Microalga Scenedesmus bajacalifornicus BBKLP-07, a new source of bioactive compounds with in vitro pharmacological applications.

Microalgae are photosynthetic eukaryotes which are primary producers in the food chain and also excellent sources for bioactive compounds such as alkaloids, flavonoids, phenols, saponins and other fine chemicals. In the present study, the microalga Scenedesmus bajacalifornicus BBKLP-07 was subjected to soxhlet extraction using solvents like chloroform, acetone, ethanol, methanol and aqueous solvents. All the solvents were tested for the presence of phytochemical constituents such as alkaloids, flavonoids, glycosides, phenols, lignin's, saponins, sterols, tannins, anthraquinone and reducing sugar using the standard procedures. Furthermore, all the crude extracts were subjected to antidiabetic, antioxidant, anti-inflammatory and antimicrobial activities. Antidiabetic activity of the microalgal extracts was observed maximum in Aqueous extract. Methanolic extracts have shown maximum antioxidant activity and chloroform extracts have exhibited highest anti-inflammatory effects. Antimicrobial activities were tested against E.coli, S, typhi, C.perfringens and B.subtilis bacteria and fungi A.niger, and C. albicans. Therefore, the green microalga Scenedesmus bajacalifornicus BBKLP-07 is a rich source of biological active compounds and nutraceuticals and can be exploited for commercial applications.

Purification and characterization of laccase from Marasmius species BBKAV79 and effective decolorization of selected textile dyes.

A novel laccase-producing white-rot fungus, Marasmius sp. BBKAV79 (Genbank Accession Number-KP455496, KP455497), was isolated and subjected to purification, characterization and dye decolorization study. The purified enzyme was obtained with a specific activity of 0.226 U mg-1 protein and a final yield of 13.5 %. The enzyme was found to be a monomeric protein with a molecular mass of ~75 kDa as estimated by non-denaturing polyacrylamide gel electrophoresis (PAGE) and further confirmed with zymogram analysis. The optimal pH and temperature of the laccase was recorded to be 5.5 and 40 °C, respectively. The metal ions Hg2+ and Ag+ were found to drastically inhibit the activity of laccase at the rate of 96.6 and 96.5 %, respectively. Nevertheless, Fe3+ was found to inhibit laccase activity at 40 %. Phenylmethanesulfonyl fluoride (PMSF) strongly inhibited the laccase activity, and additives viz, sodium dodecyl sulfate (SDS), hydrogen peroxide (H2O2) and sodium chloride (NaCl) were known to follow the earlier pattern of enzyme inhibition. The values of kinetic parameters K m and V max for purified laccase were noted at 3.03 mM and 5 µmol min-1, respectively, for guaiacol as substrate. The textile dyes were decolorized at a range of 72-76 % and 88-93 % when treated with Marasmius sp. BBKAV79 and purified laccase, respectively. Based on the outcome of the present investigation, it could be, therefore, inferred that laccase isolated from Marasmius sp. BBKAV79 effectively decolorizes the textile dyes; however, the metal ions Hg2+, Ag+ and Fe3+ and agents like PMSF, SDS, H2O2 and NaCl pose an effective inhibitory potential under specified physicochemical conditions.

Dynamics of phytoplankton in relation to physico-chemical factors of Almatti reservoir of Bijapur District, Karnataka State.

The present investigation deals with limnobiotic status of the Almatti reservoir from February, 2003 to January, 2005. The study revealed that there exists a fluctuations of the physical factors viz., rainfall, humidity, air and water temperature, pH and electrical conductivity (EC), and chemical factors viz., dissolved oxygen (DO), free carbon dioxide, total alkalinity, total hardness, calcium, magnesium, chloride, nitrate, phosphate, sulphate, bicarbonate and total dissolved solids (TDS). From the data it was also apparent that correlations between the physico-chemical factors and dynamics of phytoplankton could be seen. The simple correlation coefficient test revealed that the cyanophytes number was positively correlated with DO, nitrate, phosphate and negatively significant with total hardness, total alkalinity, EC, calcium, magnesium, sulphate, bicarbonate and TDS. They are inversely correlated with pH, chloride, rainfall and humidity. Bacillariphyceae are correlated with total alkalinity, bicarbonates, magnesium and TDS, whereas inverse correlation was found with rainfall, humidity, pH and phosphate. Desmids showed positive correlation with nitrates, while it was inversely correlated with chloride, rainfall and humidity. Dinophytes density was positively correlated with total alkalinity, EC, total hardness, calcium, bicarbonate, while it showed inverse correlation with rainfall, humidity and phosphate. However, it is obvious that the absence of significant difference between sampling stations for all these parameters in the Almatti reservoir indicated fairly homogeneous conditions and the water quality is also found to be homogeneous.

Water quality assessment of Almatti Reservoir of Bijapur (Karnataka State, India) with special reference to zooplankton.

The present investigation deals with the limnobiotic status of Almatti reservoir from February, 2003 to January, 2005. The study revealed that, the distribution and population density of zooplankton species depend upon the physico-chemical factors of the environment. Statistical analysis showed that there exists a significant relation between the biological and non-biological factors. The benthic fauna constituting the food of fish can be utilized for extensive culture operation so that the nutrients in the reservoir are not only properly cycled but also serve as a check on further eutrophication.

Carbofuran induced block of compensatory ovarian growth in hemicastrated albino mice.

Carbofuran, a systemic N-methyl carbamate pesticide was orally administered at doses of 0.4, 0.7, 1 and 1.3 mg/(kg day) to normal hemicastrated (HC) mice for 15 consecutive days to investigate the effects on compensatory ovarian growth. Sham-operated and hemicastrated control mice were administered a similar quantity of olive oil. The vaginal smear and body weight of the mice were recorded daily and mice were sacrificed on day 16. The results revealed that the HC control mice showed 54.58% increase in the ovary weight of the remaining left ovary and many follicles, corpora lutea and showed normal estrous cycle as compared to Sham-operated control mice. The remaining left ovaries of the mice treated with 1.3 mg carbofuran showed 0.82% increase in the ovary weight and significant decrease in the number of developing follicles, corpora lutea, increase in many atretic follicles, the estrous cycle and its phases were affected when compared to HC control mice. In HC mice treated with 1 mg/(kg day) carbofuran resulted in 12.63% increase in the ovary weight and showed significant decrease in follicles, corpora lutea and many increased atretic follicles. The estrous cycle was affected with significant decrease in estrus phase and increase in diestrus phase. In HC mice treated with 0.7 mg/(kg day) carbofuran showed 26.30% increase in ovary weight and showed normal number of follicles and the mice showed normal number of estrous cycles but the estrus and diestrus phases were affected significantly. However, HC mice treated with 0.4 mg/(kg day) carbofuran revealed 33.09% increase in the ovary weight with no significant change in the follicles and estrous cycle as compared with that of HC control mice. There was no significant change in the body weight gain and weight of the organs such as uterus, kidney, adrenals, liver, spleen, thymus and thyroid in all the carbofuran treated mice. These observed effects of carbofuran on ovary, follicles and estrous cycle are considered to be hormonal imbalance.

Temporal effect of carbofuran on estrous cycle compensatory ovarian hypertrophy and follicles in hemiovariectomized albino mice.

Carbofuran, a systemic N-methyl carbamate pesticide was administered orally with an effective dose of 1.3mg/kg per day to hemiovariectomized (HOVX) mice for 5, 10, and 15 days. Sham-operated and HOVX control mice were administered a similar quantity of olive oil. The vaginal smear and body weight of the mice were recorded daily and mice were sacrificed on day 16. The results of the present study indicate that there is a significant decrease in the duration of the estrus with a concomitant significant increase in the duration of the diestrus with carbofuran treatment for 10 days. In the HOVX mice treated with carbofuran for 15 days there was a significant decrease in the length of the estrous cycle and the duration of the estrus and metestrus with a concomitant significant increase in the diestrus. There was a significant increase in the ovarian weight in HOVX control mice when compared to that of the sham operated control mice. HOVX mice treated with carbofuran for 10 and 15 days showed a significant decrease in the relative ovarian weight with a concomitant inhibition of compensatory ovarian hypertrophy in HOVX mice. There was also a significant decrease in the ovarian growth rate in relation to the contralateral ovary of the same animal. There was a significant decrease in the number of small and total number of healthy follicles with a concomitant significant increase in the number of medium, large and total number of atretic follicles in HOVX mice treated with carbofuran for 10 days. There was a significant decrease in the number of small, medium, large, and total number of healthy follicles with a concomitant significant increase in the number of medium, large, and total number of atretic follicles in HOVX mice treated with carbofuran for 15 days. The administration of carbofuran in the present study showed that there is no significant change in the body and organs weight such as uterus, kidney, adrenals, liver, spleen, thymus and thyroid in all the carbofuran-treated mice. The above findings such as disruption in the estrous cycle, inhibition of compensatory ovarian hypertrophy, and follicular toxicity may be due to a direct effect on the ovary or through the hypothalamo-hypophysial-ovarian axis.

Evaluation of dimethoate toxicity on pregnancy in albino mice.

We investigated whether dimethoate, an organophosphorus insecticide, causes abortion or fetal resorption in pregnant albino mice. Graded doses of 16, 20, 24, and 28 mg/kg body weight/d were administered orally from days 7 to day 15 of pregnancy. Laparotomy was performed on day 8 of pregnancy to note the number of implantations, and the animals were autopsied on day 19. The results revealed no inhibition of pregnancy in all dimethoate treated mice relative to a suitable control group. Treatment with 24 or 28 mg dimethoate caused a significant decrease in the number of implantations, live fetuses, and corpora lutea, but a decreased percent fetal survival and increased percent post-implantation loss and gestation length were not significant when compared with control mice. In all mice treated with 28 mg dimethoate, a significant decrease occurred in the body weight of the ovaries, uterus, and liver when compared with control mice. Following treatment with 16 or 20 mg dimethoate, however, no significant change was found in body and organ weights or in the number of implantations, live fetuses, and corpora lutea, percent post-implantation loss, and fetal survival, or gestation length compared with the corresponding parameters in control mice. The results of this study clearly indicate that dimethoate does not cause abortion or fetal resorption in pregnant mice. A significant decrease in the number of corpora lutea and percent fetal survival observed at higher doses of dimethoate could be due to a toxic effect on the embryo or to a hormonal imbalance.

Morphometric analysis of follicular growth and biochemical constituents in albino rats exposed to mancozeb.

Mancozeb, an ethylenebisdithiocarbamate (EBDC), was administered orally at a dose of 700 mg/kg body weight/day to female virgin rats for 5, 10, 20, and 30 days to examine the effect on ovarian follicular development. No significant change occurred in the number of estrous cycles and the duration of proestrus, estrus, and metestrus, but mancozeb treatment for 5 days significantly increased the duration of diestrus. Mancozeb treatment for 10 days significantly increased the number of estrous cycle and the duration of estrus, with a concomitant significant increase in diestrus, but no change in proestrus and metestrus. With mancozeb treatment for 20 and 30 days, the number of estrous cycles and duration of proestrus, estrus, and metestrus were significantly decreased, with a concomitant significant increase in the duration of diestrus. Exposure of rats to mancozeb for 5 days resulted in a significant decrease in stage II and the total number of healthy follicles but no change in atretic follicles. Mancozeb treatment for 10 days resulted in a significant decrease in stages I, II, and IV and in the total number of healthy follicles. A significant increase in atretic follicles was found in rats treated with mancozeb for 20 and 30 days. No significant change occurred in body and organ weights in any group, but the thyroid weight of 20 and 30 days mancozeb-treated rats was significantly increased. The level of protein in the ovary was significantly decreased, but no change was found in the uterus and liver of mancozeb-treated animals. The level of glycogen was significantly decreased in the ovary and the uterus with mancozeb treatment, but not in the liver. With mancozeb treatment, the levels of phospholipids and neutral lipids were significantly increased in the liver but significantly decreased in the uterus. The change in the biochemical constituents of ovary, uterus, and liver was duration dependent. The results of the study thus indicate a marked disruption of the estrous cycle, pathological changes in the gonads, and organ-specific biochemical changes in rats after exposure to mancozeb.

Evaluation of dimethoate-induced implantation delay and nidation by progesterone in albino mice.

We investigated the effect of dimethoate, an organophosphorus insecticide, and progesterone on implantation. Virgin pregnant albino mice received dimethoate orally at a dose of 28 mg/kg body wt/d from days 1 to 7. Laparotomy on day 8 showed no implantation sites. Thereafter, graded doses of progesterone, 4, 9, and 12 mg/kg body wt/d, were administered up to day 15. A group of control mice received a similar quantity of distilled water. Autopsy on day 8 revealed that the control mice were pregnant, with a normal number of implantations and 8.08% pre-implantation loss, whereas treatment with dimethoate for 7 days or with dimethoate for 7 days followed by progesterone for 8 days totally abolished implantation, with a 100% pre-implantation loss. In all treated mice, a significant decrease occurred in body weight gain, as well as in the weight of the ovaries, uterus, and liver when compared with those of control mice. No significant changes were found in other organ weights (kidneys, adrenals, spleen, thymus, or thyroid). The observed effect of dimethoate could be due to an imbalance in the estrogen-progesterone ratio essential for implantation. Alternatively, dimethoate treatment could result in blastotoxicity or have an impact on the hypothalamic-pituitary axis.

Effect of edifenphos on compensatory ovarian hypertrophy, follicular kinetics and estrous cycle in hemicastrated rats.

Edifenphos, an organophosphate fungicide, was administered to hemicastrated albino rats intraperitoneally for 15 consecutive days in graded doses of 2 to 8 mg/kg body weight. In the oil-treated hemicastrated control group, ovarian weight and total number of healthy and atretic follicles were significantly higher than the same parameters in sham-operated control animals, and the estrous cycle was normal. Treatment with 2, 4, 6, or 8 mg/kg/d edifenphos significantly decreased ovarian weight (-6.75, -9.79, -18.71, and -34.13 percent, respectively) below that of the controls. Although treatment with 2 or 4 mg/kg/d edifenphos did not change the number of healthy and atretic follicles, a dosage of 4 mg/kg/d significantly decreased the number and duration of estrous cycles. Treatment with 6 or 8 mg/kg/d, however, significantly decreased the number and duration of different phases of the estrous cycle as well. In all treated groups, the weights of the thymus and uterus were significantly reduced when compared with those of hemicastrated oil-treated control animals.

Effect of endosulfan on ovarian compensatory hypertrophy in hemicastrated albino mice.

Endosulfan, a chlorinated cyclodiene insecticide, was administered orally at 1.5, 3, 6, and 9 mg/kg per day to normal hemicastrated virgin mice for 15 consecutive days to examine the effect on ovarian function. Sham-operated and hemicastrated control mice were administered a similar volume of olive oil. The vaginal smear and body weight of the mice were recorded daily and mice were sacrificed on day 16. The remaining left ovary, uterus, kidney, adrenal, liver, thymus, and thyroid were removed and weighed. The left ovary from each animal was serially sectioned and stained for histologic studies. The hemicastrated control mice had a 40.5% increase in weight of the remaining left ovary and a significant increase in healthy and atretic follicles when compared with sham-operated controls. The remaining left ovaries of mice treated with 1.5mg endosulfan had a 37.2% increase in weight with no significant difference in ovarian weight and or in the healthy and atretic follicles when compared with hemicastrated olive oil-treated controls. However, treatment with 3, 6, and 9 mg endosulfan resulted in weight increases in the remaining left ovary of only 14.9, 7.4, and 0.8% and a significant decrease in healthy follicles with a concomitant increase in the number of atretic follicles compared to the olive oil-treated controls. There was no significant change in the number of estrous cycles or the duration of each phase of the estrous cycle with 1.5 and 3mg/kg per day endosulfan. However, there was a significant decrease in the number of estrous cycles, and the duration of estrus and metestrus with a concomitant significant increase in the duration of the diestrus phase with 6 and 9 mg/kg per day endosulfan treatment when compared with hemicastrated olive oil-treated controls. There were no significant change in body weight or the weights of the uterus, kidney, adrenal, liver, thymus, or thyroid after endosulfan treatment. These observations show that endosulfan treatment caused a significant decrease in compensatory ovarian hypertrophy. An increase in the number of atretic follicles and disruption of the estrous cycle may have been due to a direct effect on the ovary or to effects on the hypothalamo-hypophysial-ovarian axis.

The anti-implantation action of endosulfan in albino mice: possible mechanisms.

Endosulfan, a chlorinated pesticide, was administered orally for 7 consecutive days to a group of virgin pregnant albino mice at doses of 1 to 4 mg/kg/d to examine its effect on implantation. For comparison, a second group of animals received 5 microg/kg/d estradiol-17beta, a third group received similar quantities of olive oil. Autopsy on day 8 revealed that the olive oil-treated mice were pregnant with a normal number of implantations and a normal duration of diestrus. Treatment with estradiol-17beta completely inhibited implantation and significantly decreased the duration of diestrus with a concomitant increase in estrus. Treatment with 1, 2, or 3 mg/kg/d endosulfan neither inhibited implantation nor significantly changed diestrus, whereas 4 mg/kg/d endosulfan completely inhibited implantation, with the uterus showing no signs of implantation. This group exhibited a significant decrease in diestrus with a concomitant increase in estrus. In endosulfan-treated mice, no significant changes occurred in body and organ weight, except for a significant increase in uterine weight in groups treated with 4 mg/kg/d endosulfan or estradiol-17beta. Because endosulfan is neither tubal locking nor causes expulsion of the blastocyst from the uterus like estradiol-17beta, the pesticide might inhibit implantation by altering the estrogen-progesterone ratio essential for implantation.

Effect of dimethoate administration schedules on compensatory ovarian hypertrophy, follicular dynamics, and estrous cycle in hemicastrated mice.

Dimethoate, a widely used organophosphate insecticide, was administered orally (28 mg/kg body weight) to hemicastrated (HC) virgin mice on day 1 and for 5, 10, and 15 days. Hemicastrated untreated control mice showed a significant increase in relative ovarian weight, with 42.74% hypertrophy and an increase in healthy and atretic follicles when compared with those of sham-operated control animals. The HC mice treated for 1 day or for 5 days showed no significant change in ovarian weight (36.64% and 25.19% hypertrophy, respectively) or in healthy and atretic follicles, when compared with HC-control mice. Treatment with dimethoate for 10 or 15 days, however, resulted in a significant decrease in ovarian weight (19.84% and 0.76% hypertrophy, respectively), a significant decrease in the number of healthy follicles, and a concomitant significant increase in the number of atretic follicles when compared with those in HC control animals. No significant change occurred in the number of estrous cycles or duration of each phase of the estrous cycle in HC mice treated with dimethoate for either 1 or 5 days. In HC mice treated with dimethoate for 10 or 15 days, however, a significant decrease in the number of estrous cycles, duration of proestrus, estrus, and metestrus, a concomitant significant increase in the diestrus phase, when compared with HC control animals. In mice receiving dimethoate for 15 days (but not for 1, 5, or 10 days) a significant decrease occurred in body weight and in the weights of the uterus, kidney, spleen, and liver when compared with the parallel weights in HC control animals. The overall findings suggest that following dimethoate treatment, a significant decrease in ovarian weight with a concomitant increase in compensatory ovarian hypertrophy and in the number of healthy follicles, with a concomitant increase in the number of atretic follicles and interrupted estrous cycles, may be due to the direct effect on the ovary or may be due to a hormonal imbalance in any stage of the hypothalamo-hypophysial ovarian axis.

Induction of gonadal toxicity to female rats after chronic exposure to mancozeb.

Mancozeb, a fungicide of ethylenebisdithiocarbamate group was orally administered at doses of 500, 600, 700 and 800 mg/kg body weight/day to normal virgin rats of Wistar strain for 30 days. The vaginal smear and body weight of the rats were recorded daily and rats were sacrificed on 31st day. Estrous cycle was effected by showing a significant decrease in the number of estrous cycle, duration of proestrus, estrus and metestrus with concomitant significant increase in the duration of diestrus in all the mancozeb treated groups when compared with controls. There were a significant decrease in the number of healthy follicles and a significant increase in the number of atretic follicles in all the mancozeb treated groups when compared with controls. The histologic observation of the ovary revealed the presence of less number of corpora lutea and the size of the ovary was also reduced in high doses of mancozeb treated rats. There was a significant increase in the thyroid weight in all the mancozeb treated rats except in 500 mg/kg/d. In rats treated with 500 mg/kg/d showed a significant increase in the level of total lipids in the liver. In rats treated with 600 mg/kg/d mancozeb showed a significant decrease in the levels of glycogen and total lipids in the uterus and total lipids in the liver. In rats treated with 700 mg/kg/d showed a significant decrease in the levels of protein in ovary, glycogen, total lipids, phospholipids and neutral lipids in the uterus and a significant increase in the levels of phospholipids, neutral lipids in the ovary and total lipids, phospholipids and neutral lipids in the liver. In rats treated with 800 mg/kg/d showed a significant decrease in the levels of protein and glycogen in the ovary and protein, glycogen, total lipids, phospholipids and neutral lipids in the uterus and a significant increase in the levels of total lipids, phospholipids and neutral lipids in the ovary and liver when compared with controls. These observed effect of mancozeb on the estrous cycle, follicles and biochemical constituents may be due to imbalance in the hormone or toxic effect.

Effect of dexamethasone on implantation and pregnancy in albino rats.

Administration of 3 mg/kg body weight of dexamethasone from day 1 or 3 to 7 of pregnancy did not prevent implantation in albino rats. But the same dose when administered from day 8 to 11 resulted in complete abortion / resorption in all rats. Administration of 2 mg / kg body weight of dexamethasone from day 8 to 11 of pregnancy held no effect on the foetal survival. The results indicate that a high dose of dexamethasone does not affect implantation but the same dose affects the more advanced stages of pregnancy.

Possible mechanisms for the anti-implantation action of dicofol in albino rats.

Dicofol, a chlorinated pesticide showing estrogenic activity, was administered orally to virgin pregnant rats at doses of 300, 400, 500, 600, or 700 mg/kg/day for 7 consecutive days to examine its effect on blastocyst implantation. One group of rats received estradiol-17beta (10 microg) for comparison and control animals received similar quantities of olive oil. Autopsy on day 8 revealed that the olive-oil treated rats were pregnant and had a normal number of implantations and a normal duration of diestrus. Treatment with estradiol-17beta completely inhibited implantation, significantly decreased the duration of diestrus with concomitant increase in estrus. Treatment with 300, 400, or 500 mg/kg/d dicofol neither inhibited implantation nor significantly changed diestrus. Treatment with 600 mg dicofol partially inhibited implantation and significantly decreased diestrus with concomitant increase in estrus. Treatment with 700 mg dicofol completely inhibited implantation, and the uterus showed placental scars. This group exhibited a significant decrease in diestrus with concomitant increase in estrus. In all dicofol-treated rats, no significant changes were found in body weight or organ, except for a significant decrease in the weight of the uterus in groups receiving either 700 mg dicofol or estradiol-17beta. The inhibitory effect of dicofol on implantation may be due to an imbalanced estrogen-progesterone ratio, essential for implantation. The pesticide is neither tubal locking nor causes expulsion of the blastocyst from the uterus like estradiol-17beta.

Effect of mancozeb on ovarian compensatory hypertrophy and biochemical constituents in hemicastrated albino rat.

Mancozeb a carbamate fungicide was administered orally at doses of 500, 600, 700, and 800 mg/kg/day to normal hemicastrated virgin rats for 15 consecutive days to examine the effect on ovarian hypertrophy. Sham-operated and hemicastrated control rats were administered a similar quantity of olive oil. The vaginal smear and body weight of the rats were recorded daily and rats were sacrificed on the Day 16. The ovary, uterus, kidney, adrenal, spleen, liver, lungs, heart, thymus, and thyroid were removed and weighed. The left ovary from each animal was serially sectioned and stained for histologic studies. The hemicastrated control rats revealed a significant increase in relative ovarian weight with 66.3% hypertrophy. Treatment with 700 and 800 mg/kg/day mancozeb revealed a decrease in ovarian hypertrophy with 28.2 and 22.8% hypertrophy, respectively. There was no significant change in the number of estrous cycles and duration of each phase of the estrous cycle with 500 mg/kg/day mancozeb. However, there was a decrease in the number of estrous cycles, duration of proestrus, estrus, and metestrus with concomitant significant increase in the duration of the diestrus phase with 600, 700, and 800 mg/kg/day mancozeb treatment. There was a significant decrease in the number of healthy follicles with concomitant increase in the number of atretic follicles at higher doses of mancozeb. There were no significant changes in the body and organ weight with 500, 600, 700, and 800 mg/kg/day of mancozeb. The levels of protein, glycogen, total lipid, phospholipid, and neutral lipid were elevated in the liver, uterus, and ovary after hemicastration. Protein, glycogen, total lipid, phospholipid, and neutral lipid were not significantly changed in the liver, uterus, and ovary after 500 mg/kg/day mancozeb. However, treatment with 600, 700, and 800 mg/kg/day mancozeb showed a significant decrease in the levels of protein, glycogen, total lipid, phospholipid, and neutral lipid in the liver, uterus, and ovary, with the exception of liver total lipid and uterine glycogen. In addition to the decrease in the compensatory ovarian hypertrophy, mancozeb treatment reduced the number of healthy follicles with a concomitant increase in the number of atretic follicles. This finding plus disruption of the estrous cycle may be due to a direct effect on the ovary or the hypothalamo-hypophysial-ovarian axis.