Inhibition of mitochondrial bioenergetics by carbaryl is only evident for higher concentrations -- Relevance for carbaryl toxicity mechanisms

Although pesticides have been useful in agriculture pest control, there is a considerable risk for human health and damage to ecosystems. Carbaryl is a carbamate often taken as a safe insecticide, although data on metabolic activities is still scarce, viz. mitochondrial toxicity. Therefore, it is the goal of this work to assay the compound on isolated mitochondria, a biochemical model already used with other pesticides. Mitochondria isolated from the livers of Wistar rats were assayed for bioenergetic parameters, namely mitochondrial respiration, membrane potential, membrane integrity and enzyme activities. For higher concentrations, it was observed that carbaryl has a depressive effect on mitochondrial respiration and on the generation of mitochondrial membrane potential, but with preservation of membrane integrity. A locus between Complex II and III appears particularly affected and the mitochondrial phosphorylation system relatively insensitive. Therefore, carbaryl inhibits mitochondrial respiration without affecting the phosphorylation complex. Carbaryl is toxic for mitochondria, although at concentrations higher as compared with other insecticide compounds. Mitochondrial toxicity should be excluded as one of the primary causes for carbaryl immediate toxicity, as concluded from the range of concentrations where carbaryl shows effective mitochondrial toxicity.     

Effects of the insect growth regulator (S)-methoprene on survival and reproduction of the freshwater cladoceran Moina macrocopa

This study examines the acute and chronic effects of the insecticide (S)-methoprene on the survival and reproduction of the freshwater cladoceran Moina macrocopa. In laboratory toxicity tests, the 24- and 48-h LC50 values for (S)-methoprene were 0.51 and 0.34 mg litre(-1), respectively. Survival, longevity and fecundity were reduced at 0.05 mg litre(-1) and higher concentrations. At 0.005 and 0.01 mg litre(-1), longevity and fecundity increased slightly as compared to control animals. If environmental concentrations of methoprene do not exceed 0.05 mg litre(-1), as is generally the case, application of this insecticide is unlikely to induce detrimental effects on natural cladoceran populations. The stimulatory effect of very low levels of methoprene on reproductive performance is consistent with the hypothesis of a regulatory role of juvenile hormone-like compounds in crustacean reproduction.     

Influence of an insect growth regulator on the larval development of an estuarine shrimp

The influence of methoprene, an insect growth regulator used in mosquito control, on larval development of the estuarine grass shrimp (Palaemonetes pugio) was examined in the laboratory. No grass shrimp larvae successfully completed metamorphosis when continuously exposed to 1000 microg methoprene litre(-1). Completion of larval metamorphosis was significantly reduced by exposure to 100 microg litre(-1) of the isomeric mixture (R,S)-methoprene but not the single isomer formulation (S)-methoprene. No statistically significant difference was revealed, however, in ability to inhibit metamorphosis between these two isomeric types across the broad range of exposure concentrations from 0.1 to 1000.0 microg litre(-1). The first two larval stages and the final premetamorphic larval stage were more sensitive to methoprene toxicity than intermediate larval stages. Methoprene exposure did not alter either the duration of total larval development or the total number of larval stages prior to metamorphosis.     

Titanium dioxide nanoparticles relieve biochemical dysfunctions of fifth-instar larvae of silkworms following exposure to phoxim insecticide

Phoxim insecticide is widely used in agriculture, which is toxic to insect pests and nontarget organisms. The phoxim poisoning is hard to prevent for silkworms. TiO₂ NPs have been widely applied in whitening, brightening foods, toothpaste or sunscreens, and orally-administered drugs. However, whether TiO₂ NPs can increase resistance of silkworm to phoxim poisoning has not been reported. The results demonstrated that added TiO₂ NPs significantly decreased reduction of protein, glucose and pyruvate contents, lactate dehydrogenase, succinate dehydrogenase and malate dehydrogenase activities, and attenuated increases of free amino acids, urea, uric acid and lactate levels, activities of protease, alanine aminotransferase and aspartate aminotransferase in the hemolymph of silkworms caused by phoxim exposure. From the present study, it is clearly evident that added TiO₂ NPs may relieve toxic impacts of phoxim insecticide on silkworm metabolism, which in turn may result in an increase in silk yield.     

Polychlorinated biphenyls and organochlorine pesticides in Baltic Sea sediments and bivalves

Insecticide resistance is an evolutionary response to insecticides and, as such, important for environmental biomonitoring and for pest management. Fitness disadvantage in the absence of insecticide is a frequent assumption in models of insecticide resistance evolution, which was observed in different insect species. Fitness studies are based in demographic performance of isolated populations without direct interaction between insecticide-resistant and -susceptible populations. Here we reported a study of direct competition following a factorial bivariate design between an insecticide-susceptible population of the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), and one of two insecticide-resistant populations - one exhibiting fitness disadvantage based on demographic studies (Juiz de Fora) and other not exhibiting it (Jacarezinho). Total number of insects, mortality by insecticide, insect body mass and respiration rate were recorded after three generations of competition. Indeed, fitness disadvantage was observed in the resistant population from Juiz de Fora, but not in the population from Jacarezinho, as expected. The higher body mass and respiration rate of the insecticide-resistant insects from Jacarezinho are probably mitigating the physiological costs associated with the insecticide resistance minimizing their fitness disadvantage, what does not take place with the insecticide-resistant insects from Juiz de Fora. These distinct responses between the insecticide-resistant populations are probably related to the length and intensity of field-selection with insecticides.     

Comparative effects of the Roundup and glyphosate on mitochondrial oxidative phosphorylation

The potential toxicity of the herbicide Roundup and its fundamental substance (glyphosate) was tested in bioenergetic functions of isolated rat liver mitochondria. Roundup stimulates succinate-supported respiration twice, with simultaneous collapse of transmembrane electrical potential, while glyphosate used in the same concentrations does not induce any significant effect. Additionally, Roundup depresses state 3 respiration by about 40%, at 15 mM, whereas uncoupled respiration in the presence of FCCP is depressed by about 50%. Depression of uncoupled respiratory activity is mediated through partial inhibition of mitochondrial complexes II and III, but not of complex IV. The phosphorylative system was affected by both a direct and an indirect effect on the F0F1 ATPase activity. The addition of uncoupled concentrations of Roundup to Ca2+-loaded mitochondria treated with Ruthenium Red resulted in non-specific membrane permeabilization, as evidenced by mitochondrial swelling in isosmotic sucrose medium. Therefore, the uncoupling of oxidative phosphorylation is also related to the non-specific membrane permeabilization induced by Roundup. Glyphosate alone does not show any relevant effect on the mitochondrial bioenergetics, in opposition to Roundup formulation products. The differences in the toxicity observed could be either attributed to some products of Roundup or to a synergic effect of glyphosate and formulation products. Bearing in mind that mitochondria is provided with a variety of bioenergetic functions mandatory for the regulation of intracellular aerobic energy production and electrolyte homeostasis, these results question the safety of Roundup on animal health.     

Acute toxicity of the insect larvicide abate® (temephos) on the fish tilapia melanopleura and the dragonfly larvae neurocordelia virginiensis.

Abstract

Acute bioassay tests on the toxic effects of the insect larvicide Abate® (temephos) on the mouth brooder cichlid fish Tilapia melanopleum and the dragonfly larvae (Odonata) Neurocordulia virginiensis were conducted in static non renewal toxicity test set ups. The 96h‐LC50 (95% confidence intervals) was 30.2 (20.5‐ 44.20) mg/L for the fish and 2.0 (1.16–2.0) mg/L for the dragonfly larvae. The dragonfly larvae were 15 times more susceptible to the larvicide than the tilapia. The calculated NOEC (No Observable Effect Concentration) was 14.1 mg/L for the fishand less than 1.0 mg/L for the insect larvae. The estimated ‘safe’ concentration of the pesticide to the fish was 3.0 mg/L and 0.2 mg/L for insect larvae. These figures are far above the concentrations approved for use in the control of mosquito larvae(0.0004–0.01 mg/L). It appears that the application of toxic levels of the insecticide for the elimination of some aquatic invertebrates may be ‘safe’ for normal survival, growth and reproduction offish and some aquatic insect larvae.     

Effect of aldicarb on the growth, respiration and (14C) - glucose metabolism of Axotobacter chroococcum BEIJ.

Aldicarb [2-methyl-2(methyl thio) propionaldehyde-O-methyl carbamoyl oxime], which forms the active ingredient of a systemic oximic carbamate insecticide, at 2 ppm level did not affect the in vitro growth and respiration of Azotobacter chroococcum Beij, while concentrations at 5 ppm and 10 ppm levels were inhibitory. The insecticide treatment at 10 ppm level suppressed the assimilation of (14C) - glucose in the whole cells and in the cellular constitutents viz., cold - TCA soluble, hot TCA soluble fractions and insoluble residue. However, the 14C - incorporation in the alcohol soluble and alcohol-ether soluble fractions was enhanced indicating that aldicarb considerably altered the glucose metabolism of the organism.     

In vitro effects of acephate on carbonic anhydrase activity in the blood and gills of rainbow trout, Salmo gairdneri

Acephate, a water-soluble organophosphate pesticide used to control terrestrial insect pests, may enter aquatic ecosystems in the course of its use and adversely affect fish populations. The in vitro effects of this insecticide on gill and red blood cell (RBC) carbonic anyhdrase (CA) activity in rainbow trout were investigated over a range of 100 mg/1 (0.55 mM) to 50,000 mg/l (273 mM) to assess the manner in which acephate might affect respiratory capacity in exposed fish. Concentrations required to produce 50% inhibition of CA activity in the gill and RBC preparations were 38,000 mg/l (207 mM) and 8,900 mg/l (48 mM) respectively. The toxic action of acephate may be related to inhibition of CA activity in the blood and gills with resultant disturbances of respiratory capacity and salt balance.     

The influence of lindane and lindane metabolites on microsomal and mitochondrial ATPase in vitro

In vitro investigations of the influence of lindane and its metabolites were performed on microsomal and mitochondrial ATPases from liver, kidney and brain of rat and mouse. The microsomal Na+-K+-ATPases in rat liver were inhibited by the tested substances. An increase of activity was observed only with 2.5 X 10(-5) M gamma-HCH. Effects on the microsomal Na+-K+-ATPase from kidney and brain of rat were also indicated. The mitochondrial enzyme in rat liver was stimulated by all the compounds tested at concentrations of 10(-4) M - 10(-2) M. The effects on mitochondrial enzymes from kidney and brain varied in dependence on the tested substances. In the microsomes and mitochondria of mouse an influence on the Na+-K+-ATPases similar to the effects on the preparations from organs of rat was evident.     

Excretion and toxicity of Cyolane in the rat

The excretion of ¹⁴C-labelled Cyolane, 24 hours after single oral application was found to be about 50% of the applied dose in urine, faeces and expired air.The activity of Acetylcholine esterase in brain, plasma and erythrocytes, and liver succinate dehydrogenase was studied for 16 weeks in 4 groups of rats after repeated orally daily doses of Cyolane, 0.9, 0.45, 0.09 and 0.045 mg/kg.It has been found that at all the dose levels there were cumulative inhibition effects in brain and blood choline esterase and liver succinate dehydrogenase till the 2nd week and then there was a recovery at low dose rates in blood choline esterase and liver succinate dehydrogenase activities. After 6 - 16 weeks the effects were nearly compensated. At higher doses almost all the rats died after 6 weeks due to the insecticide toxicity.     

Effect of Dursban 480 EC (chlorpyrifos) and Talstar 10 EC (bifenthrin) on the physiological and genetic diversity of microorganisms in soil

This investigation was undertaken to determine the impact of the insecticides Dursban 480 EC (with organophosphate compound chlorpyrifos as the active ingredient) and Talstar 10 EC (with pyrethroid bifenthrin as the active ingredient) on the respiration activity and microbial diversity in a sandy loam luvisol soil. The insecticides were applied in two doses: the maximum recommended dose for field application (15 mg kg^?1 for Dursban 480 EC and 6 mg kg^?1 for Talstar 10 EC) and a 100-fold higher dose for extrapolation of their effect. Bacterial and fungal genetic diversity was analysed in soil samples using PCR DGGE and the functional diversity (catabolic potential) was studied using BIOLOG EcoPlates at 1, 3, 7, 14, 28, 56 and 112 days after insecticide application. Five bacterial groups (α, β, γ proteobacteria, firmibacteria and actinomycetes) and five groups of fungi or fungus-like microorganisms (Ascomycota, Basidiomycota, Chytridiomycota, Oomycota and Zygomycota) were analysed using specific primer sets. This approach provides high resolution of the analysis covering majority of microorganisms in the soil. Only the high-dose Dursban 480 EC significantly changed the community of microorganisms. We observed its negative effect on α- and γ-proteobacteria, as the number of OTUs (operational taxonomic units) decreased until the end of incubation. In the β-proteobacteria group, initial increase of OTUs was followed by strong decrease. Diversity in the firmibacteria, actinomycetes and Zygomycota groups was minimally disturbed by the insecticide application. Dursban 480 EC, however, both positively and negatively affected certain species. Among negatively affected species Sphingomonas, Flavobacterium or Penicillium were detected, but Achromobacter, Luteibacter or Aspergillus were supported by applied insecticide. The analysis of BIOLOG plates using AWCD values indicated a significant increase in metabolic potential of microorganisms in the soil after the high-dose Dursban application. Analysis of respiration demonstrated high microbial activity after insecticide treatments; thus, microbial degradation was relatively fast. The half-life of the active insecticide compounds were estimated within the range of 25 to 27 days for Talstar and 6 to 11 days for Dursban and higher doses stimulated degradation. The recommended dose levels of both insecticides can be considered as safe for microbial community in the soil.     

The effects of elevated metals on benthic community metabolism in a Rocky Mountain stream

The effects of elevated metals (dissolved Zn, Mn and/or Fe) in a Rocky Mountain stream were assessed using measures of primary productivity, community respiration and water-column toxicity. Primary productivity was measured as rates of O2 evolution from natural substrates incubated in situ in closed chambers. Oxygen depletion within these chambers, when incubated in the dark, provided estimates of periphyton community respiration. Sediment community respiration on fine-grained sediments, collected and composited along each stream study reach, was measured on-site by incubating these sediments in closed chambers and measuring O2 depletion. Toxicity was measured as percent mortality of Ceriodaphnia dubia during 48 h acute tests. Gross (GPP) and net primary productivity (NPP) decreased significantly with increasing metal concentrations, from 10.88 +/- 1.46 g O2 m(-2) day(-1) to 0.83 +/- 0.20 g O2 m(-2) day(-1) and 9.85 +/- 1.43 g O2 m(-2) day(-1) to 0.81 +/- 0.20 g O2 m(-2) day(-1), respectively for the reference and most impacted site. Community respiration (CR) declined from 0.65 +/- 0.08 g O2 m(-2) day(-1) to 0.02 +/- 0.01 g O2 m(-2) day(-1) with increasing metal concentrations. Sediment community respiration (SCR) decreased from 0.26 +/- 0.02 g O2 m(-2) day(-1) to 0.01 +/- 0.01 g O2 m(-2) day(-1) at these same sites. Ceriodaphnia dubia mortality increased from 0% at the reference site to 95 +/- 5% at the most impacted sites. Net daily metabolism, quantum yield and assimilation ratio all decreased with increasing metal concentrations, suggesting that both autotrophic and heterotrophic components of the periphyton community were impaired. Overall, functional measures were able to discern sites receiving greater metal impacts from less-impacted sites, with combinations of dissolved metals explaining between 25 and 92% of the variance in the regression models. Using these regression models we were able to calculate lethal and inhibition concentrations of dissolved Zn in the Eagle River. The lethal concentration (LC50) of Zn for Ceriodaphnia dubia is 123 mg liter(-1). The concentrations of Zn which inhibited respiration (IC50) were 177 mg liter(-1) for CR and 199 mg liter(-1) for SCR. These results indicate functional measures may be as sensitive to metal concentrations as acute toxicity tests.     

Insecticide residues in the blood serum and domestic water source of cacao farmers in Southwestern Nigeria

The blood serum of cacao farmers and their domestic water sources were analyzed for insecticide residues in selected cacao growing communities of Southwestern Nigeria. The farmers were grouped into five exposure periods based on their years of involvement in insecticide application, viz, <5 years, 5-9 years, 10-14 years, 15-19 years and >20 years. The residue analyses revealed that 42 out of the 76 farmers had residues of diazinon, endosulfan, propoxur and lindane in their blood; and 47.6% out of these farmers belonged in the >20 years exposure duration period. About 34% of the farmers had diazinon with a mean concentration of 0.067 mg kg(-1), 29% endosulfan (mean=0.033 mg kg(-1)), 23% propoxur (mean=0.095 mg kg(-1)), and 17% lindane (mean=0.080 mg kg(-1)) in their blood. The residues of lindane, endosulfan and propoxur in all the exposure duration categories were found to be far below the no observable adverse effect level (NOAEL) while diazinon residues detected in the blood serum of the farmers in all the exposure duration categories exceeded the NOAEL of 0.02 mg kg(-1) for the insecticide. The study also revealed that the sources of drinking water had been contaminated with dazinon and propoxur in some of the farmers' localities; and the concentrations of the insecticides exceeded the acceptable daily intake (ADI). It is concluded that cacao farmers in Southwestern Nigeria may have been occupationally exposed due to insecticide application for mirid control in their cacao plantations; and the exposure at times is of such magnitude as to be hazardous to the farmers and their respective communities.     

Effects of carbaryl,chlorpyrifos and endosulfan on growth,reproduction and respiration of tropical epigeic earthworm,Perionyx excavatus (Perrier)

Effects of sub-lethal doses of carbaryl (1-Naphthyl-methylcarbamate), chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridinyl-phosphorothioate) and endosulfan (6,7,8,9,10,10-Hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin-3-oxide), respectively a carbamate, an organophosphate and an organochlorine insecticide on growth, reproduction and respiration of the tropical earthworm, Perionyx excavatus (Perrier) were investigated under laboratory conditions. The results showed significant reduction in biomass, production and hatching of cocoon and production of juveniles of the worms exposed to 0.75 to 3.03 mg/kg soil of carbaryl, 0.91 to 3.65 mg/kg soil of chlorpyrifos and 3.75 to 15.0 μg/kg soil of endosulfan corresponding to 12.5 to 50 % of LC₅₀ value of the respective insecticide for P. excavatus. Endosulfan was found most dangerous among the three insecticides followed by carbaryl and chlorpyrifos. There was no hatching of the worms at endosulfan treatment 5.0 μg/kg soil (25 % LC₅₀) or above while the highest dose of carbaryl and chlorpyrifos (50 % of LC₅₀) rendered respectively 87.13 and 24.84 % reductions in hatching as compared to control. Chlorpyrifos produced no change in respiration of the worms except at the highest dose, while the worms showed an increase in evolution of CO₂ at all doses of carbaryl and endosulfan. Based on the recommended agricultural dose of each insecticide, it was concluded that application of endosulfan and carbaryl was potentially dangerous to earthworms.     

Use of artificial sediments in a comparative toxicity study with larvae and postlarvae of the grass shrimp, Palaemonetes pugio

An artificial sediment was tested for use in evaluating the potential hazard of toxicants on benthic organisms. The seawater-sediment system was assessed by use of the pyrethroid insecticide, fenvalerate, as the model toxicant for testing with larvae of the grass shrimp, Palaemonetes pugio, an ecologically important estuarine species. The sediment was prepared from commercially available components, and mixed with the toxicant to provide concentrations of 1, 10 and 100 microg fenvalerate kg(-1) dry sediment in 20 ppt seawater. Sediment free of the insecticide served as the control. Throughout the study, fenvalerate was not detected in the water column, but was measured in sediment at the nominal concentration of 100 microg kg(-1). The P. pugio population was adversely affected by fenvalerate. The effect occurred at metamorphosis, when larvae changed from pelagic individuals to benthic organisms. At this period, larvae were in direct contact with sediment. A portion of the population was tolerant of the insecticide.     

Soil dissipation of juvenile hormone analog insecticide pyriproxyfen and its effect on the bacterial community

This investigation was undertaken to examine the dissipation rate of pyriproxyfen as well as the change in the soil bacterial community. Residues of pyriproxyfen were measured using high performance liquid chromatography (HPLC) and the changes in bacterial community were determined by comparing the 16S rDNA bands on patterns by denaturing gradient gel electrophoresis (DGGE). The dissipation of pyriproxyfen was affected by both the concentration applied and incubation temperature. Lower concentrations (1 mg Kg(-1)) and higher incubation temperatures (30 and 40°C) showed more rapid dissipation rates. The population of microbial community decreased rapidly after incubation with 10 mg Kg(-1) of pyriproxyfen for 91 days, indicating the toxicity of pyriproxyfen toward bacterial communities in a closed soil ecosystem. Lower concentrations of pyriproxyfen showed less toxicity toward the microbial community. From cluster analysis, the structure of the bacterial community showed roughly a 60 % similarity throughout the experiment period in the control experiment, indicating the stability within soil microbiota without chemical agitation. However, the similarity was lower than 50 % both in the one and 10 mg Kg(-1) of insecticide pyriproxyfen spiked experiment, indicating the soil bacterial community changed after the insecticide pyriproxyfen was applied.     

Effects of carbaryl, chlorpyrifos and endosulfan on growth, reproduction and respiration of tropical epigeic earthworm, Perionyx excavatus (Perrier)

Effects of sub-lethal doses of carbaryl (1-Naphthyl-methylcarbamate), chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridinyl-phosphorothioate) and endosulfan (6,7,8,9,10,10-Hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin-3-oxide), respectively a carbamate, an organophosphate and an organochlorine insecticide on growth, reproduction and respiration of the tropical earthworm, Perionyx excavatus (Perrier) were investigated under laboratory conditions. The results showed significant reduction in biomass, production and hatching of cocoon and production of juveniles of the worms exposed to 0.75 to 3.03 mg/kg soil of carbaryl, 0.91 to 3.65 mg/kg soil of chlorpyrifos and 3.75 to 15.0 μg/kg soil of endosulfan corresponding to 12.5 to 50 % of LC(50) value of the respective insecticide for P. excavatus. Endosulfan was found most dangerous among the three insecticides followed by carbaryl and chlorpyrifos. There was no hatching of the worms at endosulfan treatment 5.0 μg/kg soil (25 % LC(50)) or above while the highest dose of carbaryl and chlorpyrifos (50 % of LC(50)) rendered respectively 87.13 and 24.84 % reductions in hatching as compared to control. Chlorpyrifos produced no change in respiration of the worms except at the highest dose, while the worms showed an increase in evolution of CO(2) at all doses of carbaryl and endosulfan. Based on the recommended agricultural dose of each insecticide, it was concluded that application of endosulfan and carbaryl was potentially dangerous to earthworms.     

Growth, respiration and nitrogen content in needles of Scots pine exposed to elevated ozone and carbon dioxide in the field

Single Scots pine (Pinus sylvestris L.) trees, aged 30 years, were grown in open-top chambers and exposed to two atmospheric concentrations of ozone (O3; ambient and elevation) and carbon dioxide (CO2) as single variables or in combination for 3 years (1994-1996). Needle growth, respiration and nitrogen content were measured simultaneously over the period of needle expansion. Compared to ambient treatment (33 nmol mol(-1) O3 and 350 micromol mol(-1) CO2) doubled ambient O3 (69 nmol mol(-1)) significantly reduced the specific growth rates (SGRs) of the needles in the early stage of needle expansion and needle nitrogen concentration (N1) in the late stage, but increased apparent respiration rates (ARRs) in the late stage. Doubled ambient CO2 (about 650 micromol mol(-1)) significantly increased maximum SGR but reduced ARR and N1 in the late stage of needle expansion. The changes in ARR induced by the different treatments may be associated with treatment-induced changes in needle growth, metabolic activities and turnover of nitrogenous compounds. When ARR was partitioned into its two functional components, growth and maintenance respiration, the results showed that neither doubled ambient O3 nor doubled ambient CO2 influenced the growth respiration coefficients (Rg). However, doubled ambient O3 significantly increased the maintenance respiration coefficients (Rm) regardless of the needle development stage, while doubled ambient CO2 significantly reduced Rm only in the late stage of needle expansion. The increase in Rm under doubled ambient O3 conditions appeared to be related to an increase in metabolic activities, whereas the decrease in Rm under doubled ambient CO2 conditions may be attributed to the reduced N1 and turnover rate of nitrogenous compounds per unit. The combination of elevated O3 and CO2 had very similar effects on growth, respiration and N1 to doubled ambient O3 alone, but the interactive mechanism of the two gases is still not clear.     

Evaluation of bendiocarb cytotoxicity in mammalian and insect cell cultures

There is an increasing need for rapid and easily interpreted in vitro assays to screen for possible cytotoxicity of pesticides. The objective of this study was to investigate the effect of the carbamate insecticide bendiocarb on mammalian and insect cell cultures. The cytotoxicity of this insecticide was evaluated by cell proliferation and cellular damage was assessed by evaluation of the cytopathic effect and lactate dehydrogenase (LDH) leakage. Cells of insect origin (Sf21) were the most sensitive to bendiocarb with significant (P < 0.01) suppression of their proliferative activity ranging from 10(-1)-10(-5) M. However, significant suppression of proliferative activity was also recorded in rat liver cells (WBF344; 10(-1)-10(-3) M; P < 0.01-0.05) and rabbit kidney cells (RK13; 10(-1) M; P < 0.01). In contrast with the proliferation activity of cells, a cytopathic effect based on cellular damage and LDH leakage into the medium was observed only at the highest concentration (10(-1) M) in RK 13 and WBF344 cells, but not in the Sf21 insect cell line. Our results indicate that bendiocarb exposure caused a cell-type dependent decrease in cell proliferation; however, cell damage and LDH leakage into the medium were not present or were strongly limited, dependent on the cell phenotype. Cell proliferation was shown as a sensitive indicator for evaluation of the cytotoxic effect of bendiocarb in vitro; on the other hand, microscopic signs of cellular damage and LDH leakage were insufficient in vitro markers.