Molecular and microscopic detection of Theileria luwenshuni infection in goats in and around Guwahati of Assam, India

ABSTRACT

Theileriosis, an important tick-borne hemoprotozoan disease that can reduce production and productivity, results in severe economic loss in goat industry. Due to lack of facilities and interest of the researchers the sub-clinical form of theileriosis caused by Theileria luwenshuni was under diagnosed in Assam, India. Therefore, a study was carried out to determine the prevalence of theileriosis in goats from Kamrup (Metro) district of Assam. A total of 454 blood samples collected from goats revealed 39.87 % (181/454) infection with Theileria sp. The genomic DNA extracted from blood samples amplified for a fragment of 18S ribosomal RNA (18S rRNA) gene employing polymerase chain reaction (PCR) using Theileria genus specific primers showed distinct bands at 1098 bp in agarose gel. The resultant PCR products were sequenced and BLAST analysis indicated 99.9% homology with T. luwenshuni. Parasitological methods and PCR technique followed by sequence analysis suggest infection with T. luwenshuni was associated with theileriosis in investigated goats. The results of the present study showed PCR to be more efficient in detection of Theileria positive cases than the microscopic method.

Abbreviations: rRNA: Ribosomal ribonucleic acid; PCR: polymerase chain reaction; ml: milliliter; EDTA: Ethylene Diammine Tetraacetic Acid; °C: degree Celsius; DNA: Deoxyribonucleic acid; ME: microscopic examination; sp.: species; V-volt; µg: micro gram; TAE: Tris-acetate-EDTA; F: forward; R: reverse; NCBI: National Center for Biotechnology Information; BLAST-n Basic Local Alignment Search Tool for nucleotide.

KEYWORDS:

• Theileria luwenshuni
• blood smear
• PCR
• 18S rRNA gene
• sequencing

Acknowledgments

The authors express their sincere thanks to the Dean, Faculty of Veterinary Science, Assam Agricultural University, Khanapara and the Director of Post Graduate studies AAU, Jorhat for rendering necessary facilities to conduct the present investigation.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The authors are also highly thankful to Assam Agricultural University, Jorhat for providing funding for this project.