Ginger extract ameliorates phosphamidon induced hepatotoxicity.

Organophosphorus (OP) compounds commonly used as pesticides in agriculture cause serious health problems to living beings. The present study enumerates the ameliorating effect of ginger extract (GE) against phosphamidon (PHO, an organophosphorus insecticide) induced hepatotoxicity. GE was prepared from dried ginger and characterized for compound profile and antioxidant activity. Eight groups of albino rats (n = 6) were treated with 1/5th lethal dose of PHO for 5-20 days. Out of the treated 8 groups, 4 were simultaneously fed with GE (1 mg/kg body wt.) along with PHO. Alterations in the levels of hepatocellular oxidative stress (OS) markers in the treated groups indicated an enhanced generation of reactive oxygen species (ROS) and oxidative stress (OS). Upregulation of apoptotic markers, DNA fragmentation and appearance of apoptotic nuclei suggested induction of apoptosis in the liver cell that was found to be attenuated after GE treatment. Moreover, no toxicity and mortality was observed up to 100 mg/kg dose of GE for 30 days in the rat model studied. Thus, GE can be considered as an effective, economical and safe extract to circumvent PHO-induced hepatotoxicity.

Neurotoxic impact of organophosphate pesticide phosphomedon on the albino rat.

Organophosphate pesticide phosphomedon was exposed to albino rat at a concentration of 35 ppm in the time interval of 30, 45 and 60 days. During the exposure period neurobehavioral symptoms such as reduce food intake, weight loss, increase water intake, low defecation frequency, increase locomotion frequency at high dose were observed. Locomotion frequency were less initially but higher in increasing dose concentration. The result was also different in both the sexes. A decrease social interaction and increase force swimming with increasing dose concentration, which was not significant as comparison to control. A significant histopathological changes observed in three dose concentrations. In 30 days phosphomedon exposure the nuclear shape changes to oval, pear shaped along with fibrosis, lipidosis, 45 days exposure showed the increase number of nucleus, Chromatolysis, inflammatory nucleus, pyknosis. In 60 days test group histopathological picture showed the swelling of cell body, lipidosis, demylination, necrosis in brain cells. Over all result indicated that due to impact of O. P pesticide phosphomedon a severe histopathological changes occurs which was distinctly observed in neurobehavioural changes.

Effect of piracetam and vitamin E on phosphamidon-induced impairment of memory and oxidative stress in rats.

Organophosphate pesticides, such as phosphamidon (PHOS), have been shown to adversely affect memory and induce oxidative stress after both acute and chronic exposure. The present study was therefore designed to investigate the effects of piracetam (PIR) and vitamin E on PHOS-induced modulation of cognitive function and oxidative stress in rats. Cognitive function was assessed using step-down latency (SDL) on a passive avoidance apparatus and transfer latency (TL) on an elevated plus maze. Oxidative stress was assessed by examining the levels of malondialdehyde (MDA) and nonprotein thiols (NP-SH) in isolated homogenized whole brain samples. The results showed a significant reduction in SDL and a prolongation of TL in the PHOS (1.74 mg/kg/day per oral; p.o.)-treated group at weeks 6 and 8, as compared to the control group. Administration of PIR (600 mg/kg/day p.o.) or vitamin E (125 mg/kg/day p.o.) for 2 weeks antagonized the effect of PHOS on SDL as well as TL. PHOS per se produced a significant increase in brain MDA levels and a decrease in brain NP-SH levels, whereas administration of PIR (600 mg/kg/day p.o.) or vitamin E (125 mg/kg/day p.o.) attenuated these effects. Thus, the results of the study showed that both PIR and vitamin E attenuated the cognitive dysfunction and oxidative stress induced by PHOS in the rat brain.

Ameliorating effect of N-acetylcysteine and curcumin on pesticide-induced oxidative DNA damage in human peripheral blood mononuclear cells.

Endosulfan, malathion, and phosphamidon are widely used pesticides. Subchronic exposure to these contaminants commonly affects the central nervous system, immune, gastrointestinal, renal, and reproductive system. There effects have been attributed to increased oxidative stress. This study was conducted to examine the role of oxidative stress in genotoxicity following pesticide exposure using peripheral blood mononuclear cells (PBMC) in vitro. Further possible attenuation of genotoxicity was studied using N-acetylcysteine (NAC) and curcumin as known modulators of oxidative stress. Cultured mononuclear cells was isolated from peripheral blood of healthy volunteers, and exposed to varying concentrations of different pesticides: endosulfan, malathion, and phosphamidon for 6, 12, and 24 h. Lipid peroxidation was assessed by cellular malondialdehyde (MDA) level and DNA damage was quantified by measuring 8-hydroxy-2'-deoxyguanosine (8-OH-dG) using ELISA. Both MDA and 8-OH-dG were significantly increased in a dose-dependent manner following treatment with these pesticides. There was a significant decrease in MDA and 8-OH-dG levels in PBMC when co-treated with NAC or/and curcumin as compared to pesticide alone. These results indicate that pesticide-induced oxidative stress is probably responsible for the DNA damage, and NAC or curcumin attenuate this effect by counteracting the oxidative stress.

Assessment of phosphamidon-induced apoptosis in human peripheral blood mononuclear cells: protective effects of N-acetylcysteine and curcumin.

The molecular mechanism for noncholinergic toxicity of phosphamidon, an extensively used organophosphate pesticide, is still not clear. The aim of the present study is to find the possible molecular mechanism of this pesticide to induce apoptosis and the role of different drugs for attenuation of such effects. Human peripheral blood mononuclear cells (PBMC) were incubated with increasing concentrations of phosphamidon (0-20 µM) for 6-24 h. The MTT assay reveals that phosphamidon induces cytotoxicity in a dose-dependent manner. Cellular glutathione (GSH) is depleted in a dose-dependent manner from 55% to 70% at concentrations between 10 and 20 µM. The percentage of cells that bind to Annexin-V, which is a representative of cells either undergoing apoptosis or necrosis during 24 h incubation, increases in a dose-dependent manner. Above 5 µM, significant necrosis of cells was observed. DNA fragmentation assay revealed that at low concentration of phosphamidon (1 µM), no appreciable change in DNA fragmentation was seen; however, distinct fragmentation was observed beyond 2.5 µM. Phosphamidon was found to cause significant depletion of GSH, which correlates well with the percentage of cells undergoing apoptosis. An increasing trend in levels of cytochrome c was observed with increasing concentration of phosphamidon, indicating that the apoptotic effect of phosphamidon is mediated through cytochrome c release. Coadministration of the antioxidants N-acetylcysteine and curcumin attenuated phosphamidon-induced apoptosis. This further supports our hypothesis that oxidative stress, as indicated by GSH depletion, results in the induction of apoptosis by release of cytochrome c.

Determination of pesticides and toxic potency of rainwater samples in western Greece.

Rainwater samples from four municipalities located in Achaia Prefecture, Greece, were collected from March to September 2006. The toxic potency of pollutants present in 36 rainwater samples was tested using Daphnia pulex. The pesticide determination was conducted with GC-MS. Only phosphamidon was detected, which appeared in 52% and 13% of the rural and urban areas, respectively. The toxicity of rainwater was determined in 52% and 46.7% of the rural and urban area samples, respectively. Chemical analyses showed that in rural areas, the PO(4)(3-) ions had higher concentrations than in urban areas. On the other hand, the SO(4)(2-), NO(-)(3), and NO(-)(2) anions are more highly concentrated in urban areas. Correlation analysis proved that the toxicity of the rainwater samples is moderate, affected by the presence of the insecticide only in the rural areas. The results indicated that toxicity can be directly assessed via bioassays, even when unknown pollutants are present.

Immunotoxicity of phosphamidon following subchronic exposure in albino rats.

Effect of subchronic doses of phosphamidon exposure on humoral and cell mediated immune (CMI) responses were studied in male albino rats using SRBC, ovalbumin and KLH as antigens. Humoral immune responses were assessed by estimating antibody titre against antigen and splenic plaque forming cells (PFC) assay. CMI responses were studied by using leucocyte migration inhibition (LMI), macrophage migration inhibition (MMI) and delayed type hypersensitivity (DTH) response. Results obtained in the present study revealed marked suppression of humoral and CMI responses in a dose dependent pattern. Hence, suppression of immune responses by phosphamidon even at subchronic doses is clearly an important aspect for its safety evaluation.

Endocrine response of the freshwater teleost, Sarotherodon mossambicus (Peters) to dimecron exposure.

The endocrine response in a freshwater teleost, Sarotherodon mossambicus (Peters) under dimecron (an organophosphate pesticide) toxicity was investigated by estimating the serum levels of T3 (triiodothyronine), T4 (thyroxine), cortisol, prolactin and insulin in control and sub-lethal (0.001 ml l(-1)) dimecron-exposed fish for 1, 6, 12, 24h and 5 days. In control S. mossambicus, the serum levels of T3 ranged from 0.80+/-0.01 to 0.82+/-0.01 ng ml(-1); T4 from 2.20+/-0.01 to 2.25+/-0.01 microg dl(-1); cortisol from 8.30+/-0.03 to 8.34+/-0.01 microg dl(-1); prolactin from 1.50+/-0.01 to 1.54+/-0.01 microg ml(-1); insulin from 9.70+/-0.01 to 9.76+/-0.01 microU ml(-1) up to a maximum period of 5 days maintained in pollutant-free tap water. Exposure of fish to sub-lethal concentration of dimecron caused varying changes in the levels of serum hormones studied. Based on the results obtained, it was concluded that (i) the fish adaptively maintains a probable low metabolic rate, as indicated by the reduced levels of thyroid hormone (T3) as well as the glucocorticoid hormone (cortisol), which could be considered advantageous for the fish to indirectly reduce the toxic impact of the pesticide, (ii) the elevated levels of prolactin in the fish under pesticide stress is indicative of a possible hydromineral regulatory effect of the hormone (probably by influencing specific organs such as gills and kidney) under pesticide toxicity, (iii) the increased insulin level in the fish under pesticide stress is indicative of its role in favouring an adaptive tissue glycogenesis besides a possible increased lipogenesis to sequester the pesticide residue thereby reducing the toxic effect of the pesticide and (iv) the prolonged exposure of the fish (for 5 days) to sub-lethal dimecron appeared to exhibit a uniform recovery response in the different hormonal levels of the fish.

Genotoxic effects of some systemic pesticides: in vivo chromosomal aberrations in bone marrow cells in rats.

The genotoxic effects of five pesticides (benomyl, 2,4-D, dimecron, monocrotophos, and vitavax) were evaluated in the rat bone marrow cytogenetic assay. The spectrum of aberrations observed included chromatid breaks, chromatid fragments, ring chromosomes, dicentric chromosomes, and chromosome fragments. It was observed that 2,4-D, dimecron, and vitavax were clastogenic, but the results obtained with benomyl and monocrotophos were equivocal.

Evaluation of the teratogenic potential of phosphamidon in mice by gavage.

Phosphamidon, an organophosphate pesticide, is an established cholinesterase inhibitor. Alteration of tissue and plasma cholinesterase activity at a critical developmental period may influence cellular division and growth sufficiently to produce anatomically or functionally abnormal tissue or organ. The present study was, therefore, undertaken to evaluate the teratogenic potential of phosphamidon in pregnant Swiss albino mice, when administered at different gestational days during the period of organogenesis. The animals were sacrificed on day 18 of gestation for routine teratological examinations. It was observed that phosphamidon was more embryotoxic than teratogenic. Maximum effects were observed when administered on day 7 and day 13. Treatment on day 10 produced little effects. Repeated exposure during the organogenetic phase also produced significant adverse effects. This possibly indicates that phosphamidon is more embryotoxic during the post-implantation period (day 7) and during late organogenesis (day 13) as compared to the early organogenesis period (day 10).

Induction of sister chromatid exchanges and chromosome aberrations in vivo in Etroplus suratensis (Bloch) following exposure to organophosphorus pesticides.

The genotoxic potential of methyl parathion and phosphamidon, two commercial formulations of organophosphorus pesticides, was evaluated through induction of sister chromatid exchanges (SCE) and chromosome aberrations in fish gill tissues. Fishes exposed to the medium containing 0.05, 0.1 and 0.2 ppm of methyl parathion or 0.5, 1.0, and 2.0 ppm of phosphamidon for a duration of 96 h revealed significant increase in the number of SCE and chromosome aberrations against control values. This demonstrates the feasibility of the fish in vivo system in detecting genotoxic potential of pollutants.

Genotoxic potential of an organophosphate insecticide, phosphamidon (dimecron): an in vivo study in mice.

The genotoxic effect of phosphamidon in mice in an in vivo test system was investigated by 3 different assays: chromosomal aberration, micronucleus and sperm-shape abnormality. The chemical was administered to groups of mice via 3 routes (i.p., p.o. and s.c.), acutely in 3 dose regimens (5, 4 and 3 mg/kg) and subacutely or chronically (5 X 1 mg/kg). The animals were sacrificed at different times: after 6, 24, 48 and 120 h for chromosomal aberration, 30 h for micronucleus and 35 days for sperm-shape abnormality. Significant effects were observed in all assays. Chronic exposure to fractionated doses induced less effect than the equivalent acute dose. The results were dose- as well as time-responsive and indicated the genetic peril of phosphamidon in the present, in vivo system.

Induction of micronuclei in bone marrow by two pesticides and their differentiation with CREST staining: an in vivo study in mice.

Two pesticides, organophosphate phosphamidon (PHO) and organochlorine dieldrin (DED) were assayed by the mouse bone marrow micronucleus test, to ascertain whether they showed genotoxic activity in vivo. Two doses, sub-lethal (PHO=3 mg/kg b.wt.; DED=60 mg/kg b.wt.) and lethal (PHO=5 mg/kg b.wt.; DED=90 mg/kg b.wt.), of each substance were administered intraperitoneally to 9-10-week old CBA male mice, in acute and repeated exposure. The sub-lethal dose was also administered at two different times and twice at 24-h intervals. Both PHO and DED proved able to induce a dose-dependent increase of micronucleated polychromatic erythrocytes (PCE). The two pesticides also showed a different detoxification time. Furthermore, the CREST staining with antikinetochore antibodies allowed us to conclude that the two chemicals are clastogens.

Mutagenicity of dimecron studied in 3 mosaic assays and the sex-linked recessive lethal test in Drosophila melanogaster.

The genotoxicity of dimecron, a systemic organophosphate pesticide, has been tested in the wing, eye and germ line mosaic assays and the sex-linked recessive lethal test in Drosophila melanogaster. Larvae heterozygous for recessive marker mutations were fed the compound for various periods of time. On emergence, the wings and eyes of the adults were screened for mosaic spots and the eggs laid by the females were checked for induction of female germ line mosaicism. Dimecron is mutagenic to the somatic and germ line cells of Drosophila and induces a high frequency of sex-linked recessive lethals.

Ameliorating effects of thyroxine and atropine in phosphamidon intoxicated chick embryos.

The effects of thyroxine and atropine in ameliorating phosphamidon intoxication in chick embryos was studied. Treatment of phosphamidon significantly enhanced the mortality and abnormality rates, decreased the average body weights, and cholinesterase activity in chick embryos. When thyroxine was administered to the phosphamidon intoxicated embryos, the above parameters changed significantly, indicating an ameliorating effect of thyroxine against phosphamidon intoxication in chick embryos. The combined thyroxine and atropine therapy did not further improve the ameliorating effect. Since in many respects chick embryo development parallels that of mammalian embryos, a short-term use of thyroxine as a protective agent against organophosphate toxicity might be useful.

Individual and combined toxicity of common pesticides to teleost Puntius conchonius Hamilton.

Individual and combined toxicity of three pesticides, endosulfan, phosphamidon, and aldicarb was evaluated in P. conchonius. The 48 hr LC50 was 21.36 and 446.5 ppm respectively for endosulfan and phosphamidon. When tested jointly, 48 hr LC50 for different ratios of these pesticides were 0.332 (IE:3P), 0.224 (IE:1P), and 0.178 ppm (3E:1P). The cotoxicity coefficients for these combinations were 1793, 3986, and 10009, respectively. An equitoxic mixture of endosulfan, phosphamidon, and aldicarb yielded a 48 hr LC50 of 130.5 ppm. An enhanced toxic impact is indicated when the pesticides are present together rather than as individual compounds.

Acetylhomocysteine thiolactone protection against phosphamidon-induced alteration of regional superoxide dismutase activity in the central nervous system and its correlation with altered lipid peroxidation.

Phosphamidon intoxication (2 mg/kg body wt./day for 7 days) inhibited SOD activity, but enhanced the lipid peroxidation in various CNS regions. Administration of cithiolone (8 mg/kg body wt./day, ip for 7 days), however, elevated SOD activity and depleted lipid peroxidation. Interestingly, no significant change was observed either in SOD activity or in lipid peroxidation following simultaneous administration of phosphamidon and cithiolone.

Male reproductive toxicity of phosphamidon: histopathological changes in epididymis.

Phosphamidon, a neurotoxic insecticide, was tested for male reproductive toxicity with special reference to the epididymis. The insecticide was fed to Wistar strain male albino rat at 35 ppm concentration in drinking water ad libitum for 30 days. After vascular perfusion, thin slices of caput and cauda epididymidis were embedded in plastic, cut at 1 micron thickness and stained in toluidine blue for light microscopic observation. Principal cells of the caput epididymidis were vacuolarized and seen to pinch off fragments apically. In the proximal cauda the clear cells increased in height and in the size of the secondary lysosomal granules. In the distal cauda the clear cells appeared swollen out of proportion. Phosphamidon appears to affect the principal cells indirectly through its toxic effect on the Leydig cells; the clear cells of the cauda appear to be directly vulnerable to the toxic action of the pesticide.

Acute neurobehavioural toxicity of phosphamindon and its drug-induced alteration.

Phosphamidon, a systemic organophosphate insecticide, (1.4 mg/kg - dose 1/4th of LD50 given ip), produced several autonomic, neurological and behavioral effects in mice with peak effects being at 15 min. Similar dose in rats also abolished conditioned avoidance response. Pre-treatment with atropine, iproniazid, alpha-methyl-p-tyrosine, p-chlorophenylalanine or thiosemicarbazide reduce many of these effects. This suggests that phosphamidon toxicity involves the central cholinergic, adrenergic, serotonergic and GABAergic systems in addition to peripheral cholinergic effects.

Effect of dimecron on the blood parameters of Heteropneustes fossilis.

The effects of an organophosphate insecticide. dimecron. has been studied on certain haematological parameters, viz., haemoglobin concentration, RBC number, haematocrit, O2 carrying capacity of blood, etc. of Heteropneustes fossilis following exposures to the LC50 for 24 h and 96 h and 1/10 and 1/50 parts of 96 h LC50 for 90 days. There was a significant decrease in the Hb%, RBC number, HCt% and O2 carrying capcity of blood. But, there was significant increase in the MCH and MCV values following both acute and chronic exposures. The results indicate possible induction of anaemia in the exposed fish.