Title Identifikacija virulentnih faktora parazita malarije koji nadilaze prirodnu rezistenciju komarca
Title (english) Identification of malaria parasite virulence factors that counteract mosquito natural resistance
Author Eva Vodopija
Mentor Stephanie Blandin (mentor)
Mentor Željka Maglica (mentor)
Committee member Rozi Andretić Waldowski (predsjednik povjerenstva)
Committee member Igor Jurak (član povjerenstva)
Committee member Željka Maglica (član povjerenstva)
Granter University of Rijeka (Faculty of Biotechnology and Drug Development) Rijeka
Defense date and country 2018-09-26, Croatia
Scientific / art field, discipline and subdiscipline BIOTECHNICAL SCIENCES Biotechnology
Abstract Malaria is an infectious disease caused by Plasmodium parasites and transmitted by Anopheles mosquitoes. The ability of a given mosquito to transmit malaria parasites is determined by different factors: mosquito genetic factors (TEP1, responsible for mosquito resistance to malaria parasites), parasite genetic factors (Plasmodium falciparum Pfs47 surface protein, allows P. falciparum to by-pass the mosquito TEP1-dependent antiparasitic response) and diverse environmental factors. To identify novel parasite factors that contribute to Plasmodium infectivity in mosquitoes, the group of Stephanie Blandin has established a mutagenesis screen in the murine parasite Plasmodium berghei. They have recovered mutant parasites that are able to invade a normally fully resistant mosquito line. To identify the causative mutation(s), a virulent parasite VP1 was isolated, cloned and phenotyped (both the virulent parasite and the clones) in naturally resistant mosquitoes. During this project, we optimized the workflow for the phenotyping. We show that the optimal time for an infection that leads to high parasite loads is at 3 and 4 days post passage and that the optimal time for mosquito midgut dissections and microscopic analysis is at 10 to 12 days post infection. We also show that an antibiotic/antimycotic treatment of mosquitoes improves mosquito survival, but at the same time renders resistant mosquitoes more susceptible to infections (with both the wild type and the virulent parasites), indicating that the mosquito microbiota contributes in part to determine the resistance of resistant mosquitoes to parasites. With the intention of distinguishing virulent from avirulent virulent parasite clones, we phenotyped them in naturally resistant mosquitoes. We observed variable prevalences ranging from 2% to 38% for the different clones. Among the 12 clones, one stood out with the highest prevalence (38%) and parasite load (6), consequently being the best candidate for further exploration, despite its potentially partial phenotypic penetrance.
Abstract (croatian) Malarija je zarazna bolest uzrokovana Plasmodium parazitima, a prenose ju Anopheles komarci. Mogućnost prijenosa parazita malarije je određena različitim faktorima: genetičkim faktorima komarca (TEP1, odgovoran za rezistenciju komaraca protiv parazita malarije), genetičkim faktorima parazita (Plasmodium falciparum Pfs47 površinski protein, omogućuje P. falciparum da zaobiđe TEP1-ovisan antiparazitni odgovor) i različitim okolišnim čimbenicima. Za identifikaciju novih faktora parazita koji doprinose infektivnosti Plasmodium-a u komarcima, grupa vođena od strane Stephanie Blandin je uspostavila probir mutagenezom u murinskom parazitu Plasmodium berghei. Proizveli su mutirane parazite koji mogu napasti inače potpuno rezistentnu liniju komaraca. Kako bi identificirali ključne mutacije, virulentni parazit VP1 je izoliran, kloniran te zajedno s klonovima fenotipiziran u inače rezistentnim komarcima. Tokom ovog projekta, optimizirali smo proces fenotipiziranja. Pokazali smo da je optimalno vrijeme za infekciju koja vodi visokom broju parazita 3 i 4 dana poslije pasaža, a optimalno vrijeme za seciranje crijeva komaraca i mikroskopsku analizu istih 10 do 12 dana poslije infekcije. Isto tako, pokazali smo da antibiotski/antimikotski tretman komaraca poboljšava preživljavanje komaraca, ali u isto vrijeme čini prirodno rezistentne komarce podložnima infekciji (s parazitom divljeg tipa i virulentnim parazitom), što govori da mikrobiota u crijevima komaraca djelomično doprinosi u određivanju rezistencije rezistentnih komaraca na parazite. S namjerom razlikovanja avirulentnih i virulentnih klonova virulentnog parazita, fenotipizirali smo ih u rezistentnim komarcima. Zapazili smo varijabilne prevalencije od 2% do 38% za različite klonove. Između 12 klonova, jedan se istaknuo s najvišom prevalencijom (38%) i brojem parazita (aritmetička sredina 6) što bi indiciralo da je klon 8 najbolji kandidat za daljnje istraživanje, bez obzira na potencijalno parcijalnu penetraciju fenotipa.
Keywords
Malaria
Plasmodium berghei
Anopheles gambiae
virulence
parasitemia
Keywords (croatian)
Malarija
Plasmodium berghei
Anopheles gambiae
virulencija
parazitemija
Language english
URN:NBN urn:nbn:hr:193:204746
Study programme Title: Drug research and development Study programme type: university Study level: graduate Academic / professional title: magistar/magistra istraživanja i razvoja lijekova (magistar/magistra istraživanja i razvoja lijekova)
Type of resource Text
File origin Born digital
Access conditions Access restricted to students and staff of home institution
Terms of use
Created on 2018-10-31 08:47:10