Linkage of biomarkers along levels of biological complexity in juvenile and adult diazinon fed terrestrial isopod (Porcellio scaber, Isopoda, Crustacea)

In parallel laboratory experiments, we determined the effect of a typical representative of organophosphorous pesticides, diazinon, on AChE activity, lipid, protein and glycogen content, weight change, feeding activity and mortality of juvenile and adult terrestrial isopods Porcellio scaber (Isopoda, Crustacea). Organophosphorous pesticides (OP) are among the most extensively used pesticides, which have replaced organochlorine pesticides. OPs inhibit the enzyme acetylcholinesterase (AChE), resulting in neurotoxicity. They have more widespread effects on non-target organisms than do organochlorine pesticides. The aim of this study was to link effect of diazinon on target enzyme to energy reserves and to integrated biomarker responses in juvenile and adult P. scaber. The non-observed effect concentration (NOEC) for AChE activity after diazinon exposure in two weeks toxicity study with isopods was below 5 microg/g diazinon. There was a good agreement between concentrations at which AChE and survival were affected (10 microg/g diazinon in juveniles, 100 microg/g diazinon in adults). We revealed a link among AChE activity, protein content and mortality. Glycogen and lipid content, feeding activity and weight change were not affected in two weeks diazinon exposure up to 100 microg/g diazinon. Juveniles were affected at concentrations that were an order of magnitude lower than those provoking similar effects on adults. Recommendations are made for future toxicity studies with terrestrial isopods.     

Does monensin in chicken manure from poultry farms pose a threat to soil invertebrates?

Monensin is a carboxylic polyether ionophore used in the poultry industry as a coccidiostat. It enters the environment via manure from broiler farms. In spite of its potential presence in the environment, information concerning monensin residues in manure and soil and its toxicity to soil organisms are insufficient. In the present study, two beneficial soil invertebrate species, earthworms (Eisenia andrei) and woodlice (Porcellio scaber), were used to assess the toxicity of monensin. Animals were exposed to a range of monensin concentrations via soil or food. Earthworm reproduction was found to be the most susceptible endpoint (NOEC=3.5 mg kg(-1) dry soil; EC(50)=12.7 mg kg(-1) dry soil), while no adverse effects were recorded in isopods (NOEC?849mgkg(-1) dry soil, NOEC?357mgkg(-1) dry food). The obtained toxicity data were compared with potential concentrations of monensin in soil. In view of this, manure from broiler chickens treated with monensin at a poultry farm was sampled. According to monensin and nitrogen concentrations in the chicken manure and the degradation time of monensin, the predicted environmental concentration (PEC) was calculated. PEC of monensin is around 0.013 mg kg(-1) soil if manure is used after 3 months of composting and 0.05 mg kg(-1) soil if used without storage. Data for earthworm reproduction was used to estimate the predicted no-effect concentration (PNEC). If fresh chicken manure is applied to terrestrial ecosystems, the risk quotient (PEC/PNEC ratio) is above 1, which indicates that monensin might pose an environmental risk under certain conditions. To prevent this, it is strongly recommended to compost chicken manure for several months before using it as fertiliser.     

Bioavailability of contaminants estimated from uptake rates into soil invertebrates

It is often argued that the concentration of a pollutant inside an organism is a good indicator of its bioavailability, however, we show that the rate of uptake, not the concentration itself, is the superior predictor. In a study on zinc accumulation and toxicity to isopods (Porcellio scaber) the dietary EC(50) for the effect on body growth was rather constant and reproducible, while the internal EC(50) varied depending on the accumulation history of the animals. From the data a critical value for zinc accumulation in P. scaber was estimated as 53 microg/g/wk. We review toxicokinetic models applicable to time-series measurements of concentrations in invertebrates. The initial slope of the uptake curve is proposed as an indicator of bioavailability. To apply the dynamic concept of bioavailability in risk assessment, a set of representative organisms should be chosen and standardized protocols developed for exposure assays by which suspect soils can be evaluated.     

Toxicity of abamectin and doramectin to soil invertebrates

This study aimed at determining the toxicity of avermectins to soil invertebrates in soil and in faeces from recently treated sheep. Abamectin was more toxic than doramectin. In soil, earthworms (Eisenia andrei) were most affected with LC50s of 18 and 228 mg/kg dry soil, respectively, while LC50s were 67-111 and >300 mg/kg for springtails (Folsomia candida), isopods (Porcellio scaber) and enchytraeids (Enchytraeus crypticus). EC50s for the effect on reproduction of springtails and enchytraeids were 13 and 38 mg/kg, respectively for abamectin, and 42 and 170 mg/kg for doramectin. For earthworms, NOEC was 10 and 8.4 mg/kg for abamectin and doramectin effects on body weight. When exposed in faeces, springtails and enchytraeids gave LC50s and EC50s of 1.0-1.4 and 0.94-1.1 mg/kg dry faeces for abamectin and 2.2->2.4 mg/kg for doramectin. Earthworm reproduction was not affected. This study indicates a potential risk of avermectins for soil invertebrates colonizing faeces from recently treated sheep.     

The degradation of chlorpyrifos and diazinon in aqueous solution by ultrasonic irradiation: effect of parameters and degradation pathway

In this paper, elimination of two types of organophosphorus pesticides (OPPs), chlorpyrifos and diazinon spiked in aqueous solution by ultrasonic irradiation was investigated. Results showed that chlorpyrifos and diazinon could be effectively and rapidly degraded by ultrasonic irradiation, and the degradation of both pesticides was strongly influenced by ultrasonic power, temperature and pH value. Furthermore, two and seven products for the degradation of chlorpyrifos and diazinon formed during ultrasonic irradiation have been identified by gas chromatography-mass spectrometry, respectively. The hydrolysis, oxidation, hydroxylation, dehydration and decarboxylation were deduced to contribute to the degradation reaction and the degradation pathway for each pesticide under ultrasonic irradiation was proposed. Finally, the toxicity evaluation indicated that the toxicity decreased for diazinon solution after ultrasonic irradiation, but it increased for chlorpyrifos solution. The detoxification of OPPs by ultrasonic irradiation was discriminative.     

Electrospray Ionization Mass Spectrometry (ESI-MS) monitoring of the photolysis of diazinon in aqueous solution: Degradation route and toxicity of by-products against Artemia salina

The photolytic degradation of diazinon, an organophosphorus pesticide, in aqueous medium under assorted pH values was continuously monitored by direct infusion electrospray ionization mass spectrometry (ESI-MS). The results indicated that the UV radiation was quite efficient in promoting the pesticide degradation at the three pH levels evaluated (5, 7 and 8). The m/z of the most abundant ions observed in the mass spectra (MS), in conjunction with the fragmentation patterns of such ionic species (MS/MS data), made possible the proposition of chemical structures for the main by-products formed. As a result, routes for the photodegradation of diazinon in aqueous solution could thus be suggested. In the assays using Artemia salina (brine shrimp) it was verified that the photodegradation products exhibited much lower toxicity than the primary substrate. Aiming at mimicking the conditions ordinarily found in water treatment plants, an additional series of tests was conducted with a solution containing sodium hypochlorite and diazinon. This solution, when not exposed to UV radiation, exhibited high toxicity against the microorganisms. Under the influence of UV radiation, however, the toxicity rates decreased dramatically. This result is relevant because it points toward the confident application of UV radiation to neutralize the deleterious effects caused by diazinon (and perhaps other organophosphorus pesticides) as well as sodium hypochlorite to the environment.     

Comparative acute toxicities of selected pesticides to Anguilla anguilla.

The acute toxicities (24, 48, 72 and 96 hr) of eight pesticides to Anguilla anguilla were determined. The organochlorine pesticide, endosulfan was the most toxic, with LC50 values in the range of 0.042 to 0.041 mg/L. Endosulfan was followed in order of decreasing toxicity by diazinon, fenitrothion, chlorpyrifos, lindane, methidathion, trichlorfon and methylparathion. When fishes were exposed to the pesticides tested they exhibited signs of restlessness, erratic swimming, convulsions and difficulty in respiration. This response was more persistent in fishes exposed to organophosphorus pesticides.     

Long-term Hg pollution induced Hg tolerance in the terrestrial isopod Porcellio scaber (Isopoda, Crustacea)

The aim of our work was to assess the pollution-induced community tolerance (PICT) of isopod gut microbiota and pollution-induced isopod population tolerance (PIPT). Animals collected from a chronically Hg polluted and an unpolluted location were exposed for 14 days to 10microg Hg/g dry food under laboratory conditions. The lysosomal membrane stability, hepatopancreas epithelium thickness, feeding activity and animal bacterial gut microbiota composition were determined. The results confirm the hypothesis that the response to short-term Hg exposure differs for animals from the Hg polluted and the unpolluted field locations. The animals and their gut microbiota from the Hg polluted location were less affected by Hg in a short-term feeding experiment than those from the unpolluted environment. We discuss the pollution-induced population tolerance of isopods and their gut microbiota as a measure of effects of long-term environmental pollution. The ecological consequences of such phenomena are also discussed.     

Movement and deposition of two organophosphorus pesticides within a residence after interior and exterior applications

Post-application temporal and spatial distributions of two organophosphorus pesticides, diazinon (O,O-diethyl O-[6-methyl-2-(1-methylethyl)-4-pyrimidinyl] phosphorothioate, CAS No. 333-41-5) and chlorpyrifos [O,O-diethyl-O-(2-isopropyl-6-methyl-4-pyrimidinyl) phosphorothioate, CAS No. 2921-88-2], were monitored after homeowner applications for indoor and outdoor insect control. Samples of indoor air, vacuumable carpet dust, carpet dislodgeable residues, deposits on bare floors, table tops and dinnerware, surrogate food, and residues on children's hands and toys were taken before and up to 12 days after treatments in the family room, kitchen, and child's bedroom. Results from the study demonstrate the nature and magnitude of translocation of pesticides from the areas of application to surfaces accessible for human contact and permit comparisons of potential exposures via respiration and dermal contact/oral ingestion. Potential indoor inhalation exposures were estimated to be as high as 0.5 microg/kg/day for diazinon applied indoors and 0.05 microg/kg/day for chlorpyrifos applied to the outside perimeter of the house. While ingestion of carpet dust at the rate of 100 microg/day would have added a maximum of only approximately 0.01 microg/kg/day to the daily dose, residues found on the children's hands suggest that repeated mouthing could have contributed as much as 1-1.5 microg/kg/day. These estimates are below the U.S. Environmental Protection Agency (EPA) reference dose for chlorpyrifos, but exceed those for diazinon.     

Effects of fluoride on soil fauna mediated litter decomposition

To evaluate the effect of potentially toxic compounds on soil fauna mediated carbon and other mineral fluxes in poplar litter, a terrestrial micro-ecosystem (MES) was developed, with the isopod Porcellio scaber as a representative soil fauna species. An experiment was conducted, in which MES with isopods were compared to MES without isopods. Potassium fluoride was used as a model compound. The isopods influenced the microbial respiration negatively. No effect was found for the isopods on the nitrogen and phosphorus mineralization. After the first four weeks of the experiment the total respiration of the MES was increased and the phosphate concentration decreased at the highest fluoride concentration. After nine weeks, fluoride decreased ammonium, nitrate and phosphate concentrations in the litter. It is concluded that nitrogen and phosphorus mineralization in the MES are more sensitive to effects of fluoride that microbial respiration. The no observed effect concentrations for net mineralization of ammonium, nitrate and phosphorus were, respectively, 17, 5.3 and 53 micromol fluoride per gram dry weight of litter. Fluoride seems to be toxic for microbial processes at concentrations found in moderately polluted areas.     

Comparative acute toxicities of selected pesticides to Anguilla anguilla

Abstract

The acute toxicities (24, 48, 72 and 96 hr) of eight pesticides to Anguilla anguilla were determined. The organochlorine pesticide, endosulfan was the most toxic, with LC50 values in the range of 0.042 to 0.041 mg/L Endosulfan was followed in order of decreasing toxicity by diazinon, fenitrothion, chlorpyrifos, lmdane, methidathion, trichlorfon and methylparathion. When fishes were exposed to the pesticides tested they exhibited signs of restlessness, erratic swimming, convulsions and difficulty in respiration. This response was more persistent in fishes exposed to organophosphorus pesticides.     

Lead impact on nutrition, energy reserves, respiration and stress protein (hsp 70) level in Porcellio scaber (Isopoda) populations differently preconditioned in their habitats

The impact of lead on food consumption, energy metabolism and the stress protein (hsp 70) level was investigated in the woodlouse Porcellio scaber (Isopoda), a common representative of the saprophagous soil macrofauna. To examine possible acclimation or tolerance to lead in woodlice from a contaminated habitat, animals of two populations, one deriving from a lead-contaminated artillery range and one from an uncontaminated control stand, were exposed to a series of lead concentrations under otherwise constant laboratory conditions for a maximum of 80 days. The applied lead concentrations (at a maximum 7945 mg/kg food dry wt) did not have any significant quantitative effect on the food consumption of the isopods, although the population pre-exposed in the artillery range showed a tendency toward a higher food uptake than the control population. After 80 days of exposure, both populations showed an equal trend toward increasing their respiration as lead concentrations, that they had been fed on, were increased. Accordingly, the glycogen content of the body, in both populations, was elevated with increasing lead concentrations in the food. This effect was more pronounced in the pre-exposed isopod population than in the one from the control stand. The non-pre-exposed isopods showed a general tendency toward a lower protein content of their bodies than the pre-exposed ones, although no effect of the lead on this parameter could be statistically proven. The ability of the artillery range isopods to synthesise stress proteins (hsp 70) in response to lead contamination decreased at much lower lead concentrations in their food than in the non-pre-exposed control population, even though the artillery range isopods seemed to be equally or even slightly better equipped with energy storage products. Even though the better nutrient status of these animals might refer to some lead tolerance of the pre-exposed population, the stress protein data suggest that a metal-resistant Porcellio population did not evolve in this lead-contaminated site.     

Assessment of lead uptake in reptilian prey species

As part of an investigation determining the trophically available fraction of metals in a model terrestrial food web, i.e., invertebrate prey to Western fence lizards (Sceloporus occidentalis), we evaluated the ability of several invertebrate prey to bioaccumulate lead and to form metals-rich granules, which are hypothesized to be non-available to predators. Crickets (Acheta domestica), tenebroid beetle larvae (Tenebrio molitor), and isopods (Porcellio scaber) were selected as model prey organisms. Lack of standard exposure methodologies for these species has presented a barrier to trophic transfer evaluations, as each species has particular requirements that create challenges for designing exposure conditions. We were able to devise exposure conditions for all three species that allow long-term exposure studies. All prey organisms accumulated lead from contaminated food, and for all species the majority of the accumulated Pb was associated with the exoskeleton (>50%), with metals-rich granules accounting for most of the remaining accumulated lead.     

Sorption behaviour of acetochlor,atrazine, carbendazim,diazinon, imidacloprid and isoproturon on Hungarian agricultural soil

Laboratory studies were conducted to determine the sorption behaviour of six commonly used pesticides (acetochlor, atrazine, carbendazim, diazinon, imidacloprid and isoproturon) on Hungarian brown forest soil with clay alluviation (Luvisol) using the batch equilibrium technique. The sorption isotherms could be described by the Freundlich equation in non-linear form (n < 1) for all compounds, however in case of diazinon using the extended Freundlich equation proved to be a better approach. The adsorption constant related soil organic carbon content (Koc) calculated from Freundlich equation were 314 for acetochlor, 133 for atrazine, 2805 for carbendazim, 1589 for diazinon, 210 for imidacloprid and 174 for isoproturon. The octanol-water partition coefficients (Pow), which can be a useful parameter to predict of adsorption behaviour of a chemical on soil, and dissociation coefficients of these pesticides were calculated based on the chemical structure of them using a computerized expert system. The octanol-water partition coefficients were determined experimentally from high performance liquid chromatographic parameters as well. Good agreement was observed between experimental and the computer expert system estimated data. Computer estimated log Pow values ranged 0.5 and 3.86 for the examined pesticides, with imidacloprid and diazinon being the least and most hydrophobic respectively. Experimentally determined logPow ranged between 0.92 and 3.81 with the same tendency. It can be concluded that the Freundlich adsorption constants (Kf) are slightly related to the octanol-water partition coefficients of investigated chemicals, nevertheless no close correlation could be established because of the influence of further characteristics of solutes and soil.     

Trace level determination of selected organophosphorus pesticides and their degradation products in environmental air samples by liquid chromatography-positive ion electrospray tandem mass spectrometry

This paper describes a new analytical method for determination of organophosphorus pesticides (OPs) along with their degradation products involving liquid chromatography (LC) positive ion electrospray (ESI+) tandem mass spectrometry (MS-MS) with selective reaction monitoring (SRM). Chromatography was performed on a Gemini C6-Phenyl (150 mmx2.0 mm, 3 microm) with a gradient elution using water-methanol with 0.1% formic acid, 2 mM ammonium acetate mobile phase at a flow rate of 0.2 mL min(-1). The LC separation and MS/MS operating conditions were optimized with a total analysis time less than 40 minutes. Method detection limits of 0.1-5 microg L(-1) for selected organophosphorus pesticides (OP), OP oxon degradation products, and other degradation products: 3,5,6-trichloro-2-pyridinol (TCP); 2-isopropyl-6-methyl-4-pyrimidol (IMP); and diethyl phosphate (DEP). Some OPs such as fenchlorphos are less sensitive (MDL 30 microg L(-1)). Calibration curves were linear with coefficients of correlation better than 0.995. A three-point identification approach was adopted with area from first selective reaction monitoring (SRM) transition used for quantitative analysis, while a second SRM transition along with the ratio of areas obtained from the first to second transition are used for confirmation with sample tolerance established by the relative standard deviation of the ratio obtained from standards. This new method permitted the first known detection of OP oxon degradation products including chlorpyrifos oxon at Bratt's Lake, SK and diazinon oxon and malathion oxon at Abbotsford, BC in atmospheric samples. Atmospheric detection limits typically ranged from 0.2-10 pg m(-3).     

Comparative toxicity of chlorpyrifos, diazinon, malathion and their oxon derivatives to larval Rana boylii

Organophosphorus pesticides (OPs) are ubiquitous in the environment and are highly toxic to amphibians. They deactivate cholinesterase, resulting in neurological dysfunction. Most chemicals in this group require oxidative desulfuration to achieve their greatest cholinesterase-inhibiting potencies. Oxon derivatives are formed within liver cells but also by bacterial decay of parental pesticides. This study examines the toxicity of chlorpyrifos, malathion and diazinon and their oxons on the foothill yellow-legged frog (Rana boylii). R. boylii is exposed to agricultural pesticides in the California Central Valley. Median lethal concentrations of the parental forms during a 96 h exposure were 3.00 mg/L (24h) for chlorpyrifos, 2.14 mg/L for malathion and 7.49 mg/L for diazinon. Corresponding oxons were 10 to 100 times more toxic than their parental forms. We conclude that environmental concentrations of these pesticides can be harmful to R. boylii populations.     

Aquatic toxicity of cartap and cypermethrin to different life stages of Daphnia magna and Oryzias latipes

Cartap and cypermethrin, which are among the most widely used pesticides in many countries, are considered safe because of their low mammalian toxicity and their low persistence in the environment. However, recent findings of endocrine-disrupting effects and developmental neurotoxicity have raised concerns about the potential ecological impacts of these pesticides. We evaluated the aquatic toxicity of cartap [S,S'-(2-dimethylaminotrimethylene) bis(thiocarbamate), unspecified hydrochloride] and cypermethrin [(RS)-alpha-cyano-3-phenoxybenzyl-(1RS,3RS,1RS,3SR)-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylate], both individually and combined, on different life stages of the freshwater cladoceran Daphnia magna and a freshwater teleost, Japanese medaka (Oryzias latipes). The 96-hr Daphnia median effective concentrations (EC50s) for cartap and cypermethrin were 91.0 microg/L and 0.00061 microg/L, respectively. Rapid recovery of Daphnia was observed after short-term pulsed exposure to cartap and cypermethrin; there were no adverse effects on reproduction or survival 20 d after a 24 hr exposure to cartap up to 1240 microg/L and cypermethrin up to 1.9 microg/L. Chronic continuous exposure (for 21 d) of 7-d-old Daphnia to cypermethrin significantly reduced the intrinsic population growth rate in a concentration-dependent manner. However, because the intrinsic population growth rates were all above zero, populations did not decrease even at the highest experimental concentration of 200 ng/L. Exposure of Daphnia neonates (< 24 hr old) to cypermethrin for 21 d caused significant, sub-lethal reproduction-related problems, such as increased time to first brood, reduced brood size, and reduced total brood number, at 0.0002, 0.002, and 0.2 ng/L cypermethrin, but the intrinsic population growth rate was not significantly affected. Oryzias latipes was relatively more resistant to both pesticides. In particular, embryos appeared to be more resistant than juveniles or adults, which may be partly due to the protective role of the chorion. The incidence of larval fish deformity was significantly higher after a 96 hr exposure to as low as 250 microg/L of cartap or 40 microg/L of cypermethrin. The mixture of both compounds showed no synergistic toxicity. The extremely high acute-to-chronic ratio suggests that the standard acute lethal toxicity assessment might not reflect the true environmental hazards of these frequently used pesticides. Ecological hazard assessments of long-term low dose or pulsed exposures to cartap and cypermethrin may reveal more realistic consequences of these compounds in surface water.     

In-vivo interaction of some organophosphorus insecticides with different biochemical targets in white rats

A comparative study between five organophosphorus insecticides: Leptophos, EPN, Cyanofenphos, Chlorpyrifos and Diazinon, was carried out for acute oral toxicity to white rats and for their in vivo interaction at 1/10th of LD50 doses with the activity of six serum enzymes after 4 wks from oral administration. Leptophos, Chlorpyrifos and diazinon exerted significant inhibition particularly to glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), glutamyltransferase (GT) and lactate dehydrogenase (LDH). Adding ascorbic acid in the diet at 0.5% enhanced the acute oral toxicity of leptophos, chlorpyrifos and diazinon. For all the compounds, presence of ascorbic acid protected a number of the monitored serum enzymes from being inhibited except for leptophos. Ascorbic acid caused hypoglycemia with sublethal doses of leptophos, chlorpyrifos, and diazinon. The synergist piperonyl butoxide alone at 750 mg/kg dose inhibited the activity of the six serum enzymes. Presence of ascorbic acid in the diet intensified the inhibitory effect of piperonyl butoxide to all enzymes except for GOT.     

Factors determining amounts of fluoride in woodlice Oniscus asellus and Porcellio scaber, litter and soil near an aluminium reduction plant

Amounts of fluoride were measured in whole woodlice Oniscus asellus and Porcellio scaber, together with associated soil and litter, from 17 sites near an aluminium reduction plant at Holyhead, Anglesey, in North Wales. There was a strong positive correlation between amounts of fluoride in woodlice, soil and litter in all comparisons (P<0.02). Highest and lowest fluoride concentrations in woodlice differed by a factor of >50. Where both woodlice species occurred together at the more polluted sites, fluoride levels in Oniscus asellus were generally twice as high as those found in Porcellio scaber. At any given site, amounts of environmental fluoride were determined partly by distance and direction from the pot-rooms of the plant (a function of prevailing winds), modified by the presence of trees which acted as collectors for gaseous and particulate fluoride.     

Bioaccumulation in Porcellio scaber (Crustacea, Isopoda) as a measure of the EDTA remediation efficiency of metal-polluted soil

Leaching using EDTA applied to a Pb, Zn and Cd polluted soil significantly reduced soil metal concentrations and the pool of metals in labile soil fractions. Metal mobility (Toxicity Characteristic Leaching Procedure), phytoavailability (diethylenetriaminepentaacetic acid extraction) and human oral-bioavailability (Physiologically Based Extraction Test) were reduced by 85-92%, 68-91% and 88-95%, respectively. The metal accumulation capacity of the terrestrial isopod Porcellio scaber (Crustacea) was used as in vivo assay of metal bioavailability, before and after soil remediation. After feeding on metal contaminated soil for two weeks, P. scaber accumulated Pb, Zn and Cd in a concentration dependent manner. The amounts of accumulated metals were, however, higher than expected on the basis of extraction (in vitro) tests. The combined results of chemical extractions and the in vivo test with P. scaber provide a more relevant picture of the availability stripping of metals after soil remediation.