Altered quantities and in vivo activities of cholinesterase from Daphnia magna in sub-lethal exposure to organophosphorus insecticides
Highlights
► ChE-derived antigen was utilized to measure content of ChE in Daphnia. ► Up to 68.0 percent promotion in content of ChE was found in long-term exposure to OPs. ► Inherent activity was more sensitive in detection of long-term stress of OPs. ► Inherent activity recovered faster than the rate of dissipation of OPs.
Introduction
Anticholinesterases such as organophosphorus (OPs) and carbamate (CBs) are among the most commonly used pesticides for insect pests management. Despite the high efficiencies, they tend to be toxic to non-target organisms such as fish and many species of aquatic invertebrates. It is known that the principal mechanism by which OPs and CBs elicit their toxicity comes down to the inhibition of cholinesterase (ChE) (Carvalho et al., 2003). In this sense, a number of studies had been performed to urge the enzyme as a biomarker for prediction of anticholinesterases (Sanchez-Hernandez et al., 1998, Stien et al., 1998, Cajaraville et al., 2000, Den Besten et al., 2001, Abdel-Halim et al., 2006).
Anticholinesterases may affect ChE in multi aspects in vivo. It is common to see promotion in terms of activity of the enzyme in exposures (Frawley et al., 1952, Frawley and Fuyat, 1957, Genina, 1974, Hackenberger et al., 2008), a phenomenon that can be regard as a type of hormesis featured by conversion from low-dose stimulation to high-dose inhibition (Calabrese, 2002, Chapman, 2002). This promotion is probably result from acceleration in terms of synthesis of the enzyme in order to rectify functional disable in condition that part of the enzyme was inhibited. The point is that the anticholinesterase- caused promotion oftentimes disturbs the detection of “actual” inhibition of the enzyme so as to bring about difficulty for determining exposure of anticholinesterases in field (Khattab et al., 1994, Khattab and Ali, 2007). It is necessary, therefore, to develop method for measuring relative content of ChE so as to scale the “actual” activity of the enzyme in exposures. Enzyme-linked immunosorbent assay (ELISA), which combined efficiency and specificity, is capable of achieving the task.
Daphnia magna is a species of zooplankton that belongs to Phylum of Arthropoda, Order of Crustacea, and Family of Daphniidae. It consumes algae and other small-sized phytoplankton and in turn serves as prey for freshwater fish and macro-sized aquatic invertebrates. Due to its world-wide distribution and its adaptability to laboratory culture, Daphnia has long been employed as representative species for testing of chemicals (OECD (Organisation for Economic Co-operation and Development), 2004, OECD (Organisation for Economic Co-operation and Development), 2008, USEPA (U.S. Environmental Protection Agency), 1996a, USEPA (U.S. Environmental Protection Agency), 1996b). Some papers had been published to reveal relationship between in vivo activity of ChE and ambient concentrations of anticholinesterases (Barata et al., 2001, Printes and Callaghan, 2003, Carvalho et al., 2003, Barata et al., 2004, Damásio et al., 2007, Printes et al., 2008). Results of the studies confirmed that, to behave the same as fish and many other aquatic organisms, Daphnia had its enzyme obviously inhibited at concentrations adjacent to the lethal ones, conflict results were obtained, however, in case that the Daphnia were exposed to concentrations far below the lethal ones.
Triazophos and chlorpyrifos are two kinds of OPs commonly used nowadays in China and southeast of Asia. Considering the physico-chemical properties and application rates, there is large possibility for them to enter water-sediment systems and impose acute and/or chronic hazard to non-target aquatic organisms. A former study indicated a maximal of 76.5 percent and 60.5 percent promotion in terms activity of the ChE, as the Daphnia were exposed, in semi-static condition, to either triazophos or chlorpyrifos for 21 day (Li and Tan, 2010). This suggested accelerated synthesis and consequent promotion in quantity of the enzyme.
In a study conducted with Japanese quail Coturnix coturnix, a maximal of 40 percent promotion in quantity of immunoreactive ChE was detected in serum in case the bird was orally administered with monocrotophos of 20 mg kg−1 (Khattab et al., 1994). Another study conducted with red-legged partridges Alectoris rufa cross revealed a maximal of 26.5 percent rise in quantity of immunoreactive ChE in serum in case the bird was orally administered with malathion of 167 mg kg−1 (Khattab and Ali, 2007). Up to of 50 percent increase in quantity of the enzyme was detected in brain of Psudorasbora parva in case the fish were exposed to sub-lethal concentrations of triazophos (Li et al., 2005). It is logical, therefore, to infer that anticholinesterases such as triazophos and chlorpyrifos could result in promotion in terms of quantity of the ChE in Daphnia.
A type of antigen that was formerly developed by purified ChE from Daphnia was utilized in this study for measuring content of ChE in Daphnia being exposed for 21 d to various concentrations of either of triazophos or chlorpyrifos. The aim of this study was: (1) to clarify extent and pattern of promotion in terms of quantity of ChE in Dphnia along with time and concentrations, (2) to explore relationship between in vivo activity of ChE and ambient concentrations of anticholinesterases. Result of the study would help to learn advantages and disadvantages of in vivo ChE in detecting sub-lethal and/or long-term exposure of anticholinesterases and would finally benefit in situ application of the biomarker.
Section snippets
Reagents
Triazophos (with purity of 96.6 percent) and chlorpyrifos (with purity of 99.5 percent) were obtained from the National Standards Company (Beijing, China). Horseradish peroxidase (HRP) labeled goat anti-mouse immunoglobulin G (IgG) with the enzyme of immunoassay grade purity, Coomassie Brilliant Blue G-250, propionylthiocholine iodide, and bovine serum albumin (BSA, with MW of 67,000) were obtained from Sigma-Aldrich® (Steinheim, Germany). Tramethylbenzidine (TMB), which consists of solution A
Acute bioassays
In acute bioassays, the 48h EC50 were found to be 12.92 μg L−1 and 7.12 μg L−1, respectively, with 95 percent confidence limit of 10.55–16.23 μg L−1 and 5.79–9.07 μgL−1 for triazophos and chlorpyrifos, respectively (Table 1). This indicated that the Daphnia were more sensitive to chlorpyrifos than to triazophos. Nominal concentrations employed in 21 d exposure were thus selected to be 0.06, 0.35, and 2.10 μg L−1 for triazophos, and they were selected to be 0.03, 0.20, and 1.19 μg L−1 for chlorpyrifos. The
Acute toxicities
Previous studies indicated that the 48 h EC50 of chlorpyrifos for Daphnia magna was 0.74 μg L−1 (Palma et al., 2008), 0.8 μg L−1 (Robinson, 1999), 0.19 μg L−1 (Kikuchi et al., 2000), and 3.17 pM (i.e. 1.11 μg L−1) (Printes and Callaghan, 2003), respectively, which was in some degree lower that derived from this study (i.e. 7.12 μg L−1). Not much data is published for triazophos except a paper in which the 48 h EC50 was reported to be 13.8 μg L11 (Tan et al., 2004). According to Barata et al. (2001), a clone
Conclusions
In case the Daphnia.magna were exposed to either triazophos or chlorpyrifos at nominal concentration of 1/6EC50, 1/36 EC50, and 1/216 EC50, inherent activity of the ChE, i.e. the activity that was scaled with immunoreactive protein, was found inhibited while the immureactive content of the enzyme was found increased. The increase resulted in obvious promotion in terms of total activity of ChE at different combinations of time and concentration. Result of the study indicated that the inherent
Acknowledgments
This work was financially supported by Department of Education of Zhejiang Province, PR China (Natural Science Project, no. 20070138).
References (46)
- et al.
Organophosphorus pollutants (OPP) in aquatic environment at Damietta Governorate, Egypt: Implications for monitoring and biomarker responses
Chemosphere
(2006) - et al.
Biochemical factors contributing to response variation among resistant and sensitive clones of Daphnia magna Straus exposed to ethyl parathion
Ecotoxicol. Environ. Saf.
(2001) - et al.
Role of B-esterases in assessing toxicity of organophosphorus (chlorpyrifos, malathion) and carbamate (carbofuran) pesticides to Daphnia magna
Aquat. Toxicol.
(2004) Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein dye binding
Analyt. Biochem.
(1976)- et al.
The use of biomarkers to assess the impact of pollution in coastal environments of the Iberian Peninsula: a practical approach
Sci. Total Environ.
(2000) Hormesis: changing view of the dose-response, a personal account of the history and current status
Mut. Res.
(2002)- et al.
Use of atropine-treated Daphnia magna survival for detection of environmental contamination by acetylcholinesterase inhibitor
Ecotoxicol. Environ. Saf.
(2003) Ecological risk assessment (ERA) and hormesis
Sci. Total Environ.
(2002)- et al.
Biochemical mechanisms of resistance in Daphnia magna exposed to the insecticide fenitrothion
Chemosphere
(2007) - et al.
Bioaccumulation and biomarkers in the sea star Asterias rubens (Echinodermata: Asteroidea): a North Sea field study
Marine Environ. Res.
(2001)
Trace nutrient deficiency in Daphnia magna cultured in standard medium for toxicity testing: Effects of the optimization of culture conditions on life history parameters of Daphnia magna
Wat. Res.
Effect of temephos on cholinesterase activity in the earthworm Eisenia fetida (Oligochaeta, Lumbricidae)
Ecotoxicol. Environ. Saf.
Anticholinesterase induce multigenic transcriptional feedback response suppressing cholinergic neurotransmission
Chemico-Biological Interact
Immunoassays for avian butyrylcholinesterase: Implications for ecotoxicological testing and clinical biomonitoring
Environ. Toxicol. Pharmacol.
Screening of organophosphate insecticide pollution in water by using Daphnia magna
Ecotoxicol. Environ. Saf.
Protection against soman or VX poisoning by human butyrylcholinesterase in guinea pigs and cynomolgus monkeys
Chemico-Biological Interact
The toxicity of pesticides and organics to mysid shrimps can be predicted from daphnia spp. Toxicity data
Wat. Res.
Clonal variation in acetylcholinesterase biomarkers and life history traits following OP exposure in Daphnia magna
Ecotoxicol. Environ. Saf.
Use of biochemical biomarkers as a screening tool to focus the chemical monitoring of organic pollutants in the Biobio river basin (Chile)
Chemosphere
Evaluation of biomarkers in caged fishes and mussels to assess the quality of waters in a bay of the NW Mediterranean Sea
Environ. Pollution
How many genes encode cholinesterase in arthropods?
Pestic. Biochem. Physiol
Genetic variation in the response of Daphnia to toxic substances: Implications for Risk Assessment
Characterization of cholinesterases from Daphnia magna Straus and their inhibition by zinc
Bull. Environ. Contam. Toxicol.
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