title: Biomarkers of waterborne copper exposure in the Neotropical fish Prochilodus lineatus.
abstract: The main goal of the present study was to investigate the effects of acute exposure to copper (Cu) using a Neotropical freshwater fish as sentinel species through multi biomarkers analysis at different biological levels. Juveniles of Prochilodus lineatus were kept under control condition (no Cu addition in the water) or exposed to environmentally relevant concentrations of waterborne Cu (5, 9 and 20μgL(-1)) for 96h. These concentrations were selected to bracket the current Brazilian water quality criteria for Cu in fresh water (9 and 13μgL(-1) dissolved copper). Endpoints analyzed included ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity, reduced glutathione (GSH) and metallothionein-like protein (MT) concentration, lipid peroxidation (LPO) level, tissue damage index, and incidence of free melano-macrophages (FMM) and melano-macrophage centers (MMC) in the liver. They also included DNA damage (frequency of nucleoids per comet class, number of damaged nucleoids per fish and DNA damage score) in erythrocytes, as well as muscle and brain acetylcholinesterase (AChE) activity and behavioral parameters (swimming distance and velocity, time spent swimming and swimming activity in the upper and lower layers of the water column). Fish exposed to any of the Cu concentrations tested showed increased liver MT concentration and LPO level, higher number of damaged nucleoids in erythrocytes per fish, and inhibited muscle AChE activity. Also, increased liver SOD activity was observed in fish exposed to 9 and 20μgL(-1) Cu. Fish exposed to 5 and 9μgL(-1) Cu spent lower amount of time swimming. Fish exposed to 9μgL(-1) Cu showed increased swimming distance and velocity while those exposed to 20μgL(-1) Cu had lower swimming distance and velocity, as well as, spent less time swimming in the lower layer of the water column when compared to those kept under control condition. These findings indicate that Cu exposure at environmentally relevant concentrations (below or close to the current Brazilian water quality criteria) induced significant biological (histological, biochemical and genetic) and ecological (swimming and exploratory abilities) damages in the Neotropical fish P. lineatus. They also suggest that MT concentration, DNA damage (comet assay), LPO (TBARS method), SOD and AChE activity, together with swimming behavior analyses are potential biomarkers to assess and monitor areas impacted by Cu in fresh water.
authors.affiliation: Departamento de Ciências Fisiológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil. Electronic address: julisimonato@gmail.com.
authors.forename: Juliana D
authors.initials: JD
authors.lastname: Simonato
authors.affiliation: Departamento de Biologia Celular e Departamento de Farmacologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil.
authors.forename: Maritana
authors.initials: M
authors.lastname: Mela
authors.affiliation: Departamento de Biologia Celular e Departamento de Farmacologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil.
authors.forename: Halina B
authors.initials: HB
authors.lastname: Doria
authors.affiliation: Departamento de Biologia Celular e Departamento de Farmacologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil.
authors.forename: Izonete C
authors.initials: IC
authors.lastname: Guiloski
authors.affiliation: Departamento de Biologia Celular e Departamento de Farmacologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil.
authors.forename: Marco A F
authors.initials: MAF
authors.lastname: Randi
authors.affiliation: Departamento de Zoologia, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil.
authors.forename: Paulo S M
authors.initials: PSM
authors.lastname: Carvalho
authors.affiliation: Departamento de Ciências Fisiológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
authors.forename: Paulo C
authors.initials: PC
authors.lastname: Meletti
authors.affiliation: Departamento de Biologia Celular e Departamento de Farmacologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil.
authors.forename: Helena C
authors.initials: HC
authors.lastname: Silva de Assis
authors.affiliation: Instituto de Ciências Biológicas, Universidade Federal de Rio Grande, Rio Grande, Rio Grande do Sul, Brazil.
authors.forename: Adalto
authors.initials: A
authors.lastname: Bianchini
authors.affiliation: Departamento de Ciências Fisiológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
authors.forename: Claudia B R
authors.initials: CBR
authors.lastname: Martinez
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D001522:Behavior, Animal; D015415:Biomarkers / Q000378:metabolism*; D001938:Brazil; D002374:Catalase / Q000378:metabolism; D003300:Copper / Q000032:analysis / Q000633:toxicity*; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D004781:Environmental Exposure / Q000032:analysis*; D004912:Erythrocytes / Q000378:metabolism; D005399:Fishes / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D013550:Swimming; D014329:Tropical Climate; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D015415:Biomarkers; D014874:Water Pollutants, Chemical; D003300:Copper; D002374:Catalase; D005979:Glutathione Peroxidase; D019363:Cytochrome P-450 CYP1A1; D013482:Superoxide Dismutase; D005982:Glutathione Transferase; D000110:Acetylcholinesterase; D005978:Glutathione
keywords: Acetylcholinesterase; Behavior; Genotoxicity; Histopathology; Metallothionein; Oxidative stress