The relationship between mirex-induced liver enlargement and the adrenal glands

A single oral dose of mirex given to male rats produced a doubling in liver mass (liver weight/body weight) within 3 days. The response was both dose and time related. There was an accompanying increase in adrenal gland weight (adrenal weight/body weight). In adrenalectomized animals given a single oral dose of mirex, the increase in liver weight/body weight response was drastically reduced. Adrenalectomized animals given corticosterone supplements showed a similar liver weight increase when given mirex as did intact mirex dosed animals. These results suggest a relationship between mirex-induced liver enlargement and adrenal gland secretions.     

The in vivo effect of mirex on soluble hepatic enzymes in the rat

Adult male and female rats fed dietary mirex in concentrations of 10, 20, 30, 40, and 50 ppm for 4 weeks exhibited significant decreases in liver levels of lactic dehydrogenase, malic dehydrogenase, sorbitol dehydrogenase, glutamic oxaloacetic transaminase, and glutamic pyruvic transaminase as compared to control levels. Enzyme losses were cytoplasmic and decreased in magnitude with increased time of exposure and dietary mirex concentrations. Serum sorbitol dehydrogenase levels were increased above control levels during the first week of mirex exposure while hepatic sorbitol dehydrogenase levels were concomitantly decreased below the control levels. Hepatic glutamic oxaloacetic transaminase levels were significantly decreased during the first week for all dietary mirex concentrations as were the other enzymes at the 40 and 50 ppm mirex concentrations. The magnitude of the enzyme decreases in female rat tissue was consistently lower than enzyme decreases in the male rat tissue fed equivalent dietary mirex dosages. Liver weights for male rats increased significantly at all dietary mirex levels during week one. However, only animals receiving the 40 and 50 ppm dietary mirex levels showed significant liver weight increases at the end of week 4. Female rat liver weights were increased at the 30 and 40 ppm mirex dietary levels after 4 weeks. No significant differences in body weights were observed for any dietary level of mirex. Mirex hepatic residues appeared to be equal for both sexes. No gross histological alterations were observed following treatment.     

The effect of α-hexachlorocyclohexane on liver growth in intact and adrenalectomized rats

α-Hexachlorocyclohexane was administered as a single oral dose (100 mg/kg body wt) to both intact and adrenalectomized male Sprague-Dawley rats. In the intact rats there was a peak in [³H]thymidine incorporation into DNA at 30 hr and an increase in relative liver weight (liver weight/body wt × 100) which peaked at 60 hr post α-HCH dose. However, in α-HCH-dosed adrenalectomized rats [³H]thymidine incorporation into DNA remained elevated throughout the study period and relative liver weight was not significantly increased. In corticosterone-supplemented α-HCH-dosed Adx rats, [³H]thymidine incorporation into DNA was significantly reduced and RLW was increased comparable to that seen in intact animals. These results suggest that α-HCH-induced liver growth is mediated by corticosterone.     

Mirex residues in bobwhite quail after aerial application of bait for fire ant control, South Carolina--1975-76

Mirex, the organochlorine compound used for control of the imported fire ant (Solenopsis invicta Buren), was applied aerially under supervision of the South Carolina Plant Pest Regulatory Service in October 1975 to a game management area in Hampton County, S.C. Influenced by recent reports indicating that low levels of mirex were toxic to certain nontarget organisms in laboratory studies, authors initiated a program for monitoring mirex residues in bobwhite quail (Colinus virginianus). Pretreatment residues were recorded on a dry-weight basis in bobwhite quail breast and adipose tissue; conversion factors for determining wet-weight concentrations are approximately as follows: fat, 0.77; and breast, 0.29. Residues ranged from 0.000-0.178 ppm and 0.247-2.763 ppm, respectively. Mirex residues in quail adipose tissue showed up to five-fold increase within the first month after treatment and declined thereafter. A residue peak was noticed the spring following mirex treatment, corresponding with insect emergence. Mirex residues in quail collected in summer 1976 following a fall bait application showed slightly higher residue levels than had birds taken in summer 1975; however, little, if any, human food chain contamination would result in the consumption of birds with residue levels found in this study.     

Mirex residues in wildlife and soils, Hawaiian pineapple-growing areas--1972-74.

A monitoring program was conducted in the pineapple-growing areas of Hawaii from 1972 to 1974 to survey mirex residues in sediments, soils, and aquatic and terrestrial wildlife. Residues in pineapple field soils ranged from 3 to 18 ppb 9 months after mirex had been applied. No residues were found in the sediments. Only 8 fish of 110 aquatic animals sampled contained mirex; these levels were low and ranged from 3 to 7 ppb. Mirex residues in birds ranged from undetectable to 10 ppm; residues in rodents were quite variable, but in terms of the geometric mean, the amounts in the Polynesian rat decreased with time from 1,270 to 56 ppb. Similarly, values for the roof rat ranged from 666 to 17 ppb. The geometric mean for residues in mongooses decreased from 2,200 ppb immediately after application to 238 ppb 39 weeks later. Aerial application of mirex to the pineapple fields did not contaminate the marine environment of Hawaii and no evidence of mirex residue buildup in the aquatic food chain was apparent. Mirex accumulation in terrestrial biota was temporary; there was no definitive indication of permanent accumulation in the wildlife of the areas studied.     

Mirex residues in nontarget organisms after application of experimental baits for fire ant control, southwest Georgia--1971-72.

Mirex, the only compound approved for control of the red imported fire ant (Solenopsis invicta) and the black imported fire ant (Solenopsis richteri), is normally applied at a rate of 1.40 kg/ha. (1.25 lb/acre). Influenced by recent studies showing that low levels of mirex are toxic to certain nontarget organisms, particularly estuarine species, authors report here on a monitoring study of mirex in three large treatment areas of southwest Georgia. Four formulations of bait were applied aerially in 1971-72. Low-level residues were observed in small terrestrial vertebrates and invertebrates and in fresh-water inhabitants. Levels detected were about the same for all baits. Maximum residues were detected 1-3 months after treatment and gradually declined to low levels of 0.02-1.16 ppm 1 year after treatment.     

Modulation of Mouse P450 Isoforms CYP1A2, CYP2B10, CYP2E1, and CYP3A by the Environmental Chemicals Mirex, 2,2-Bis(p-chlorophenyl)-1,1-dichloroethylene, Vinclozolin, and Flutamide

Several environmental chemicals are disruptive to the reproductive and endocrine systems of many species, including humans. Mechanisms for endocrine disruption are presently under scrutiny. Xenobiotic inducible mammalian cytochrome P450 (CYP) enzymes metabolize a variety of substrates including environmental chemicals, pesticides, and drugs. The metabolism, and thus the effect, of endogenous chemicals including steroid hormones, vitamins, etc. that are transformed by CYP enzymes can be influenced by environmental exposure to CYP-inducing chemicals. This study demonstrated that structurally diverse environmental chemicals including mirex, 2,2-Bis(p-chlorophenyl)-1,1-dichloroethylene (DDE), vinclozolin, and flutamide are capable of inducing several mouse liver CYP isozymes. As demonstrated by Western blotting, mirex induced CYP1A2, 2B10, 2E1, and 3A and vinclozolin induced 1A2 and 2B10. The only isoforms significantly induced by DDE and flutamide were 3A and 1A2, respectively. Since some of these isoforms are known to be involved in metabolism of endogenous hormones, we also studied the effects of these CYP inducers on testosterone metabolism and seminal vesicle weights. Mirex and DDE treatments had profound effects on the metabolism of testosterone, resulting in 2.5- to 3-fold more hydroxylated products than controls. Lesser, but significant, increases in specific metabolites of testosterone were also observed following treatment with vinclozolin and flutamide. Seminal vesicle weights were lower for all treatment groups except DDE. Results of this study demonstrate that, due to their CYP-inducing potential, these chemicals may significantly impact testosterone metabolism and this may be a contributing factor in their antiandrogenic effects.     

A study of liver function in rats with mirex-induced enlarged livers

The functional capacity of a mirex-induced, enlarged liver was studied in rats. The tests used were sulfobromophthalein clearance, hepatic cytochrome P-450 concentration, serum total protein concentration and electrophoretic pattern, serum total lipid concentration, serum glucose concentration, and the liver response to epinephrine. There was no indication of a loss of functional capacity in the enlarged liver. Sulfobromophthalein clearance and microsomal cytochrome P-450 concentration indicated an increase in total liver functional capacity. We conclude that mirex is not a direct hepatotoxin producing generalized parenchymal cell damage.     

Fate of endosulfan in rats and toxicological considerations of apolar metabolites

[¹⁴C]Endosulfan, α or β isomers separately, was administered to rats as a single oral dose and as a dietary supplement for 14 days. No appreciable differences were observed in the fate of the two isomers. Five days after the single dose, 75% of the dose had been voided in the feces and 13% in the urine. Of the total radiocarbon consumed in the diet after 14 days, 56% had been eliminated in the feces and 8% in the urine. Bile collection studies showed that up to 47% of a single oral dose was eliminated from the liver via this route; enterohepatic circulation was not apparent. Maximum [¹⁴C]endosulfan equivalents in body tissue occurred in the kidney and liver, 3 and 1 ppm, respectively, after 14 days of feeding 5 ppm of endosulfan. Apolar metabolites in the excreta and/or tissues were a minor portion of the total residues and consisted of the sulfate, diol, α-hydroxy ether, lactone, and ether derivatives of endosulfan. The sulfate was slightly more toxic to mice than endosulfan, while the other products were less toxic. Neither endosulfan nor its metabolites were active in the Salmonella mutagenicity test. Endosulfan in the diet of rats for 28 days at 50 ppm did not induce liver oxidase enzymes, alter liver or kidney weights, or influence the rate of weight gain of the animals.     

Comparative tissue distribution of mirex and chlordecone in fetal and neonatal rats

The transport of mirex and chlordecone (Kepone) across the placenta during late gestation and through the milk during lactation was investigated in the rat. In the placental transport study, doses of 5 mg/kg were administered on Day 15, 18, or 20 of gestation and animals were killed 4, 24, or 48 hr after treatment. Both compounds crossed the placenta and were present in the fetus at all examination times. Maternal tissue levels exceeded fetal tissue levels by a factor of 4 to 5. Slightly higher levels of chlordecone as compared to mirex were found in maternal and fetal tissues. No effects of gestational age at time of treatment or of position of the fetus in the uterus were seen. In the lactation study, doses of 1 or 10 mg/kg/day were administered on Days 2–5 postpartum and pups were killed at intervals up to 12 days after treatment. The secretion of milk appeared to be a major route of elimination for both pesticides for nursing females, and the greater amount of mirex excreted via the milk as compared with chlordecone is in agreement with differences in their reported octanol-water partition coefficients. Initially, mirex entered the milk more rapidly than chlordecone. After cessation of treatment, mirex milk levels fell quickly, but chlordecone levels remained fairly constant. In the pups, mirex tissue levels paralleled milk levels; chlordecone levels, however, continued to increase in the tissues throughout the observation period.     

Biochemical and physiological investigations into the delayed toxicity produced by O,O,S-trimethyl phosphorothioate in rats

Oral administration of O,O,S-trimethyl phosphorothioate (OOS), an impurity in several technical organophosphorus insecticides, causes delayed toxicity in rats with death occurring up to 28 days after the treatment. The oral LD₅₀ was determined to be 60 mg/kg. The effect of a single nonlethal dose of OOS (20 mg/kg) on in vivo protein synthesis in different organs was determined by measurement of the incorporation of [¹⁴C]leucine at 6 hr to 28 days after treatment. As early as 6 hr after OOS treatment the incorporation of [¹⁴C]leucine into the liver, lung, thymus, kidney, and spleen was elevated and remained elevated for up to 7 days. With the exception of the lung, organ weights were significantly decreased during the same time period. On Day 28 after treatment, the amount of [¹⁴C]leucine incorporation had decreased to the control level in all of the organs studied. Treatment with OOS at 20 mg/kg caused a significant increase in hematocrit on Days 3,5, and 7, and as early as 6 hr after treatment at 60 mg/kg. The clinical biochemistry of plasma indicated that there was no significant change from control values in the glutamic pyruvic transaminase, glutamic oxalic transaminase, lactate dehydrogenase, or alkaline phosphatase activities with the 20 mg/kg dose. The analysis of the intermediary metabolites indicated that the redox state of cytosol was more reduced on Day 5, whereas that of mitochondria was not affected by OOS. Data obtained at selected times after oral administration of a 60 mg/kg dose of OOS and that obtained from animals starved for 3 days are also discussed.     

Residues of zeta‐cypermethrin in bovine tissues and milk following pour‐on and spray application

The depletion of zeta‐cypermethrin residues in bovine tissues and milk was studied. Beef cattle were treated three times at 3‐week intervals with 1 ml 10 kg^?1 body weight of a 25 g litre^?1 or 50 g litre^?1 pour‐on formulation (2.5 and 5.0 mg zeta‐cypermethrin kg^?1 body weight) or 100 mg kg^?1 spray to simulate a likely worst‐case treatment regime. Friesian and Jersey dairy cows were treated once with 2.5 mg zeta‐cypermethrin kg^?1 in a pour‐on formulation. Muscle, liver and kidney residue concentrations were generally less than the limit of detection (LOD = 0.01 mg kg^?1). Residues in renal‐fat and back‐fat samples from animals treated with 2.5 mg kg^?1 all exceeded the limit of quantitation (LOQ = 0.05 mg kg^?1), peaking at 10 days after treatment. Only two of five kidney fat samples were above the LOQ after 34 days, but none of the back‐fat samples exceeded the LOQ at 28 days after treatment. Following spray treatments, fat residues were detectable in some animals but were below the LOQ at all sampling intervals. Zeta‐cypermethrin was quantifiable (LOQ = 0.01 mg kg^?1) in only one whole‐milk sample from the Friesian cows (0.015 mg kg^?1, 2 days after treatment). In whole milk from Jersey cows, the mean concentration of zeta‐cypermethrin peaked 1 day after treatment, at 0.015 mg kg^?1, and the highest individual sample concentration was 0.025 mg kg^?1 at 3 days after treatment. Residues in milk were not quantifiable beginning 4 days after treatment. The mean concentrations of zeta‐cypermethrin in milk fat from Friesian and Jersey cows peaked two days after treatment at 0.197 mg kg^?1 and 0.377 mg kg^?1, respectively, and the highest individual sample concentrations were 2 days after treatment at 0.47 mg kg^?1 and 0.98 mg kg^?1, respectively. © 2001 Society of Chemical Industry     

Oxidative damage, biochemical and histopathological alterations in rats exposed to chlorpyrifos and the antioxidant role of zinc

The protective effects of zinc on liver and kidney injury induced by chlorpyrifos (CPF) were investigated in rats. Male and female rats were orally administered CPF at a dose of 6.75 mg kg⁻¹ body weight for 28 consecutive days. An additional CPF group received zinc (227 mg l⁻¹) in drinking water throughout the experimental duration. Two groups more served as controls. Administration of CPF resulted in a significant increase in serum lipid peroxidation (LPO) level, while induced significant decreases in the activities of plasma superoxide dismutase (SOD), glutathione-S-transferase (GST) and serum acetylcholinesterase (AChE) either in male or female rats. Similarly, a significant increase in the levels of various serum marker enzymes [e.g. aminotransferases (AST and ALT), lactate dehydrogenase (LDH) and gamma glutamyl transferase (GGT)] and increase the level of total protein, uric acid and creatinine. In contrast, co-administration of zinc to CPF-treated animals restored most of these biochemical parameters to within normal levels. In case of AChE, supplementation of zinc showed little alteration in the activity of this enzyme especially in male rats treated with CPF. CPF caused histopathological change in liver and kidneys of male and female rats. However, zinc administration to CPF-treated animals resulted in overall improvement in liver and kidneys damage, emphasizing its antioxidant role. In light of the available data, it can deduce that CPF-induced lipid peroxidation, oxidative stress, liver and kidneys damage in male and female rats, and conjunction supplementation of zinc has resulted in pronounced ameliorating effect.     

The dietary toxicity of flocoumafen to hens: Elimination and accumulation following repeated oral administration

In a dietary toxicity study, laying hens received a diet containing the rodenticide flocoumafen at concentrations of 1.5, 5, 10 and 50 mg kg^?1 for five consecutive days. The LC₅₀ at termination following a 28-day observation period was 16.4 mg kg^?1. Livers of birds which received doses of flocoumafen between 5 and 50 mg kg^?1 had concentrations of flocoumafen (1.5 nmol g^?1) that were independent of dose. The data indicate the presence in hen liver of a saturable high-affinity flocoumafen binding site with similar characteristics and capacity to that of the quail and rat. Residues of flocoumafen in samples of breast and leg muscle were low in all exposure groups. Higher, dose-related residues were found in samples of abdominal fat and skin-associated fat and there was a clear demonstration of the transfer of dose-related residues into eggs. In a separate study in which hens were dosed with [¹⁴C]flocoumafen for five consecutive days at a daily rate of 1 and 4 mg kg^?1 body weight, the majority (68 %) of the daily radioactive dose was eliminated over the following 24 hours via excreta. Residues in liver at death or when killed accounted for < 1 % of the cumulative administered radioactivity. Residues in eggs were located primarily in the yolk with maximum concentrations 1.0 mg kg^?1 or 0.18% of the low dose; 2.1 mg kg^?1 or 0.06% of the high dose as [¹⁴C]flocoumafen equivalents were observed at 10 days after start of dosing. Some 40 % of the total activity in the yolk was unchanged flocoumafen.     

Effects of dimethoate (O,O-dimethyl S-methyl carbamoyl methyl phosphorodithioate) and Ethanol in antioxidant status of liver and kidney of experimental mice

Organophosphorus insecticides and ethanol individually cause free radical production induced by oxidative stress and alter the antioxidants and scavengers of free radicals. The present study indicates the effect caused by dimethoate in combination with ethanol on antioxidant status in mice. Daily, dimethoate at a dose of 18 mg/kg body weight and ethanol at 1 g/kg body weight were orally administered concurrently in a subacute study for 14 days. After the experimental period, the liver and kidney homogenates were analysed for various antioxidant enzymes. The results compared with dimethoate alone treated control indicated an increase in hepatic cytochrome P450 and lipid peroxidation. Decrease in superoxide dismutase, glutathione peroxidase, glutathione reductase, glutathione-S-transferase, and glutathione in liver was observed. In kidney, decrease in CAT, SOD, GR, GST, and GSH was observed. Acetyl cholinesterase activity of RBC was increased. No significant change was observed in catalase in liver and glutathione peroxidase in kidney. The results of the study allow us to hypothesize that dimethoate along with ethanol disturbs the antioxidant status.     

Influences of sub-acute exposure to paraquat on cytochrome P450 3A2 expression in rat liver and lungs

This study was designed to investigate the possible effects of paraquat sub-acute poisoning on cytochrome P450 3A2 expression in the liver and lungs. Twenty adult male rats (150-200 g) were exposed either against saline normal as control group or various doses of paraquat (3.5, 7 and 10 mg/kg, s.c.) as test groups for 7 consecutive days. Paraquat-exposed animals showed loss of body weight and elevation in serum levels of alkaline phosphates and alanine aminotransferase in a dose-dependent fashion. Moreover, animals in the test groups demonstrated a significant (P < 0.05) increase of malondialdehyde content in both the liver and lung tissues. Ultimately, by using RT-PCR method it became clear that 7 days exposure to paraquat resulted in a total suppression of cytochrome P450 3A2 at the mRNA level in the lungs. By contrast, a considerable up regulation of the same gene occurred in the liver. This data suggest that paraquat not only affect the lungs as a main target tissue but also up regulates the predominant cytochrome P450 gene in the liver which may induce detoxification processes.     

Sequential changes in lactate,isocitrate, and malate dehydrogenases in mice exposed to technical grade hexachlorocyclohexane (BHC) and their possible relationship to liver tumors

Hexachlorocyclohexane (BHC) was fed at a 500-ppm-dose level in diet to pure inbred Swiss mice for 2, 4, 6, and 8 months. Later BHC was discontinued for 4 months and subsequently the animals were refed BHC for 1 month. The protein, lactate dehydrogenase (E.C.1.1.1.27), isocitrate dehydrogenase (E.C.1.1.1.42), and malate dehydrogenase (E.C.1.1.1.37) were studied from liver. The liver weight as percentage of body weight was also determined. The results showed an increase in the liver weight/body weight ratio and a decrease in protein and all three enzymes after BHC feeding for the different time intervals. The discontinuation of BHC in the diet resulted in the reversion of values of the above parameters toward the normal and after refeeding BHC for 1 month, the decreased values, as seen after initial BHC feeding, were again observed. This indicates that the changes in the values of lactate dehydrogenase, isocitrate dehydrogenase, and malate dehydrogenase are associated with hexachlorocyclohexane feeding in the diet. The significance of these changes is discussed.     

Hormonal responses and tolerance to cold of female quail following parathion ingestion

Thirty-week-old female bobwhite quail (Colinus virgininus), maintained at 26 ± 1°C, were provided diets containing 0,25, or 100 ppm parathion ad libitum. After 10 days, birds were exposed to mild cold (6 ± 1°C) for 4, 8, 12, 24, or 48 hr. Brain acetylcholinesterase activity was inhibited in a dose-dependent manner in birds receiving 25 and 100 ppm parathion. Body weight, egg production, and plasma luteinizing hormone and progesterone concentrations were reduced in birds receiving 100 ppm parathion compared with other groups. Cold exposure did not alter plasma corticosterone levels in the 0- and 25-ppm parathion groups, but a two- to fivefold elevation of plasma corticosterone was observed in birds fed 100 ppm parathion. These findings indicate that (i) short-term ingestion of parathion can impair reproduction possibly by altering gonadotropin or steroid secretion, and (ii) tolerance to cold may be reduced following ingestion of this organophosphate.     

Dose-dependent sub-chronic toxicity of diethyl phthalate in female Swiss mice

The experiment was conducted to study the after effects of administering DEP at different doses to female Swiss mice for a period of 90 days. Group I mice were fed on normal diet and water ad libitum. Group II mice were maintained on normal diet mixed with corn oil at 8.25 mg/kg of the diet/day as oil control. Group III, IV and V mice were given diethyl phthalate dissolved in corn oil mixed with the diet at 10, 25 and 50 mg/kg of the diet/day, which is approximately equal to 1.25, 3.125 and 6.25 mg/kg body weight/day. A significant dose dependent increase was observed in serum acid phosphatase (ACP) whereas, serum and liver triglycerides levels showed a significant increase only in the high-dose treated group. Significant dose-dependent increase in serum aspartate and alanine aminotransferase (AST and ALT) and liver glycogen was observed. Serum lactate dehydrogenase (LDH) was significantly increased only in 25 and 50 ppm DEP-treated mice. Liver cholesterol was significantly increased in all the treated groups. Liver histology by light microscopy showed intracellular vacuolations in all the treated groups which was much more evident in the 25 and 50 ppm DEP-treated mice while hepatocellular degeneration and hypertrophy of the hepatocytes was evident in 50 ppm DEP-treated mice. Proliferation of mitochondria and peroxisomes was evident in the electron micrographs of the 10 ppm DEP-treated mice while 25 and 50 ppm DEP-treated mice showed increase in lipid droplets and severe mitochondrial proliferation.     

The potential use of corticosteroid hormones in rodent biocontrol

The feasibility of immunosuppression by corticosteroid hormones for the biocontrol of vertebrate pests was studied using water-soluble derivatives of prednisolone and dexamethasone administered by intubation to the levant vole (Microtus guentheri). Effect of treatment was determined through changes in body weight, histological changes in the spleen and bone marrow, and total and differential counts of white blood cells. Dexamethasone had a more drastic effect than prednisolone. In a group treated daily with 100 mg dexamethasone/kg body weight, signs of illness appeared in about 40% of the voles and death occurred in 18% of them 6–7 days after the start of the treatment. There was a loss of up to 22% in body weight, marked atrophy of the germinal center, corona and perifollicular zone of the splenic lymph nodules, but no effect on the bone marrow constitution. A similar treatment with prednisolone produced no external signs of illness or change in body weight, yet 17% of the voles died 8–10 days after the start of the treatment, and partial atrophy in the splenic lymph nodules occurred in 50% of the animals. In a group treated with 100 mg dexamethasone/kg once every 3 days, from which blood samples were taken repeatedly from the same eye, mortality was 100%, with death occurring 3–12 days after start of the treatment. Differential white blood cell counts indicated depletion of lymphocytes and monocytes, and increase in the number of granulocytes in the peripheral blood. Results of preference tests showed that prebaited voles do not reject wheat grain baits treated with a dexamethasone solution in water.