Discipline: Biological Sciences
Montessa Mitchell - Tuskegee University
Co-Author(s): Norimar Pastrana-Negron, Tuskegee University, Tuskegee, AL; Abdelrahman Mohammed, Tuskegee University, Tuskegee, AL; P. Gopal Reddy, Tuskegee University, Tuskegee, AL; Solomon Odemuyiwa, Tuskegee University, Tuskegee, AL.
Hepatitis C Virus (HCV) is estimated to have infected 170 million people worldwide. Certain genotypes of the virus have the propensity to cause liver cirrhosis which can lead to liver cancer. There is still difficulty associated with cultivating HCV in vitro and in vivo. In 2012, a new virus closely related to HCV was found in horses. Since there is no immunologically competent small animal model, the equine hepaciviridae (eHcV) is a candidate for a natural history and model of the virus. Cataloguing the similarities and differences between HCV and eHcV is important in establishing this large animal model. The hypothesis was that approximately 3% of the isolates would be positive (a comparable amount to what has been reported globally) and they would cluster closely to other eHcV isolates reported on GenBank. Two hundred eighty (280) blood samples collected from horses of the East Alabama area were screened for eHcV using nested PCR. Viral RNA was extracted from the serum, before being reverse transcribed. The RNA samples analyzed with primers that targeted a 300 base pair fragment in the highly conserved region of NS5B showed 5.3% samples as positive. These samples were purified, and the cDNA was sequenced. Sequences were aligned using ClustalW in the MEGA7 program and phylogenetic trees were constructed. When aligned with other samples worldwide, eHcV isolates from East Alabama showed a relation to those from other parts of the world. These results indicate that eHcV is prevalent in healthy horses in East Alabama. Clustering of the isolates despite geographical location implies a novel epidemiological model of emergence and spread of a blood borne pathogen in a companion/livestock animal.Not Submitted
Funder Acknowledgement(s): This study was supported in part by a grant from USDA/NIFA Capacity Building Grant # 2015-38820-24399.
Faculty Advisor: P. Gopal Reddy, firstname.lastname@example.org
Role: I processed 133 samples by isolating RNA from serum, performed RT-PCR, screened for positive samples using gel electrophoresis, and extracted the cDNA from positive bands. Additionally, I performed computer analysis on the resulting sequence data, including BLAST, Clustal W alignment, and MEGA 7 phylogenetic tree construction.