In the past few decades, extensive application of azoxystrobin has led to great concern regarding its adverse effects on aquatic organisms. The objective of the present study was to evaluate the reproductive toxicity of azoxystrobin to zebrafish. After adult zebrafish of both sexes were exposed to 2, 20 and 200 μg/L azoxystrobin for 21 days, egg production, the fertilization rate, the gonadosomatic index (GSI) and hepatosomatic index (HSI), 17β-estradiol (E2), testosterone (T) and vitellogenin (Vtg) concentrations, and histological alterations in the gonads and livers were measured. Meanwhile, expression alterations of genes encoding gonadotropins and gonadotropin receptors (fshb, lhb, fshr and lhr), steroid hormone receptors (era, er2b and ar), steroidogenic enzymes (cyp11a, cyp11b, cyp17, cyp19a, cyp19b, hsd3b and hsd17b) in the hypothalamic-pituitary-gonad (HPG) axis and vitellogenin (vtg1 and vtg2) in the livers were also investigated. The results showed that reduced egg production and fertilization rates were observed at 200 μg/L azoxystrobin. In female zebrafish, reduced E2 and Vtg concentrations, decreased GSI, increased T concentrations, and histological alterations in the ovaries and livers were observed at 200 μg/L azoxystrobin, along with significant down-regulation of lhb, cyp19b, lhr, cyp19a, vtg1 and vtg2, and up-regulation of cyp17, hsd3b and hsd17b. In male zebrafish, increased E2 and Vtg concentrations, reduced T concentration and GSI, and histological alterations in the testes and livers were observed after exposure to 20 and 200 μg/L azoxystrobin, along with significant up-regulations of cyp19b, cyp11a, cyp17, cyp19a, hsd3b and hsd17b, vtg1 and vtg2. Moreover, cyp11a, hsd3b, cyp19a, vtg1 and vtg2 in male zebrafish were significantly up-regulated after treatment with 2 μg/L azoxystrobin. The results of the present study indicate that azoxystrobin led to reproductive toxicity in zebrafish and male zebrafish were more sensitive to azoxystrobin than female zebrafish.

2,4-Dichloro-6-nitrophenol (DCNP) is an environmental transformation product of 2,4-dichlorophenol that has been identified as widespread in effluent wastewater, but little is known about its toxicity because this compound is not regulated. Therefore, to investigate the endocrine disruption potency of DCNP in Chinese rare minnows (Gobiocypris rarus), adult and juvenile fish were exposed to various concentrations of DCNP (2, 20, and 200 μg/L) for 28 d. After 28 d exposure, the plasma vitellogenin (VTG) levels were reduced in females while increased in males and juvenile fish considerably, as compared with the control. These results suggested that DCNP affects the HPG-axis in a sex-dependent way. Testosterone (T) levels in the plasma were significantly lower in adult and juvenile fish and were accompanied by an increased estradiol (E2)/T ratio. Histopathological observation revealed hypertrophy of the hepatocytes and nuclear pyknosis in the liver, the inhibition of spermatogenesis in the testes, and the degeneration of oocytes in the ovaries after DCNP exposure. The expression pattern of selected genes indicated that the nuclear receptor, steroidogenesis and gonadotropin regulation pathways were perturbed after DCNP exposure. Above all, our results demonstrated that DCNP clearly had anti-androgenic activity in both adult and juvenile fish and can therefore be considered as an endocrine-disrupting chemical.

Fluoride, a ubiquitous environmental contaminant, is known to impair testicular functions and fertility; however the underlying mechanisms remain obscure. In this study, we used a rat model to mimic human exposure and sought to investigate the roles of apoptosis and autophagy in testicular toxicity of fluoride. Sprague–Dawley rats were developmentally exposed to 25, 50, or 100 mg/L sodium fluoride (NaF) via drinking water from pre-pregnancy to post-puberty, and then the testes of offspring were excised on postnatal day 56. Our results demonstrated that developmental NaF exposure induced an enhanced testicular apoptosis, as manifested by a series of hallmarks such as caspase-3 activation, chromatin condensation and DNA fragmentation. Further study revealed that fluoride exposure elicited significant elevations in the levels of cell surface death receptor Fas with a parallel increase in cytoplasmic cytochrome c, indicating the involvement of both extrinsic and intrinsic apoptotic pathways. Intriguingly, fluoride treatment also simultaneously increased the number of autophagosomes and the levels of autophagy marker LC3-II but not Beclin1. Unexpectedly, the expression of p62, a substrate that is degraded by autophagy, was also significantly elevated, suggesting that the accumulated autophagosomes resulted from impaired autophagy degradation rather than increased formation. Importantly, these were associated with marked histopathological lesions including spermatogenic failure and germ cell loss, along with severe ultrastructural abnormalities in testes. Taken together, our findings provide deeper insights into roles of excessive apoptosis and defective autophagy in the aggravation of testicular damage, which could contribute to a better understanding of fluoride-induced male reproductive toxicity.

Benzo[a]pyrene (B[a]P) is an environmental toxicant and endocrine disruptor. Therefore, the aim of the present study was to investigate the toxicity of B[a]P in testis of rats and also to study the role of silymarin and thymoquinone (TQ) as natural antioxidants in the alleviation of such toxicity. Data of the present study showed that levels of testosterone, estrogen and progesterone were significantly decreased after treatment of rats with B[a]P. In addition, B[a]P caused downregulation of the expressions of steroidogenic enzymes including CYP17A1 and CP19A1, and decreased the activity of 17-β hydroxysteroid dehydrogenase (17β-HSD). Moreover, B[a]P decreased the activities of antioxidant enzymes including catalase (CAT), glutathione peroxidase (GPX) and superoxide dismutase (SOD), and significantly increased free radicals levels in testis of male rats. However, pretreatment of rats with silymarin prior to administration of B[a]P was found to restore the level of free radicals, antioxidant status, and activities of steroidogenic enzymes to their normal levels in testicular tissues. Moreover, histopathological finding showed that silymarin recovered the abnormalities occurred in tubules caused by B[a] P in testis of rats. On the other hand, TQ showed pro-oxidant effects and did not ameliorate the toxic effects of B[a] P on the testicular tissue since it decreased antioxidant enzymes activities and inhibited the protein expression of CYP11A1 and CYP21A2 compared to control rats. Moreover, TQ decreased the levels of testosterone, estrogen, and progesterone either in the presence or absence of B[a]P. It is concluded that B[a]P decreased testosterone levels, inhibited antioxidant enzymes activities, caused downregulation of CYP isozymes involved in steroidogenesis, and increased free radical levels in testis. Moreover, silymarin was more effective than TQ in restoring organism health and alleviating the deleterious effects caused by B[a]P in the testis of rats. Due to its negative impact, it is highly recommended to limit the use of TQ as a dietary supplement since millions of people in the Middle East are using it to improve their health.

Bombyx mori L. (B. mori) were exposed to cadmium chloride (CdCl₂) incorporated in an artificial diet (0, 6.25, 12.5, 25, and 50 mg kg⁻¹) throughout the larval stage. Changes in malondialdehyde (MDA) and reduced glutathione (GSH) contents and activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), as well as their corresponding messenger RNA (mRNA) levels in the testes of the fifth instar larvae were evaluated. Additionally, spermatozoon deformation in the testes was examined. Upon Cd treatment, the MDA content in the testes was significantly increased in a concentration-dependent manner. Cd-exposed larvae had increased levels of glutathione. Pearson’s correlation analysis revealed that SOD and CAT activities were positively correlated (R ² = 0.605, P = 0.017). The changing trends in the mRNA levels of these enzymes were not always consistent with those of enzymatic activities. Alterations in GSH-Px activities and mRNA levels were positively correlated (R ² = 0.771, P < 0.01). Morphological analysis revealed that Cd deformed and affected the maturation of spermatozoa. Our results collectively support a relationship between Cd and alterations in the levels of antioxidant enzymes in B. mori testes.

In this study, the effects of cypermethrin (CYP), amitraz (AMT) and combined cypermethrin-amitraz (CYP-AMT) on some serum biochemical, oxidative stress and drug-metabolising parameters were investigated in male Wistar albino rats. CYP, AMT and combined CYP-AMT were administered at doses of 80 mg kg⁻¹ bw⁻¹ of CYP and 170 mg kg⁻¹ bw⁻¹ of AMT for 1 day (single dose), and at doses of 12 mg kg⁻¹ bw⁻¹ of CYP and 25 mg kg⁻¹ bw⁻¹ of AMT for 40 days by oral gavage. Oxidative stress (malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and glucose-6-phosphate dehydrogenase (G6PD)), serum biochemical (glucose, triglyceride, cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), blood urea nitrogen (BUN), creatinine, asparatate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total protein, albumin) in blood/tissues (liver, kidney, brain, spleen and testis) and hepatic drug-metabolising (cytochrome P450 2E1 (CYP2E1), NADH-cytochrome b₅ reductase (CYPb5), NADPH-cytochrome c reductase/NADPH cytocrome P450 reductase (CYTC), glutathione S-transferase (GST), glutathione (GSH)) parameters were measured in liver samples taken on days 1 and 40. In result, it was determined that CYP, AMT and their combinations led to significant changes in the parameters investigated, and it was ascertained that long-term exposure to insecticides and the administration of insecticide combinations produced greater toxic effects in comparison with the administration of insecticides alone.

Thyroid hormones play a fundamental role in the regulation of metabolism of almost all mammalian tissue including the reproductive system. Hyperthyroidism in early life may cause delayed sexual maturation, although physical development is normal and skeletal growth may be accelerated. Hyperthyroidism after puberty influences reproductive functions and increases testosterone level. The aim of this work is to study the effect of induced hyperthyroidism by L-thyroxine sodium administration on the testis of rats and to evaluate the ameliorating role of different antioxidants as ascorbic acid and folic acid on the hyperthyroid state via the assessment of different biochemical markers, histopathological and immunochemical sections. DNA analysis of the D1 deiodinase was performed to determine genetic mutation due to hyperthyroidism. The results showed partially disrupted in the measured biochemical parameters and spermatogenesis in hyperthyroid rats. Post-administration of both folic and ascorbic acids together in hyperthyroid rats showed the best ameliorating effects on the thyroid hormones, testosterone, testicular GGT and ALP, and all oxidative stress markers. There is no genetic mutations that occurred in D1 deiodinase due to hyperthyroidism. These findings were indicated by the proliferating cell nuclear antigen (PCNA) studies of testes.

Organophosphates (OPs) like dimethoate (DMT), are pesticides used worldwide, which can affect both animals and human. Whereas their toxicity is due to acetylcholinesterase inhibition, their secondary toxic effects have been related to free oxygen radical biosynthesis. The present study was designed to investigate the reprotoxic effects of DMT and the protective role of N-acetylcysteine (NAC) in male rat. DMT (20 mg/ kg/body weight) was administered daily to rats via gavage in corn oil and NAC (2 g/l) was added to drinking water for 30 days. Rats were sacrificed on the 30th day, 2 h after the last administration. Markers of testis injury (steroidogenesis impairment) and oxidative stress (lipid peroxidation, reduced glutathione, and antioxidant status) were assessed. In DMT-exposed rats, the serum level of testosterone was decreased. Further, a significant increase in lipid peroxidation level and a significant decrease in the activities of antioxidant enzymes were observed in the testis of rats during DMT intoxication. Real-time PCR (RT-PCR) analysis demonstrated a decrease in messenger RNA (mRNA) levels for testicular steroidogenic acute regulatory StAR protein, cytochrome P450scc, 3β-hydroxysteroid dehydrogenase (3β-HSD), and 17β hydroxysteroid dehydrogenase (17β-HSD) in the testis after DMT exposure. No significant changes in the oxidative stress status and selected reproductive variables were observed on CTN group, whereas NAC restored the oxidative stress and the steroidogenesis on NAC group. Dimethoate induces reprotoxicity and oxidative stress. N-acetylcysteine showed therapeutic recovery effects against dimethoate toxicity.

Natural population of Bufo melanostictus in response to environmental cues shows several physiologic changes such as reproductive activity, hibernation, aestivation and metabolic depression in different seasons. We investigated the effects of seasonal fluctuations on oxidative stress (OS) physiology biomarkers, such as endogenous (ELPx) and induced (ILPx) lipid peroxidation, front-line redox regulatory enzymes (superoxide dismutase: SOD and catalase) and two non-enzyme antioxidant metabolites (ascorbic acid and reduced glutathione) in liver, gonad and cerebral hemisphere of toads collected from the Bhubaneswar area of India, where temperature fluctuates considerably rising to the highest in summer (∼46 °C) and being lowest in winter (<10 °C). Soil and air of the sampling site, although varying seasonally, were mostly found to be unpolluted, except for suspended particulate matter and respiratory particulate matter that were above recommended value. The magnitude of both ELPx and ILPx levels in most of the tissues, for example, ELPx in liver, cerebral hemisphere and ovary, and ILPx in liver of males and ovary, were found to be higher in rainy season in comparison to the other seasons. Nevertheless, levels of both ELPx and ILPx were low in testes in rainy season in comparison to the other two seasons. No correlation was observed between temperature and the studied OS parameters except a positive correlation with SOD and negative correlations with non-enzymatic small redox regulatory molecules in some selected tissues. Conversely, discriminant function analysis reveals a clear impact of the changing season on OS physiology of the toad. It implies that season considerably modulates OS physiology which be a reflection of the toads to abiotic pollutants alone and/or as results of metabolic changes under hibernation, aestivation and due to reproductive activities. Therefore, seasonal changes in OS physiological responses in poikilothermic models especially in toads must be cautiously used as indicators to assess environmental impact, mainly soil pollution. Results of the present study may be used as baseline data for any future analyses of the physiological impacts of environmental changes using toads as model organism.

This study evaluated the protective and antioxidant potential of Omega3 fatty acids (FAs) against aluminum intoxicated male albino rats. Twenty-four male albino rats were divided into four equal groups: group I served as control; group II (Omega3-treated) received Omega3 FAs 1000 mg/kg bwt/day orally; group III (aluminum-treated) received aluminum chloride 100 mg/kg bwt/day orally and group IV (aluminum + Omega3-treated) received aluminum chloride 100 mg/kg bwt/day and Omega3 FAs 1000 mg/kg bwt/day orally. Treatments lasted for 4 weeks. Results indicate that administration of aluminum chloride showed non-significant changes in serum alanine aminotransferase, urea, and creatinine levels, a significant increase in serum aspartate aminotransferase and malondialdehyde as well as a significant reduction in serum-reduced glutathione levels. Aluminum treatment induced histopathological alterations in the liver, kidney, brain, testes, and epididymis. Omega3 FAs supplementation improved the serum parameters, enhanced endogenous antioxidant status, reduced lipid peroxidation, and ameliorated the intensity of the histopathological lesions. These findings reveal that Omega3 FAs supplementation can lighten the toxic effects of aluminum through their antioxidant and free radical-scavenging effects.