Toxicity of carbaryl, diquat dibromide, and fluoranthene, individually and in mixture, to larval grass shrimp, Palaemonetes pugio

This study examined the toxicity of two pesticides (carbaryl and diquat dibromide) and one polycyclic aromatic hydrocarbon (fluoranthene), both singly and in mixture, to grass shrimp larvae (Palaemonetes pugio). These three chemicals are all present in coastal environments and can easily enter estuarine ecosystems. Fluoranthene was the most toxic chemical with a 96-h LC50 value of 32.45 microg/L, followed by carbaryl (43.02 microg/L) and diquat dibromide (1624 microg/L). In the chemical mixture tests, the binary carbaryl/diquat dibromide mixture and the ternary carbaryl/diquat dibromide/fluoranthene mixture had additive results.     

Mixture Toxicity of the Antifouling Compound Irgarol to the Marine Phytoplankton Species Dunaliella tertiolecta

This study examined the toxicity of irgarol, individually and in binary mixtures with three other pesticides (the fungicide chlorothalonil, and the herbicides atrazine and 2,4-D), to the marine phytoplankton species Dunaliella tertiolecta. Standard 96-h static algal bioassays were used to determine pesticide effects on population growth rate. Irgarol significantly inhibited D. tertiolecta growth rate at concentrations ≥ 0.27 μ g/L. Irgarol was significantly more toxic to D. tertiolecta than the other pesticides tested (irgarol 96 h EC₅₀ = 0.7 μ g/L; chlorothalonil 96 h EC₅₀ = 64 μ g/L; atrazine 96 h EC₅₀ = 69 μ g/L; 2,4-D 96 h EC₅₀ = 45,000 μ g/L). Irgarol in mixture with chlorothalonil exhibited synergistic toxicity to D. tertiolecta, with the mixture being approximately 1.5 times more toxic than the individual compounds. Irgarol and atrazine, both triazine herbicides, were additive in mixture. The toxicity threshold of 2,4-D was much greater than typical environmental levels and would not be expected to influence irgarol toxicity. Based on these interactions, overlap of certain pesticide applications in the coastal zone may increase the toxicological risk to resident phytoplankton populations.     

Lethal and sublethal effects of the pyrethroid,bifenthrin, on grass shrimp (Palaemonetes pugio) and sheepshead minnow (Cyprinodon variegatus)

This study investigated the lethal and sublethal effects of the pyrethroid insecticide bifenthrin on adult and larval grass shrimp, Palaemonetes pugio, and adult sheepshead minnows, Cyprinodon variegatus. The effects were determined by conducting 96-h aqueous static renewal tests and 24-h static tests with sediment. Oxidative stress biomarkers, lipid peroxidation, glutathione, and catalase were also assessed. The 96-h aqueous LC₅₀ value for adult shrimp was 0.020 μ g/L (95% CI: 0.015–0.025 μ g/L) and for larval shrimp was 0.013 μ g/L (95% CI: 0.011–0.016 μ g/L). The 96-h aqueous LC₅₀ for adult sheepshead minnow was 19.806 μ g/L (95% CI: 11.886–47.250 μ g/L). The 24-h sediment LC₅₀ for adult shrimp was 0.339 μ g/L (95% CI: 0.291–0.381 μ g/L) and for larval shrimp was 0.210 μ g/L (95% CI: 0.096–0.393 μ g/L). The oxidative stress assays showed some increasing trends toward physiological stress with increased bifenthrin concentrations but they were largely inconclusive. Given the sensitivity of grass shrimp to this compound in laboratory bioassays, additional work will be needed to determine if these exposure levels are environmentally relevant.     

Toxicity of the pyrethroid pesticide fenvalerate to freshwater catfish clarias gariepinus: Lethality,biochemical effects and role of dietary ascorbic acid

Static bioassays were made in the laboratory to determine lethal concentration of the pyrethroid pesticide fenvalerate [(RS)-alpha-cyano-3-phenoxybenzyl (RS)-2-(4-chlorophenyl)-3-methylbutyrate] for the freshwater catfish Clarias gariepinus and effects of sublethal concentrations of the pesticide on some biochemical parameters of the fish. For exposure periods of 24 to 96 h, LC₅₀ values of fenvalerate ranged from 5.83–4.76 μ g/L and 4.24–2.94 μ g/L, respectively for water and acetone soluble fenvalerate. Two sublethal concentrations of fenvalerate were used in the bioassays for biochemical parameters: 2.1 μ g/L for 24 h and 1.4 μ g/L for 96 h exposure, both concentrations representing 50% of LC₅₀ value of acetone soluble fenvalerate for the respective exposure period. Hepatosomatic index, liver glycogen, alkaline phosphatase of liver and ascorbic acid of blood, liver, and kidney decreased while haemoglobin (Hb) %, plasma glucose levels and acid phosphatase level of liver increased after 24 h exposure to 2.1 μ g/L fenvalerate. Longer exposure (96 h) to even a lower concentration (1.4 μ g/L) of fenvalerate resulted in reduction of all the parameters (except Hb %) tested as compared with control. Fish previously fed for 60 days with a diet supplemented by a high level of ascorbic acid (100 mg/100 g diet) could reverse most of the effects caused by 24 h exposure to 2.1 μ g/L fenvalerate. A lower level of ascorbic acid (50 mg/ 100 g diet) supplement could not influence these effects of fenvalerate. Even the higher dose of ascorbic acid supplementation (100 mg/100 g diet) could not relieve the stress parameters, except for Hb% and HSI, when the pesticide was applied at 1.4 μ g/L for a longer time period (96 h).     

The effect of pesticides and metals on acetylcholinesterase (AChE) in various tissues of blue mussel (Mytilus trossulus L.) in short-term in vivo exposures at different temperatures

The purpose of the study was to assess the impact of short-term exposure to selected toxicants as well as metal accumulation upon acetylcholinesterase (AChE) in the blue mussel, Mytilus trossulus L., in laboratory in vivo experiments. Mussels were exposed for up to 48 hours to a mixture of copper (Cu^{2 +}, 400 μ g L^?1) and cadmium (Cd^{2 +}, 200 μ g L^?1), to dichlorvos (DDVP, 100 μ g L^?1), and to carbaryl (100 μ g L^?1) at two temperatures: 5°C and 20°C. Samples were collected after 0, 6, 12, 24, and 48 hours of exposure, and AChE activity and metal concentration (where applicable) were analysed in gills, digestive gland, mantle+muscles, and the whole soft tissue. Very strong AChE inhibition was observed in response to the dichlorvos treatment, mainly in gills. Carbaryl and the metals caused a short-term inhibition effect. Considerable differences in AChE activity between the two temperatures were noticed. In particular, the metals were accumulated much faster at 20°C than at 5°C, especially in gills. No correlation between AChE activity and metal concentration was found. Gills turned out to be the optimal tissue for AChE activity analysis in short-term studies.     

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.     

Acute and chronic toxicity of organophosphate monocrotophos to Daphnia magna

The acute and chronic toxicity of monocrotophos (MCP), the binary joint toxicity of MCP and bifenthrin (BF), and sodium dodecyl benzene sulfonate (SDBS) to Daphnia magna (D. magna) was evaluated. The 24 h-median effective concentration (24 h-EC₅₀) and 48 h-median lethal concentration (48 h-LC₅₀) of MCP towards D. magna were 161 and 388 μ g/L, respectively. In addition, the lowest-observed effective concentration (LOEC) and non-observed effective concentration (NOEC) of MCP to D. magna were 10 and 5 μ g/L, respectively. Furthermore, the chronic value (ChV) of MCP against D. magna was 7 μ g/L and the acute chronic ratio (ACR) was 55. The number of offspring per female and the intrinsic rate of natural increase (r) were identified as the parameters that were most sensitive to MCP. In addition, toxic unit (TU) analysis was employed to evaluate the joint toxicities. The calculated TU_mix values of binary equitoxic mixtures of MCP + BF and MCP + SDBS were 1.47 and 1.63, respectively, which suggests that both equitoxic mixtures exert a limited antagonistic effect. The results of this study revealed that the toxic threshold of MCP towards D. magna is higher than its reported highest residue (4 μ g/L) in the ordinary aquatic environment, and that concurrent exposure to BF or SDBS may exert a slight antagonistic effect.     

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 μ g/L and 0.00061 μ g/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 μ g/L and cypermethrin up to 1.9 μ g/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 μ g/L of cartap or 40 μ g/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.     

Diquat associated with copper sources for algae control: Efficacy and ecotoxicology

The aims of this research were to evaluate the efficacy of copper oxychloride (CuCl₂.3Cu(OH)₂), copper hydroxide (Cu(OH)₂) and diquat (1.1′-ethylene-2.2′-bipyridyldiylium dibromide), isolated and in association with 0.1% of both copper sources, in the control of the unicellular algae Ankistrodesmus gracilis and the filamentous algae Pithophora kewesis, and to determine the acute toxicity of the tested chemicals in Hyphressobrycon eques, Pomacea canaliculata, Lemna minor and Azolla caroliniana. The efficacy was estimated by the methods of chlorophyll a and pheophytin a readings, changed into growth inhibition percentage. Both algae were exposed to the following concentrations: 0.2; 0.4; 0.8; 1.2 mg L^?1 of diquat and its association with the copper sources; and 0.1; 0.3; 0.5; 0.7; 1.0 and 1.5 mg L^?1 in the isolated applications of copper hydroxide and copper oxychloride. An untreated control was kept. The acute toxicity was estimatedby 50% lethal concentration (LC50). The copper sources were effective for A. gracilis control, at rates as high as 0.1 mg L^?1 (>95% efficacy). Isolated diquat and its association with copper hydroxide were both effective at rates as high as 0.4 mg L^?1, with 95 and 88% control efficacy, respectively. The copper oxychloride was effective at 0.2 mg L^?1, with 93% efficacy. None of the tested chemicals and associations was effective on P. kewesis control. The most sensitive non target organism to the tested chemicals was L. minor; the less sensitive was H. eques.     

Manure placement depth impacts on crop yields and N retained in soil

The objective of this study was to determine the impact of manure placement depth on crop yield and N retention in soil. Experimental treatments were deep manure injection (45 cm), shallow manure injection (15 cm), and conventional fertilizer-based management with at least three replications per site. Water infiltration, and changes in soil N and P amounts were measured for up to 30 months and crop yield monitored for three seasons following initial treatment. Deep and shallow manure injections differed in soil inorganic N distributions. For example, in the manure slot the spring following application, NO₃-N in the surface 60 cm was higher (p < .01) when injected 15 cm (21.4 μ g/g) into the soil than 45 cm (11.7 μ g/g), whereas NH₄-N had opposite results with shallow injection having less (p = 0.045) NH₄-N (102 μ g/g) than deep (133 μ g/g) injection. In the fall one year after the manure was applied, NO₃-N and NH₄-N were lower (p = 0.001) in the shallow injection than the deep injection. The net impact of manure placement on total N was that deep injection had 31, 59, and 44 more kg N ha^{? 1} than the shallow injection treatment 12, 18, and 30 months after application, respectively. Deep manure injection did not impact soybean (Glycine max L.) yield, however corn (Zea mays L.) yield increased if N was limiting. The higher corn yield in the deep injected treatment was attributed to increased N use efficiency. Higher inorganic N amounts in the deep injection treatment were attributed to reduced N losses through ammonia volatilization, leaching, or denitrification. Results suggest that deep manure placement in glacial till soil may be considered a technique to increase energy, N use efficiency, and maintain surface and ground water quality. However, this technique may not work in glacial outwash soils due to the inability to inject into a rocky subsurface.     

Fungicidal Activity of Artemisia herba alba Asso (Asteraceae)

The antifungal activity of Artemisia herba alba was found to be associated with two major volatile compounds isolated from the fresh leaves of the plant. Carvone and piperitone were isolated and identified by GC/MS, GC/IR, and NMR spectroscopy. Antifungal activity was measured against Penicillium citrinum (ATCC 10499) and Mucora rouxii (ATCC 24905). The antifungal activity (IC₅₀) of the purified compounds was estimated to be 5 μ g/ml, 2 μ g/ml against Penicillium citrinum and 7 μ g/ml, 1.5 μ g/ml against Mucora rouxii carvone and piperitone, respectively.     

Inhibitory Effects on Esterase Enzymes Buche and Ache in Rainbow Trout (Oncorhyncus mykiss) Produced by the Slow Release Insecticide Chitosan Diethyl Phosphate

The study on the toxicity of chitosan diethyl phosphate (ChDP), a controlled release insecticide, on the activities of butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE) in rainbow trout exposed to this pesticide was carried out. It was found that ChDP reduced BuChE activity in O. mykiss by a factor of eight at 6 days, with high fluctuation to the end of the exposition time at 12 days. The in vitro analysis of brain AChE treated with ChDP and Phenamiphos showed that it was competitively inhibited by both organophosphates. The values obtained for K_m and V_max for the AChE-ChDP (K_m: 21.23 μ M; V_max: 43.10 μ mol/min/g) and AChE-Phenamiphos (K_m: 38.62 μ M; V_max: 38.91 μ mol/min/g) systems were relatively low compared to values of the AChE (control) system (K_m: 62.99 μ M; V_max: 63.29 μ mol/min/g). Results reported in this study confirmed that chitosan diethyl phosphate performs similarly to organophosphate pesticides, producing inhibition in cholinesterases in rainbow trout.     

Comparative acute toxicity of two phytosanitary molecules,lambda-cyhalothrin and acetamiprid,on Nile Tilapia (Oreochromis Niloticus) juveniles

Abstract

This study aimed to compare the toxicity for fish of two active ingredients (lambda-cyhalothrin-20?g L^?1, a pyrethroid, and acetamiprid-15?g L^?1, a neonicotinoid) which are components of a commercial insecticide (Acer 35 EC) used in cotton crop in many West African countries. The juveniles of Oreochromis niloticus (4.01?±?0.34?g, mean body weight) were exposed for 96?h to increasing concentrations of active ingredients (lambda-cyhalothrin and acetamiprid) or a mixture similar to Acer 35 EC (composed by 20?g of chemical compound lambda-cyhalothrin and 15?g of acetamiprid dissolved in 1?L of acetone). The experiments were carried out under controlled conditions in aquaria according to OECD Guidelines. During the experiments, the behavioral responses (loss of balance, color change, hyperactivity, etc.) that usually precede death were observed in exposed fish. Mortalities were recorded in each aquarium and the LC₅₀-96h of each chemical was determined. The LC₅₀-96h obtained were respectively 0.1268, 0.0029, 182.9 and 0.5685?ppm for Acer 35 EC, lambda-cyhalothrin, acetamiprid and mixture. All insecticides used in this study had profound impact on Nile tilapia behavior which may confirm the neurotoxicity of each single active compound as well as of their mixture.     

Commercial trials evaluating the novel use of ethyl formate for in-transit fumigation of shipping containers

Sediment samples from 25 sites in 17 rivers of the Pantanal (Brazil) were analyzed with the objective of evaluating pesticide contamination in sediments. Samples were extracted with an acetone, ethylacetate, and water mixture 2:2:1 (v/v/v). The extract was purified by flash chromatography with aluminum oxide and florisil. A multiresidue gas chromatography-mass spectrometry method was applied to monitor 23 pesticides of different chemical classes (organochlorine, organophosphorus, triazines, anilides and pyrethroids) with some of their degradation products. Compounds identified in sediment samples included λ -cyhalothrin (1.0 to 5.0 μ g kg^{? 1}), p,p′-DDT (3.6 μ g kg^{? 1}), deltamethrin (20.0 μ g kg^{? 1}) and permethrin (1.0 to 7.0 μ g kg^{? 1}).     

Tylosin Sorption to Silty Clay Loam Soils,Swine Manure,and Sand

The objectives of this study were to assess sorption and desorption of tylosin, a macrolide antimicrobial chemical used in swine, cattle, and poultry production, in three silty clay loam soils of South Dakota and compare soil sorption to sand and manure sorption. The silty clay loam soils, from a toposequence in eastern South Dakota, standardized sand samples, and swine manure were used in 24-h batch sorption studies with tylosin concentrations ranging from 25 to 232 μ mole/L. Desorption from soil was conducted over a four-day period. Partition coefficients, based on the Freundlich isotherm (K _f ) or K _d values, were calculated. K _f values for the silty clay loams were similar, not influenced by landscape position, and averaged 1350 with isotherm slopes ranging from 0.85 to 0.93. K _f values for sand were dependent on solution/sand ratios and pH, ranging from 1.4 to 25.1. K _d values of manure were dependent on the solution type and ranged from 840 L/kg with urine to about 175 L/kg when sorbed from water. Desorption of tylosin from each soil over the four-day period was < 0.2% of the amount added. The soils' high K _f values and low desorption amounts suggest that once tylosin is in these soils, leaching to lower depths may not occur. However, this does not preclude runoff with soil eroded particles. If tylosin reaches a sand aquifer, through bypass flow or other mechanism(s), movement in the aquifer most likely would occur.     

An evaluation of the cytochrome P450 inhibition potential of selected pesticides in human hepatic microsomes

The goal of this work was to study the ability of 18 pesticides to inhibit selective model activities for all major xenobiotic-metabolizing enzymes, namely CYP1A1/2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1 and 3A4. Generally organophosphorus insecticides were the most potent and extensive inhibitors, especially towards CYP1A1/2 (IC₅₀ values of chlorpyrifos, fenitrothion and profenofos ～3 μ M), CYP2B6 (IC₅₀ values of chlorpyrifos and fenitrothion 2.5 μ M), CYP2C8 (fenitrothion 4.3 μ M), CYP2C9 (fenitrothion and malathion 4.8 and 2.5 μ M, respectively), CYP2D6 (chlorpyrifos and phenthoate ～ 3 μ M) and CYP3A4 (chlorpyrifos, fenitrothion and phenthoate 3–4 μ M). Otherwise there were quite considerable differences in potency and extent of inhibition between different organophosphates. Pyrethroids were in general very weak or inactive. Deltamethrin and fenvalerate were potent inhibitors of CYP2D6 (IC₅₀ values of ～ 3 μ M) while lambda-cyhalothrin potently inhibited both CYP2D6 and CYP3A4-mediated activities (IC₅₀'s about 3–4 μ M). Some pesticides caused relatively potent inhibitions sporadically (carbendazim, CYP2D6, IC₅₀ = 12 μ M; atrazine, CYP3A4, IC₅₀ = 2.8 μ M; glyphosate, CYP2C9, IC₅₀ = 3.7 μ M; hexaflumuron, IC₅₀ = 6.0 μ M). With the exceptions of alpha-cypermethrin, cypermethrin, isoproturon, carbaryl and abamectin, most pesticides inhibited relatively potently at least one CYP-selective activity, which may have relevance for potential interactions in occupational exposures and for further studies on the CYP-associated metabolism of respective pesticides.     

粉煤灰沸石负载Ce3+/TiO2光催化降解水中的菲和荧蒽

以钛酸四丁酯为前驱体，粉煤灰合成沸石为载体，采用溶胶-凝胶方法，在低温条件下制备了稀土铈掺杂的TiO₂光催化剂。利用SEM-EDS、XRD、FTIR对催化剂进行了分析和表征。以高压汞灯为灯源，对多环芳烃菲、荧蒽的降解进行了研究。实验考查了稀土铈掺杂质量分数、催化剂用量、溶液pH、目标物初始质量浓度等因素对光催化降解的影响，研究了其光降解动力学。结果表明，当稀土铈含量为0.5%，催化剂用量为3 g/L，pH偏碱性时，催化效果最佳。光催化反应符合Langmuir-Hinshelwood 动力学规律，菲、荧蒽的降解过程符合一级反应动力学，反应速率常数分别为0.0126 min^-1，0.0099 min^-1。     

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.     

Soluble esterases as biomarkers of neurotoxic compounds in the widespread serpulid Ficopomatus enigmaticus (Fauvel, 1923)

Abstract

The characterization of soluble cholinesterases (ChEs) together with carboxylesterases (CEs) in Ficopomatus enigmaticus as suitable biomarkers of neurotoxicity was the main aim of this study. ChEs of F. enigmaticus were characterized considering enzymatic activity, substrate affinity (acetyl-, butyryl-, propionylthiocholine), kinetic parameters (K_m and V_max) and in vitro response to model inhibitors (eserine hemisulfate, iso-OMPA, BW284C51), and carbamates (carbofuran, methomyl, aldicarb, and carbaryl). CEs were characterized based on enzymatic activity, kinetic parameters and in vitro response to carbamates (carbofuran, methomyl, aldicarb, and carbaryl). Results showed that cholinesterases from F. enigmaticus showed a substrate preference for acetylthiocholine followed by propionylthiocholine; butyrylthioline was not hydrolyzed differently from other Annelida species. CE activity was in the same range of cholinesterase activity with acetylthiocholine as substrate; the enzyme activity showed high affinity for the substrate p-nytrophenyl butyrate. Carbamates inhibited ChE activity with propionylthiocholine as substrate to a higher extent than with acetylthiocoline. Also CE activity was inhibited by all tested carbamates except carbaryl. In vitro data highlighted the presence of active forms of ChEs and CEs in F. enigmaticus that could potentially be inhibited by pesticides at environmentally relevant concentration.     

Development of IC-ELISA for detection of organophosphorus pesticides in water

Diethyl (carboxymethyl) phosphonate (DECP) was used as the hapten to develop an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) for detecting organophosphorus pesticides (OPs). Conjugator of DECP with bovin serum albumin (BSA) was used as the immunogen for producing the polyclonal antibodies (PcAbs). Three antisera were obtained after the immune procedure. Characterization studies of the PcAbs indicated that the titer of antiserum-1 was highest in 3 antisera, and antiserum-1 had high affinity and specificity to the parathion, dichlorvos and pirimiphos. The IC-ELISA showed an IC₅₀ of 0.428 μ g/mL with a detection limit of 0.0125 μ g/mL to parathion. The assay also indicated that the IC₅₀ values of pirimiphos and dichlorvos were 0.331 μ g/mL and 1.25 μ g/mL respectively, and the detection limits of pirimiphos and dichlorvos were 0.0116 μ g/mL and 0.048 μ g/mL respectively. Recoveries of parathion, pirimiphos and dichlorvos spiked into water samples ranged from 90% to 160%. The results indicated that the ELISA could be a convenient and supplemental analytical tool for monitoring OPs residues in environmental water samples.