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Metagenomska analiza kožne mikrobiote črne podvrste človeške ribice
ID Pajek Arambašič, Neža (Author), ID Kostanjšek, Rok (Mentor) More about this mentor... This link opens in a new window, ID Gostinčar, Cene (Co-mentor)

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Abstract
Endemit belokranjskega krasa, črna podvrsta človeške ribice (Proteus anguinus parkelj Sket & Arntzen 1994), je eden najbolj ogroženih vretenčarjev v Evropi. Kostanjšek in sod. (2019) so pri kožni mikrobioti osebka črne podvrste opazili prevlado enterobakterij in aeromonad, kar bi lahko nakazovalo na obremenitev okolja s fekalno mikrobioto. V nalogi smo zato želeli preveriti sestavo kožne mikrobiote pri večjem številu osebkov črne podvrste. Z analizo sekvenciranih pomnožkov taksonomskih označevalcev na nivoju bakterij, gliv, arhej in evkariontov smo izvedli razširjeno analizo kožne mikrobiote štirih osebkov črne podvrste in devetih belih osebkov človeških ribic iz dveh filogenetskih linij, linije reke Ljubljanice in dolenjske linije. Pri črni podvrsti smo rod Aeromonas zasledili le pri enem osebku in to v zelo nizki relativni številčnosti (< 1%), zato domnevamo, da je pri črnem osebku predhodne raziskave šlo za osamljen primer disbioze kožne mikrobiote. S statističnimi testi diverzitete beta (PERMANOVA, ANOSIM) smo pokazali, da filogenetska linija gostitelja lahko pojasni del variance bakterijskih združb, ne pa tudi združb ostalih preiskovanih taksonomskih skupin. Naša analiza v primerjavi z dosedanjimi tovrstnimi raziskavami prvič vključuje tudi glive, arheje in evkarionte. Poleg tega je dodana vrednost naše raziskave vzpostavitev bioinformatskega cevovoda za analizo pomnožkov taksonomskih označevalcev z uporabo paketa Qiime2 v jeziku Python v obliki zvezkov Jupyter Notebook.

Language:Slovenian
Keywords:kožna mikrobiota, Proteus anguinus parkelj, bioinformatika, QIIME2, DNA metabarkodiranje
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Place of publishing:Ljubljana
Publisher:[N. P. Arambašič ]
Year:2022
PID:20.500.12556/RUL-143596 This link opens in a new window
UDC:579
COBISS.SI-ID:137393411 This link opens in a new window
Publication date in RUL:30.12.2022
Views:481
Downloads:72
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Secondary language

Language:English
Title:Metagenomic analysis of skin microbiota in the black olm subspecies
Abstract:
One of the most endangered vertebrates in Europe, the black olm subspecies (Proteus anguinus parkelj Sket & Arntzen 1994), is endemic to only a few square kilometers large area of the karst region of Bela krajina in Southeast Slovenia. Kostanjšek et al. (2019) discovered that the normal bacterial cutaneous microbiota of the black olm specimen was almost entirely replaced by enterobacteria and aeromonads, which could indicate fecal microbiota pollution of its natural habitat. The thesis aimed to analyze the composition of the skin microbiota on a larger sample of the black olm subspecies. We analyzed the amplicon sequencing data of cutaneous bacteria, fungi, archaea, and eukaryotes in four specimens of the black olm subspecies and nine white olm specimens from the phylogenetic lineage of Ljubljanica river and Dolenjska lineage. Since the Aeromonas genus was present in only one black olm specimen and in very low relative abundance (< 1%), we conclude that the prevalence of fecal bacteria in the skin microbiota of the black specimen of preliminary research was an isolated case of dysbiosis. While beta diversity statistical tests (PERMANOVA, ANOSIM) showed that the host's phylogenetic lineage could explain a part of bacterial skin community variance, the same was not true for other investigated taxonomic groups. Our analysis is the first black olm skin microbiota study to examine the diversity of fungi, archaea, and eukaryotes. Further added value of our work is a pipeline for sequencing data analysis using Qiime2, written in Python in the form of Jupyter Notebooks.

Keywords:skin microbiota, Proteus anguinus parkelj, bioinformatics, QIIME2, DNA metabarcoding

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