Determination of pesticides in postmortem blood and bone marrow of pesticide treated rabbits.

Forensic toxicological analyses have traditionally focused on the use of blood, body fluids, and certain organs in examinations of deaths due to intoxication. However, in some situations, putrefaction and contamination make proper sampling from tissues impossible, such as in exhumation cases. In these cases, bone marrow might be useful as an alternative specimen since it is a potential depot for drugs. This study aims to determine pesticides in postmortem and putrefied bone marrow of pesticide treated rabbits, so as to reveal the diagnostic value of toxicological analysis of bone marrow in exhumation cases. Out of thirteen rabbits, a 110 mg/kg dose of endosulfan was orally given to six through a gavage tool, and a 2500 mg/kg dose of diazinon was given to six using the same method. One rabbit was not treated with anything and served as a control sample. Venous blood, liver, lung, kidney, brain, and bone marrow samples were collected just after spontaneous death or cervical dislocation. After this, the rabbits were buried in soil. All of them were exhumed 1 month later, and putrefied viscera and bone marrow were sampled. Blood and tissue samples underwent solvent extraction and solid phase extraction, and then the samples were analyzed by GC-MS. Mean residue levels of diazinon in early postmortem samples were 85 mg/kg, 71 mg/kg, 23 mg/kg, 21 mg/kg, 19 mg/kg, and 0.4 mg/l in the liver, bone marrow, kidney, lung, brain, and blood, respectively. Mean residue levels of diazinon in the putrefied body were 3327 mg/kg in putrefied viscera and 1783 mg/kg in the bone marrow. Mean residue levels of endosulfan isomers and metabolites in early postmortem samples (blood, liver, lung, kidney, brain, and bone marrow) were 0.46 mg/kg (endosulfan sulfate), 0.32 mg/kg (alpha and beta isomers of endosulfan), and 0.14 mg/kg (endosulfan ether) while the same levels were 0.26 mg/kg (endosulfan sulfate), 0.24 mg/kg (alpha and beta isomers of endosulfan), and 0.1 mg/kg (endosulfan ether) in putrefied samples (putrefied bone marrow and putrefied viscera). Based on these experimental results, it can be concluded that cause of death can be determined as acute pesticide poisoning by toxicological analysis of samples from bone marrow and putrefied viscera in exhumation cases.

Liquid chromatography-tandem mass spectrometry method for the simultaneous quantitative determination of the organophosphorus pesticides dimethoate, fenthion, diazinon and chlorpyrifos in human blood.

Simultaneous determination of the organophosphorus pesticides dimethoate, fenthion, diazinon and chlorpyrifos in human blood by HPLC-tandem mass spectrometry was developed and validated. The pesticides were extracted by a simple one-step protein precipitation procedure. Chromatography was performed on a Luna C(18) (30mmx2.0mm, 3microm) column, using a step-gradient at a flow rate of 0.4ml/min. The assay was linear from 0.5 to 100ng/ml (r(2)>0.992, n=24) for all pesticides. The inter- and intra-day accuracy and precision for the method was 96.6-106.1% and <10%, respectively. The lower limit of quantification was 0.5ng/ml. In conclusion, the method described displays analytical performance characteristics that are suitable for the quantification of these pesticides in cases of acute poisoning.

Metoclopramide protection of diazinon-induced toxicosis in chickens.

The efficacy of metoclopramide for preventing organophosphate insecticide-induced (diazinon) toxicosis was evaluated in 7~14 days old chicks. Injection of metoclopramide at 25 mg/kg, s.c. 15 min before diazinon increased the oral 24 h median lethal dose of the insecticide in the chicks by 80%. Metoclopramide alone inhibited the in vitro and in vivo plasma and whole brain cholinesterase activities of the chicks. Metoclopramide pretreatment at 100 mg/kg, s.c. reduced the extent of cholinesterase inhibition that was caused by diazinon (10 mg/kg, p.o.) in the plasma and whole brain by 24% and 7%, respectively. Diazinon at 10 mg/kg, p.o. produced signs of cholinergic toxicosis in the chicks, and these signs included salivation, lacrimation, gasping and convulsions within 2 h, and the 2-h and 24-h lethalities were 88 and 100%, respectively. Metoclopramide at the dose rates of 12.5, 25, 50, 100 and 200 mg/kg, s.c. given 15 min before diazinon (10 mg/kg, p.o.) variably decreased the occurrence of toxic manifestations in the chicks. The highest dose of metoclopramide (200 mg/kg, s.c.) reduced the 2-h and 24-h lethality of diazinon to 75% each and it reduced the overall toxicity score of diazinon by 32%. The data suggest that metoclopramide pretreatment only partially protected chicks against the acute toxicity of diazinon.

Diazinon toxicity in sheep and cattle arising from the misuse of unlicensed and out-of-date products.

Two incidents involving the misuse of unlabelled or out-of-date diazinon products are described. In the first incident, 81 of a group of 210 five-month-old lambs died within 12 hours of being dipped with the contents of a partly used tin of 15-year-old sheep dip. Analysis showed that the diazinon had decomposed by hydrolysis into a range of breakdown products, including tetraethyldithiopyrophosphate (sulfotepp) and monothiono-tetraethylpyrophosphate (monothiono-TEPP), which are much more toxic to mammals than diazinon. In the second incident, four yearling bulls were affected within 48 hours of being treated with a liquid that the farmer believed to be a lice treatment but was actually old diazinon sheep dip. Three of the bulls recovered but one died four days after the treatment. Analysis of the dip confirmed the presence of diazinon but in this incident no breakdown products were detected.

Inhibición de colinesterasa plasmática en el zorzal pardo (Turdus grayi), expuesto a diazinón en cultivos de papaya maradol en Yucatán, México / Plasma cholinesterase inhibition in the clay-colored robin (Turdus grayi) exposed to diazinon in maradol papaya crops in Yucatan, Mexico

El uso de plaguicidas organofosforados en la agricultura tiene como consecuencia que las aves que forrajean en los cultivos se puedan intoxicar con estas sustancias. Los efectos que se han observado en las aves debido a intoxicación por plaguicidas son variados, desde efectos en el comportamiento y la reproducción hasta la muerte. Uno de los insecticidas ampliamente utilizado en el cultivo de papaya maradol es el diazinón, el cual ha ocasionado diversos casos de intoxicación y muerte en aves silvestres. El objetivo del presente estudio fue el de evaluar el impacto de la aplicación de diazinón en cultivos de papaya en la actividad de la colinesterasa plasmática del zorzal pardo (Turdus grayi). Se capturaron individuos de zorzal pardo forrajeando en un cultivo de papaya al día siguiente de haber sido fumigado con diazinón a una dosis de 1,5 kg/ha en el mes de marzo y mayo respectivamente. A estas aves se les tomó una muestra de sangre de la vena braquial y se determinó la actividad enzimática en el suero. Las muestras de suero de las aves utilizadas como control se tomaron en el mismo horario y se analizaron de la misma manera. La actividad enzimática de los machos fue mayor que la de las hembras (53,52%) y la inhibición promedio de la colinesterasa fue de 49,43%. La inhibición de colinesterasa fue mayor en el mes mayo que en marzo debido probablemente a la exposición e ingestión más continua del insecticida a través del alimento y posible absorción a través de la piel. Este grado de inhibición enzimática es posible que afecte el comportamiento del zorzal pardo y en casos severos podría ocasionarle la muerte

The use of organophosphorous pesticides in agriculture can result in intoxication of birds foraging in sprayed crops. Effects on birds resulting from pesticide intoxication are varied and include behavioral and reproductive effects, including death. One widely used insecticide in Maradol papaya crops is diazinon which has been associated with various incidents of intoxication and death of wild birds. The objective of this study was to evaluate the impact of diazinon application to papaya crops on plasma cholinesterase activity of the clay-colored robin (Turdus grayi). We captured clay-colored robins foraging in a papaya crop the following day after the field had been sprayed with diazinon at a dose of 1.5 kg/ha during March and May, respectively. We took a blood sample from the brachialis vein of the birds captured and measured plasma enzymatic activity. The plasma samples from birds used as controls were taken during the same time period and were analyzed in a similar way. Enzymatic activity of males was greater than that of females (53,52%) and mean cholinesterase inhibition was 49.43%. Cholinesterase inhibition was greater during May than in March probably due to more continuous exposure and ingestion of the insecticide through food and possible absorption through the skin. This degree of enzymatic inhibition is possibly affecting the behavior of the clay-colored robin and could result in death in severe cases

Organophosphate poisoning in pregnancy: a case report.

A 42-year-old pregnant woman (26 weeks of gestation, G(4)P(0+3)) presented at the emergency department with a two-hour history of dizziness, blurred vision and repeated vomiting. These symptoms started during the use of an undiluted insecticide liquid (diazinon 60 EC) while cleaning a small non-aired bathroom. After clinical and laboratory confirmation for organophosphate poisoning (plasma pseudocholinesterase levels 161 U/l), treatment with atropine and pralidoxime was started. She recovered within 7 days and delivered a healthy baby 12 weeks later (Apgar score 9 and 10) by elective cesarean section. The child showed no signs or symptoms of organophospate, atropine or pralidoxime exposure.

Diazinon-induced endocrine disruption in bluegill sunfish, Lepomis macrochirus.

An attempt to reveal endocrine disruption in bluegill fish (Lepomis macrochirus) upon exposure to diazinon was made. Fish blood serum was collected and estradiol levels were measured following times of exposure to 60 microg/L of diazinon. Samples were drawn from the fish during each exposure period of 24, 48, 72 and 96 h and 1 and 2 weeks. The sample estradiol levels were compared to those of a control. Ovarian tissue was examined. As necrosis increased within the ovaries, estradiol blood serum levels altered from those of the control sample (101 pg/mL). Histopathology within the follicles became apparent as exposure time to diazinon increased. The changes in the ovarian follicles can be correlated to the estradiol levels in the blood. Mean estradiol level for the control group was 101.53 pg/mL. Estradiol levels from the trial groups varied from those of the control (24 h, undetectable; 48 h, 74.04 pg/mL; 72 h, 57.16 pg/mL; 96 h; 39.15 pg/mL; 1 week, 66.50 pg/mL; 2 weeks, undetectable). It seems that the initial shock experienced by the fish after 24 h of exposure resulted in an undetectable level of estradiol. The level of estradiol initially ascended, but it began to decline and after 1 week of exposure it ascended a little and became undetectable after 2 weeks. Nevertheless, throughout the exposure period the level of estradiol was lower than normal. Severe damage to the mature oocytes may be the reason for undetectable levels of estradiol after 2 weeks of exposure.

Pesticides and lung cancer risk in the agricultural health study cohort.

The authors examined the relation between 50 widely used agricultural pesticides and lung cancer incidence in the Agricultural Health Study, a prospective cohort study of 57,284 pesticide applicators and 32,333 spouses of farmer applicators with no prior history of lung cancer. Self-administered questionnaires were completed at enrollment (1993-1997). Cancer incidence was determined through population-based cancer registries from enrollment through December 31, 2001. A lung cancer standardized incidence ratio of 0.44 (95% confidence interval: 0.39, 0.49) was observed overall, due in large part to a low cigarette smoking prevalence. Two widely used herbicides, metolachlor and pendimethalin (for low-exposed groups to four higher exposure categories: odds ratio (OR) = 1.0, 1.6, 1.2, 5.0; p(trend) = 0.0002; and OR = 1.0, 1.6, 2.1, 4.4; p(trend) = 0.003, respectively), and two widely used insecticides, chlorpyrifos and diazinon (OR = 1.0, 1.1, 1.7, 1.9; p(trend) = 0.03; and OR = 1.0, 1.6, 2.7, 3.7; p(trend) = 0.04, respectively), showed some evidence of exposure response for lung cancer. These excesses could not be explained by previously identified lung cancer risk factors. The usage levels in this cohort are considerably higher than those typically experienced by the general population. An excess risk among spouses directly exposed to pesticides could not be evaluated at this time.

Health effects of diazinon on a family.

There is increasing evidence of permanent sequalae from acute organophosphate poisoning. We report on accidental diazinon overexposure with acute organophosphate poisoning through cutaneous absorption and inhalation followed by persistent neurological effects. In addition, we observed skeletal and endocrine effects likely attributable to the diazinon poisoning. A family of seven was exposed to diazinon in June 1999 over a two-day period. The pesticide company mistakenly used diazinon to heavily spray the inside of the home instead of permethrin. The applicator applied the pesticide over the entire surface of the floor, carpeting, furniture, and clothing in closets to eradicate an infestation of fleas. Acute symptoms in the family members included headaches, nausea, skin irritation, runny nose, and vomiting. The family was first evaluated at 3 months and then 3 years after the acute poisoning. There were persisting neurological symptoms of memory loss, decreased concentration, irritability, and personality changes of varying degrees in all family members. Objective neurological findings of impaired balance, reaction time, color vision, slotted pegboards and trials making were present in the three older children who could be tested. Neuropsychological evaluation revealed evidence of organic brain dysfunction in all seven family members. Bone growth difficulties are present in four of five children. One child has delayed menarche.

Determination of solid-liquid partition coefficients (K(d)) for diazinon, propetamphos and cis-permethrin: implications for sheep dip disposal.

Two groups of chemicals are currently licensed for use in sheep dip products in the UK. These are organophosphate (OP) insecticides and synthetic pyrethroid (SP) insecticides. SPs are deemed to be less toxic to human health than OPs, although they are approximately 100 times more toxic to some elements of the aquatic environment. Three insecticides were selected for experimental investigation: diazinon, propetamphos (OPs) and cis-permethrin (SP), representative of the active ingredients used in sheep dip formulations, with additional uses in insect control in crops, and for domestic control of flies, mosquitoes, cockroaches, lice, ticks and spiders. The UK Government has recently reviewed agricultural practices relating to the disposal of used sheep dip, because the constituent insecticides are frequently detected in UK watercourses and the presence of these compounds is a severe hazard to the aquatic environment. Standard batch sorption experiments were carried out to investigate insecticide partitioning from water to soil, and the relationship between sorption and soil organic carbon content is discussed. Sorption isotherms and K(d) values showed that cis-permethrin adsorption was fastest on all five soils investigated, exhibiting the greatest total partitioning to the soil phase (83.8-94.8%) and high resistance to desorption. In comparison, the OP insecticides exhibited moderately strong soil adsorption as evidenced by their K(d) coefficients (diazinon K(d) 12-35 and propetamphos K(d) 9-60), with low sorption reversibility (< 15%). Calculation of a hydrological retardation factor in a scenario representative of a typical UK environment suggested that SP insecticides such as cis-permethrin will not migrate in the soil profile due to their virtual immobility and strong soil retention, and thus waste sheep dip disposal to agricultural land should not pose a risk to aquatic life if applied with appropriate controls.

Gastric outlet obstruction as a late complication of ingestion of diazinon.

Acute gastrointestinal symptoms are known to occur with organophosphorus compounds poisoning. Delayed complication in the form of gastric outlet obstruction has not been reported to date. We report gastric outlet obstruction developing after diazinon ingestion in a young girl. She responded to endoscopic balloon dilatation.

Supranuclear gaze palsy and opsoclonus after Diazinon poisoning.

A 52 year old man developed a supranuclear gaze palsy and opsoclonus after Diazinon poisoning. The diagnosis was confirmed by low plasma and red blood cell cholinesterase activity and urine mass spectroscopy. Saccadic control may be mediated in part by acetylcholine. Opsoclonus in the setting of organophosphate intoxication may occur as a result of cholinergic excess which overactivates the fastigial nuclei.

Histological examination of sublethal effects of diazinon on ovary of bluegill, Lepomis macrochirus.

The effects of the insecticide, diazinon (an organophosphorous compound), on the ovaries of bluegill (Lepomis macrohirus) were studied. Histological preparations of bluegill ovarian tissue was examined at 24, 48, 72 and 96 h and 1, 2 and 3 weeks following exposure to sublethal doses of diazinon (60 microg/l). The control contained primary follicles with an intact ovigerous lamellae and tunica albuginea. The control also revealed well developed stage IV oocytes with properly distributed provitelline nucleoli. After 24 h of exposure to diazinon, primary follicles began to show adhesion and cytoplasmic retraction in oocyte II occured. Cytoplasmic degeneration and additional adhesion and more retraction were visible at 48 h in oocyte II; 72 h brought forth adhesion, retraction and cytoplamic expulsion from oocyte IV. The number of atretic oocytes increased. Damages to the oocyte IV started to occur after 72 h of exposure. Cytoplasmic retraction and clumping was more visible at 96 h in oocyte IV. Partial destruction of the ovigerous lamellae and vitellogenic membrane occurred after 1 week. Two weeks continued to reveal destruction of follicles. Severe damage of the ovigerous lamellae, increased intrafollicular spaces, vacuolated cytoplasm, extrusion of karyoplasm and necrosis in the cytoplasm were most evident following 3 weeks of exposure. The ovarian wall became frayed and broken. Additionally, a marked increase of atretic follicles, shrinkage, and embedded nucleoli into the surrounding cytoplasm in oocyte II, III and IV were observed at week 3. This study revealed that oocytes at their different stages of maturation get affected differently at various exposure. Based on observations of the ovarian tissue compositional and structural changes following given exposure times, it becomes evident that consistent sublethal doses of diazinon can and will alter microscopic anatomy of the fish ovary.

Skin diseases and enzymatic antioxidant activity among workers exposed to pesticides.

In this study, 150 workers exposed to pesticides and 50 healthy control subjects were given clinical and dermatological examinations, patch tests, tests of liver and renal function, complete blood count, blood sugar and urinalysis. Activity of the antioxidant enzymes superoxide dismutase, glutathione peroxidase and glutathione reductase was also evaluated. Dermatological findings were positive in 78%, 76% and 54% of workers exposed to organophosphates, pyrethroids and carbamate pesticides respectively. The patch test was positive in 70% of workers exposed to pyrethroids and 64% exposed to carbamate pesticides. Liver enzyme levels were generally increased in workers while antioxidant enzyme activity was significantly decreased in all workers compared with the controls.

Agricultural use of organophosphate pesticides and the risk of non-Hodgkin's lymphoma among male farmers (United States).

OBJECTIVE: Data from three population-based case-control studies conducted in Kansas, Nebraska, Iowa, and Minnesota were pooled to evaluate the relationship between the use of organophosphate pesticides and non-Hodgkin's lymphoma (NHL) among white male farmers. METHODS: The data set included 748 cases of non-Hodgkin's lymphoma and 2236 population-based controls. Telephone or in-person interviews were utilized to obtain information on the use of pesticides. Odds ratios (OR) adjusted for age, state of residence, and respondent status, as well as other pesticide use where appropriate, were estimated by logistic regression. RESULTS: Use of organophosphate pesticides was associated with a statistically significant 50% increased risk of NHL, but direct interviews showed a significantly lower risk (OR = 1.2) than proxy interviews (OR = 3.0). Among direct interviews the risk of small lymphocytic lymphoma increased with diazinon use (OR = 2.8), after adjustment for other pesticide exposures. CONCLUSIONS: Although we found associations between the risk of NHL and several groupings and specific organophosphate pesticides, larger risks from proxy respondents complicate interpretation. Associations, however, between reported use of diazinon and NHL, particularly diffuse and small lymphocytic lymphoma, among subjects providing direct interviews are not easily discounted.