Genotoksični učinak sedimenta Riječkog zaljeva na staničnu kulturu ribljih hepatocita PLHC-1 mjeren Komet testom

master's thesis

• Title (english): Genotoxic effect of Rijeka Bay sediments on PLHC-1 fish hepatoma cell culture assessed by the Comet assay
• Author: Sonja Kralj
• Mentor: Goran Klobučar (mentor)
• Mentor: Maja Šrut (pomoć pri izradi, neposredni voditelj)
• Committee member: Goran Klobučar (predsjednik povjerenstva)
• Committee member: Zdravko Dolenec (član povjerenstva)
• Committee member: Mirjana Pavlica (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2010, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Brojni toksični onečišćivači antropogenog porijekla mogu se akumulirati u morskim sedimentima i kasnije prijeći u vodene organizme te izazvali toksični učinak. Procjena genotoksičnog učinka onečišćenih sedimenata na staničnim kulturama može poslužiti kao rani pokazatelj negativnih promjena u okolišu. U ovom je radu genotoksičnost uzoraka sedimenata prikupljenih na šest postaja različitog stupnja onečišćenja u Riječkom zaljevu istražena komet testom na staničnoj liniji ribljih hepatocita PLHC-1. PLHC-1 stanice bile su izložene različitim koncentracijama organskih ekstrakata sedimenta tijekom 24 h. Svih šest uzoraka sedimenta pokazali su genotoksični učinak ovisan o koncentraciji. Kako bi se odredilo koja je grupa onečišćivača većinom odgovorna za opaženu genotoksičnost, sediment s najvećom genotoksičnosti dodatno je ekstrahiran s tri otapala različitog stupnja polarnosti. Nepolarna otapala (cikloheksan, diklormetan) pokazala su veću genotoksičnost nego polarno otapalo (metanol), dok je najveću genotoksičnost pokazao sediment ekstrahiran smjesom otapala diklormetan-metanolom koja ekstrahira široki spektar onečišćivača. Komet test na staničnoj kulturi PLHC-1 pokazao se kao prikladna metoda procjene genotoksičnosti sedimenata.

Abstract (english)

Numerous toxic anthropogenic pollutants can accumulate in marine sediments and, later on, enter the aquatic organisms and cause toxic effects. Assessment of genotoxic effects of contaminated sediments using cell culture can serve as an early indicator of negative alterations in the environment. In this study, the genotoxicity of sediment samples collected at the six sites of different pollution intensity in the Rijeka Bay was assessed by the Comet assay on PLHC-1 fish hepatoma cell line. PLHC-1 cells were exposed to different concentrations of organic sediment extracts for 24 h. All of the six sediment samples showed a concentration dependent genotoxic effect. In order to determine which group of pollutants is mostly responsible for the observed genotoxicity, the sediment that demonstrated highest genotoxicity was additionally extracted with three solvents which differ by the degree of polarity. Nonpolar solvents (cyclohexane, dichloromethane) revealed higher genotoxicity than the polar solvent (methanol), while the highest genotoxicity was obtained by the sediment extracted using a solvent mixture dichloromethane-methanol which extracts a wide range of pollutants. Comet assay on PLHC-1 cell culture proved to be an advantageous method of sediments genotoxicity assessment.

• Keywords: sediment; genotoksičnost; komet test; PLHC-1 stanična linija
• Keywords (english): sediments; genotoxicity; Comet assay; PLHC-1 cell line
• Language: croatian
• URN:NBN: urn:nbn:hr:217:658898
• Study programme: Title: Biology; specializations in: Ecology; Course: Ecology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra biologije (magistar/magistra biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2018-03-27 10:09:18