Clinical and epizootiological examination of equine piroplasmosis on the central balkan region and its effect on host DNA damage

Abstract

Equine piroplasmosis is a significant vector-prone disease with a high degree of widespread and economic impact on horse industry throughout the world. The causative agents of equine piroplasmosis are protozoas Theileria equi and Babesia caballi. The disease can be present subclinical in horses, but also it can be accompanied by a serious clinical picture. The goal of this doctoral dissertation was the comprehensive clinical and epizootiological examination of the equine piroplasmosis on the central Balkan region, where we would investigate the presence of parasites by molecular detection methods, to follow the haematological and biochemical parameters, the parameters of oxidative stress and at the end, the degree of host cells DNA damage due to parasites presence. In order to gain insight into the prevalence of the disease, we conducted a molecular epidemiological study involving 165 horses, from eight locations from three countries (Serbia, Montenegro, Bosnia and Herzegovina). First, we did a PCR method for the detection of a 450 bp long segment of the 18S rRNA gene which is present in all Theileria and Babesia species. For the definitive species identification, a multiplex PCR was done. In order to confirm obtained results, sequencing of positive samples was made. The overall prevalence of T. equi and B. caballi in the study area was 32.1% and 1.8% respectively. In Serbia a prevalence of 40.2% was established for T. equi and 0.6% for B. caballi, in BiH 16.7% for T. equi and 4.2% for B. caballi, and in Montenegro 8.3% for T. equi and 8.3% for B. caballi. Results of the sequence analysis demonstrated 100% correspondence to multiplex PCR diagnosed Babesia species...

Piroplazmoza konja predstavlja značajno vektorski prenosivo oboljenje sa velikom raširenošću i ekonomskim uticajem na konjarstvo širom sveta. Uzročnici piroplazmoze konja su protozoe Theileria equi i Babesia caballi. Oboljenje može biti prisutno kod konja i supklinički, ali može biti praćeno i ozbiljnom kliničkom slikom. Cilj ove doktorske disertacije je bio opsežno kliničko- epizootiološko ispitivanje priplazmoze konja na prostoru centralnog Balkana, gde bi ispitivali prisustvo parazita molekularnim metodama detekcije, pratili hematološke i biohemijske parametre, parametre oksidativnog stresa i na kraju stepen oštećenja DNK ćelija domaćina usled prisustva parazita. U cilju sticanja uvida u prevalenciju oboljenja sproveli smo molekularnu epizootiološku studiju koja je uključivala 165 konja, sa osam lokaliteta sa prostora tri države (Srbija, Crna Gora, Bosna i Hercegovina). Prvo smo uradili PCR metodu za detekciju 450 bp dugog segmenta 18S rRNK gena koji je prisutan kod svih Theileria i Babesia vrsta. Za definitivnu detekciju vrste je urađen multipleks PCR. Da bi potvrdili dobijene rezultate urađeno je sekvenciranje pozitivnih uzoraka. Ukupna prevalencija T. equi i B. caballi na ispitivanom prostoru iznosila je 32,1% i 1,8%, respektivno. U Srbiji je ustanovljena prevalencija od 40,2% za T. equi i 0,6% za B. caballi, u BiH 16,7% za T. equi i 4,2% za B. caballi, a u Crnoj Gori 8,3% za T. equi i 8,3% za B. caballi. Rezultati sekvenciranja su potvrdili 100% podudarnost sa vrstama ustanovljenim na osnovu metode multipleks PCR...