The effect of five organophosphorus insecticides on survival and temperature tolerance in the copepod, Macrocyclops albidus (Copepoda: Cyclopidae)

Macrocyclops albidus were exposed for 24 hours to ABATE®, fenthion, malathion, methyl parathion, and chlorpyrifos at concentrations of 1, 5, 10, 25, and 50 parts per billion (ppb). Exposure to chlorpyrifos resulted in the greatest mortality to M. albidus; concentrations of 25 and 50 ppb caused 100% mortality. The thermal tolerance was lowered significantly by all chemicals at concentrations from 1 to 50 ppb.     

The effects of sublethal concentrations of five organophosphorus insecticides on temperature tolerance, reflexes, and orientation in Gambusia affinis affinis (Pisces: Poeciliidae)

Gambusia affinis were exposed to the insecticides ABATE®, fenthion, chlorpyrifos, methyl parathion, and malathion for 24 h at concentrations of 1, 5, and 10 ppb and for 48 h at 5 ppb. The thermal tolerance was lowered significantly for male and female G. affinis by the toxicants methyl parathion and chlorpyrifos at 5 and 10 ppb with 24 h exposure. Activity was restricted with fish exposed to methyl parathion and chlorpyrifos at concentrations of 10 ppb for 24 h and at 5 ppb for 48 h.     

Lethal and sublethal effects of selected insecticides and an insect growth regulator on the boll weevil (Coleoptera: Curculionidae) ectoparasitoid Catolaccus grandis (Hymenoptera: Pteromalidae)

A laboratory culture of Catolaccus grandis (Burks), an ectoparasitoid of the boll weevil, Anthonomus grandis grandis Boheman, was exposed to lethal and sublethal doses of insecticides and an insect growth regulator using a spray chamber bioassay. Materials tested were azinphos-methyl, endosulfan, fipronil, malathion, cyfluthrin, dimethoate, spinosad, methyl parathion, acephate, oxamyl, and tebufenozide. At full rates, spinosad was significantly less toxic to female C. grandis than other treatments except endosulfan. Fipronil and malathion were significantly more toxic to females than other treatments. Most of the chemicals tested were highly toxic to male C. grandis; spinosad was least toxic. At reduced rates, most of 4 selected chemicals tested were low in toxicity to C. grandis; however, a reduced rate of malathion was significantly more toxic to females than other treatments. No C. grandis pupae developed from parasitism during a 24-h treatment period with malathion or spinosad. The sex ratio of progeny from sprayed adults appeared to be unaffected by the treatments.     

Toxicological studies of pesticides on cytoplasmic streaming in Nitella

In the present study, changes in velocity of cytoplasmic streaming in the giant internodal cells of Nitella for varying concentration of the pesticides, 2,4-D, dieldrin, malathion, methyl parathion and endosulfan, were measured. Marked decrease in the velocity of cytoplasmic streaming was found at the concentrations of 0.01, 0.1, 1, 10 and 100mM. Dieldrin was the most toxic to all the pesticides investigated, followed by methyl parathion, endosulfan, malathion and 2,4-D. Threshold values for dieldrin, methylparathion, endosulfan, malathion and 2,4-D as indicated by the onset of decrease in the normal cytoplasmic streaming velocity were less than 6.25 x 10(-6), 2.5 x 10(-5), 5 x 10(-5), 5 x 10(-5) and 1.25 x 10(-5)M respectively. Cessation of streaming was noticed above 1mM in dieldrin and above 10mM when exposed to methylparathion and endosulfan. Cessation of streaming was not seen up to 100mM concentration of 2,4-D and malathion.     

Comparative susceptibility of sweetpotato weevil (Coleoptera: Brentidae) to selected insecticides

The response of sweetpotato weevil, Cylas formicarius (F.) (Coleoptera: Brentidae), to insecticides used for its control was tested in laboratory bioassays. A glass vial bioassay technique was used to determine the susceptibility of two cohorts of sweetpotato weevil to selected insecticides. Vials were treated with methyl parathion, bifenthrin, cyfluthrin, carbaryl, and phosmet. Sweetpotato weevils demonstrated a mortality response to increasing concentrations of all insecticides tested, and our results indicated decreases in susceptibility of the Louisiana cohort of sweetpotato weevil compared with the Texas cohort for all insecticides tested. Methyl parathion was the most toxic chemical tested for both cohorts, followed by the pyrethroids, cyfluthrin and bifenthrin. Phosmet exhibited moderate toxicity compared with other chemicals tested, whereas sweetpotato weevils were least susceptible to carbaryl. Significant differences in lethal concentration (LC)50 and LC90 values for cyfluthrin and bifenthrin, the LC50 values for methyl parathion and phosmet, and the LC90 values for carbaryl were observed between the two cohorts. This study documents baseline toxicological data for five insecticides in two populations of sweetpotato weevil and demonstrates that susceptibility to all insecticides tested is lower for the Louisiana population compared with the Texas population.     

Microbial cleavage of various organophosphorus insecticides.

Bacteria able to utilize Aspon, Azodrin, Dasanit, diazinon, malathion, Orthene, parathion, Trithion, dimethoate, Dylox, methyl parathion, and Vapona as sole phosphorus sources were isolated from soil and sewage. Individual isolates used from 3 to 10 of these insecticides as sole phosphorus sources. The extent of growth of two Pseudomonas strains in media containing diazinon and malathion was in the range expected from the amount of insecticide supplied, and their proliferation resulted in disappearance of the chemical. Resting cells of the pseudomonads derived from cultures grown on diazinon or malathion but not orthophosphate caused extensive destruction of these two organophosphates in the presence or absence of chloramphenicol. Extracts of the two bacteria derived from organophosphate-grown cultures catalyzed the disappearance of Aspon, Azodrin, Dasanit, diazinon, malathion, Orthene, parathion, and Trithion but not dimethoate, Dylox, methyl parathion, and Vapona. Results from gas chromatographic analysis suggested that the extracts formed dimethyl phosphate from azodrin, dimethyl phosphorodithioate from malathion, diethyl phosphorodithioate from Trithion, and diethyl phosphorothioate from Dasanit, diazinon, and parathion. Dimethyl phosphate, dimethyl phosphorothioate , dimethyl phosphorodithioate, diethyl phosphate, and diethyl phosphorothioate were not used by the pseudomonads as sole phosphorus sources.     

Microbial cleavage of various organophosphorus insecticides.

Bacteria able to utilize Aspon, Azodrin, Dasanit, diazinon, malathion, Orthene, parathion, Trithion, dimethoate, Dylox, methyl parathion, and Vapona as sole phosphorus sources were isolated from soil and sewage. Individual isolates used from 3 to 10 of these insecticides as sole phosphorus sources. The extent of growth of two Pseudomonas strains in media containing diazinon and malathion was in the range expected from the amount of insecticide supplied, and their proliferation resulted in disappearance of the chemical. Resting cells of the pseudomonads derived from cultures grown on diazinon or malathion but not orthophosphate caused extensive destruction of these two organophosphates in the presence or absence of chloramphenicol. Extracts of the two bacteria derived from organophosphate-grown cultures catalyzed the disappearance of Aspon, Azodrin, Dasanit, diazinon, malathion, Orthene, parathion, and Trithion but not dimethoate, Dylox, methyl parathion, and Vapona. Results from gas chromatographic analysis suggested that the extracts formed dimethyl phosphate from azodrin, dimethyl phosphorodithioate from malathion, diethyl phosphorodithioate from Trithion, and diethyl phosphorothioate from Dasanit, diazinon, and parathion. Dimethyl phosphate, dimethyl phosphorothioate , dimethyl phosphorodithioate, diethyl phosphate, and diethyl phosphorothioate were not used by the pseudomonads as sole phosphorus sources.     

Genotoxicity studies with four organophosphorus insecticides using the unstable white-zeste system of Drosophila melanogaster

The present study was carried out to evaluate the suitability of the unstable white-zeste system in Drosophila melanogaster by testing 4 organophosphorus insecticides for potential genotoxic activity: dimethoate, fenitrothion, malathion, and methyl parathion. In view of the high sensitivity to insecticides of the unstable zeste strain used in this assay and the negative results obtained in this work, the white-zeste system does not appear to be sufficiently accurate for the evaluation of the mutagenic potential of specifically toxic chemicals, like insecticides.     

Blood gene expression markers to detect and distinguish target organ toxicity

The purpose of this study was to investigate whether the expression of specific genes in peripheral blood can be used as surrogate marker(s) to detect and distinguish target organ toxicity induced by chemicals in rats. Rats were intraperitoneally administered a single, acute dose of a well-established hepatotoxic (acetaminophen) or a neurotoxic (methyl parathion) chemical. Administration of acetaminophen (AP) in the rats resulted in hepatotoxicity as evidenced from elevated blood transaminase activities. Similarly, administration of methyl parathion (MP) resulted in neurotoxicity in the rats as evidenced from the inhibition of acetyl cholinesterase activity in their blood. Administration of either chemical also resulted in mild hematotoxicity in the rats. Microarray analysis of the global gene expression profile of rat blood identified distinct gene expression markers capable of detecting and distinguishing hepatotoxicity and neurotoxicity induced by AP and MP, respectively. Differential expressions of the marker genes for hepatotoxicity and neurotoxicity were detectable in the blood earlier than the appearance of the commonly used clinical markers (serum transaminases and acetyl cholinesterase). The ability of the marker genes to detect hepatotoxicity and neurotoxicity was further confirmed using the blood samples of rats administered additional hepatotoxic (thioacetamide, dimethylnitrobenzene, and carbon tetrachloride) or neurotoxic (ethyl parathion and malathion) chemicals. In summary, our results demonstrated that blood gene expression markers can detect and distinguish target organ toxicity non-invasively.     

In vitro breakage of plasmid DNA by mutagens and pesticides.

Covalently closed circular molecules of the colicinogenic plasmid E1 can serve as sensitive indicators for detecting in vitro breakage of DNA. After these molecules are radioactively labeled and purified by cesium chloride density-gradient centrifugation, they are incubated with the compounds to be tested. Samples are analyzed on alkaline sucrose gradients to determine the fraction of unbroken molecules and a breakage rate is calculated. Positive results were obtained for all three mutagenic alkylating agents (MMS, EMS, and MNNG) and of the 11 pesticides tested, dexon, dichlorvos, malathion, and methyl parathion induced breaks in molecules at a rate significantly greater than the controls.     

Comparative susceptibility of western corn rootworm (Coleoptera: Chrysomelidae) adults to selected insecticides in Kansas

Susceptibility of adult populations of the western corn rootworm, Diabrotica virgifera virgifera LeConte, to several insecticides was evaluated in seven Kansas counties, including Dickinson, Ford, Finney, Pottawatomie, Republic, Riley, and Stevens, between 1996 and 2002. All populations surveyed were highly susceptible to methyl parathion with the largest difference in susceptibility of only three-fold based on 16 complete bioassays for the populations from six counties over a 5-yr period. Noticeable decreases in carbaryl susceptibility were found in populations collected from Republic County between 1997 and 2001 when the cucurbitacin-carbaryl-based bait SLAM was widely used as an areawide management approach for adult corn rootworm control. However, the lowered carbaryl susceptibility returned to previous levels 1 yr after the use of SLAM was halted in the managed (treated) cornfields. This change implies possible dispersal of insects into the relatively small managed area from surrounding untreated cornfields and / or some fitness cost associated with carbaryl resistance within the population. Relative susceptibility of western corn rootworm adults also was evaluated for seven commonly used insecticides, including bifenthrin, carbaryl, chlorpyrifos, cypermethrin, fipronil, malathion, and methyl parathion. They were tested with corn rootworm adults collected from a single cornfield. Methyl parathion and bifenthrin were highly toxic to corn rootworm adults, and cypermethrin, chlorpyrifos, carbaryl, and malathion were only slightly less toxic. Although fipronil was highly toxic to adult rootworms, its activity was much slower than that of other insecticides. Thus, bifenthrin and methyl parathion were among the most effective in killing corn rootworm adults.     

Characterizing response behavior of medaka (Oryzias latipes) under chemical stress based on self-organizing map and filtering by integration

Behavioral responses (BRs) of medaka (Oryzias latipes) were observed after exposure to low concentrations (0.1 TU (Toxic Unit, TU), 1 TU, 5 TU, and 10 TU) of trichlorfon, parathion and malathion. Overall response patterns of test organisms were reflected from surface shapes of BS (Behavior Strength) values in 3-D: parathion appeared to be most variable in presenting response behaviors whereas trichlorfon showed relatively simple response patterns. The self-organizing map (SOM) addressed the time and toxic effects efficiently. An evident circadian rhythm observed in the control diminished at a low concentration of toxic unit, and variability of toxic effects was accordingly observed according to chemicals and concentrations. Subsequently filtering by integration was conducted to time series BS values. The highly fluctuating nature of original BS values was filtered efficiently to produce linear fitting closely. Slopes of regression decreased monotonically as toxic concentrations increased. Residual curves of integral BS values from linear fitting were further used for determining different BS phases proposed by empirical observations; the positive and negative phases were in accordance with acclimation, adjustment and toxic effects in behavior response modes. According to inclination and declination periods observed in residual curves, new states of test organisms were further defined to present intoxicating and recovering tendencies Profiles based on residual curves of integral BS values were able to show landscape of response patterns across toxic concentrations in different chemicals. Computational methods for defining behavior states provide an objective ground for analyzing complex stress response and could be suitable in referencing toxic behavior modes of test organisms quantitatively.     

Interaction of insecticides with lipid membranes.

The permeability of liposome membranes is increased by organophosphorus and organochlorinated insecticides at concentrations of 10(-5)--10(-4) M. The order of effectiveness is similar to the toxicity of the compounds to mammals, and is the following for permeation of non-electrolytes and for valinomycin-induced permeation of K+: parathion greater than 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (DDT) approximately aldrin greater than malathion greater than lindane. The degree of effectiveness for X-537A-induced permeation of Ca2+ was the following: aldrin greater than or equal to DDT greater than parathion greater than malathion greater than lindane. The organophosphorus compound, ethyl azinphos (10(-4) M), dramatically increases the permeability of liposome membranes to all the tested substances, probably as a consequence of surfactant effects. Some organochlorinated insecticides appear to react with cation ionophores and modulate their motion across lipid membranes. It is suggested that the insecticides may exert some of their toxic actions by modifying certain mechanisms in the cell membrane.     

Frequencies of chromosomal aberrations in smokers exposed to pesticides in cotton fields

Blood samples were collected from 50 smokers who were exposed to the pesticides DDT, BHC, endosulfan, malathion, methyl parathion, monocrotophos, quinolphos, dimethoate, phosphomidon, cypermethrin and fenvelrate. Samples were also collected from 20 non-smokers (control I) and 27 smokers (control II) who were unexposed to pesticides. Control II showed a significant increase in chromosomal aberrations when compared to control I. There was a significant increase in total chromosomal aberrations in smokers exposed to pesticides when compared to unexposed populations.     

Threat-sensitive Predator Avoidance by Larval Pacific Treefrogs (Amphibia, Hylidae)

According to the threat-sensitive predator avoidance hypothesis, the intensity of a prey animal's antipredator response should reflect its vulnerability to a specific predator. In laboratory experiments, we observed the intensity of antipredator responses of Pacific treefrog ( Hyla regilla ) tadpoles to stimuli from caged larval northwestern salamander ( Ambystoma gracile ) predators. We varied the sizes of the tadpoles relative to the salamanders in an attempt to create differences in vulnerability of tadpoles to the salamander predators. After documenting the response of the tadpoles to the caged predator, we tested the tadpole's vulnerability to the predator by releasing the tadpole with the predator. We observed that as the relative size of the tadpoles to the caged salamanders increased, the antipredator response of the tadpoles decreased. These changes in behaviour closely mirrored changes in actual vulnerability to the predator. Our results provide experimental support for the threat-sensitive predator avoidance hypothesis.     

The effects of acute and developmental temperature on burst swimming speed and myofibrillar ATPase activity in tadpoles of the Pacific tree frog, Hyla regilla

The effects of acute and developmental temperature on maximum burst swimming speed, body size, and myofibrillar ATPase activity were assessed in tadpoles of the Pacific tree frog, Hyla regilla. Tadpoles from field-collected egg masses were reared in the laboratory at 15 degrees (cool) and 25 degrees C (warm). Body size, maximum burst swimming speed from 5 degrees to 35 degrees C, and tail myofibrillar ATPase activity at 15 degrees and 25 degrees C were measured at a single developmental stage. Burst speed of both groups of tadpoles was strongly affected by test temperature (P<0. 001). Performance maxima spanned test temperatures of 15 degrees -25 degrees C for the cool group and 15 degrees -30 degrees C for the warm group. Burst speed also depended on developmental temperature (P<0.001), even after accounting for variation in body size. At most test temperatures, the cool-reared tadpoles swam faster than the warm-reared tadpoles. Myofibrillar ATPase activity was affected by test temperature (P<0.001). Like swimming speed, enzyme activity was greater in the cool-reared tadpoles than in the warm-reared tadpoles, a difference that was significant when assayed at 15 degrees C (P<0. 01). These results suggest a mechanism for developmental temperature effects on locomotor performance observed in other taxa.     

Effects of Adaptation on Biodegradation Rates in Sediment/Water Cores from Estuarine and Freshwater Environments

Experiments were devised to determine whether exposure to xenobiotics would cause microbial populations to degrade the compounds more rapidly during subsequent exposures. Studies were done with water/sediment systems (ecocores) taken from a salt marsh and a river. Systems were tested for adaptation to the model compounds methyl parathion and p-nitrophenol. ¹⁴CO₂ released from radioactive parent compounds was used as a measure of mineralization. River populations preexposed to p-nitrophenol at concentrations as low as 60 μg/liter degraded the nitrophenol much faster than did control populations. River populations preexposed to methyl parathion also adapted to degrade the pesticide more rapidly, but higher concentrations were required. Salt marsh populations did not adapt to degrade methyl parathion. p-Nitrophenol-degrading bacteria were isolated from river samples but not from salt marsh samples. Numbers of nitrophenol-degrading bacteria increased 4 to 5 orders of magnitude during adaptation. Results indicate that the ability of populations to adapt depends on the presence of specific microorganisms. Biodegradation rates in laboratory systems can be affected by concentration and prior exposure; therefore, adaptation must be considered when such systems are used to predict the fate of xenobiotics in the environment.     

Pharmacokinetics of methyl parathion: a comparison following single intravenous,oral or dermal administration

Assessment of the risks posed by the residential use of methyl parathion requires an understanding of its pharmacokinetics after different routes of exposure. Thus, studies were performed using adult female rats to define the pharmacokinetic parameters for methyl parathion after intravenous injection and to apply the described model to an examination of its pharmacokinetics after single oral or dermal exposure. The pharmacokinetics of methyl parathion after intravenous administration (1.5 mg/kg) were best described by a three-compartment model; the apparent volume of the central compartment was 1.45 liters/kg, clearance was 1.85 liters/h/kg and the terminal half-life was 6.6 h with an elimination constant of 0.50 h(-1). The apparent oral absorption coefficient for methyl parathion (1.5 mg/kg) was 1.24 h(-1), and its oral bioavailability was approximately 20%. The latter likely includes a significant first pass effect. Concentrations of methyl parathion increased during the initial 10-60 min and then declined during the next 15-36 h. After dermal administration (6.25-25 mg/kg), methyl parathion concentrations peaked within 12-26 h and then declined dose dependently. The apparent dermal absorption coefficient was approximately 0.41 h(-1), and only two pharmacokinetic compartments could be distinguished. In conclusion, the pharmacokinetics of methyl parathion are complex and route dependent. Also, dermal exposure, because of sustained methyl parathion concentrations, may pose the greatest risk.     

Hydrolysis of organophosphate insecticides by an immobilized-enzyme system.

An enzyme preparation that could detoxify parathion and eight other organophosphate pesticides was covalently bound to either porous glass or porous silica beads. This immobilized-enzyme system was examined for its use in detoxification of pesticides in production wastewaters. The kinetics of parathion hydrolysis were examined at flow rates up to 96 liter/hr and at influent substrate concentrations ranging from 10--250 mg/liter. The enzyme reactor was able to hydrolyze 95% or more of the parathion added to industrial wastewaters generated during its production, thus reducing the effluent parathion concentration to below 500 ppb. Laboratory continuous-flow experiments were conducted for 70 days with industrial wastewater and indicated no loss in immobilized-enzyme activity. The influence of pH, temperature, solvents, and detergents on enzyme stability and activity and enzyme reactor kinetics will be discussed.     

Parathion and methyl parathion toxicity and metabolism in piperonyl butoxide and diethyl maleate pretreated mice

Pretreatment of male mice with piperonyl butoxide, 400 mg/kg 1 h before challenge with insecticides, resulted in a 40-fold antagonism of the acute i.p. toxicity of methyl parathion but potentiated the toxicity of parathion two-fold. Piperonyl butoxide had no effect on the toxicity of the oxygen analogs of these insecticides, methyl paraoxon and paraoxon. Diethyl maleate (1 ml/kg) depleted liver glutathione by 80% after one hour, potentiated the toxicity of both methyl parathion and methyl paraoxon, and partially counteracted the protective effect of piperonyl butoxide on methyl parathion toxicity. Piperonyl butoxide delayed the onset of brain cholinesterase inhibition by parathion. Studies of the metabolism of the insecticides by liver homogenates in vitro demonstrated that piperonyl butoxide inhibited both the oxidative formation of the oxygen analogs (activation) and oxidative cleavage to p-nitrophenol and dialkylphosphorothioic acid (detoxification). While parathion metabolism was mostly oxidative, methyl parathion metabolism appeared to be predominantly via glutathione-dependent enzymes. Studies of in vitro distribution of the insecticides demonstrated that piperonyl butoxide pretreatment resulted in elevated tissue concentrations of parathion and methyl parathion; however, the rate constant for elimination from plasma for both insecticides was unaffected by piperonyl butoxide. The overall rate of metabolism of methyl parathion in vivo was approximately twice that of parathion. These results suggest that during piperonyl butoxide inhibition of oxidative activation and cleavage, methyl parathion detoxification continues through uninhibited glutathione-dependent pathways of metabolism. The net result is a reduction in the acute toxicity of methyl parathion. Lack of an effective alternate pathway of detoxification may explain the delayed but greater toxicity of parathion in piperonyl butoxide pretreated mice.