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Molecular genetic identification of Lotmaria passim Schwarz, 2014 trypanosome and the analysis of its impact on the health of bee colonies and economic effects in apiculture.

dc.contributor.advisorStevanović, Jevrosima
dc.contributor.otherMirilović, Milorad
dc.contributor.otherStanimirović, Zoran
dc.contributor.otherAleksić, Nevenka
dc.contributor.otherĆirković, Dragan
dc.creatorVejnović, Branislav
dc.date.accessioned2019-10-17T11:02:01Z
dc.date.available2019-10-17T11:02:01Z
dc.date.available2020-07-03T09:28:34Z
dc.date.issued2019-03-14
dc.identifier.urihttp://eteze.bg.ac.rs/application/showtheses?thesesId=6881
dc.identifier.urihttp://nardus.mpn.gov.rs/handle/123456789/11424
dc.identifier.urihttps://fedorabg.bg.ac.rs/fedora/get/o:20112/bdef:Content/download
dc.identifier.urihttp://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=51497231
dc.description.abstractZbog nepostojanja molekularne metode za determinaciju vrsta pčelinjih tripanozoma Crithidia mellificae i Lotmaria passim bez naknadnog sekvencioniranja, definisan je cilј ovog rada, a to je molekularna identifikacija tripanozome L. passim i ispitivanje njenog uticaja na zdravlјe pčelinjih zajednica i ekonomske efekte u pčelastvu. Razvijeni su specijes-specifični prajmeri za PCR detekciju C. mellificae i L. passim kao i real-time PCR metoda za detekciju i kvantifikaciju L. passim. Uzorci pčela koji su sakupljani u Srbiji u devetogodišnjem periodu (2007-2015) retrospektivno su analizirani kako bi se utvrdila prevalencija ovog eukariotskog parazita molekulanrnom analizom DNA. Kako bi se utvrdila godišnja dinamika L. passim, njen uticaj na oksidativni stres, kao i na ekonomsku dobit formirane su dve grupe košnica. Istraživanje je sprovedeno na ukupno dvadeset Langstroth košnica, 10 košnica koje su bile L. passim-pozitivne i 10 košnica koje su bile L. passim-negativne, a koje su se nalazile na dva različita pčelinjaka. Tokom trajanja eksperimenta (od marta 2016. do marta 2017. godine) svakog meseca izuzev zimskih meseci (oktobar-februar) iz svake košnice je uzorkovano najmanje 120 živih odraslih pčela sa leta košnice (za detekciju i kvantifikaciju L. passim i Nosema spp), a u junu i septembru 20 živih odraslih pčela za potrebe analize parametara oksidativnog stresa. Razvijeni su specijes-specifični prajmeri za PCR detekciju tripanozoma vrste C. mellificae i L. passim, koji su validirani i u uslovima mešovite infekcije, kao i infekcije tripanozomom bumbara Crithidia bombi. Infekcija sa N. apis i N. ceranaesu analizirane korišćenjem PCR. Tehnikom real-time PCR je utvrđen nivo infekcije vrstom L. passim kod L. passim-pozitivnih društva, dok je brojanjem spora određen nivo infekcije sa N. ceranae u svim društvima. Analizirani su uzorci pčela iz 162 društva (18 iz svake godine), koji su prikupljeni sa 57 različitih lokacija. L. passim je detektovana u uzorcima koji su poticali iz svake godine i kod prosečno 62,3% (godišnja prevalencija od 38,9% do 83,3%)...sr
dc.description.abstractDue to the absence of molecular methods intended for species-specific identification of bee trypanosomes Crithidia mellificae and Lotmaria passim without further sequencing, the goal of this thesis has been defined: molecular identification of L. passim trypanosome and the research into its impact on the colony health and economic effects in apiculture. Species-specific primers for PCR detection of C. mellificae and L. passim as well as a real-time PCR method for the detection and quantification of L. passim were developed. Archival bee samples collected in Serbia during a nine-year period (2007-2015) were analysed retrospectively to assess the prevalence of this eukaryotic parasite by molecular DNA analysis. To determine the annual dynamic of L. passim infection, and its influence on oxidative stress and the economic income two apiaries were chosen. Ten hives from each made up the two experimental groups, the L. passim-positive from the one and the L. passim-negative from another. During the experimental period (from March 2016 to March 2017) each month with the exception of winter months (October-February) from every hive 120 live adult bees were collected from the hive entrance for the detection and quantification of L. passim and Nosema spp. In addition, in June and September 20 live adult bees were collected in the same manner for oxidative stress analysis. Species-specific primers for PCR detection of the two trypanosomatids, C. mellificae and L. passim, were developed and validated for target specificity under single and mixed-species conditions as well as against the bumblebee trypanosomatid Crithidia bombi. Infections with N. apis and N. ceranae were analysed using the PCR assay. Infection levels with L. passim in L. passim-positive colonies were detected with the real-time PCR, and the N. ceranae infection level by spore counting in all of the colonies. A total of 162 colonies (18 originating from each year), collected from 57 locations were analysed. L. passim was detected in samples provided each year in 62.3% of samples (ranging between 38.9% and 83.3%)...en
dc.formatapplication/pdf
dc.languagesr
dc.publisherУниверзитет у Београду, Факултет ветеринарске медицинеsr
dc.relationinfo:eu-repo/grantAgreement/MESTD/Integrated and Interdisciplinary Research (IIR or III)/46002/RS//
dc.rightsopenAccessen
dc.sourceУниверзитет у Београдуsr
dc.subjectApis melliferasr
dc.subjectApis melliferaen
dc.subjecttripanozomesr
dc.subjectmikrosporidijesr
dc.subjectPCRsr
dc.subjectreal-time PCRsr
dc.subjectmolekularna detekcijasr
dc.subjectanaliza troškova i dobitisr
dc.subjecttrypanosomatidaeen
dc.subjectmicrosporidiaen
dc.subjectPCRen
dc.subjectreal-time PCRen
dc.subjectmolecular detectionen
dc.subjectcost-benefit analysisen
dc.titleMolekularno-genetička identifikacija tripanozome Lotmaria passim Schwarz, 2014 i analiza njenih efekata na zdravlje pčelinjih zajednica i ekonomske efekte u pčelarstvusr
dc.title.alternativeMolecular genetic identification of Lotmaria passim Schwarz, 2014 trypanosome and the analysis of its impact on the health of bee colonies and economic effects in apiculture.en
dc.typedoctoralThesis
dc.rights.licenseBY-NC-ND
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/19418/Disertacija.pdf
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/19419/Referat.pdf


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