levels.

Protective Effects of Quercetin on Clothianidin-Induced Liver Damage in the Rat Model.

Clothianidin (CTD) is a member of the neonicotinoid group of insecticides. This study was performed to determine the effect of quercetin on clothianidin-induced liver injury (CTD) in rats. Rats were randomly assigned to a normal control (saline), a CTD control-treated group (20 mg/kg) every 3 days for 21 days, and CTD + quercetin-treated groups (2.5, 5, and 10 mg/kg) for 35 days intraperitoneally. Enzyme activity, including alanine aminotransferase (ALT) and aspartate aminotransferase (AST), was measured by spectrophotometry in serum samples by an automatic biochemical analyzer using commercial kits. Total antioxidant capacity (TAC), malondialdehyde (MDA), and nitrate-nitrite were measured in homogeneous liver tissue samples of animals. A significant increase in ALT and AST enzyme activity was observed in the CTD group in comparison with that of the control groups. In the clothianidin + quercetin (10 mg/kg) group, the ALT and AST enzyme levels decreased compared to the clothianidin control group significantly (P < 0.05). The MDA value of the liver increased in the clothianidin-treated group compared to that of the control groups (P < 0.05). Decreased tissue TAC level was observed in the CTD-treated group in comparison with that of the control groups (P < 0.05). The MDA level of the liver decreased in the clothianidin + quercetin (10 mg/kg) group compared to that of the CTD control group (P < 0.05). Quercetin significantly raised the level of TAC in the liver tissue of the clothianidin + quercetin (10 mg/kg) treated group compared to that of the clothianidin control group (P < 0.05). Liver nitrate-nitrite measurement showed a significant increase in the clothianidin group compared to that of the normal control group (P < 0.05). Nitrate-nitrite level in the liver was decreased in clothianidin + quercetin (10 mg/kg) compared to that of the clothianidin control group significantly (P < 0.05). Histopathological investigation revealed that contact to the CTD induced tissue disorganization and inflammatory cell infiltration, but minor histopathological alterations in the liver tissues of rats treated with CTD and quercetin (10 mg/kg) were detected.

Ethephon causes reproductive malfunction in adult male mice: Histological and biochemical evidence.

Ethephon (C2H6ClO3P; ETP), an organophosphorus pesticide regulating plant growth, is widely used for early ripening of fruits and vegetables in agriculture. The aim of this study was to investigate the effects of ETP on histomorphometrical and biochemical parameters in mouse testicular tissue. In this study, 90 adult male mice were randomly divided into six equal groups (n = 15). The ETP was administered orally at different doses (120, 240 and 480 mg kg-1) daily for 35 days. Untreated control, sham (received only normal saline) and neostigmine bromide-treated (positive control; 0.10 mg kg-1 orally; once per week) groups were also considered. Following 35 days, animals were euthanized and testicle and serum samples were taken. Accordingly, blood and serum acetylcholinesterase (AChE), catalase, total antioxidant capacity (TAC) and malondialdehyde (MDA) levels as well as histomorphometrical changes of testicles were investigated. The ETP-administered animals represented a significant reduction in AChE, TAC and catalase levels and remarkable increment in MDA content. A marked reduction was also seen in the germinal epithelium height, connective tissue thickness, seminiferous tubules diameter and Leydig cell number as well as spermiogenesis and Sertoli cell indices in ETP-treated mice compared to control ones. Similar findings were found in neostigmine bromide-treated animals. In conclusion, the ETP significantly affects the serum and blood anti-oxidant statuses and results in severe histological damages both at germ and somatic cell levels, suggesting its hematotoxic and reprotoxic characteristic.

Ameliorative role of trans-ferulic acid on induced oxidative toxicity of rooster semen by ß-cyfluthrin during low temperature liquid storage.

Current study was designed to evaluate the effects of ß-cyfluthrin, as a toxicant substance, and trans-ferulic acid (trans-FA), as a protective agent, on different parameters of rooster semen upon liquid storage. For this purpose, semen samples of roosters (Ross 308, n = 10, 32-wk-old) were collected twice a week. Good quality samples (≥70% progressive motion) were diluted, pooled and then divided for the purposes of the study. In the first experiment, motility of spermatozoa was evaluated following exposure to different concentrations of ß-cyfluthrin (1, 2.5, 5, 10, 20, 40, and 80 µM) at 0, 24, and 48 h of storage. In the second experiment, constant doses of ß-cyfluthrin (10 µM) alone or in combination with trans-FA (10, 25 mM) were assessed on motility and viability of spermatozoa at 0, 24, and 48 h time points. Moreover, amounts of malondialdehyde (MDA), total antioxidant capacity (TAC), total nitrate-nitrite, total hydroperoxide (HPO), and activity of superoxide dismutase (SOD) were evaluated in the homogenate of spermatozoa-diluent at studied time points. Results of the first experiment showed that amounts of ß-cyfluthrin greater than 5 µM, significantly reduced the motility of spermatozoa at 24 and 48 h of storage (P < 0.05). The second experiment demonstrated that, trans-FA especially at 10, 25 mM doses restored the motility and viability of spermatozoa compared to ß-cyfluthrin treated group (P < 0.05). Amounts of MDA (10, 25 mM), hydroperoxide (10, 25, and 50 mM), and nitrate-nitrite (10, 25, and 50 mM) were lower and TAC (10 and 25 mM) were greater in trans-FA + ß-cyfluthrin treated groups compared to ß-cyfluthrin alone treated samples (P < 0.05). However, activity of SOD did not show significant changes by the treatment (P > 0.05). It seems that trans-FA could ameliorate toxic effect of ß-cyfluthrin via reduction of peroxidative (as evident by measurement of MDA) and nitrosative (as evident by measurement of nitrate-nitrite) reactions over cold preservation of rooster semen.

Quercetin attenuates H2 O2 -induced toxicity of rooster semen during liquid storage at 4°C.

Current study was carried out to examine the protective effects of quercetin against toxicity induced by hydrogen peroxide in rooster semen in vitro. Semen samples were collected from ten roosters (Ross 308 broiler breeder males, 32 weeks old) twice a week by abdominal massage method. Samples with ≥70% progressive motility were selected, pooled, diluted and used for the study. Experimental groups consisted of negative control, control that received solvent of quercetin, H2 O2 (40 µM) and combination groups which incubated with constant dose of H2 O2 (40 µM) plus various levels of quercetin (20, 40 and 80 µM). Measurement of total hydroperoxide (HPO), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (TAC) and superoxide dismutase activity as well as routine sperm tests were done at 0, 24 and 48 hr of storage at 4°C. Results revealed that exposure to hydrogen peroxide significantly increased HPO (138.43 ± 7.32 vs. 66.08 ± 3.97 µmol/g protein), MDA (7.21 ± 0.08 vs. 5.71 ± 2.16 µmol/g protein) and NO (0.367 ± 0.013 vs. 0.215 ± 0.011 µmol/g protein) levels and decreased sperm progressive motility (27.28 ± 1.21 vs. 47.49 ± 1.29%), and amounts of TAC (11.49 ± 0.39 vs. 15.70 ± 0.79 mmol/g protein) compared to control at 24 hr (p < 0.05). Changes at mentioned variables were repeated at 48 hr of storage. Also, co-administration of quercetin (especially at 40 and 80 µM) with hydrogen peroxide restored the toxic effects of hydrogen peroxide on rooster semen parameters such as primary and secondary lipid peroxidative indicators and other evaluated variables. The study concluded that rooster semen enrichment with quercetin would protect lipid peroxidative and nitrosative hydrogen peroxide-mediated damage during cold liquid storage of rooster semen.

Evaluation of linoleic acid on lipid peroxidative/antioxidative parameters, motility and viability of rooster spermatozoa during cold storage.

Sustainable reduction in semen quality due to detrimental effects of primary and secondary peroxidative products was occurred during liquid storage. The objective of the current experiment was to explore the influence of bovine serum albumin conjugated linoleic acid (LA) on the rooster spermatozoa routine tests and lipid peroxidative/antioxidative levels stored at 4 °C over 48 h. For this purpose, collected ejaculates (≥ 80% progressive motile spermatozoa) pooled and extended with the phosphate buffer medium without (control) or enriched with different amounts of LA (0.125, 0.25 or 0.50 mM). Contents of total antioxidant status (TAS) and thiobarbituric reactive substances (TBARS) were measured separately in the medium and spermatozoa, as well as percent of viability and motility at 0, 24 and 48 h intervals. Viability was not affected by treatment during the study intervals (P > 0.05). While, higher motility was recorded in LA 0.50 mM group (77.98 ± 1.89 and 57.02 ± 2.45) compared to the control group (68.78 ± 1.29 and 45.09 ± 1.86) at 24 and 48 h, respectively (P < 0.03). Amounts of TBARS in medium and spermatozoa were lower in LA 0.25 and 0.50 groups compared to the control at 48 h (P < 0.01). Moreover, TAS levels of medium and spermatozoa were lower in control samples compared to LA treated groups at 48 h (P < 0.03). Because of the ability of the LA to lowering the quantities of lipid peroxidation index and improving motility especially at 0.5 mM levels, it can be proposed as an additive during liquid storage of rooster semen.

Kinetin improves motility, viability and antioxidative parameters of ram semen during storage at refrigerator temperature.

The present experiment was conducted to evaluate the effect of kinetin on ram semen quality during cold storage. Ejaculates were collected using an artificial vagina from five Qezel rams. Ejaculates which met the criteria (volume of 0.75-2 ml; minimum spermatozoa concentration of 2.5 × 109 spermatozoa/ml and forward progressive motility of 80%), were pooled, diluted with extender without kinetin (control) or enriched with 25 (K 25), 50 (K 50), 100 (K 100) and 200 (K 200) µM kinetin at a final concentration of 500 × 106 spermatozoa per mL. Spermatozoa motion characteristics were evaluated by computer-assisted sperm analysis. In addition, percent of viability (spermatozoa showing no color was considered to be alive) and spermatozoa with intact plasma membrane (spermatozoa with curled/swollen tail was considered healthy) were determined. Moreover, amounts of malondialdehyde (MDA), total antioxidant activity (AOA), nitric oxide (NO) and superoxide dismutase (SOD) activity were determined in the seminal plasma and spermatozoa at 0, 24, 48 and 72 h of storage. Higher percent of total and forward progressive motility was observed in K 25, K 50 and K 100 groups compared to control group at 72 h of storage (P < 0.001). Moreover, K 25 (78.61 ± 1.11%), K 50 (82.46 ± 1.08%) and K 100 (82.96 ± 1.49%) groups showed higher percentage of viability compared to control (72.38 ± 1.49%) at 72 h of storage (P < 0.05). Semen enrichment with kinetin resulted in the higher percent of intact plasma membrane of spermatozoa at 48 and 72 h (P < 0.001). Amounts of MDA were lower and amounts of AOA were higher in K 50 and K 100 groups compared to control at 48 and 72 h (P < 0.05). There were no significant differences in NO levels and SOD activities of seminal plasma and spermatozoa among groups during the experiment. The present experiment revealed that kinetin at proper concentration could enhances spermatozoa kinematics, viability, spermatozoa plasma membrane functionality and amounts of AOA and reduces the level of lipid peroxidation during chilled storage of ram semen.