Abstract
Problem statement: Equine Infectious Anemia (EIA) is a chronic, relapsing infectious disease of horses caused by Equine Infectious Anemia Virus (EIAV). The objective of this experiment was compared EIAV effectiveness detection in Peripheral Blood Mononuclear Cells (PBMCs) and Bronchoalveolar wash (BAW) from naturally infected horses. Approach: Fifty seven DNA samples isolated from PBMCs and BAW of naturally infected animals were used for a nested PCR amplifying a 408 bp gag gene fragment region. Equine Dermal cells (ED) infected and non infected by EIAV was used as nested PCR negative and positive control. Horses’ blood samples were firstly tested as positive in Agar Gel Immunodiffusion Assay (AGID). Results: Results demonstrated 89% (51/57) of gag gene amplification in PBMCs samples and only 47% (27/57) of gag gene amplification in BAW samples. Conclusion: The nested PCR assay used in the present study detected more EIAV positive samples in PBMC than in BAW, indicating PBMCs as a reliable source for EIAV diagnosis.
Highlights
Equine Infectious Anemia (EIA) is a horse disease caused by Equine Infectious Anemia Virus (EIAV), a retrovirus transmitted by bloodsucking insects
Preliminary data indicate that infection of blood monocytes by EIAV causes a non-productive infection and that differentiation of the infected monocytes into macrophages is necessary to trigger viral replication (Maury, 1994; Sellon et al, 1996) and macrophages are the major cell type found in bronchoalveolar wash samples from equids (Ainsworth et al, 2002; 2003; Dyer et al, 1983)
A nested Polymerase Chain Reaction amplifying a region of the gag gene of Equine Infectious Anemia Virus (EIAV) was developed for comparing the virus detection effectiveness in Peripheral Blood Mononuclear Cells (PBMC) and Bronchoalveolar Wash (BAW) samples from 57 naturally infected horses accredited by the Ministry of Agriculture in Minas Gerais state, Brazil
Summary
Equine Infectious Anemia (EIA) is a horse disease caused by Equine Infectious Anemia Virus (EIAV), a retrovirus transmitted by bloodsucking insects. The control of EIA is made through the identification of infected animals by the serological tests ELISA and IDGA (Alvarez et al, 2007), but many factors contribute to inconsistent results with these tests that require confirmation using more specific techniques (Alvarez et al, 2007). Preliminary data indicate that infection of blood monocytes by EIAV causes a non-productive infection and that differentiation of the infected monocytes into macrophages is necessary to trigger viral replication (Maury, 1994; Sellon et al, 1996) and macrophages are the major cell type found in bronchoalveolar wash samples from equids (Ainsworth et al, 2002; 2003; Dyer et al, 1983). Some studies have found that the latent infection of mononuclear cells in blood is an important mechanism for viral persistence and dissemination (Gendelman et al, 1985; 1986)
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