Molecular and Phylogenetic analysis revealed new genotypes of Theileria annulata parasites from India

Abstract

BackgroundTick borne diseases impinge cattle worldwide causing mortality and resulting in huge economic losses. Theileriosis is one of the important tick borne diseases mainly caused by Theileria annulata and one of the commonly occurring infections among the livestock. T. annulata causes immense loss to the livestock industry and therefore, efficacious eradication and control strategies are needed for the control of the disease. Genetic diversity among T. annulata parasites is another important aspect which is overlooked in India. Thus, the present study aims to evaluate the prevalence along with genetic diversity and phylogeny of the prevailing T. annulata population of India.MethodsGenomic DNA was extracted from cattle blood samples (n = 862) from different regions of Andhra Pradesh. Molecular diagnosis using T. annulata 18S rRNA based PCR was performed to detect parasites in cattle. Further, 18S rRNA gene was cloned and sequenced to determine similarity and diversity from the known T. annulata sequences.ResultsWe observed an overall prevalence rate of 32.40 % T. annulata infection in Andhra Pradesh based on PCR assay. The sequence analysis revealed novel genotypes among the T. annulata strains from India. Thirteen strains showed closed proximity with a strain from China whereas one Indian strain showed similarity with a South African strain [Theileria sp (buffalo)] based on phylogenetic analysis. Nucleotide heterogeneity of the 18S rRNA sequence among the strains examined varied from 0.1 to 8.6 % when compared with the published strains.ConclusionThe present study provides us with the molecular prevalence of theileriosis, and will support the accomplishment of actions or in design of strategy to control theileriosis transmission to cattle. Additionally, it highlights the emergence of strains with novel genotypes from India.

A Real-Time PCR based assay for determining parasite to host ratio and parasitaemia in the clinical samples of Bovine Theileriosis

“…Genomic DNA was isolated from 2 ml of blood samples using previously published DNA isolation protocol 1 . Quality and integrity of the DNA were checked using Nanodrop and by running the genomic DNA on 0.8% agarose gel.…”

“…Bovine theileriosis is caused by an apicomplexan parasite Theileria spp . which is an important tick-borne disease of livestock 1 . Theileria annulata and Theileria parva are the two economically important species responsible for livestock morbidity and mortality worldwide 2 – 4 .…”

Potential Sabotage of Host Cell Physiology by Apicomplexan Parasites for Their Survival Benefits

“…Theileria annulata and T. parva cause tropical theileriosis and east coast fever, respectively, in ruminants predominantly in cattle causing enormous economic loss to the livestock industry ( 106 , 107 ). Tick vector transmits the parasite upon feeding on animal through the saliva ( 108 – 110 ).…”

Molecular surveillance of Theileria parasites of livestock in Oman

“…PCR 159 amplification, cloning and sequencing were carried out as described previously 160 (George et al, 2015). Alignment and analysis of the 18S rRNA sequences was 161 performed using BioEdit version 7.2.5 software (Hall, 1999) Hoogstraal (1956) and Walker et al, (2003 (Fig 1).…”