Ecogenotoxicological evaluation of surface water quality with comet assay on different tissues of european chub (Squalius cephalus L.)

Intenzivni procesi proizvodnje i oslobađanje velikog broja hemijskih agenasa, kao i metala u spoljašnju sredinu, najvećim delom u rečne tokove, dovela je do potrebe za primenom testova koji će detektovati njihov genotoksični potencijal. Mnogi akvatični organizmi, među kojima bitno mesto zauzimaju ribe, imaju sposobnost bioakumulacije genotoksičnih agenasa u tkivima, te se mogu koristiti kao bioindikatori u ekogenotoksikološkim studijama. Ribe se smatraju za jedne od značajnijih indikatora za procenu nivoa metala u slatkovodnim sistemima, jer su u vrhu lanca ishrane sa mogućnošću akumulacije visokih koncentracija pojedinih metala. Procena genotoksičnosti površinskih voda vršena je na osnovu detekcije oštećenja DNK molekula alkalnim komet testom, testom elektroforeze pojedinačnih ćelija, (engl. Single Cell Gel Electroforesis, SCGE), u krvi, jetri i škrgama klena (Squalius cephalus L.). Rangiranje parametara za procenu DNK oštećenja: dužina repa komete (TL), intenzitet repa komete (TI) i „Olive tail moment” (OTM), urađeno je metodom SRD (engl. Sum of Ranking Differences). Paralelno sa komet testom, u jetri i škrgama, pratila se i koncentracija metala i metaloida pomoću induktivno spregnute plazme – optičko emisione spektrometrije (ICP - OES). Analiza metala i metaloida je urađena i u mišićima i gonadama istom metodom. Procena ekološkog statusa površinskih voda određivala se na dve akumulacije, Garaši i Zlatar (Uvac) i dve reke, Peštan i Beljanica tokom 2011/2012 godine. Procena kvaliteta vode određena je na osnovu fizičko-hemijskih i mikrobioloških parametara. Analizirano je 7 grupa mikroorganizama: ukupni koliformi, fekalni koliformi, fekalne enterokoke, aerobni mezofili, aerobni heterotrofi, heterotrofi i oligotrofi. Poreklo zagađenja dobijeno je na osnovu odnosa fekalnih koliforma i fekalnih enterokoka F/E, a status samoprečišćavanja dobijen je iz odnosa heterotrofa i oligotrofa O/H. Integrisani odgovor biomarkera (IBR) je korišćen kao pokazatelj odnosa različitih grupa mikroorganizama tokom višemesečnog monitoringa...Intensive production and release of a large number of chemical agents, as well as metals into the environment, mainly in river flows, has led to the need for tests which can determine their genotoxic potential. Many aquatic organisms, especially fish, have a potential for bioaccumulation of genotoxic agents in the tissues, and hence can be used as bioindicators in ecogenotoxicological studies. Fish are considered one of the most important indicators for assessing the levels of metals in freshwater systems, because they are at the top of the food chain with the ability to accumulate high concentrations of certain metals. Assessment of surface water genotoxicity was carried out based on the detection of DNA damage with alkaline comet assay or single cell gel electrophoresis (SCGE) in the blood, liver and gills of chub (Squalius cephalus L.). Ranking of parameters for assessing the DNA damage: comet tail length (TL), the intensity of the tail of the comet (TI) and "Olive tail moment" (OTM), was done by the SRD (Sum of Ranking Differences) method. In parallel with the comet assay in liver and gills, inductively coupled plasma - optical emission spectrometry (ICP - OES) was used to follow the concentration of metals and metalloids. The same method was used for the analysis of metals and metalloids in the muscles and gonads. Assessment of the ecological status of surface waters was conducted at two reservoirs, Garaši and Zlatar (Uvac), and two rivers Peštan and Beljanica, during the 2011/2012 year. Water quality assessment is based on the physicochemical and microbiological parameters. Microbiological analysis included: total coliforms, fecal coliforms, fecal enterococci, aerobic mesophile, aerobic heterotrophs, heterotrophs and oligotrophs. The type of contamination was determined by the ratio of fecal coliforms and fecal enterococci F/E, and the status of self-purification is obtained from the ratio of heterotrophs and oligotrophs O/H. Integrated biomarker response (IBR) has been used as an indicator of the relationship between different groups of microorganisms during the months of monitoring..