Cholinesterase activity on Echinogammarus meridionalis (Pinkster) and Atyaephyra desmarestii (Millet): characterisation and in vivo effects of copper and zinc
- 298 Downloads
Metals are released into freshwater ecosystems from natural and anthropogenic sources, compromising their structural and functional equilibrium. As early warning tools, cholinesterases (ChEs) are usually used to assess the effects of organophosphate and carbamate pesticides, but are also known to be inhibited by metals. The objectives of this work were to characterise the activity of ChE present in the amphipod Echinogammarus meridionalis and the shrimp Atyaephyra desmarestii and to evaluate the in vivo effects of the metals copper and zinc in their ChE activity. To achieve this, firstly the activity of ChE forms were characterised using different in vitro assays with substrates and selective inhibitors. Then, the in vivo effects of 48 h exposures to increasing concentrations of copper and zinc on ChE activity were determined. The ChE form present in both species was acetylcholinesterase (AChE) since both revealed preference for the acetylthiocholine iodide substrate, total inhibition with eserine, the inhibitor of ChEs, and with 1,5-bis(4-allyldimethylammoniumphenyl)-pentan-3-one dibromide, the specific inhibitor of AChE, and presented insensitivity to iso-OMPA, a specific inhibitor of butyrylcholinesterase. The activity of ChEs was inhibited by zinc exposures in the amphipod species, but was not affected by copper. Exposure to copper and zinc did not affect ChEs activity in the shrimp at the concentrations tested. This work is a relevant contribution as foundation for the use of AChE in freshwater crustaceans in further studies including biomonitoring campaigns in different contamination scenarios.
KeywordsAcetylcholinesterase Amphipod Crustacean Essential metals Neurotoxicity Shrimp
The work of C. Quintaneiro was funded by FCT-Portuguese Foundation for Science and Technology through a doctoral and pos-doctoral fellowships (SFRH/BD/28705/2006 and SFRH/BPD/89951/2012) and M. Monteiro by a pos-doctoral fellowship (SFRH/BPD/45911/2008). We are very extremely grateful to Sr. Eduardo from Quinta do Alcaide who provided the access to Lena River sampling site.
Conflict of interest
The authors declare that they have no conflict of interest.
- Chuiko GM (2000) Comparative study of acetylcholinesterase and butyrylcholinesterase in brain and serum of several freshwater fish: specific activities and in vitro inhibition by DDVP, an organophosphorus pesticide. Comp Biochem Physiol C 127(3):233–242Google Scholar
- Eto M (1974) Organophosphorus pesticides; organic and biological chemistry. CRC Press, OhioGoogle Scholar
- Ferreira NGC, Rosário F, Domingues I, Calhôa CF, Soares AMVM, Loureiro S (2010) Acetylcholinesterase characterization in the terrestrial isopod Porcellionides pruinosus. In: Hamamura N, Suzuki S, Mendo S, Barroso CM, Iwata H, Tanabe S (eds) Interdisciplinary Studies on Environmental Chemistry—Biological Responses to Contaminants. TERRAPUB, pp. 227–236Google Scholar
- Frasco MF, Colletier JP, Weik M, Carvalho F, Guilhermino L, Stojan J, Fournier D (2007) Mechanisms of cholinesterase inhibition by inorganic mercury. Fed Eur Biochem Soc J 274(7):1849–1861Google Scholar
- Gagnaire B, Geffard O, Xuereb B, Margoum C, Garric J (2008) Cholinesterase activities as potential biomarkers: characterization in two freshwater snails, Potamopyrgus antipodarum (Mollusca, Hydrobiidae, Smith 1889) and Valvata piscinalis (Mollusca, Valvatidae, Müller 1774). Chemosphere 71(3):553–560CrossRefGoogle Scholar
- Garric J, Gagnaire B, Geffard O, Xuereb B, Margoum C (2008) Cholinesterase activities as potential biomarkers: characterization in two freshwater snails, Potamopyrgus antipodarum (Mollusca, Hydrobiidae, Smith 1889) and Valvata piscinalis (Mollusca, Valvatidae, Muller 1774). Chemosphere 71(3):553–560CrossRefGoogle Scholar
- Key PB, Fulton MH, Harman-Fetcho JA, McConnell LL (2003) Acetylcholinesterase activity in grass shrimp and aqueous pesticide levels from South Florida drainage canals. Arch Environ Contam Toxicol 45(3):371–377Google Scholar
- Lionetto MG, Caricato R, Giordano ME, Pascariello MF, Marinosci L, Schettino T (2003) Integrated use of biomarkers (acetylcholinesterase and antioxidant enzymes activities) in Mytilus galloprovincialis and Mullus barbatus in an Italian coastal marine area. Mar Pollut Bull 46(3):324–330CrossRefGoogle Scholar
- Macedo-Sousa JA, Gerhardt A, Brett CMA, Nogueira AJA, Soares AMVM (2008) Behavioural responses of indigenous benthic invertebrates (Echinogammarus meridionalis, Hydropsyche pellucidula and Choroterpes picteti) to a pulse of acid mine drainage: a laboratorial study. Environ Pollut 156(3):966–973CrossRefGoogle Scholar
- Monserrat JM, Bianchini A (1998) Some kinetic and toxicological characteristics of thoracic ganglia cholinesterase of Chasmagnathus granulata (Decapoda, Grapsidae). Comp Biochem Physiol C 120(2):193–199Google Scholar
- Peakal DB (ed) (1992) Animal biomarkers as pollution indicators. Chapman & Hall, LondonGoogle Scholar
- Quintaneiro C, Ranville J, Nogueira AJA (2013a) Effects of essential metals in two freshwater detritivores species: biochemical approach. Ecotoxicology (submitted)Google Scholar
- Quintaneiro C, Ranville J, Nogueira AJA (2013b) Physiological effects of essential metals on two detritivores: Atyaephyra desmarestii (Millet) and Echinogammarus meridionalis (Pinkster). Arch Environ Contam Toxicol (submitted)Google Scholar
- Sanchez-Hernandez JC (2006) Ecotoxicological perspectives of B-esterases in the assessment of pesticide contamination. In: Plattenberg RH (ed) Environmental pollution: new research. Nova Science, New York, pp 1–45Google Scholar
- Suresh A, Sivaramakrishna B, Victoriamma PC, Radhakrishnaiah K (1992) Comparative study on the inhibition of acetylcholinesterase activity in the freshwater fish Cyprinus carpio by mercury and zinc. Biochem Int 26(2):367–375Google Scholar
- Wilkinson GN (1961) Statistical estimations in enzyme kinetics. Biochem J 80:324–332Google Scholar