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Prevalenca borelij lymske borelioze v malih sesalcih
ID Cvetko, David (Author), ID Ružić Sabljić, Eva (Mentor) More about this mentor... This link opens in a new window, ID Cerar Kišek, Tjaša (Comentor)

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Abstract
V Sloveniji je lymska borelioza najpogostejša bolezen, katere povzročitelje prenašajo klopi. Povzročajo jo predstavniki kompleksa Borrelia burgdorferi sensu lato. Borelije za uspešno razmnoževanje potrebujejo kompetentne gostitelje. Pri nas so to mali sesalci, ptiči in kuščarji. Opravili smo epidemiološko raziskavo prekuženosti malih sesalcev z Borrelia burgdorferi sensu lato in Borrelia miyamotoi. Prisotnost borelij lymske borelioze smo preverili v srcih in pljučih testiranih živali. S PCR v realnem času smo dokazovali prisotnost gena ospA v vzorcih. Odkrili smo, da je okuženih 20,25 % malih sesalcev. Manjšo diagnostično občutljivost je pokazala metoda kultivacije, s katero smo odkrili prisotnost B. burgdorferi sensu lato pri 6,01 % malih sesalcev. Pričakovano je bila prevalenca B. miyamotoi mnogo manjša. Z molekularnimi metodami in sekvenciranjem smo pri malih sesalcih odkrili 0,61 % prevalenco B. miyamotoi. Želeli smo natančneje identificirati osamljene borelije lymske borelioze, zato smo izolirane seve genotipizirali z metodo polimorfizma dolžin restrikcijskih fragmentov celotnega genoma (angl. large restriction fragment pattern, MluI-LRFP) in metodo tipizacije na osnovi multilokusnih zaporedji (angl. multilocus sequence typing, MLST). Z metodo MluI-LRFP smo ugotovili, da vsi izolirani sevi borelij lymske borelioze pripadajo podtipu Mla1 vrste Borrelia afzelii. Boljšo sposobnost ločevanja znotraj vrste omogoča MLST. Z MLST smo odkrili tri različne sekvenčne tipe. Vsi trije sekvenčni tipi pripadajo B. afzelii in so že bili odkriti v Sloveniji. Naše ugotovitve potrjujejo uspešno razmnoževanje in kroženje borelij med gostitelji, prenašalci in ljudmi.

Language:Slovenian
Keywords:Borrelia burgdorferi sensu lato, Borrelia miyamotoi, prevalenca, mali sesalci, lymska borelioza, molekularne metode, kultivacija, MLST, MluI, LRFP
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Publisher:[D. Cvetko]
Year:2018
PID:20.500.12556/RUL-102769 This link opens in a new window
UDC:579.8+579.25:577.2.086:616.993
COBISS.SI-ID:4941688 This link opens in a new window
Publication date in RUL:08.09.2018
Views:2047
Downloads:405
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Secondary language

Language:English
Title:Prevalence of Borrelia burgdorferi sensu lato in small mammals
Abstract:
Lyme borreliosis is the most common tick-borne disease in Slovenia. Lyme borreliosis is caused by a group of related spirochaetes named Borrelia burgdorferi sensu lato. In Slovenia the main competent reservoirs are small mammals, birds and lizards, which are crucial for successful maintenance and transmission for borrelia. The aim of the present study was to perform an epidemiological survey to determine the prevalence of B. burgdorferi sensu lato and Borrelia miyamotoi in small mammals. Heart and lungs was examined with molecular methods and culture techniques. DNA of B. burgdorferi sensu lato was detected in 20,25 % rodents, using real time PCR, targeting gene ospA. B. burgdorferi sensu lato was isolated from 6,01 % rodents. Molecular methods were more sensitive than culture techniques. B. miyamotoi DNA was detected in 0,61 % rodents, using molecular methods and confirmed with sequencing. Identification of isolated Lyme borrelia was performed on limited number of samples with large restriction fragment pattern (MluI-LRFP) and multilocus sequence typing (MLST). MluI-LRFP revealed all isolates as Borrelia afzelii Mla1. According to our results MLST has greater discriminatory power. MLST analysis revealed three distinct sequence types, all belonging to B. afzelii species and all three already known in Slovenia. Our findings confirm successful reproduction and transmission between reservoir, vectors and humans.

Keywords:Borrelia burgdorferi sensu lato, Borrelia miyamotoi, prevalence, small mammals, lyme borreliosis, molecular methods, cultivation, MLST, MluI, LRFP

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