Parazitološka i molekularna ispitivanja genotipova i haplotipova metacestoda Echinococcus granulosus sensu lato i epizootiološke karakteristike hidatidoze kod različitih vrsta životinja
Parasitological and molecular examination of genotypes and haplotypes of metacestoda Echinococcus granulosus sensu lato and epizootiological characteristics of hydatidosis in different species of animals
Докторанд
Debeljak, Zoran R.Ментор
Kulišić, ZoranЧланови комисије
Radojičić, SonjaTeodorović, Vlado
Ilić, Tamara
Džamić, Aleksandar
Метаподаци
Приказ свих података о дисертацијиСажетак
Sprovedenim istraživanjima definisane su patomorfološke, morfometrijske,
parazitološke karakteristike metacestoda Echinococcus granulosus sensu lato kao i
epizootiološke karakteristike hidatidoze kod 96 životinja (49 domacih svinja, 2 divlje
svinje, 25 goveda i 20 ovaca), poreklom sa teritorije razlicitih epizootioloških podrucja
(20 opština) Republike Srbije. Molekularnim ispitivanjima pregledan je materijal
poreklom od 52 životinje (15 goveda, 18 ovaca, 17 domacih i 2 divlje svinje).
Karakteristike ustanovljenih hidatidnih cista u potpunosti su odgovarale nacinu gajenja i
starosti pregledanih životinja. Fertilne ciste su ustanovljene kod 61,9% ovaca, 22,4%
svinja i 8,0% goveda. Samo kod goveda sve fertilne ciste (100%) su bile vijabilne.
Degenerativne ciste su ustanovljene kod svinja i ovaca, a kalcifikovane samo kod ovaca.
Identifikacija vrste, genotipova i haplotipova metacestoda E. granulosus sensu lato, u
uzorcima hidatidnih cista radena je analizom DNK sekvence mitohondrijalnog ...citohrom
C-oksidaze 1 (cox1) gena. Korišcen je protokol MI-05 Evropske referentne laboratorije za
parazite u Rimu (EU RFLP, 2010), a u okviru ispitivanja radene su: lancana reakcija
polimeraze, sekvenciranje i haplotipizacija. U okviru E. granulosus s.l. determinisane su
2 vrste: E. granulosus s.s. i E. canadensis, 4 genotipa (G1, G2, G3 i G7) i 7 haplotipova
(Hap 1 - Hap 7). Kod domacih svinja ustanovljen je E. canadensis, genotip G7, kod
goveda 2 genotipa E. granulosus s.s., (G1 i G3) i 5 haplotipova, kod ovaca prisustvo sva
tri genotipa E. granulosus s.s., (G1, G2 i G3) i 4 haplotipa (Hap 2, Hap 3, Hap 5 i Hap
7), a kod 2 ovce ustanovljena je mešovita infekcija, na nivou genotipa i haplotipa. Od 7
ustanovljenih haplotipova (Hap 1 - Hap 7), dva haplotipa (Hap 2 i Hap 5) su dominantni
kod 40,4% životinja i zajednicki su za goveda i ovce. Sekvence dobijene kod goveda i
ovaca (koje pripadaju haplotipu 5 - genotip G1), 100% su identicne sa haplotipom koji
je globalno rasprostranjen u svetu. S obzirom na razlicite biološke i epizootiološke
karakteristike pojedinih genotipova i haplotipova E. granulosus s.l., njihova identifikacija i
definisanje prevalencije predstavljaju znacajan doprinos poznavanju epizootioloških i
molekularnih karakteristika hidatidoze kod domacih životinja na širem prostoru
Republike Srbije i važni su za razumevanje biološkog ciklusa parazita i predlog mera
kontrole.
According to conducted researches we defined histological, morphometric and
parasitological characteristics of metacestoda Ecchinococcus granulosus sensu lato as
well as epizootiological characteristics of hydatiosis in 96 animals ( 49 domestic pigs, 2
wild boars, 25 cattle and 20 sheep) originating from the territory of different
epizootiological areas ( 20 municipalities) of the Republic of Serbia. By molecular
analysis, examined material originated from 52 animals (15 cattle, 18 sheep, 17
domestic pigs and 2 wild boars). The characteristics of established hydatid cysts
completely fit the manner of cultivation and the age of examined animals. Fertile cysts
are established in 61.9% of sheep, 22.4% of pigs and 0.8% of cattle. Only in cattle, all
fertile cysts ( 100%) were viable. Degenerative cysts are established in pigs and sheep,
while calcified are found only in sheep. Identification of the species, genotypes and
haplotypes of metacestoda E. granulosus sensu lato in samples of hyda...tid cyst was
performed by analyzing DNA sequence of mitochondrial cytochrome C- oxidase 1 (cox
1) gene. We used MI-05 protocol of European Reference Laboratory for Parasites in
Rome (EU RFLP, 2010) and within examination we performed: polymerase chain
reaction, sequencing and haplo standardization. Within the scope of E. granulosus s.l.
we determined 2 species: E. granulosus s.s. and E. canadensis, 4 genotypes (G1, G2,
G3 and G7) and 7 haplotypes (Hap1-Hap7). In domestic pigs we established E.
canadensis, genotype G7; in cattle we established 2 genotypes E. granulosus s.s. (G1
and G3) and 4 haplotypes; in sheep there is a presence of all three genotypes: E.
granulosus s.s. (G1,G2 and G3) and 4 haplotypes (Hap 2, Hap 3, Hap 5 and Hap 7). In
two sheep we established mixed infections at the level of genotype and haplotype.
From 7 established haplotypes (Hap1-Hap7), two haplotypes (Hap 2 and Hap 5) are
dominant at 40,4% of animals and are common for both cattle and sheep. The sequences
we got in cattle and sheep (that belong to haplotype 5 - genotype1) are 100% identical
with the haplotype which is globally prevalent. Considering different biological and
epizootiological characteristics of certain genotypes and haplotypes of E. granulosus
s.l., their identification and definition of prevalence represent a significant contribution
to the knowledge of epizootiological and molecular characteristics of hydatidosis in
domestic animals in the wider area of the Republic of Serbia and are important for
understanding the biological cycle of the parasite and proposed control measures.