Exposure to manganese during sertoli cell formation and proliferation disturbs early testicular development in rats.

Manganese (Mn) is a metal and important micronutrient. However, exposure to supraphysiological levels of Mn, which occur through fungicides, atmospheric emissions, drainages, and spills, has been related to health risks, including morphometric changes in the male reproductive organs and impairment on gametogenesis and sperm quality, impacting the fertile ability of adult animals. Despite the relevance of the fetal/perinatal period for toxicological studies on Mn, previous data only deal with the physical and neurological development of the offspring, without mentioning their reproductive development. The present study investigated whether exposure to Mn during fetal/perinatal phase, specifically during the period of formation and proliferation of Sertoli cells, impairs the reproductive development of male offspring in early postnatal life. Therefore, pregnant Wistar rats were randomly distributed into 3 experimental groups: Ctl (received saline solution), Mn-9 (received 9 mg/kg of MnCl2), and Mn-90 (received 90 mg/kg of MnCl2). The female rats received the experimental treatment by gavage from gestational day 13 to lactational day 15, i.e., postnatal day (PND) 15 of the pups. Oxidative damage to the genetic material of germ and Sertoli cells, together with a decrease in connexin 43 immunolabeling were observed in the testis of male pups evaluated at PND 15. In addition, an increase in the seminiferous tubules presenting slight epithelium vacuolization and cells with eosinophilic cytoplasm were observed, without apparent epididymal changes. In conclusion, it was demonstrated that Mn perturbed the initial testicular development by altering Sertoli cell integrity through oxidative insult, which may compromise the spermatogenesis in the long-term.

Brazilian green propolis extracts modulate cholesterol homeostasis in a preclinical guinea pig model: an in vitro and in vivo study.

Green propolis produced by Apis melífera bees, having Baccharis dracunculifolia D.C. (Asteraceae) as the primary botanical source, has been used in traditional medicine to treat numerous disorders. However, studies evaluating propolis' potential in treating cardiovascular diseases via its effects on cholesterol metabolism are lacking. Therefore, this study investigated the effects of green propolis extracts on lipid metabolism in hypercholesterolemic guinea pigs. Chemical characterization of ethanolic extracts of green propolis samples was undertaken using HPLC. The in vitro characterization included an evaluation of the antioxidant capacity of the hydroalcoholic extract of green propolis (DPPH and FRAP assays) and its ability to act as an inhibitor of the HMG-CoA reductase enzyme. In vivo, we investigated the effect of the hydroalcoholic extract of green propolis on lipid metabolism in hypercholesterolemic guinea pigs. Results obtained validated previous reports of significant antioxidant activity. HPLC analysis confirmed that coumaric acid, artepillin C, and baccharin were the most common and abundant compounds in green propolis samples among the studied compounds. Furthermore, the compounds in these extracts acted as effective HMG-CoA reductase inhibitors in vitro. In vivo assays demonstrated that a hypercholesterolemic diet significantly reduced serum levels of the HDL cholesterol fraction. Simvastatin and propolis hydroalcoholic extracts promoted a significant increase in HDL cholesterol, suggesting that these extracts can improve the serum lipid profile of hypercholesterolemic guinea pigs. Results obtained in this study provide a perspective on the possible hypocholesterolemic effect of green propolis, suggesting that it can improve the serum lipid profile in hypercholesterolemic guinea pigs.

Reproductive toxicity in male juvenile rats: Antagonistic effects between isolated agrochemicals and in binary or ternary combinations.

The management of agrochemicals in Brazilian agriculture impacts global environmental sustainability and food security, since this country is one of the major agro-food exporters in the world. Acephate, carbendazim, and dithiocarbamates (DTCs) such as mancozeb, are among the most detected agrochemicals in Brazilian agro-food products, occurring in combination in several crops, especially in fruit cultures. The present study evaluated the impact of the exposure to isolated agrochemicals and all the combined possible mixtures (binary and ternary forms) on the reproductive parameters of male juvenile rats, known to be a vulnerable biological system and developmental window. Data were analyzed using Generalized Linear Models (GzLM), considering each agrochemical as an independent factor. The study revealed higher reproductive toxicity exerted by isolated agrochemicals when compared to the combined treatments, which exhibited mostly an antagonistic effect. Results suggest endocrine disruptive effects of each one separately on the weight of reproductive organs and testicular histomorphometry, besides changes in testicular SOD activity. The full factorial experimental design employed here allowed us to conclude that it is not possible to scale-up the effects of the isolated treatments to the mixtures, showing how difficult it is to know beforehand the response and cross-talk among the multiple physiological mechanisms disturbed by complex mixtures. Considering that food products are shared on a global scale and that some of these three agrochemicals have already been prohibited in EU countries, the consumption of some Brazilian products puts global human health at risk, that of children.

Biochemical and histopathological responses in peripubertal male rats exposed to agrochemicals isolated or in combination: A multivariate data analysis study.

The widespread use of agrochemicals results in the exposure of the general human population, including children, to several of these chemicals simultaneously. In the present preclinical study, it was investigated the hepatic damages caused by exposure to acephate, carbendazim and mancozeb when administered alone or in different combinations (binary and ternary). Juvenile male Wistar rats were exposed to agrochemicals from post-natal day 53, by gavage. The doses of agrochemicals applied here were determined from previous studies whose results showed no signs of systemic toxicity. All exposures provoked a significant increase in DNA damage (except for acephate alone) and activation of the xenobiotic biotransformation system (except for the ternary mixture). Interestingly, the ternary mixture did not exhibit an exacerbation in adverse effects caused by agrochemicals isolated or in binary combination, even though they are sharing genotoxicity damage induction as a common toxicity pathway. Conversely, some effects observed for isolated or binary combinations of agrochemicals were not observed for ternary combination, suggesting a chemical interaction that could imply antagonism character. Using a multivariate data analysis approach, exposure to isolated agrochemicals were related to a group of adverse effects characterized by hepatic lesion and the attempt of the tissue to mobilize defense cells and increase mitotic rates to minimize damages. Binary mixtures also share similarities in relation to the effects they exhibited, mainly a moderate to high increase in the GST activity and in histopathological alterations suggesting that binary combinations trigger an increased response of the mechanism of xenobiotics biotransformation. Together, obtained results bring important insights regarding adverse effects and possible interaction of the three agrochemicals whose residues are commonly detected in agro-food products.

Food-triad: An index for sustainable consumption.

This study proposes an index for food labeling in order to promote sustainable consumption. The index is calculated by ranking multiple features from the environmental, health and nutritional dimensions of the target product in relation to a pre-set reference value; the obtained scores from each dimension are plotted in a radar chart resulting in a triangular area. An increase in area represents a greater impact. As examples, tuna and the potato-based foods at three different processing levels (in natura or minimally processed, processed and ultra-processed) were analyzed. For both cases, the index increases according to the processing grades and has proved to be capable of expressing in numbers and graphically a wide range of environmental, nutritional and health issues.

Prepubertal exposure to low doses of sodium arsenite impairs spermatogenesis and epididymal histophysiology in rats.

For the first time, juvenile toxicity of inorganic arsenic (As) was investigated in male rats, focusing on reproductive effects. As is a metalloid naturally occurring in the environment, being the inorganic forms the most toxics. Contaminated drinking water and agricultural products are the main prospectors of intoxication for general population. In the present study, Wistar male rats (21 days old) were distributed into three groups (n = 10/group): control (received vehicle-filtered drinking water), As1 (received AsNaO2 at 0.01 mg L-1 ) and As2 (received AsNaO2 at 10 mg L-1 ). The animals were euthanized on PND 53. Testicular damages increased in As1 and As2 compared to control (ie, presence of vacuolization, acidophilic cells, and epithelium degeneration). Testicular interstitium of As1 and As2 presented fluid's increase and intense inflammatory infiltration. In the epididymis there was reduction of sperm amount in the lumen, besides epithelium areas presenting cribriform aspect in As1 and As2, exfoliation of cells in the light (in As1) and vacuoles (in As2). In epididymis interstitium, inflammatory infiltrates were observed in initial segment of As1 and As2. AsNaO2 changed immunolabeling pattern for androgen receptor in epididymis of As2, although serum testosterone levels was statistically comparable to control. Mass spectrometry revealed higher As concentrations in testis and epididymis of As2 compared to As1 and Control. These results indicate compromise of spermatogenesis and epididymal histophysiology in AsNaO2 -treated animals, possibly impairing sperm quality and fertility in long-term, even at low levels of exposure. Investigations about the reversibility of reproductive damages are necessary to better understand the mechanisms of As reproductive toxicity.

Could male reproductive system be the main target of subchronic exposure to manganese in adult animals?

Manganese (Mn) is one of the most common chemical elements on Earth and an essential micronutrient in animal organism. However, in supraphysiological levels and long-term exposures, it is a potential toxicant. Although nervous system is the most studied in relation to Mn toxicity, other tissues can have their function impaired by Mn in high doses. The present study investigated the possible adverse effects of subchronic exposure to supraphysiologic level of Mn (5 mg/kg or 15 mg/kg, intraperitoneally) on reproductive, neurobehavioral, renal and hepatic parameters of male rats. For the first time, the vulnerability of these parameters to Mn was concomitantly investigated. While our results demonstrate that Mn treatments were not sufficient to produce a marked effect of neurotoxic, hepatotoxic or renal toxicity in adult rats, we found typical indicators of reproductive toxicity such as histopathological changes (major in testes and epididymis) and impaired sperm concentration and quality. Mn, under these experimental conditions, seems to exert reproductive toxicity by different testicular mechanisms, i.e. direct and indirect action on germ cells. On the other hand, exposure to Mn did not change the pattern of cognitive and emotional behaviors and the histological organization of kidneys of experimental rats. The liver showed a weight increasement and hidropic degeneration, probable due to the detoxification overload. In summary, for the first time it was demonstrated that adult male reproductive system was more sensitive to Mn toxicity than nervous, hepatic and renal systems, although nervous system is known as the main target tissue of this metal.

Effects of Anticancer Drugs in Reproductive Parameters of Juvenile Male Animals and Role of Protective Agents.

Nowadays, the advances in knowledge about oncologic treatment have led to an increase in survival rate for cancer patients and, consequently, a growing concern about the adverse effects of treatment in medium and long term, in order to ensure the future quality of life. For male patients in reproductive age or younger, one of the key concerns after cancer therapy is their ability to father children, since anticancer drugs exert cytotoxic effects on germ cells. Considering the incidence of cancer in children and adolescents and the vulnerability of these developmental phases to chemical injuries, this review is an attempt to highlight the importance of juvenile experimental models to test new anticancer drugs and agents with protective action. There is a relative scarcity of studies investigating the effects of chemotherapy in juvenile animals and an urgent need for further information. As far as this review was able to recover, available data about reproductive toxicology related to peripubertal treatment with anticancer drugs includes only the following pharmaceuticals: toposide, doxorrubicin, cisplatin, ciclophosphamide, cytarabine, flutamide and procarbazine. Together with the evaluation of adverse effects of anticancer drugs, is necessary to investigate possible protective agents to be pre-, co-, or post administrated with chemotherapy. Modern technologies and increasing knowledge about the cancer biology have allowed studies of new chemotherapy strategies, more effective and selective. Many of these compounds are derived from toxins and metabolites of microorganisms, plants, and animals, being a number of them isolated from marine sources, a relatively unexplored environment. Investment in research programs in bioprospecting, especially in marine environments, and pharmaceutical field, including toxicology risk evaluation, are crucial to discovery and improve new anticancer treatments.

Sperm quality and fertility in rats after prenatal exposure to low doses of TCDD: A three-generation study.

Exposure to Tetrachlorodibenzo-p-dioxin (TCDD) in male rats promotes, decreased sperm concentration, alterations in motility and in sperm transit time. We evaluated the effect transgenerational of in utero exposure to low doses TCDD in the sperm quality. Pregnant rats (F0) were exposed to 0.1; 0.5 and 1.0µg of TCDD, on gestational day 15, coincides with the end of most organogenesis in the fetus. Adult male offspring (F1, F2 and F3 generation) were investigated for fertility after artificial insemination in utero. After collection of the uterus and ovaries, the numbers of corpora lutea and implants were determined. TCDD provoked alterations in sperm morphology and diminution in serum testosterone levels and sperm transit time in the cauda epididymis. The fertility significantly decreased in all the generations, at least at one dose. In conclusion, TCDD exposure decreases rat sperm quality and fertility in adult male offspring and this effects persist into the next generation.

Absence of effects on the rat sperm quality after subacute exposure to low doses of fungicide prochloraz.

Prochloraz (PCZ) is a fungicide and androgen-receptor antagonist used worldwide in horticulture and agriculture. Pre- and perinatal exposure to this pesticide during sexual differentiation is deleterious for male offspring. Since data on the effects of PCZ on epididymal functions are scarce, and because sperm maturation occurs in this organ, the present investigation aimed to determine whether low PCZ doses administered to rats during the phase of sperm transit through the epididymis might affect the morphophysiology of this organ and sperm quality. Adult male Wistar rats were assigned to 4 different groups: 0 (control, vehicle) or 10, 15, or 30 mg/kg bw/d PCZ diluted in corn oil administered orally for 4 consecutive days. Morphofunctional parameters of the male reproductive tract, hormone concentrations, sperm evaluations, and fertility and histopathologic analysis of testis and epididymis were assessed. There were no statistically significant differences between treated and control groups in relation to all evaluated parameters. Data demonstrated show that PCZ exposure for a brief 4-d exposure and low doses did not produce reproductive toxicity or compromise sperm quality in adult rats.

The male peripubertal phase as a developmental window for reproductive toxicology studies.

The normal development of the male reproductive system can be divided into five phases: fetal, neonatal, childhood, puberty and adulthood. Childhood/peripuberty has yet been relatively little studied. Chemical insults during the peripubertal phase may result in adverse consequences that may be already visible during puberty as well as during later adult life. This occurs because endocrine disruptors often interfere in the developmental programming. The most important is to note that children are not just little adults and should be particularly investigated. The aim of this review is to discuss the recent literature (2000-2013) on male reproductive aspects in prepubertal toxicity assays, focusing on experimental in vivo studies, establishing a comparative analysis between the design, endpoints, results and consequent conclusion. The studies discussed in the present review were selected based on the period of exposure. Only studies with post-lactational exposures were included. 33 papers were included using rats, mice, rabbits or pigs as experimental model. There is a relative scarcity of studies investigating animals in development and thus an urgent need for further studies in order to evaluate the possible persistent effects on fertility and other reproductive parameters at adulthood. Another point is the lack of studies with chemical mixtures, an imminent problem in modern society. It is vital to consider the refinement of alternative methods and the experimental designs and endpoints to improve the scientific knowledge in this area.

Morphologic and biomechanical changes of thoracic and abdominal aorta in a rat model of cigarette smoke exposure.

BACKGROUND: Smoking is the most relevant environmental factor that affects the development of aortic aneurysm. Smokers have elevated levels of elastase activity in the arterial wall, which leads to weakening of the aorta. The aim of this study was to verify whether cigarette smoke exposure itself is capable of altering the aortic wall. METHODS: Forty-eight Wistar rats were divided into 2-, 4-, and 6-month experimental periods and into 2 groups: smokers (submitted to smoke exposure at a rate of 40 cigarettes/day) and nonsmokers. At the end of the experimental periods, the aortas were removed and cross-sectioned to obtain histologic specimens for light microscopic and morphometric analyses. The remaining longitudinal segments were stretched to rupture and mechanical parameters were determined. RESULTS: A degenerative process (i.e., a reduction in elastic fibers, the loss of lamellar arrangement, and a reduction of smooth muscle cells) was observed, and this effect was proportional in intensity to the period of tobacco exposure. We observed a progressive reduction in the yield point of the thoracic aorta over time (P < 0.05). There was a decrease in stiffness (P < 0.05) and in failure load (P < 0.05) at 6 months in the abdominal aorta of rats in the smoking group. CONCLUSIONS: Chronic exposure to tobacco smoke can affect the mechanical properties of the aorta and can also provoke substantial structural changes of the arterial wall.

Sperm quality in adult male rats exposed to cadmium in utero and lactation.

Recent studies indicate that several anomalies of the male reproductive system may be produced by acute or chronic exposure to chemical substances released into the environment, attributed to increased industrial development. Among these substances are trace metals such as cadmium (Cd). The aim of this study was to assess reproductive parameters in adult male rats whose mothers were exposed to Cd during pregnancy and lactation. For this, pregnant rats were divided into two experimental groups: treated rats, which received ad libitum cadmium acetate (CdAc) solution in distilled water (10 mg Cd/L), and control rats, which received sodium acetate (NaAc) solution in distilled water (equimolar to the CdAc). The results showed that the exposure to Cd in utero and through lactation adversely affected sperm quality of adult rats, as evidenced by compromised sperm morphology and motility and increased rate of cell death in testis.

Reproductive physiology, and physical and sexual development of female offspring born to diabetic dams.

OBJECTIVES: The objective of this study was to evaluate physical and sexual development and reproductive physiology in female rat offspring that developed in hyperglycemia conditions in utero and during lactation. MATERIALS AND METHODS: Maternal diabetes was induced in female rats by a single IV injection of streptozotocin before mating. Female offspring development was evaluated by means of the following parameters: physical development; age of vaginal opening and first estrus; weight and histological evaluation of uterus and ovaries; duration of the estrous cycle, sexual behavior, and fertility after natural mating. RESULTS: In the female offspring, maternal diabetes caused delays in initial physical development; diminution in ovary weight and number of follicles; and inferior reproductive performance compared with the control group. CONCLUSIONS: The exposure to hyperglycemia in uterus and during lactation caused delays in physical and sexual development, and affected the reproductive physiology of female rats negatively.

Morphometric-stereological and functional epididymal alterations and a decrease in fertility in rats treated with finasteride and after a 30-day post-treatment recovery period.

OBJECTIVE: To evaluate morphometric-stereological changes in the epididymal caput, sperm quality, and fertility parameters in rats treated with finasteride and after a 30-day post-treatment recovery period. DESIGN: Experimental study in a research laboratory. SETTING: Reproductive biology research laboratory. ANIMAL(S): Male and female Sprague Dawley rats. INTERVENTION(S): Treatment with finasteride (5 mg/kg/day) for 56 days followed by 30 days without treatment. MAIN OUTCOME MEASURE(S): Serum hormone analyses, morphometric-stereological and ultrastructural evaluation of the epididymal caput, sperm transit time, natural mating, in utero insemination, sperm membrane integrity, and fertility parameters. RESULT(S): Serum dihydrotestosterone levels in the finasteride group decreased by ~40% compared with that of control rats. Ultrastructural analysis revealed significant reductions in several morphometric-stereological parameters of the epididymal caput. All parameters recovered significantly in the post-treatment period. There was no alteration in daily sperm production in the finasteride group. However, significant reductions in sperm transit time, motility, sperm membrane integrity, and fertility parameters were observed in rats treated with finasteride. CONCLUSION(S): Treatment with finasteride caused morphometric-stereological and functional changes in the epididymis and in sperm function that led to a reduction in fertility parameters. A 30-day post-treatment recovery period was insufficient to restore normal sperm motility, sperm transit time, and some fertility parameters.

Reproductive physiology, and physical and sexual development of female offspring born to diabetic dams / Desenvolvimento físico, sexual e fisiologia reprodutiva da prole feminina de ratas diabéticas

OBJECTIVES: The objective of this study was to evaluate physical and sexual development and reproductive physiology in female rat offspring that developed in hyperglycemia conditions in utero and during lactation. MATERIALS AND METHODS: Maternal diabetes was induced in female rats by a single IV injection of streptozotocin before mating. Female offspring development was evaluated by means of the following parameters: physical development; age of vaginal opening and first estrus; weight and histological evaluation of uterus and ovaries; duration of the estrous cycle, sexual behavior, and fertility after natural mating. RESULTS: In the female offspring, maternal diabetes caused delays in initial physical development; diminution in ovary weight and number of follicles; and inferior reproductive performance compared with the control group. CONCLUSIONS: The exposure to hyperglycemia in uterus and during lactation caused delays in physical and sexual development, and affected the reproductive physiology of female rats negatively.

OBJETIVOS: O objetivo deste estudo foi avaliar o desenvolvimento físico e sexual e a fisiologia reprodutiva de ratas que se desenvolveram em condições hiperglicêmicas in utero e lactação. MATERIAIS E METODOS: Para induzir o diabetes nas ratas, foi utilizada estreptozotocina em dose única via intravenosa antes do acasalamento. A prole feminina foi avaliada por meio dos seguintes parâmetros: o desenvolvimento físico; a idade de abertura vaginal e do primeiro estro, peso e avaliação histológica do útero e ovários; a duração do ciclo estral, o comportamento sexual e a fertilidade após acasalamentos naturais. RESULTADOS: O diabetes materno provocou, na prole feminina, retardo no desenvolvimento físico; diminuição do peso dos ovários e do número de folículos; a performance reprodutiva foi inferior à do grupo controle. CONCLUSÕES: Concluiu-se que a exposição aos meios intrauterino e lactacional hiperglicêmicos provocou retardo no desenvolvimento físico e sexual e prejudicou a fisiologia reprodutiva de ratas.

Impairment on sperm quality and fertility of adult rats after antiandrogen exposure during prepuberty.

This study evaluated the effects of antiandrogen exposure during the prepubertal period on reproductive development and reproductive competence in adults. Male rats were divided into two groups: flutamide, receiving 25 mg/kg/day of flutamide by oral gavage and control, receiving vehicle daily. Dosing continued from PND 21 to 44, and animals were killed on PND 50 or PND 75-80. The epididymis, prostate, vas deferens and seminal vesicle weights were lower in Flutamide group on PND 50, while on PND 80 only seminal vesicle weight was reduced. Fertility assessed by IUI revealed a decrease in the fertility potential in the flutamide-treated adults. Flutamide accelerated sperm transit time through the epididymis, impairing sperm motility and storage. A quantitative analysis of the cauda sperm membrane proteome revealed a few significant changes in protein expression. Thus, exposure to flutamide during the prepubertal period compromises the function of the epididymis along with epididymal sperm quality at adulthood.

Assessment of female reproductive endpoints in Sprague-Dawley rats developmentally exposed to Diuron: potential ovary toxicity.

BACKGROUND: Diuron is widely used in agriculture but its deleterious effects on the reproductive system and mammary gland are still poorly understood. This study evaluated whether early-life-stage exposure to Diuron alters puberty onset or susceptibility to mammary carcinogenesis in female Sprague-Dawley rats. METHODS AND RESULTS: Pregnant rats received basal diet or diet containing Diuron at 500, 750, and 1,250 ppm, from gestational day 12 to the end of lactation (postnatal day 21 [PND21]). After weaning, female offspring continued receiving basal diet or diet containing Diuron until PND 51. At PND 51, female Sprague-Dawley offspring received a single dose of 50 mg/kg b.w. of 7,12-dimethylbenz(a)anthracene (DMBA) for initiation of mammary carcinogenesis. The animals were sacrificed on PND 51, 75, and 226 to 233 (week 25) for mammary gland morphology, reproductive organs and tumor analysis, respectively. There were no significant differences among groups on vaginal opening, estrous cycle, mammary morphology, or carcinogenesis. However, reductions in ovary weight and corpora lutea were observed at PND 75 in the group treated with Diuron at 1,250 ppm. CONCLUSIONS: The findings suggesting that Diuron exposure (1,250 ppm) may have been potentially toxic to the ovaries.

Decreased sperm motility in rats orally exposed to single or mixed pesticides.

The Brazilian Agency of Sanitary Vigilance (ANVISA) conducted a study that demonstrated the presence of residues of several pesticides in fresh fruits and vegetables that were available for purchase by the general populace. In order to evaluate potential adverse health effects of low-level exposure to agrochemicals, the reproductive toxicity of the pesticides dicofol, dichlorvos, permethrin, endosulfan, and dieldrin was evaluated in rats dosed with these chemicals individually or as mixtures. Sixty male Lewis rats (6 wk old, 200 x g) were randomly allocated to 8 groups: (1) control group, received basal diet; (2) 5 groups designated a to e received the diet containing each pesticide individually, at the respective effective doses: lowest-observed-adverse-effect level (LOAEL) for dieldrin and endosulfan, lowest-observed-effect level (LOEL) for dicofol, and lowest effect level (LEL) for dichlorvos and permethrin, respectively, depending on the published data; (3) effective dose group, which received a mixture of pesticides added to basal diet at the respective doses reported to produce adverse effects; and (4) low dose group, which received a pesticide mixture added to the basal diet, where each pesticide was at its no-observed-effect level (NOEL). After 8 wk of treatment, reproductive parameters were evaluated. Sperm morphology, daily sperm production (DSP), sperm transit time through the epididymis, hormonal levels, and histopathological evaluation of testis and epididymis did not differ significantly among the groups. However, sperm motility was significantly decreased in animals that received a mixture of dieldrin, endosulfan, dicofol, dichlorvos, and permethrin, as well as in the group receiving dicofol alone. Exposure to the individual pesticides endosulfan, dichlorvos, and permethrin did not markedly affect sperm motility. The impairment of sperm motility in the mixture of pesticides at the NOEL level indicates that reproductive effects not seen with individual pesticides may occur in presence of several pesticides due to an additive effect. However, the pesticide mixtures did not appear to affect DSP or spermatogenesis despite reduced sperm motility.