Cholinesterase characterization in Corbicula fluminea and effects of relevant environmental contaminants: a pesticide (chlorfenvinphos) and a detergent (SDS)

This study characterizes the cholinesterasic forms present in the bivalve Corbicula fluminea, through the analysis of differential affinity towards various substrates and inhibitors. This approach also permitted the validation of cholinesterase inhibition in the mentioned species for monitoring purposes by evaluating the inhibitory activity of a detergent (SDS) and a pesticide (chlorfenvinphos), both in vitro and in vivo. The cholinesterasic form present in the tissues of C. fluminea showed intermediate properties in terms of its relation with substrates and inhibitors. Furthermore, it was not inhibited by the detergent SDS under any of the test conditions, but was clearly inhibited by the selected organophosphate. The data show clearly that the main cholinesterasic form in C. fluminea is responsive to the organophosphate chlorfenvinphos, thus supporting the use of this biomarker for the environmental monitoring of specific contaminants such as anticholinesterasic pesticides. Despite not being commonly attained in the wild, the levels of chlorfenvinphos required to elicit a significant ChE inhibition were only one order of magnitude above actual values of aquatic contamination by this pesticide.     

Biota-sediment metal accumulation and human health risk assessment of freshwater bivalve Corbicula fluminea in Dongting Lake,China

In this study, we analyzed the concentrations of metals in sediments and Corbicula fluminea in China’s Dongting Lake to assess the suitability of C. fluminea as an effective biomonitor of metal contamination in sediments and food safety. We analyzed the biota-accumulation capacity by calculating the biota-sediment accumulation factor (BSAF) and assessed the potential human health risk of metals exposure via consumption of C. fluminea using the target hazard quotient (THQ) and total target hazard quotient (TTHQ). The results showed that the average concentrations of As (31.93 mg kg⁻¹), Cd (5.54 mg kg⁻¹), Cr (105.50 mg kg⁻¹), Cu (32.53 mg kg⁻¹), and Zn (207.89 mg kg⁻¹) in sediments were higher than their respective standard set by the General Administration of Quality Supervision, Inspection and Quarantine of the People’s Republic of China. The sediment metals, which were mainly anthropogenic in origin, were at high levels and pose a relatively high ecological risk. Cadium (Cd) showed very high potential ecological risk levels and should be included in the prior pollutants list in the studied area. The mean levels of As (0.81 mg kg⁻¹) in C. fluminea were 1.62-times higher than that set by the National Health Commission of the People’s Republic of China. BSAF values of the soft tissues of C. fluminea were between 0.05 and 2.14, with higher values for Cu (2.14), Cd (1.77), Zn (1.60), and Ni (1.27). Soft tissues of C. fluminea were able to reflect spatial differences in Sr within sediments around Dongting Lake. The results indicated that C. fluminea could be an potential biomonitor for sediment metals assessment in biomonitoring programs, especially for Cu, Cd, Zn, Ni, and Sr. The mean values for THQ and TTHQ of all the analyzed metals were below 1.0, indicating that the intake of metals via comsumption of C. fluminea does not result in an appreciable risk to human health.

     

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.     

Distribution and bioaccumulation of organochlorine pesticides in surface sediments and benthic organisms from Taihu Lake, China

The spatial distribution and bioaccumulation of organochlorine pesticides (OCPs) in surface sediments and benthic organisms from Taihu Lake were studied. OCPs were detected in all sediment samples with total concentrations ranging from 4.22 to 461 ng g⁻¹ dry weight (dw). The ratios of certain metabolites to their parent compounds indicated there are still new inputs of parent DDT (dichlorodiphenyltrichloroethane) to Taihu Lake, while the highest residues of HCHs (hexachlorocyclohexanes) mainly came from earlier usage and fresh γ-HCH (lindane). No positive correlation was found between the distribution of OCPs and organic matter contents in sediments. Concentrations of OCPs and lipids in typical large benthic organisms, Bellamya aeruginosa (B. aeruginosa) and Corbicula fluminea (C. fluminea), increased with body weight. HCHs, DDTs, chlordanes and heptachlors were the dominant compounds detected in organisms and C. fluminea accumulated much more OCP than B. aeruginosa. Higher values of biota-sediment accumulation factor (BSAF) were detected in C. fluminea, which was both affected by biological characteristics of the organisms and physicochemical properties of the compounds.     

Tissue distribution, isozyme abundance and sensitivity to chlorpyrifos-oxon of carboxylesterases in the earthworm Lumbricus terrestris

A laboratory-based study was conducted to determine the basal carboxylesterase (CbE) activity in different tissues of the earthworm Lumbricus terrestris, and its sensitivity to the organophosphate (OP) pesticide chlorpyrifos-oxon (CPx). Carboxylesterase activity was found in the pharynx, crop, gizzard, anterior intestine, wall muscle and reproductive tissues of L. terrestris, and multiple tissue-specific isozymes were observed by native gel electrophoresis. Esterase activity and sensitivity to CPx inhibition varied on a tissue- and substrate-specific basis, suggesting isoforms-specific selectivity to OP-mediated inhibition. Three practical issues are recommended for the use of earthworm CbE activity as a biomarker of pesticide exposure: (i) CbE should be measured using several routine substrates, (ii) it should be determined in selected tissues instead of whole organism homogenate, and (iii) earthworm CbE activity should be used in conjuncture with other common biomarkers (e.g., ChE) within a multibiomarker approach to assess field exposure of OPs, and potentially other agrochemicals.     

Insecticides biomarker responses on a freshwater fish Corydoras paleatus (Pisces: Callichthyidae)

This study was undertaken to investigate the effects of sublethal concentration of three different classes of insecticides (carbamate, organophosphate, and pyrethroid compounds) on the freshwater fish Corydoras paleatus. For this purpose, fish were exposed for 96 hours to commercial pesticides. Different biomarkers were analyzed as levels of lipid peroxidation (LPO), piscine micronucleus test, and enzymatic activities of catalase (CAT), glutathione S-transferase (GST), and acetylcholinesterase (AChE). The brain AChE was inhibited with carbaryl and methyl parathion, but no inhibition was observed with deltamethrin. The insecticides did not cause oxidative stress or genotoxic effects at the tested concentrations. Further studies are needed to elucidate the biotransformation of Corydoras paleatus insecticides and a possible resistance mechanism.     

Surface and ground waters characterization in Tuscany (Italy) by using algal bioassay and pesticide determinations: comparative evaluation of the results and hazard assessment of the pesticides impact on primary productivity

Algal bioassays and pesticide analysis were joined in Tuscany regional monitoring of surface and ground waters. Results were compared. Waters affected algal growth in any of the following ways: stimulation, inhibition, stimulation at low concentrations and inhibition at higher concentrations. Most of ground waters inhibited algal growth whereas only a little percentage were contaminated with pesticide. Most of surface waters stimulated algal growth and a higher percentage were contaminated with pesticides. In waters with inhibitory effect toxicological parameters (EC50, EC25, EC10 and NOEC) were measured and compared; EC25 values were very close to NOEC values. Hazard to aquatic primary productivity for each pesticide found during chemical monitoring was evaluated; for this aim the margin of safety (MOS) was calculated by using algal toxicity and the pesticide concentration measured.     

Plasma esterases in the tegu lizard Tupinambis merianae (Reptilia, Teiidae): impact of developmental stage, sex, and organophosphorus in vitro exposure

Purpose  

In this study, we determined normal serum butyrylcholinesterase (BChE) and carboxylesterase (CbE) activities in Tupinambis merianae in order to obtain reference values for organophosphorus pesticide monitoring.     

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 Km and Vmax for the AChE-ChDP (Km: 21.23 microM; Vmax: 43.10 micromol/min/g) and AChE-Phenamiphos (Km: 38.62 microM; Vmax: 38.91 micromol/min/g) systems were relatively low compared to values of the AChE (control) system (Km: 62.99 microM; Vmax: 63.29 micromol/min/g). Results reported in this study confirmed that chitosan diethyl phosphate performs similarly to organophosphate pesticides, producing inhibition in cholinesterases in rainbow trout.     

Effects of synthetic pyrethroids and methidation on activities of some digestive enzymes in carp (Cyprinus carpio L.)

Abstract

The effects of pyrethroid pesticides (deltamethrin, permethrin and cypermethrin) and an organophosphate ester (methidation) on the activities of carp trypsin, α‐chymotrypsin, carboxypeptidase A and lipase were studied. The enzymes were isolated from the gastrointestinal tract and the effects of the pesticides were investigated during incubation for 5 min. The activity of trypsin was influenced only slightly by the presence of deltamethrin and methidation, whereas permethrin and cypermethrin caused significant inhibition. The pyrethroid pesticides at lower concentrations resulted in a slight activation of α‐chymotrypsin. Methidation inhibited the α‐chymotrypsin activity by about 20%. These pesticides modified the lipase activity to a lesser extent; the highest inhibition was measured with cypermethrin. The carboxypeptidase A activity was inhibited by both pyrethroid pesticides and methidation. The results suggest that these pesticides might interact with the active conformation of the studied hydrolytic enzymes, resulting in changes in their activities.     

Using enriched stable isotope technique to study Cu bioaccumulation and bioavailability in Corbicula fluminea from Taihu Lake,China

In this study, we measured trace metals (Cd, Cr, Cu, Ni, Pb, and Zn) in water and sediment from representative sites of Taihu Lake, with focus on the analysis of trace metal accumulation in Corbicula fluminea (bivalve). The results showed that the quality of water in Taihu Lake was generally good and the correlation was not found between Cu bioaccumulation in C. fluminea and the concentration in water and sediment. Thus, using the stable isotope tracer method, we studied Cu uptake from the water phase, the assimilation of Cu from the food phase, and the efflux of Cu in vivo by C. fluminea. The result revealed that this species exhibited a relatively lower efflux rate constant of Cu compared with other zoobenthos species. Using a simple bioenergetics-based kinetic model, Cu concentrations in the C. fluminea were calculated with the measured efflux rate. We put forward a novel method, which was taking the influence of biological kinetic on metal bioaccumulation into account to explain the field survey data.     

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.     

Inhibition, recovery and oxime-induced reactivation of muscle esterases following chlorpyrifos exposure in the earthworm Lumbricus terrestris

Assessment of wildlife exposure to organophosphorus (OP) pesticides generally involves the measurement of cholinesterase (ChE) inhibition, and complementary biomarkers (or related endpoints) are rarely included. Herein, we investigated the time course inhibition and recovery of ChE and carboxylesterase (CE) activities in the earthworm Lumbricus terrestris exposed to chlorpyrifos, and the ability of oximes to reactivate the phosphorylated ChE activity. Results indicated that these esterase activities are a suitable multibiomarker scheme for monitoring OP exposure due to their high sensitivity to OP inhibition and slow recovery to full activity levels following pesticide exposure. Moreover, oximes reactivated the inhibited ChE activity of the earthworms exposed to 12 and 48 mg kg⁻¹ chlorpyrifos during the first week following pesticide exposure. This methodology is useful for providing evidence for OP-mediated ChE inhibition in individuals with a short history of OP exposure (≤1 week); resulting a valuable approach for assessing multiple OP exposure episodes in the field.     

Solubility and adsorption behaviors of chlorpyriphos-methyl in the presence of surfactants

In the present work changes in the adsorption of the pesticide chlorpyrifos-methyl (CLP-m) on soil colloids induced by application of surfactants were determined using a batch equilibrium method. The surfactants used were sodium dodecyl sulphate (SDS), Tween 20, and dihexadecyldimethylammonium bromide (DHAB). The adsorption isotherms of CLP-m in aqueous medium and in surfactant solutions at concentration equal to the critical micelle concentration (CMC) fitted the Freunlich adsorption equation generally with R² values greater than 0.96. While the addition of SDS and DHAB decreased the pesticide adsorption, the addition of Tween 20 increased the pesticide adsorption. The increases or decreases in the adsorption in the experiment revealed that the behavior of CLP-m in soil water-systems mainly depends on the type of surfactant. Moreover water solubility of CLP-m changes by the three surfactants below and above their CMC were studied. While the solubility of CLP-m was enhanced by SDS both below and above the CMC, the solubility of the pesticide was enhanced by DHAB only above the CMC. Tween 20 did not influence the solubility of CLP-m.     

Alterations in liver marker enzymes and recovery ability of cyprinids as indicators of aquatic pollution

Information on the detection of pesticides in fresh water Lakes of Kashmir (Dal and Mansbal) through GC–MS technique is scarce, and also the recovery in biochemical parameters (AST, ALT and ALP) of fish after transferring them to clean media has not been reported yet. The water samples were collected from three sites and analyzed for their pesticide profile by dispersive liquid–liquid micro extraction (DLLME) followed by GC–MS. Influence of pesticides on liver marker enzymes of Cyprinus carpio and Carassius carassius was also investigated. The results obtained showed the presence of three main pesticides viz. chlorpyrifros, dimethoate and dichlorvos in waters of Dal Lake whereas no pesticide was detected in waters of Mansbal. The higher values for AST, ALT and ALP activities and decrease in protein content were obtained in the samples from the Dal Lake compared with those from Mansbal Lake (p < 0.05). These data when compared with the values found in C. carpio and C. carassius from both the Dal Lake and Mansbal Lake kept under laboratory conditions after 15, 30, 45 and 60 days of maintenance in clean media found that during depuration, all the enzyme activities came down significantly (p < 0.05) indicating the compensatory response by the fish against the pesticide stress. Therefore, these parameters could be used as indicators of pesticide pollution in aquatic organisms and were recommended for environmental monitoring for investigating the mechanism involved in the recovery pattern.

     

Influence of atrazine on the mineralisation of n-dodecylbenzene-sulfonate-14C in soil

Under the influence of 2,5, or 10 μg/g atrazine, the degradation of 10 μg/g of the detergent n-dodecylbenzenesulfonate-¹⁴C to ¹⁴CO₂ in an Alfisol soil is inhibited for about 12 days, then stimulated. The overall influence of atrazine after 38 days is a slight inhibition of ¹⁴CO₂-formation.     

Determination of organochlorine and organophosphate pesticide residues in fruits,vegetables and sediments

Abstract

A method is described for the determination of organochlorine and organophosphate pesticide residues in fruits, vegetables and sediments. The concentrated solvent extract was sealed in a polymeric membrane tube, dialysed in cyclohexane and the solvent replaced with hexane. The organophosphates were analysed on a specific thermionic detector without further clean‐up. For the organochlorine pesticides the extract was eluted through 3 g of alumina and analysed on GC/ECD. The clean‐up for sediment extract was carried out on a10 g alumina column with 100 mL hexane containing 5% acetone and the eluate was concentrated to 5 mL.

The detection limit for organophosphates on a 40 g sample and a final volume of 10 mL was on the average 0.01 mg/kg. The detection limit for organochlorine pesticides, with the final volume of 25 mL, was 0.005 mg/kg for all pesticides except for p,p'‐DDT and endosulfan sulphate, which was 0.01 mg/kg.

The detection limit for oganochlorine pesticides in sediment, with the final volume of 2 mL, was less than 1 μg/kg and for organophosphate pesticides less than 10 μg/kg when the final volume was made to 0.5 mL. At the detection limits the method produced a very high coefficient of variation for both organochlorine and organophosphate pesticides.     

Effects of dimethoate on rotifer Brachionus calyciflorus using multigeneration toxicity tests

As important members of the zooplankton community and sources of food for fish, rotifers are used extensively in ecotoxicological research to assess the health of the environment and safety of compounds. However, most rotifer toxicity tests are only conducted using rotifer neonates derived from unexposed mothers, thus ignoring the potential transfer of contaminants from mother to offspring. To understand better the mother to offspring exposure, a multigenerational study was conducted using three successive generations (F₀, F₁ and F₂) of the common freshwater rotifer Brachionus calyciflorus to investigate the toxic effects of the widely used organophosphate pesticide, dimethoate (O, O-dimethyl S-methylcarbamoylmethyl phosphorodithioate). When the F₀ generation was exposed to five pesticide concentrations, the population growth rate (r) displayed symptoms of hormesis, characterized by the conversion of low-concentration stimulation to high-concentration inhibition. Despite this observation, the exposure to any given concentration of dimethoate reduced the population growth rates of the F₁ and F₂ generation rotifers. Significant differences existed between the F₀, F₁ and F₂ rotifers for the population growth rate under dimethoate stress: F₂ individuals were more sensitive than F₁, whereas the F₁ individuals were more sensitive than F₀. The results indicated that the parental exposure to a given toxic stress could result in increased sensitivity and decreased fitness in the offspring. This study illustrates the utility of multigenerational toxicity tests, which may better reflect and more accurately predict the effects of long-term pesticide exposure to aquatic organisms at the population level.     

Enhanced visual detection of pesticides using gold nanoparticles

The presence of parts per billion (ppb) levels of chlorpyrifos (O,O-Diethyl-O-(3,5,6-trichloro-2-pyridyl) phosphorothioate) and malathion (S-1,2-bis(ethoxycarbonyl) ethyl O,O-dimethyl phosphorodithioate), two common pesticides found in the surface waters of developing countries, have been visually detected using gold nanoparticles. Visual detection of the presence of pesticide is possible when the color change occurring by the adsorption of pesticides on gold nanoparticles is enhanced by sodium sulfate. The method presented here is simple and there is no need of sample preparation or preconcentration. The response occurs within seconds and the color change is very clear. The detection is possible if chlorpyrifos and malathion are present up to a concentration of 20 and 100 ppb, respectively. The method shows great potential for on-site pesticide monitoring. The method is also applicable as a qualitative technique for the performance evaluation of various household water filters, which claim pesticide removal.     

Combined effects of deltamethrin, temperature and salinity on oxidative stress biomarkers and acetylcholinesterase activity in the black tiger shrimp (Penaeus monodon)

This study aimed to investigate the interactions of two abiotic factors (temperature and salinity) and deltamethrin (pyrethroid pesticide) exposure on some oxidative stress biomarkers as well as on acetylcholinesterase activity (AChE) in hepatopancreas, gills and muscle of black tiger shrimp (Penaeus monodon). A combination of three temperatures (24, 29 and 34 °C), two salinities (15 and 25 ppt), and the absence or presence of 0.1 μg L⁻¹ deltamethrin was applied on shrimp during 4 d under laboratory conditions. Lipid peroxidation level (LPO) and glutathione S-transferase activity (GST) were not affected by combined effect of temperature, salinity and deltamethrin in any of the studied tissues. Deltamethrin impaired other tested oxidative stress biomarkers, i.e. total glutathione (tGSH), catalase (CAT), glutathione peroxidase (GPx). tGSH level significantly increased in hepatopancreas due to deltamethrin exposure mainly at 34 °C, while pesticide effects on tGSH and CAT activity in gills were influenced by both temperature and salinity. In addition, GPx activity in hepatopancreas decreased after deltamethrin treatment mainly at 24 °C. Finally, AChE in muscle was strongly inhibited by deltamethrin at all tested temperatures and salinities. These novel findings demonstrate that interactions between abiotic factors and a commonly used pesticide exposure should be taken into account when analyzing some widespread biomarkers in black tiger shrimp.