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Določanje mikrobnih združb na mikroplastiki v morski vodi in v školjkah
ID Koroša, Vanja (Author), ID Klančnik, Anja (Mentor) More about this mentor... This link opens in a new window, ID Kovac Viršek, Manca (Comentor)

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Abstract
Sredozemsko morje predstavlja eno najbolj prizadetih območij onesnaženja s plastičnimi polimeri. Mikroplastiko lahko zaradi majhnosti zaužijejo morski organizmi, ki so del prehrane ljudi. Posledično predstavljajo delci mikroplastike tveganje za prenos kemijskih in mikrobioloških onesnaževal. V sklopu naloge smo vzorčili morsko vodo in školjke na treh lokacijah na slovenski obali. Iz vzorcev morske vode in školjk smo izolirali mikroplastiko, izolirali DNA z namenom določanja mikrobnih združb na delcih ter določili kemijsko strukturo delcev. Osredotočili smo se izključno na delce v obliki fragmentov, ki so poleg filamentov, kroglic in pen ena izmed oblik mikroplastike. Iz vode smo vzorce mikroplastike pridobili s filtracijo, iz školjk pa s sekcijo školjk in vizualnim pregledom s stereomikroskopom. Zaradi majhnosti vzorcev smo za čim večjo količino DNA najprej s testnimi vzorci mikroplastike optimizirali izolacijo DNA, pri čemer smo spreminjali korake za mehansko razgradnjo celic ter komercialne kite za izolacijo. Z metagenomsko analizo bakterijskega gena za 16S rRNA izbranih delcev mikroplastike smo ugotovili, da so bile na mikroplastiki najpogosteje prisotne bakterije debel Proteobacteria, Cyanobacteria, Firmicutes, Actinobacteria in Bacteroidetes, ki se poleg ostalih debel v povprečju uvrščajo v preko 400 različnih vrst bakterij. Nekatere izmed bakterij, izoliranih iz školjk, so bili človeški patogeni. Vseh rezultatov metagenomske analize nismo mogli povezati z mikroplastiko zaradi nezmožnosti določitve polimerne sestave nekaterih delcev. S pomočjo spektrokopije FTIR smo določili kemijsko sestavo delcev in za pet izmed 23 vzorcev potrdili, da gre za polimerne materiale iz polivinil klorida, polietilena, akrilonitril butadien stirena in polipropilena. Za preostale delce nismo mogli z gotovostjo dokazati, da so iz polimernih materialov.

Language:Slovenian
Keywords:mikroplastika, morsko okolje, Slovenija, školjke, mikrobne združbe, optimizacija, izolacija DNA, spektroskopija, metagenomska analiza, sekcija
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Publisher:[V. Koroša]
Year:2024
PID:20.500.12556/RUL-156221 This link opens in a new window
UDC:504.5:678.5/.7:579.6:614.3
COBISS.SI-ID:195561987 This link opens in a new window
Publication date in RUL:15.05.2024
Views:132
Downloads:310
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Secondary language

Language:English
Title:Determination of microbial communities on microplastics in marine environment and shellfish
Abstract:
The Mediterranean Sea is one of the most affected areas by plastic pollution. Because of their small size, microplastics can be ingested by marine organisms that are part of the human diet. This poses the risk of transporting of chemical and microbiological contaminants. In the present study, we isolated DNA to determine the microbial communities on microplastics in seawater and shellfish on the Slovenian coast, as well as the chemical structure of the particles. We focused primarily on fragments, which are one of the types of microplastics besides filaments, beads and foams . Microplastic samples were obtained from water by filtration, and from mussels by cutting the mussels open and visual inspection with a stereomicroscope. Due to the small size of the samples, we attempted to optimize DNA isolation by adding steps to mechanically disrupt the cells to maximize DNA extraction with simulated microplastic samples. Metagenomic analysis of the 16S rRNA gene showed that the most abundant bacteria on microplastics were dominated by the strains Proteobacteria, Cyanobacteria, Firmicutes, Actinobacteria and Bacteroidetes. The bacterial communities found on microplastics belong to over 400 different species in average. Some of the bacteria isolated from shellfish are human pathogens. We cannot link all the results of the metagenomic analysis with microplastics because it is not possible to determine the polymer composition of all the particles. FTIR spectroscopy was used to determine the chemical composition of the particles. The polymer composition was confirmed for 5 of the total 23 samples. The polymers identified in the samples were polyvinyl chloride, polyethylene, acrylonitrile butadiene styrene and polypropylene.

Keywords:microplastics, marine environment, Slovenia, shellfish, microbial communities, optimization, DNA isolation, spectroscopy, metagenomic analysis, section

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