Acute Chlorpyrifos Exposure Research Articles

Multi-Biomarker Assessment in Common Carp (Cyprinus carpio, Linnaeus 1758) Liver after Acute Chlorpyrifos Exposure

The excessive use of pesticides at different stages of crop production can pose a great danger to the aquatic environment, and particularly to fish. The purpose of the present work was to assess the negative effects of chlorpyrifos (CPF) on the liver histological architecture and the activities of marker enzymes in common carp (Cyprinus carpio Linnaeus, 1758), by applying a multi-biomarker technique. The tested insecticide is categorized as a priority pollutant in surface waters in terms of Directive 2013/39/EU. The carps were exposed to different and environmentally relevant CPF concentrations for 72 h (a short-term acute experiment). The results showed that the tested insecticide alters the liver histological structure, causing degenerative lesions, such as granular and vacuolar degeneration; necrobiotic alterations and necrosis, as well as changes in the circulatory system. In addition, CPF induces changes in the enzymatic activity of lactate dehydrogenase (LDH), aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), cholinesterase (ChE), glutathione peroxidase (GPx) and catalase (CAT). The results from such experimental set ups could be successfully used in the legislation related to the protection of water bodies from contamination, in areas with intensive application of plant protection products used in agricultural practices, and also in implementing the Water Frame Directive by using multi-biomarker approaches.

Long term compulsivity on the 5-choice serial reaction time task after acute Chlorpyrifos exposure

Pesticide exposure has been associated with neuropsychological and psychiatric impairments and neurodegenerative disorders. Pesticide exposure commonly causes a deficit in inhibitory control behaviours. In the present study, we investigated whether acute exposure to organophosphate (OP) chlorpyrifos (CPF) is related to long-term lack of inhibitory control; we also examined the possible neurochemical basis of this association. Lister Hooded rats were exposed to an acute dose of CPF (250mg/kg). Seven months later, we tested inhibitory control with the 5-choice serial reaction time task (5-CSRTT). We manipulated the baseline conditions of this task and also systemically pre-administered d-amphetamine, quinpirole, dizocilpine (MK-801) or ketanserin. We also analysed the post-mortem baseline levels of monoamines and amino acids in different brain regions. On the 5-CSRT task, CPF-exposed rats showed elevated perseverative responses that persisted across manipulation of baseline conditions of the task and under most of the pharmacological challenges tested. Only d-amphetamine induced a dose-dependent amelioration of the increased perseverative responses in the CPF group. The CPF group also exhibited increased levels of dopamine metabolism in the hippocampus and decreased levels of gamma-aminobutyric acid (GABA) and glutamate in the striatum compared to the vehicle group. These findings suggest that CPF induced a long-term compulsivity that was apparent in the 5-CSRT task and associated with changes in monoaminergic and amino acid brain systems of inhibitory control function. Exposure to high doses of OP should be taken into account in studies of environmental causes for neurodegenerative, neuropsychological and neuropsychiatric disorders.

Diphenyl diselenide attenuates hepatic and hematologic toxicity induced by chlorpyrifos acute exposure in rats

In this study, we investigated the effect of diphenyl diselenide [(PhSe)(2)] on chlorpyrifos (CPF)-induced hepatic and hematologic toxicity in rats. Rats were pre-treated with (PhSe)(2) (5 mg/kg) via the oral route (oral gavage) once a day for 7 days. On the eighth and ninth days, rats were treated with (PhSe)(2) (5 mg/kg) 30 min prior to CPF (50 mg/kg, by subcutaneous route). The aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase activities were determined in plasma of rats. Lipid peroxidation, protein carbonyl, and non-protein thiol levels as well as catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and gluthatione S-transferase activities were determined in livers of rats. Hematological parameters were also determined. The results showed that CPF caused hepatic oxidative damage, as demonstrated by an increase in lipid peroxidation and protein carbonyl levels which was associated with a decrease in antioxidant defenses. CPF exposure caused a reduction in the leukocyte, indicating hematologic toxicity. (PhSe)(2) was effective in attenuating these toxic effects caused by CPF exposure in rats. The results indicated that (PhSe)(2) was effective in protecting the hepatic and hematologic toxicity induced by acute CPF exposure in rats.

Short-Term Sensorimotor and Cognitive Changes Induced by Acute Chlorpyrifos Exposure in Wistar Rats: Ameliorative Effect of Vitamin E

Short-Term Sensorimotor and Cognitive Changes Induced by Acute Chlorpyrifos Exposure in Wistar Rats: Ameliorative Effect of Vitamin E

Amyloid β Peptide Levels Increase in Brain of AβPP Swedish Mice after Exposure to Chlorpyrifos

Chlorpyrifos (CPF) is an organophosphate pesticide widely used in intensive agriculture. Various studies have demonstrated delayed neurotoxic effects in adult mammals after acute CPF exposure. This pesticide induces oxidative stress and neuronal damage, which suggests a possible relationship between CPF exposure and Alzheimer's disease (AD). In the present study, we examined in a mice model of AD, long-term changes in the behavior and brain levels of amyloid β after acute CPF exposure. Fifty mg/kg of CPF were subcutaneously injected to Tg2576 (Tg) mice carrying the Swedish amyloid-β protein precursor (AβPP) mutation for AD. General status, body weight, acetyl cholinesterase (AChE) inhibition, and behavioral changes were assessed. Amyloid β fragment (1-40 and 1-42) levels were also measured in the cortical and hippocampal brain regions. A significant and transient decrease in body weight was observed 72 hr after treatment, while no autonomic effects were noted. Motor activity was decreased in Tg mice seven months after CPF treatment. Acquisition learning in a water maze task was not affected, but retention was ameliorated in CPF-exposed Tg mice. Amyloid β levels increased in the brains of treated Tg mice eight months after CPF exposure. The results of this study show that some behavioral changes persisted or emerged months after acute CPF exposure, while amyloid β levels increased. These findings raise concern about the risk of developing neurodegenerative diseases following moderate exposure to CPF in vulnerable subjects.

Dose-related gene expression changes in forebrain following acute, low-level chlorpyrifos exposure in neonatal rats

Chlorpyrifos (CPF) is a widely used organophosphorus insecticide (OP) and putative developmental neurotoxicant in humans. The acute toxicity of CPF is elicited by acetylcholinesterase (AChE) inhibition. We characterized dose-related (0.1, 0.5, 1 and 2 mg/kg) gene expression profiles and changes in cell signaling pathways 24 h following acute CPF exposure in 7-day-old rats. Microarray experiments indicated that approximately 9% of the 44,000 genes were differentially expressed following either one of the four CPF dosages studied (546, 505, 522, and 3,066 genes with 0.1, 0.5, 1.0 and 2.0 mg/kg CPF). Genes were grouped according to dose-related expression patterns using K-means clustering while gene networks and canonical pathways were evaluated using Ingenuity Pathway Analysis®. Twenty clusters were identified and differential expression of selected genes was verified by RT-PCR. The four largest clusters (each containing from 276 to 905 genes) constituted over 50% of all differentially expressed genes and exhibited up-regulation following exposure to the highest dosage (2 mg/kg CPF). The total number of gene networks affected by CPF also rose sharply with the highest dosage of CPF (18, 16, 18 and 50 with 0.1, 0.5, 1 and 2 mg/kg CPF). Forebrain cholinesterase (ChE) activity was significantly reduced (26%) only in the highest dosage group. Based on magnitude of dose-related changes in differentially expressed genes, relative numbers of gene clusters and signaling networks affected, and forebrain ChE inhibition only at 2 mg/kg CPF, we focused subsequent analyses on this treatment group. Six canonical pathways were identified that were significantly affected by 2 mg/kg CPF (MAPK, oxidative stress, NFΚB, mitochondrial dysfunction, arylhydrocarbon receptor and adrenergic receptor signaling). Evaluation of different cellular functions of the differentially expressed genes suggested changes related to olfactory receptors, cell adhesion/migration, synapse/synaptic transmission and transcription/translation. Nine genes were differentially affected in all four CPF dosing groups. We conclude that the most robust, consistent changes in differential gene expression in neonatal forebrain across a range of acute CPF dosages occurred at an exposure level associated with the classical marker of OP toxicity, AChE inhibition. Disruption of multiple cellular pathways, in particular cell adhesion, may contribute to the developmental neurotoxicity potential of this pesticide.

Ameliorative effects of vitamin C on short-term sensorimotor and cognitive changes induced by acute chlorpyrifos exposure in Wistar rats

Human and experimental animal studies have shown long- and short-term neurological sequelae following acute organophosphate (OP) exposure. Although the main molecular mechanism of OP neurotoxiicty involves acetylcholinesterase (AChE) inhibition, studies have also implicated the induction of oxidative stress. The present study was therefore aimed at evaluating the effect of acute chlorpyrifos (CPF) exposure on short-term sensorimotor and cognitive changes in Wistar rats, the role of brain lipoperoxidative changes and the effect of pretreatment with vitamin C. Twenty-eight rats divided into four groups of seven rats in each group served as subjects for this study. Rats in group I were given soya oil (2 ml/kg) while those in group II were dosed vitamin C (100 mg/kg). Group III were administered CPF only (42.5 mg/kg ∼50% of LD(50)), while group IV were pretreated with vitamin C (100 mg/kg) and then exposed to CPF (42.5 mg/kg), 30 min later. The regimens were administered once orally and the animals were examined for clinical signs, death and subjected to periodic neurobehavioral evaluation for motor strength, coordinated gait, neuromuscular coordination, learning and memory. At the end of 8 weeks of evaluation, the rats were sacrificed and the brain tissue evaluated for AChE activity and malonaldehyde (MDA) concentration, as an index of lipoperoxidative changes. The rats administered with CPF showed deficits in motor strength, coordinated gaits, neuromuscular coordination, learning and memory, slight decrease in AChE activity and an increase in brain MDA concentration. Pretreatment with vitamin C improved the neurobehavioral deficits and AChE activity, and caused a significant reduction in MDA concentration. In conclusion, the study has demonstrated that acute CPF exposure in Wistar rats caused short-term impairment in sensorimotor and cognitive functions partly due to brain lipoperoxidative changes, which were ameliorated by pretreatment with vitamin C.

Long-term monoamine changes in the striatum and nucleus accumbens after acute chlorpyrifos exposure

This study examined the time-course effects (2, 7, 14 and 30 days) of acute chlorpyrifos (CPF) intoxication (250 mg/kg, s.c.) on monoamine systems and acetylcholinesterase (AChE) activity in the striatum and nucleus accumbens of adult male rats. We show that CPF produced significant long-term inhibition of AChE activity in the striatum and nucleus accumbens. In the striatum, CPF intoxication resulted in changes in dopamine (DA) metabolism after 2 days and changes in serotonin (5-HT) turnover after 7 and 15 days. Significant decreases in monoamine content including norepinephrine (NE), DA, 5-HT and their metabolites were found in the nucleus accumbens 30 days after CPF intoxication. These results suggest that acute exposure to CPF induces long-term changes in the monoamine systems (NE, DA and 5-HT) in adult animals. The lack of correlation between regional AChE activity and neurochemical outcomes points to independent mechanisms.

Effects of acute chlorpyrifos exposure on in vivo acetylcholine accumulation in rat striatum

This study examined the acute effects of chlorpyrifos (CPF) on cholinesterase inhibition and acetylcholine levels in the striatum of freely moving rats using in vivo microdialysis. Adult, male Sprague-Dawley rats were treated with vehicle (peanut oil, 2 ml/kg) or CPF (84, 156 or 279 mg/kg, sc) and functional signs of toxicity, body weight and motor activity recorded. Microdialysis was conducted at 1, 4 and 7 days after CPF exposure for measurement of acetylcholine levels in striatum. Rats were then sacrificed and the contralateral striatum and diaphragm were collected for biochemical measurements. Few overt signs of cholinergic toxicity were noted in any rats. Body weight gain was significantly affected in the high-dose (279 mg/kg) group only, while motor activity (nocturnal rearing) was significantly reduced in all CPF-treated groups at one day (84 mg/kg) or from 1-4 days (156 and 279 mg/kg) after dosing. Cholinesterase activities in both diaphragm and striatum were markedly inhibited (50-92%) in a time-dependent manner, but there were relatively minimal dose-related changes. In contrast, time- and dose-dependent changes in striatal acetylcholine levels were noted, with significantly higher levels noted in the high-dose group compared to other groups. Maximal increases in striatal acetylcholine levels were observed at 4-7 days after dosing (84 mg/kg, 7-9-fold; 156 mg/kg, 10-13-fold; 279 mg/kg, 35-57-fold). Substantially higher acetylcholine levels were noted when an exogenous cholinesterase inhibitor was included in the perfusion buffer, but CPF treatment-related differences were substantially lower in magnitude under those conditions. The results suggest that marked differences in acetylcholine accumulation can occur with dosages of CPF eliciting relatively similar degrees of cholinesterase inhibition. Furthermore, the minimal expression of classic signs of cholinergic toxicity in the presence of extensive brain acetylcholine accumulation suggests that some compensatory process(es) downstream from synaptic neurotransmitter accumulation limits the expression of toxicity following acute CPF exposure.

Age-related Effects of Chlorpyrifos on Acetylcholine Release in Rat Brain

Chlorpyrifos (CPF) is an organophosphorus insecticide that elicits toxicity through inhibition of acetylcholinesterase (AChE). Young animals are markedly more sensitive than adults to the acute toxicity of CPF. We evaluated acetylcholine (ACh) release and its muscarinic receptor-mediated regulation (i.e. muscarinic autoreceptor function, MAF) during maturation as a possible contributing factor to age-related differences in sensitivity. Cortical and striatal slices were prelabeled with [ 3H ]choline chloride, superfused in the presence or absence of the anticholinesterase physostigmine (PHY, 20 μM) and stimulated twice (S1 and S2) with a high concentration of potassium chloride (20 mM). Depolarization-stimulated ACh release (DSAR) was lowest in neonatal, intermediate in juvenile and markedly higher in adult tissues. MAF was not detectable in tissues from neonatal rats but was present in juvenile and adult tissues. ACh release and MAF were studied at 4, 24 and 96 h following oral exposure to CPF (0, 0.5 or 1×LD 10). In general, 40–60% and 80–90% maximal AChE inhibition followed exposure to the respective 0.5 and 1×LD 10 dosages. DSAR was decreased in neonatal cortex 1 day after LD 10 exposure but increased in juvenile striatum 1 day after LD 10 treatment. In adults, DSAR was reduced at 4 and 24 h after exposure, but increased 96 h after CPF exposure. In juveniles, MAF was reduced in both brain regions at 24 h after 0.5LD 10 exposure and at 24 and 96 h after LD 10 exposure in cortex. A later reduction in MAF was noted in adult tissues (i.e. only at 96 h after LD 10 treatment). Together, the results suggest that ACh release dynamics in brain vary markedly during postnatal maturation and that acute CPF exposure can alter ACh release in an age-related manner. The functional status of presynaptic processes regulating neurotransmitter release may contribute to age-related neurotoxicity elicited by high-dose exposures to chlorpyrifos.