Impaired terrestrial and arboreal locomotor performance in the western fence lizard (Sceloporus occidentalis) after exposure to an AChE-inhibiting pesticide

We examined the effects of a commonly used AChE-inhibiting pesticide on terrestrial and arboreal sprint performance, important traits for predator avoidance and prey capture, in the western fence lizard (Sceloporus occidentalis). Lizards were exposed to carbaryl (2.5, 25, and 250 microg/g) and were raced before and 4, 24, and 96 h after dosing. In the terrestrial setting, exposure to low concentrations of carbaryl had stimulatory effects on performance, but exposure to the highest concentration was inhibitory. No stimulatory effects of carbaryl were noted in the arboreal environment and performance in lizards was reduced after exposure to both the medium and highest dose of carbaryl. Our findings suggest that acute exposure to high concentrations of carbaryl can have important sublethal consequences on fitness-related traits in reptiles and that arboreal locomotor performance is a more sensitive indicator of AChE-inhibiting pesticide poisoning than terrestrial locomotor performance.     

Histopathological effects of carbaryl on testes of snake-eyed lizard, Ophisops elegans

Background, aim and scope  

Due to their ecological niche and insectivore nature, lizards are of increased risk of exposure to pesticides in agricultural areas. In addition to their potential direct effects on non-target species, insecticides can also result in indirect impacts on lizard population by reducing their food source. Carbaryl is a common insecticide that is widely used in areas of Turkey that are home to a variety of reptiles. However, to date, little is known about the potential effects of the exposure of reptiles such as lizards or snakes to this pesticide. The aim of the study was to investigate toxic effects of carbaryl on the testes of snake-eyed lizard, Ophisops elegans that is common to regions in Turkey where Carbaryl is applied and that it can be easily cultured in the laboratory.     

Effects of repeated application of carbaryl on zooplankton communities in experimental ponds with or without the predator Chaoborus

Zooplankton communities with or without Chaoborus larvae were established in outdoor experimental ponds, to which the insecticide carbaryl was applied repeatedly at 10 or 100 microg litre(-1). In the ponds without Chaoborus, Cladocera dominated, but the species composition differed among the treatments. A large-sized Cladocera Daphnia galeata dominated the controls, the medium-sized cladocerans Diaphanosoma brachyurum and Moina micrura became dominant in the low-dose treatment, and the small-sized Bosmina fatalis increased in the high-dose treatment. These results indicated differential sensitivity to carbaryl among the cladocerans, and that smaller Cladocera were more tolerant of the chemical than the larger one. In the ponds with Chaoborus, rotifers dominated the zooplankton, probably because Chaoborus released rotifers from competition with cladocerans and calanoid copepods, which were eliminated by the Chaoborus predation. No effects of low-dose carbaryl treatment were detected on zooplankton communities in the Chaoborus ponds. The dominance by rotifers, organisms tolerant to carbaryl, minimized the effects. Thus, Chaoborus altered the zooplankton community responses to the chemical application by changing community structure. Repeated application of high-dose carbaryl did affect the rotifer community, decreasing the dominance of Polyarthra trigla and increasing that of Keratella valga. These rotifer species may differ in their sensitivity to carbaryl.     

Carbaryl-A selective genotoxicant

Mutagenic effects of carbaryl, a contact insecticide with slight systemic properties, have been investigated employing histidine reversion assay in Salmonella typhimurium strains and in vivo chromosomal aberrations in root meristems of Allium cepa. A detailed investigation revealed that carbaryl did not enhance significantly the frequency of histidine revertants in any of the strains of Salmonella i.e. frameshift mutagen tester (TA98), base pair substitution tester strain (TA1535) and ochre mutant strain (TA102). The supplementation with S9 mix did not have any appreciable effect. S14 prepared from wheat seedlings also did not enhance the reversion frequency significantly. However, carbaryl induced both clastogenic and physiological types of chromosomal aberration. The spectrum of chromosomal aberrations included c-mitosis, stickiness, vagrant chromosomes, polyploidy multi-polarity, delayed anaphases, end to end joining of chromosomes, chromosome breaks, ring chromosomes and anaphase bridges. The frequency of chromosomal aberrations was reduced by transferring the carbaryl treated bulbs to distilled water for 24 and 48 h. Similarly, recovery in the mitotic index was noticed by such transfer. The differences between the two assays may be attributed to differences in the metabolism of the test organisms.     

Effects of long- and short-term exposure to carbaryl on survival, growth and reproduction of Daphnia ambigua

In laboratory experiments a cladoceran, Daphnia ambigua, was exposed for six instars to the insecticide carbaryl at five different concentrations (1, 2, 3, 4, 5 microg litre(-1)) or received short-term (10-h) exposure to 5 microg carbaryl litre(-1) at different life stages of the animal. In the long-term exposure experiments carbaryl at 1 microg litre(-1) did not have any detectable effects on the animals, whereas 2 microg litre(-1) reduced the survival, growth and reproduction of the animals, and higher concentrations killed all the animals examined before they grew to the fifth instar. These results indicate that the range of sublethal concentrations of carbaryl for D. ambigua is narrow. In the short-term exposure experiments the animals did not suffer any damage when they were exposed to the chemical during the egg stage. However, their growth and egg production were reduced when the short-term exposure was conducted during the life stages from the final embryo stage to the third instar. The effect was most marked when the animals were treated during the first instar, indicating that this is the stage when the animals are most sensitive to the chemical.     

Nondestructive indices of trace element exposure in squamate reptiles

Compared with birds, mammals, fish, and even amphibians, very little is known about the effects of contaminants on reptiles. Recent evidence that many reptile populations may be declining has stimulated demand for toxicological studies of reptiles as well as development of nondestructive sampling techniques useful for assessing and monitoring contaminant exposure. The current study experimentally evaluated the utility of shed skins, tail clips, and blood samples as nondestructive indices of trace element exposure in banded water snakes, Nerodia fasciata. For 13.5 months, snakes were either fed fish from a coal ash-contaminated site or uncontaminated food from a reference site. Snakes fed contaminated prey accumulated As, Cd, Se, Sr, and V in various organs (i.e. liver, kidney, and/or gonads). Moreover, non-parametric discriminant function analysis revealed that snakes could be placed in two groups that reliably reflected their experimental diet based upon Se, Sr, and As concentrations in tail clips, blood, and/or shed skins. We suggest that nondestructive sampling techniques, particularly analyses of blood and tail clips, may be easily applied in evaluations of contaminant exposure in the field and laboratory and may prevent excessive destructive sampling of potentially threatened reptile species.     

Predictability by Box-Behnken model for carbaryl adsorption by soils of Indian origin

This study was undertaken to investigate the adsorption capacity of carbaryl on four Indian soils with different physiochemical properties. A batch adsorption study was carried out in order to evaluate the maximum adsorption capacity of carbaryl using a Response Surface Methodology (RSM). The effects of operating parameter such as initial carbaryl concentration (1–20 mgL^?1), adsorbent dosage (0.5–6 g) and contact time (10–180 min) were examined. The proposed quadratic model for Box-Behnken design fits very well to the experimental data because it may be used to navigate design space according to ANOVA results. The regression co-efficient (R²) of the models developed and the results of validation experiments conducted at optimal conditions strongly suggests that the predicted values are in good agreement with experimental results. Contour and response surface plots are used to determine the interactions effects of main factors and optimal conditions of the process. The experiment can be utilized as a guideline for better understanding of carbaryl adsorption onto soil under different operating conditions. The results show that the forest soil is most efficient in binding carbaryl (Sevin) than the other types of soil tested.     

Acute and chronic ecotoxicity of carbaryl with a battery of aquatic bioassays

The ecotoxic effects of carbaryl (carbamate insecticide) were investigated with a battery of four aquatic bioassays. The nominal effective concentrations immobilizing 50% of Daphnia magna (EC50) after 24 and 48 h were 12.76 and 7.47 µg L^?1, respectively. After 21 days of exposure of D. magna, LOECs (lowest observed effect concentrations) for cumulative molts and the number of neonates per surviving adult were observed at carbaryl concentration of 0.4 µg L^?1. An increase of embryo deformities (curved or unextended shell spines) was observed at 1.8 and 3.7 µg L^?1, revealing that carbaryl could act as an endocrine disruptor in D. magna. Other bioassays of the tested battery were less sensitive: the IC50-72h and IC10-72h of the algae Pseudokirchneriella subcapitata were 5.96 and 2.87 mg L^?1, respectively. The LC50-6d of the ostracod Heterocypris incongruens was 4.84 mg L^?1. A growth inhibition of H. incongruens was registered after carbaryl exposure and the IC20-6d was 1.29 mg L^?1. Our results suggest that the daphnid test sensitivity was better than other used tests. Moreover, carbaryl has harmful and toxic effects on tested species because it acts at low concentrations on diverse life history traits of species and induce embryo deformities in crustaceans.     

The excretion of pesticides in saliva and its value in assessing exposure

The possibility that pesticides are secreted in the saliva and that this secretion can be used to assess exposure to pesticides is discussed. Both Kepone and carbaryl were tested through a series of in vitro studies and in vivo experiments including both acute and subchronic studies. In the in vivo studies, both Kepone and carbaryl were detected in the saliva in the first twenty-four hours, but seldom thereafter. Analysis of urinary samples indicated a rapid clearance of the substances from the animals. In vitro uptake and efflux studies suggested that movement of both substances was by passive diffusion and reached a plateau after 60 minutes of incubation. Efflux rates were unaffected by experimental conditions, but were ten times faster for carbaryl.     

Absorption of current use pesticides by snapping turtle (Chelydra serpentina) eggs in treated soil

Reptiles often breed within agricultural and urban environments that receive frequent pesticide use. Consequently, their eggs and thus developing embryos may be exposed to pesticides. Our objectives were to determine (i) if turtle eggs are capable of absorbing pesticides from treated soil, and (ii) if pesticide absorption rates can be predicted by their chemical and physical properties. Snapping turtle (Chelydra serpentina) eggs were incubated in soil that was treated with 10 pesticides (atrazine, simazine, metolachlor, azinphos-methyl, dimethoate, chlorpyrifos, carbaryl, endosulfan (I and II), captan, and chlorothalonil). There were two treatments, consisting of pesticides applied at application rate equivalents of 1.92 or 19.2 kg a.i/ha. Eggs were removed after one and eight days of exposure and analyzed for pesticides using gas chromatography coupled with a mass selective detector (GC-MSD) or high performance liquid chromatography (HPLC). Absorption of pesticides in eggs from soil increased with both magnitude and duration of exposure. Of the 10 pesticides, atrazine and metolachlor generally had the greatest absorption, while azinphos-methyl had the lowest. Chlorothalonil was below detection limits at both exposure rates. Our preliminary model suggests that pesticides having the highest absorption into eggs tended to have both low sorption to organic carbon or lipids, and high water solubility. For pesticides with high water solubility, high vapor pressure may also increase absorption. As our model is preliminary, confirmatory studies are needed to elucidate pesticide absorption in turtle eggs and the potential risk they may pose to embryonic development.     

Distribution and persistence of carbaryl in some terrestrial and aquatic components of a forest environment

Abstract

An oil‐based formulation of carbaryl (1‐naphthyl N‐methyl‐carbamate) (Sevin‐2‐Oil) was applied twice by a fixed‐wing aircraft at a dosage rate of 280 g of A.I./ha/application to a coniferous forest near Allardville, New Brunswick. The highest concentrations of the chemical in fir foliage, litter and forest soil 1 h after application were respectively 4.20, 1.21 and 0.59 ppm (fresh weight). The residues dissipated rapidly and the DT50 values obtained from the depletion curves were 2.3 d for foliage and 1.5 d for litter and soil samples. Very low levels (<0.1 ppm) of carbaryl persisted in foliage and litter beyond the 10 d sampling period. The maximum residue level found in stream water was 0.314 ppm and more than 50% of it had dissipated within 1 h. Low but detectable levels (0.001 ppm) of the chemical persisted in water until the end of the 10 d sampling period. Sediment samples contained a maximum level of 0.04 ppm, which dissipated below the detection limit within 5 h. Brook trout and slimy sculpins captured in the stream 1 d after the spray contained on average about 0.04 ppm of carbaryl and none of it was found in 3 d postspray samples.     

The use of active metabolism and swimming activity to evaluate the toxicity of dodecyl benzene sodium sulfonate (LAS-C12) on the Mugil platanus (Mullet) according to temperature and salinity.

Active metabolism and swimming activity were used to study the effects of dodecyl benzene sodium sulfonate (LAS-C12) in Mugil platanus, a species traditionally considered as estuarine. The effects of exposure to different concentrations of LAS-C12 (0.0, 1.0, 2.5, and 5.0 mg/L) on the active metabolism and swimming activity of Mugil platanus were evaluated. The active metabolism and swimming activity were estimated through experiments conducted on each of 9 possible combinations of three temperatures (35, 20, and 15 degrees C) and three salinities (35, 20, and 5 per thousand). The results show that the active metabolism increases according to the LAS-C12 concentration in all temperatures and salinities studied. At the highest tested concentration (5.0 mg/L), the active metabolism was 111%; 84.8 and 105% higher than the control, at 35 per thousand salinity at the three temperatures. However, the swimming activity decreased according to the LAS-C12 concentration in all temperatures and salinities studied. At the highest tested concentration, the swimming activity was 78.6, 73.6, and 78.7% less than the control, at 25 degrees C at the three salinities. The active metabolism and swimming activity averages, achieved in the different salinities studied, were not significantly different, as a result of the LAS-C12 concentration.     

Acute effects of copper and mercury on the estuarine fish Pomatoschistus microps: Linking biomarkers to behaviour

The main objective of the present study was to investigate possible links between biomarkers and swimming performance in the estuarine fish Pomatoschistus microps acutely exposed to metals (copper and mercury). In independent bioassays, P. microps juveniles were individually exposed for 96 h to sub-lethal concentrations of copper or mercury. At the end of the assays, swimming performance of fish was measured using a device specially developed for epibenthic fish (SPEDE). Furthermore, the following biomarkers were measured: lipid peroxidation (LPO) and the activity of the enzymes acetylcholinesterase (AChE), lactate dehydrogenase (LDH), glutathione S-transferases (GST), 7-ethoxyresorufin-O-deethylase (EROD), superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPx). LC₅₀s of copper and mercury (dissolved throughout metal concentrations) at 96 h were 568 μg L⁻¹ and 62 μg L⁻¹, respectively. Significant and concentration-dependent effects of both metals on swimming resistance and covered distance against water flow were found at concentrations equal or higher than 50 μg L⁻¹ for copper and 3 μg L⁻¹ for mercury (dissolved throughout metal concentrations). These results indicate that SPEDE was efficacious to quantify behavioural alterations in the epibenthic fish P. microps at ecologically relevant concentrations. Significant alterations by both metals on biomarkers were found including: inhibition of AChE and EROD activities, induction of LDH, GST and anti-oxidant enzymes, and increased LPO levels, with LOEC values ranging from 25 to 200 μg L⁻¹ for copper and from 3 to 25 μg L⁻¹ for mercury (dissolved throughout metal concentrations). Furthermore, significant and positive correlations were found between some biomarkers (AChE and EROD) and behavioural parameters, while negative correlations were found for others (LPO, anti-oxidant enzymes and LDH) suggesting that disruption of cholinergic function through AChE inhibition, decreased detoxification capability due to EROD inhibition, additional energetic demands to face chemical stress, and oxidative stress and damage may contribute to decrease the swimming performance of fish. Since a reduced swimming capability of fish may reduce their ability to capture preys, avoid predators, and interfere with social and reproductive behaviour, the exposure of P. microps to copper and/or mercury concentrations similar to those tested here may decrease the fitness of wild populations of this species.     

Responses of spruce seedlings (Picea abies) to exhaust gas under laboratory conditions--I. Plant-insect interactions

The effects of motor vehicle exhaust gas on Norway spruce seedlings (Picea abies (L.) Karst) and plant-insect interaction of spruce shoot aphid (Cinara pilicornis Hartig) was studied. The exhaust gas concentrations in the fumigation chambers were monitored and controlled by measuring the concentration of nitrogen oxides (NO(x)) with a computer aided feedback system. The concentrations of major exhaust gas components (black carbon [BC], fine particles, VOCs and carbonyl compounds) in the chamber air were also measured. Responses of Norway spruce seedlings to a 2 and 3-week exhaust gas exposure and subsequent performance of spruce shoot aphid were studied using realistic exposure regimes; 50, 100 and 200 ppb NO(x). The feedback control system based on NO(x) concentrations proved an adequate and practical means for controlling the concentration of exhaust gases and studying plant responses in controlled environment chambers. The exhaust exposure resulted in increased concentrations of proline, glutamine, threonine, aspartic acid, glycine and phenylalanine and decreased concentration of arginine, serine, alanine and glycine in young needles. No changes in soluble N concentrations were observed. The results are interpreted as a stress response rather than use of NO(x) as a nitrogen source. No changes in total phenolics and only transient changes in some individual terpene concentrations were detected. The exhaust gas exposure stressed the exposed seedlings, but had no significant effect on N metabolism or the production of defence chemicals. Aphid performance was not significantly affected. Soluble N, secondary metabolism and aphid performance were not sensitive to exhaust gas exposure during shoot elongation in Norway spruce.     

Effect of carbaryl (carbamate insecticide) on acetylcholinesterase activity of two strains of Daphnia magna (Crustacea,Cladocera)

The present study was designed to investigate the effect of carbaryl (carbamate insecticide) on the acetylcholinesterase activity in two strains (same clone A) of the crustacean cladoceran Daphnia magna. Four carbaryl concentrations (0.4, 0.9, 1.8 and 3.7 µg L^?1) were compared against control AChE activity. Our results showed that after 48 h of carbaryl exposure, all treatments induced a significant decrease of AChE activities whatever the two considered strains. However, different responses were registered in terms of lowest observed effect concentrations (LOEC: 0.4 µg L^?1 for strain 1 and 0.9 µg L^?1 for strains 2) revealing differences in sensitivity among the two tested strains of D. magna. These results suggest that after carbaryl exposure, the AChE activity responses can be also used as a biomarker of susceptibility. Moreover, our results show that strain1 is less sensitive than strain 2 in terms of IC50-48 h of AChE activity. Comparing the EC50-48 h of standard ecotoxicity test and IC50-48 h of AChE inhibition, there is the same order of sensitivity with both strains.     

Rapid-behaviour responses as a reliable indicator of estrogenic chemical toxicity in zebrafish juveniles

Whereas biochemical and molecular parameters have been well recognised as important “signposts” of individual disturbance to endocrine disrupting chemical’s (EDCs) exposure, behavioural endpoints are yet greatly overlooked as a routine tool in environmental risk assessment of EDCs. However, life histories are intimately associated with numerous inter- and intra-specific interactions, which invariably depend on the performance of effective behaviours. Within fish species, one of the most important factors influencing energy turnover earlier in the development is locomotor activity. This essential trait reflects the organism’s ability to generate and coordinate the metabolic energy required for both reproductive and non-reproductive behaviours. Inappropriate movement responses due to toxic effects of contaminants may ultimately impact important ecological variables.Therefore, in the present study, the swimming bursts of zebrafish juveniles exposed for 40 d to the synthetic estrogen ethinylestradiol (EE₂), tested at environmentally relevant concentrations (nominal concentrations of 0.5, 1 and 2 ng L⁻¹), were investigated in order to address the potential of rapid-behaviour patterns as an effective response indicator of estrogenic endocrine disrupting chemical’s exposure. This synthetic estrogen was selected due to its high prevalence in aquatic ecosystems, ability to mimic natural estrogens and proven record of causing negative effects in fish reproduction. The behavioural responses were compared with established endpoints used in the screening of EE₂ effects at adulthood. Results indicate that zebrafish juveniles’ swimming activity was significantly decreased upon EE₂ exposure. Since reduced locomotion of zebrafish may impact foraging, predator avoidance, drift and transport, and even interfere with social and reproductive behaviours, a fitness decline of wild fish populations can ultimately be hypothesized. Furthermore, behavioural endpoints were found to display higher sensitivity to EE₂ than either vitellogenin gene induction or reproductive parameters determined at adulthood.Overall, the findings of this work not only demonstrate the power of high-throughput behavioural responses, able to act as sensitive early warning signals of EDC exposure, but also highlight the potential of behavioural endpoints in providing a more comprehensive and non-invasive measure of EDC’s exposure.     

In vitro effects of pesticides and metals on the activity of acetylcholinesterase (AChE) from different tissues of the blue mussel,Mytilus trossulus L.

The objective of this study was to conduct a comprehensive comparison of the effect on acetylcholinesterase (AChE) from various tissues of Mytilus trossulus caused by in vitro exposure to several pesticides and metals, because results available in the literature are inconsistent, difficult to compare, and sometimes contradict each other. For this purpose, fraction S10 extracted from gills, digestive gland, mantle and muscles, and the whole soft tissue of the mussel was exposed to several pesticides (dichlorvos, chlorpyrifos, fenitrothion, carbofuran and carbaryl) and metals (Cu, Zn, Cd, Hg, Pb) at a wide range of concentrations. AChE was inhibited in 50% or more in all the tissues exposed to dichlorvos, Cu, Hg and the mixture of Cu+Cd, and in some tissues exposed to carbaryl and carbofuran. The IC₅₀ was calculated where possible. No inhibition was found in the case of chlorpyrifos, Cd, Pb, and Zn.     

Metabolism of 14C‐carbaryl and 14C‐1‐naphthol in moist and flooded soils

Abstract

The metabolism of ¹⁴C‐carbaryl and ¹⁴C‐1‐naphthol in moist and flooded soils was studied in a continuous flow‐through system over a period of 28 days permitting ¹⁴C‐mass balance. The percent distribution of radiocarbon in organic volatiles, carbon dioxide, extractable and non‐extractable (bound) fractions of soils were determined. Organic volatiles could not be detected in both carbaryl and 1‐naphthol treated soils. More of ¹⁴CO₂ (25.6%) was evolved from moist than flooded soil (15.1%) treated with carbaryl. However, the mineralization of ¹⁴C‐1‐naphthol was negligible. The extractable radiocarbon was more in flooded soil (28.9%) than moist soil (5.5%) from carbaryl treatment. Less than one percent was present as parent compound, whereas carbaryl was mainly metabolized to 5‐hydroxy carbaryl in moist soil and to 4‐ and 5‐hydroxy carbaryl in flooded soil. The extractable radiocarbon amounted to 18.2 and 24.3% in moist and flooded soils respectively and the parent compound was less than one percent with 1‐naphthol treatment. Most of the radiocarbon was found as soil bound residues; the formation being more with 1‐naphthol than carbaryl. Humin fraction of the soil organic matter contributed most to soil bound residues of both carbaryl and 1‐naphthol.     

Bioaccumulation of polychlorinated biphenyls in ranid frogs and northern water snakes from a hazardous waste site and a contaminated watershed

Livers of bullfrogs (Rana catesbeiana) from a polychlorinated biphenyl (PCB) contaminated watershed and hazardous waste site located in Pickens County, South Carolina, contained significantly higher concentrations of PCBs (2.33 and 2.26 ppm, respectively) than those from a reference site (0.05 ppm). Green frogs (R. clamitans) from the two contaminated sites also accumulated higher levels of PCBs (2.37 and 3.88 ppm, respectively) than those from the reference site (0.02 ppm). No temporal variation was observed in PCB concentrations of bullfrogs or green frogs from the contaminated sites between 1992 and 1993. Levels of PCBs in the livers of northern water snakes (Nerodia sipedon) were significantly higher in snakes from the contaminated watershed (13.70 ppm) than in those from the waste site (2.29 ppm) and two reference sites (2.50 and 1.23 ppm). When compared to frogs, significantly higher bioaccumulation occurred in water snakes from the contaminated watershed. No significant differences in PCB levels were found with respect to sex or body size (snout-vent length (SVL) or body mass) for frogs or snakes. PCBs were detected also in eggs of both frogs and snakes. Results of this study provide baseline data and document the bioaccumulation of PCB residues in frog and snake tissues; however, the significance of these tissue residues to reproduction, survival, growth/development, and population dynamics in contaminated habitats is unknown.     

Tube-immunoassay for rapid detection of carbaryl residues in agricultural products

An antibody-based rapid, quantitative, and qualitative tube enzyme-linked immunosorbent assay (tube-ELISA) was developed and used to determine carbaryl (1-naphthyl methylcarbamate) residues in agricultural products (apple, Chinese cabbage, rice, and barley). The tube-ELISA is a competitive immunoassay in which the antibody is coated in the polystyrene tube, with a dynamic range between 0.7 and 46.3 microg kg(-1). Carbaryl was extracted from each agricultural sample by hand-shaking with methanol and examined for application to on-site analysis. After the liquid extraction, the sample extracts diluted with buffer were analyzed by rapid tube-ELISA directly. The overall test time was around 15-30 min, including sample preparation and assay performance. The results obtained from tube-ELISA correlated well with high-performance liquid chromatography (R2 > 0.9). The study shows that tube-ELISA is useful as a quality control tool and can be used to quantitatively detect carbaryl as well.