Académique Documents
Professionnel Documents
Culture Documents
Aquatic Toxicology
journal homepage: www.elsevier.com/locate/aquatox
CIIMAR Centro Interdisciplinar de Investigaco Marinha e Ambiental, Universidade do Porto, Porto, Portugal
CESAM Centro de Estudos do Ambiente e do Mar, Departamento de Biologia, Universidade de Aveiro, Aveiro, Portugal
ICBAS Instituto de Cincias Biomdicas Abel Salazar, Departamento de Estudos de Populaces, Universidade do Porto, Porto, Portugal
a r t i c l e
i n f o
Article history:
Received 19 February 2012
Received in revised form 9 August 2012
Accepted 14 August 2012
Keywords:
Swimming velocity
Cholinesterase
Palaemon serratus
Deltamethrin
Ecological relevant endpoint
a b s t r a c t
The main purpose of the present study was to investigate the effects of deltamethrin on biomarkers and
behavior of Palaemon serratus (common prawn), since this attempt to link different levels of biological
organization will allow determining which biomarkers might be ecologically relevant and will be useful
to complement the information about the effects of pesticides by using behavioral parameters. Therefore,
parameters of liver antioxidant status, energy metabolism and neurotransmission were determined in
different tissues of the common prawn and used to assess the effects at sub-individual level, whereas
swimming velocity was used to assess the effects at the individual level. It was also investigated if the
swimming velocity can be used as an endpoint in ecotoxicology bioassays and if it can be as sensitive as
biomarker endpoints.
Swimming velocity was signicantly reduced in prawns exposed to deltamethrin, showing a lowest
observed effect (LOEC) of 0.6 ng L1 . Eye acetylcholinesterase (AChE) activity was signicantly increased
in prawns exposed to 0.6, 1.2 and 2.4 ng L1 deltamethrin, whereas muscle cholinesterase (ChE) activity
was signicantly increased in prawns exposed to 19 and 39 ng L1 . On the other hand, lactate dehydrogenase (LDH) activity was signicantly increased in muscle of prawns exposed to 0.6, 1.2, 2.4,
4.9 ng L1 deltamethrin, showing that organisms were requiring additional energy, but probably using it
for detoxication processes rather than locomotion, since swimming velocity was inhibited. Glutathione
S-transferase (GST) activity was signicantly increased in the digestive gland of common prawn exposed
to 19 and 39 ng L1 deltamethrin. Catalase (CAT) activity was signicantly increased in digestive gland of
prawn exposed to 19 ng L1 deltamethrin. However, CAT activity decreased in digestive gland of prawn
exposed to 39 ng L1 , suggesting an antioxidant defense system failure concomitant with high levels of
lipid peroxidation. Thus, global results showed that decreased swimming velocity was not associated
with cholinesterase inhibition. In fact, the impairment of swimming velocity may be due to allocation
of energy for detoxication and antioxidant protection instead of swimming activity. The present study
showed that swimming velocity could be used as an ecologically relevant tool and a sensitive endpoint
to assess and complement the study of pesticide effects on marine organisms.
2012 Elsevier B.V. All rights reserved.
1. Introduction
Pyrethroids are a class of neurotoxic pesticides, the use of which
has been continuously increasing during the last two decades
(Amweg et al., 2005; Oros and Werner, 2005; Meacham et al., 2008;
Wolansky and Harrill, 2008). These compounds are derivatives and
synthetic analogues of natural pyrethrins (Soderlund et al., 2002;
Corresponding author at: CIIMAR Centro Interdisciplinar de Investigaco Marinha e Ambiental, Laboratrio de Ecotoxicologia e Ecologia, Universidade do Porto,
Rua dos Bragas, 289, 4050-123, Porto, Portugal. Tel.: +351 223401828.
E-mail addresses: colivei@ciimar.up.pt, cristianav.oliveira@gmail.com
(C. Oliveira).
0166-445X/$ see front matter 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.aquatox.2012.08.010
Hossain et al., 2005; Wolansky and Harrill, 2008), and can be categorized in two types, based on structure-activity and symptomology
(Soderlund et al., 2002; Nasuti et al., 2003; Hossain et al., 2005;
Meacham et al., 2008; Anadn et al., 2009). Nevertheless, both
types act mainly on the nervous system, especially on the voltagedependent sodium channels of excitable membranes, inducing a
prolongation of the sodium current during excitation caused by
membrane depolarization (Narahashi, 1992).
Wolansky and Harrill (2008) have reviewed a large number of
studies showing behavioral alterations after pyrethroid exposure.
Since most of them were performed with mammals, there is a need
to evaluate the effects of these pesticides in aquatic organisms, as
pyrethroids may be spread to aquatic environments by agricultural
and urban runoff (Wang et al., 2009). In fact, these compounds are
210
211
212
60
50
70
40
30
20
10
19.5
39
6
5
4
3
2
1
0
0
0
0.6
0'
1.2
2.4
4.9
9.8
19.5
39
0.6
0'
1.2
2.4
4.9
9.8
Fig. 2. Eye AChE (A) and muscle ChE (B) activities of P. serratus exposed to 0.639 ng L1 deltamethrin for 96 h. Values are the mean of 10 prawns per treatment with the
corresponding standard error bars. 0 and 0 represents control only with saltwater and control with solvent (control + DMSO), respectively. * indicates statistically signicant
differences (p < 0.05, Dunnett test) relative to control (control + DMSO).
reported for Paratya australiensis exposed to deltamethrin in articial saltwater at different salinities (about 35 ng L1 ) (Thomas
et al., 2008). Moreover, Viran et al. (2003) observed that the 48 h
LC50 value for guppies (Poecilia reticulata) exposed to deltamethrin
was 5130 ng L1 and Velsek et al. (2007) observed that the
96 h LC50 value for rainbow trout (Oncorhynchus mykiss) larvae and fry exposed to deltamethrin was 1170 and 1700 ng L1 ,
respectively. Thus, the current results show that P. serratus is
3.5
40
*
*
30
25
20
15
10
B
3
CAT activity
(nmol mg-1 protein-1 )
GST activity
(nmol mg-1 protein-1 )
35
2.5
2
1.5
1
0.5
0
0
0'
0.6
1.2
2.4
4.9
9.8
19.5
39
0'
800
2.4
4.9
9.8
19.5
39
700
LPO
nmol TBARS g wt-1
5
GPx activity
(nmol mg-1 protein-1 )
1.2
0.6
4
3
2
600
500
400
300
200
1
100
0
0
0'
0.6
1.2
2.4
4.9
9.8
19.5
39
0
0
0'
0.6
1.2
2.4
4.9
9.8
19.5
39
Fig. 3. Digestive gland GST (A), CAT (B), and GPx (C) activities and LPO levels (D) of P. serratus exposed to 0.639 ng L1 deltamethrin for 96 h. Values are the mean of 10
prawns per treatment with the corresponding standard error bars. 0 and 0 represents control only with saltwater and control with solvent (control + DMSO), respectively. *
indicates statistically signicant differences (p < 0.05, Dunnett test) relative to control (control + DMSO).
35
14
*
*
30
12
Muscle IDH activity
(nmol mg-1 protein-1)
40
213
25
20
15
10
10
8
6
4
2
5
0
0
0'
0.6
1.2
2.4
4.9
9.8
19.5
39
0'
L-1 )
0.6
1.2
2.4
4.9
9.8
19.5
39
Fig. 4. Muscle LDH (A) and IDH (B) activities of P. serratus exposed to 0.639 ng L1 deltamethrin for 96 h. Values are the mean of 10 prawns per treatment with the
corresponding standard error bars. 0 and 0 represents control only with saltwater and control with solvent (control + DMSO), respectively. * indicates statistically signicant
differences (p < 0.05, Dunnett test) relative to control (control + DMSO).
1000 ng L-1 Lost of movement coordination of Oncorhynchus mykiss juveniles (Velek et al., 2007)
1000 ng L-1 Significantly lower values of plasma glucose, alanine aminotransferase, cholinesterase
and significantly higher values of erythrocyte count, haemoglobin content, haematocrit and plasma
total protein, albumins, ammonia, aspartate aminotransferase, creatinekinase and calcium in
Oncorhynchus mykiss juveniles (Velek et al., 2007)
750 ng L-1 Significant increase of LPO levels and GSH content in liver, kidneys and gills; significant
induction of GST and GPx activities in liver and kidneys, and inhibition in gills; and significant
inhibition of CAT activity in liver, kidneys and gills in Channa punctatus (Sayeed et al., 2003)
135 to 212 ng L-1 LC50 (96 h) in Melanotaenia duboulayi exposed in water at different salinities
(Thomas et al., 2008)
100 ng L-1 Increase of tGSH content and CAT activity and inhibition of hepatopancreas GPx and
muscle AChE activities in Penaeus monodon. No significant effects on LPO levels neither on GST
activity (Tu et al., 2012)
100 ng L-1 Significantly increase od LPO levels in gills, however no significant effects on CAT, GPX
and GST activities (Dorts et al., 2009)
48 ng L-1 LC50 (96 h) in Palaemon serratus (in the present study)
39 ng L-1 Statistically significant increase of MDA content and GSH levels in Tinca tinca brain during
the recovery time, after 60 days of exposure (Hernndez-Moreno et al., 2010)
39 ng L-1 Significant increase of LPO levels in Palaemon serratus (in the present study)
32 to 37 ng L-1 LC50 (96h) in Paratya australiensis exposed in water at different salinities (Thomas et
al., 2008)
20 to 29 ng L-1 LC50 (96h) in Ceriodaphnia cf. dubia exposed in water at different salinities (Thomas
et al., 2008)
19.5 ng L-1 Significantly increase of muscle ChE activity and digestive gland GST and CAT
activities in Palaemon serratus (in the present study)
6 ng L-1 Significantly inhibition of Palaemon serratus swimming velocity and significantly
increase of eye AChE and muscle LDH activities (in the present study)
Fig. 5. Sensitivity of biomarkers and behavioral responses to deltamethrin exposure compared to lethal effects.
214
5. Conclusion
The swimming velocity was decreased in prawns exposed to
deltamethrin. The impairment of swimming velocity was not concomitant with the inhibition of AChE and ChE activities, showing
that behavioral impairment is not always explained by neurological disturbances. In addition, swimming velocity was still inhibited
despite the production of additional energy, suggesting an allocation of energy for detoxication and antioxidant protection.
Swimming velocity is essential for the performance and survival of
prawns and should be considered as an ecologically relevant endpoint, since it can be linked with tness-related parameters, such
Acknowledgements
This study was supported by the Portuguese Fundaco para a
Cincia e a Tecnologia (FCT), FEDER and European social funds of
MCTES (POPH-QREN-Tipology 4.2).
References
Abele, D., Vzquez Medina, J.P., Zenteno-Savin, T., 2011. Introduction to oxidative
stress in aquatic ecosystems. In: Abele, D., Vzquez Medina, J.P., Zenteno-Savin
(Eds.), Oxidative stress in aquatic ecosystem. Wiley-Blackwell, p. 548 pp.
Almeida, J., Oliveira, C., Gravato, C., Guilhermino, L., 2010. Linking behavioural alterations with biomarkers responses in the European seabass Dicentrarchus labrax
L. exposed to the organophosphate pesticide fenitrothion. Ecotoxicology 19,
13691381.
Almeida, J., Gravato, C., Guilhermino, L., 2012. Challenges in assessing the toxic
effects of polycyclic aromatic hydrocarbons to marine organisms: a case study
on the acute toxicity of pyrene to the European seabass (Dicentrarchus labrax L.).
Chemosphere 86, 926937.
Amiard-Triquet, C., 2009. Behavioral disturbances: the missing link between suborganismal and supra-organismal responses to stress? Prospects based on
aquatic research. Human and Ecological Risk Assessment 15, 87110.
Amweg, E.L., Weston, D.P., Ureda, N.M., 2005. Use and toxicity of pyrethroid pesticides in the Central Valley, California, USA. Environmental Toxicology and
Chemistry 24, 966972.
215
Frasco, M.F., Fournier, D., Carvalho, F., Guilhermino, L., 2006. Cholinesterase from the
common prawn (Palaemon serratus) eyes: catalytic properties and sensitivity to
organophosphate and cabamate compounds. Aquatic Toxicology 77, 412421.
Garca-de la Parra, L.M., Bautista-Covarrubias, J.C., Rivera-de la Rosa, N.,
Betancourt-Lozano, M., Guilhermino, L., 2006. Effects of methamidophos on
acetylcholinesterase activity, behavior and feeding rate of the white shrimp
(Litopenaeus vannamei). Ecotoxicology and Environment Safety 65, 372380.
Gravato, C., Guilhermino, L., 2009. Effects of benzo(a)pyrene on seabass (Dicentrrchus labrax L.): biomarkers, growth and behavior. Human and Ecological Risk
Assessment 15, 121137.
Guilhermino, L., Lopes, M.C., Carvalho, A.P., Soares, A.M.V.M., 1996. Acetylcholinesterase activity in juveniles of Daphnia magna Straus. Bulletin of
Environment Contamination and Toxicology 57, 979985.
Habig, W.H., Pabst, M.J., Jacoby, W.B., 1974. Glutathione S-transferases. The rst
enzymatic step in mercapturic acid formation. Journal of Biological Chemistry
249, 71307139.
Hernndez-Moreno, Soler, F., Mguez, M.P., Prez-Lpez, M., 2010. Brain acetylcholinesterase, malondialdehyde and reduced glutathione as biomarkers of
continuous exposure of tench, Tinca tinca, to carbofuran or deltamethrin. Science
of the Total Environment 408, 49764983.
Hossain, M.M., Suzuki, T., Sato, I., Takewaki, T., Suzuki, K., Kobayashi, H., 2004. The
modultory effect of pyrethroids on acetylcholine release in the hippocampus of
freely moving rats. NeuroToxicology 25, 825833.
Hossain, M.M., Suzuki, T., Sato, I., Takewaki, T., Suzuki, K., Kobayashi, H., 2005. Neuromechanical effects of pyrethroids, allethrin, cyhalothrin and deltamethrin on
the cholinergic processes in rat brain. Life Sciences 77, 795807.
Karasu Benli, A.C., Selvi, M., Sarikaya, R., Erkoc, F., Kocak, O., 2009. Acute toxicity
of deltamethrin on Nile Tilapia (Oreochoromis niloticus L., 1758) larvae and fry.
Gazi University Journal of Science 22, 14.
Lee, S.M., Koh, H.-J., Park, D.L., Song, B.J., Huh, T.L., 2002. Cytosolic NADP+-dependent
isocitrate dehydrogenase status modulates oxidative damage to cells. Free Radical Biology and Medicine 32, 11851196.
Lima, I., Moreira, S.M., Osten, J.R., Soares, A.M.V.M., Guilhermino, L., 2007. Biochemical responses of the marine mussel Mytilus galloprovincialis to petrochemical
environmental contamination along the North-western coast of Portugal.
Chemosphere 66, 12301242.
Livingstone, D.R., 2001. Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Marine Pollution Bulletin 42,
656666.
Meacham, C.A., Brodfuehrer, P.D., Watkins, J.A., Shafer, T.J., 2008. Developmentallyregulated sodium channel subunits are differentially sensitive to -cyano
containing pyrethroids. Toxicology and Applied Pharmacology 231, 273281.
Mohammad, A., Ranjbar, A., Shadnia, S., Nikfar, S., Rezaie, A., 2004. Pesticides and
oxidative stress: a review. Medical Science Monitor 10, 141147.
Mouneyrac, C., Leung, P.T.Y., Leung, K.M.Y., 2011. Costs of tolerance. In: AmiardTriquet, C., Rainbow, P.S., Romeo, M. (Eds.), Tolerance to Environmental
Contaminants. Environmental and Ecological Risk Assessment Series. CRC Press,
Boca Raton, p. 446.
Narahashi, T., 1992. Nerve membrane Na+ channels as target of insecticides. Trends
in Pharmacological Sciences 13, 236241.
Nasuti, C., Cantalamessa, F., Falcioni, G., Gabbianelli, R., 2003. Different effects of
type I and type II pyrethroids on erythrocyte plasma membrane properties and
enzymatic activity in rats. Toxicology 191, 233244.
Nrum, U., Friberg, N., Jensen, M.R., Pedersen, J.M., Bjerregaard, P., 2010. Behavioural
changes in three species of freshwater macroinvertebrates exposed to the
pyrethroid lambda-cyhalothrin: laboratory and stream microcosm studies.
Aquatic Toxicology 98, 328335.
Ohkawa, H., 1979. Assay for lipid peroxides in animal tissues by thiobarbituric acid
reaction. Analytical Biochemistry 95, 351358.
Oros, D.R., Werner, I., 2005. Pyrethroid insecticides: an analysis of use patterns,
distributions, potential toxicity and fate in the Sacramento-San Joaquin Delta
and Central valley. White paper for the Interagency Ecological Program. SFEI
contribution 415. San Francisco Estuary Institute, Oakland, CA.
Plaut, I., 2001. Critical swimming speed: its ecological relevance. Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 131,
4150.
Rao, V.J., 2006. Sublethal effects of an organophosphorous insecticide (RPR-II)
on biochemical parameters of tilapia, Oreochromis mossambicus. Comparative Biochemistry and Physiology. Toxicology & Pharmacology: CBP 143,
492498.
Regoli, F., Benedetti, M., Giuliani, M.E., 2011. Antioxidant defenses and acquisition
of tolerance to chemical stress. In: Amiard-Triquet, C., Rainbow, P.S., Romeo, M.
(Eds.), Tolerance to Environmental Contaminants. Environmental and Ecological
Risk Assessment Series. CRC Press, Boca Raton, p. 446.
Roast, S.D., Widdows, J., Jones, M.B., 2000a. Disruption of swimming in the hyperbenthic mysid Neomysis integer (Peracarida: Mysidacea) by the organophosphate
pesticide chlorpyrifos. Aquatic Toxicology 47, 227241.
Roast, S.D., Thompson, R.S., Donkin, P., Widdows, J., Jones, M.B., 2000b. Mysids and
trace metals: disruption of swimming as a behavioural indicator of environmental contamination. Marine Environment Research 50, 107112.
Roast, S.D., Widdows, J., Jones, M.B., 2001. Impairment of mysid (Neomysis integer) swimming ability: an environmentally realistic assessment of the impact of
cadmium exposure. Aquatic Toxicology 52, 217227.
Snchez-Fortn, S., Barahona, M.V., 2005. Comparative study on the environmental
risk induced by several pyrethroids in estuarine and freshwater invertebrate
organisms. Chemosphere 59, 553559.
216
Sancho, E., Ferrando, M.D., Andreu, E., 1997. Response and recovery of brain
acetylcholinesterase activity in the European eel. Anguilla anguilla, exposed to
fenitrothion. Ecotoxicology and Environment Safety 38, 205209.
Sancho, E., Ferrando, M.D., Andreu, E., 1998. In vivo inhibition of AchE activity in the
European eel Anguilla anguilla exposed to technical grade fenitrothion. Comparative Biochemistry and Physiology. Toxicology & Pharmacology: CBP 120,
389395.
Sayeed, I., Parvez, S., Pandey, S., Bin-Hafeez, B., Haque, R., Raisuddin, S., 2003. Oxidative stress biomarkers of exposure to deltamethrin in freshwater sh, Channa
punctatus Bloch. Ecotoxicology and Environment Safety 56, 295301.
Soderlund, D.M., Clark, J.M., Sheets, L.P., Mullin, L.S., Piccirillo, V.J., Sargent, D.,
Stevens, J.T., Weiner, M.L., 2002. Mechanisms of pyrethroid neurotoxicity: implications for cumulative risk assessment. Toxicology 171, 359.
Thomas, C.R., Hose, G.C., Warne, M.St.J., Lim, R.P., 2008. Effects of river water and
salinity on the toxicity of deltamethrin to freshwater shrimp, cladoceran and
sh. Archives of Environment Contamination and Toxicology 55, 610618.
Torres, M.A., Testa, C.P., Gaspari, C., Masutti, M.B., Panitz, C.M., Curi-Pedrosa, R., de
Almeida, E.A., Di Mascio, P., Filho, D.W., 2002. Oxidative stress in the mussel
Mytella guyanensis from polluted mangroves on Santa Catarina Island, Brazil.
Marine Pollution Bulletin 44, 923932.
Tu, H.T., Silvestre, F., Meulder, B., Thome, J.P., Phuong, N.T., Kestemont, P., 2012.
Combined effects of deltamethrin, temperature and salinity on oxidative stress
biomarkers and acetylcholinesterase activity in the black tiger shrimp (Penaeus
monodon). Chemosphere 86, 8391.
Tuzmen, N., Candan, N., Kaya, E., Demiryas, N., 2008. Biochemical effects of chlorpyrifos and deltamethrin on altered antioxidative defense mechanisms and lipid
peroxidation in rat liver. Cell Biochemistry and Function 26, 119124.
van der Oost, R., Beyer, J., Vermeulen, N.P.E., 2003. Fish bioaccumulation and
biomarkers in environmental risk assessment: a review. Environmental Toxicology and Pharmacology 13, 57149.
Vassault, A., 1983. Lactate dehydrogenase. In: Bergmyer, M.O. (Ed.), Methods of
Enzymatic Analysis Enzymes: Oxireductases, Transferases. Academic Press, New
York, pp. 118126.
Velsek, J., Jurckov, J., Dobskov, R., Svobodov, Z., Piackov, V., Mchov, J.,
L., 2007. Effects of deltamethrin on rainbow trout (Oncorhynchus
Novotny,
Environmental
Toxicology
and
Pharmacology
23,
297
mykiss).
301.
Vijayavel, K., Balasubramanian, M.P., 2009. Effect of fenvalerate on oxidative stress
biomarkers in the brackish water prawn Penaeus monodon. Pesticide Biochemistry and Physiology 95, 113116.
Viran, R., Erkoc, F.., Polat, H., Kocak, O., 2003. Investigation of acute toxicity of
deltamethrin on guppies (Poecilia reticulata). Ecotoxicology and Environment
Safety 55, 8285.
Wang, C., Chen, F., Zhang, Q., 2009. Chronic toxicity and cytotoxicity of synthetic
pyrethroid insecticide cis-bifenthrin. Journal of Environmental Sciences 21,
17101715.
Wang, X., Zhai, W., 1998. Cellular and biochemical effects in the bronchoalveolar
lavage uid of the rats exposed to fenvalerate. Chinese Journal of Pharmacology
and Toxicology 2, 271276.
Wolansky, M.J., Harrill, J.A., 2008. Neurobehavioral toxicology of pyrethroid insecticides in adult animals: a critical review. Neurotoxicology and Teratology 30,
5578.
Yousef, M.I., Awad, T.I., Mohamed, E.H., 2006. Deltamethrin-induced oxidative
damage and biochemical alterations in rat and its attenuation by vitamin E.
Toxicology 227, 240247.
Zhang, P., Zhang, X., Li, J., Huang, G., 2006. Swimming ability and physiological response to swimming fatigue in whiteleg shrimp, Litopenaeus vannamei.
Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 145, 2632.