Serum samples collected from 547 equids in the Pantanal region of Brazil were evaluated for antibodies to Equine Infectious Anemia Virus (EIAV) by the agar gel immunodiffusion test. Risk factors associated with EIAV seropositivity were evaluated and spatial dependence investigated using a Spatial Lag Model. EIAV prevalence on farms in the Pantanal was 52.0% (13/25) with adjusted prevalence between equids of 31.5% (17.4-48.8% 95% CI). Intra-herd prevalence ranged from 5.0 to 77.0%. Statistical analysis demonstrated that farms and animals in regularly flooded areas had respectively 60 and 146 fold higher chance to be sero-positive than farms and animals located in non-flooded areas. Spatial Lag Model results were generally consistent with this conclusion although there was a negative spatial correlation between farms located within in regularly inundated regions, suggesting that other factors, such as management practices, probably play a significant role in transmission of EIAV. Equids with clinical signs were 3.74-fold more likely to be sero-positive than those without clinical signs. The results of this work reveal a high prevalence of EIAV in the Pantanal area of Brazil demonstrating that equids reared in this region are at great risk of infection.
The aim of the study was to evaluate exposure of equids to rickettsial agents (Rickettsia rickettsii, Rickettsia parkeri, 'Candidatus Rickettsia amblyommii', Rickettsia rhipicephali, and Rickettsia bellii) and rickettsial infection in ticks of a Pantanal region of Brazil. Sera of 547 equids (500 horses and 47 donkeys) were evaluated by indirect immunofluorescence assay. In total, 665 adults and 106 nymphal pools of Amblyomma cajennense F. sensu lato, 10 Dermacentor nitens Neumann ticks, and 88 larval pools of Amblyomma sp. were tested by polymerase chain reaction (PCR). Overall, 337 (61.6%) equids were reactive (titer ≥64) to at least one antigen of Rickettsia spp. The prevalence values for Rickettsia were 66%, and the highest endpoint titers were observed for 'Ca. R. amblyommii'. By PCR 3 (0.45%) A. cajennense s.l. females were positive for 'Ca. R. amblyommii'. Minimum infection rates of 0.75% for nymphs and 0.34% for larvae were calculated. Positive samples of ticks have had a fragment of the 16S mitochondrial rRNA gene sequenced and sequences showed 99% identity to Amblyomma sculptum Berlese. This study reports a wide exposure of equids to Rickettsia agents, and PCR evidence of infection with 'Ca. R. amblyommii', for the first time, in A. sculptum.
Equine infectious anemia virus (EIAV) is a persistent lentivirus that causes equine infectious anemia (EIA). In Brazil, EIAV is endemic in the Pantanal region, and euthanasia is not mandatory in this area. All of the complete genomic sequences from field viruses are from North America, Asia, and Europe, and only proviral genomic sequences are available. Sequences from Brazilian EIAV are currently available only for gag and LTR regions. Thus, the present study aimed for the first time to sequence the entire EIAV genomic RNA in naturally infected horses from an endemic area in Brazil. RNA in plasma from naturally infected horses was used for next-generation sequencing (NGS), and gaps were filled using Sanger sequencing methodology. Complete viral genomes of EIAV from two horses were obtained and annotated (Access Number: MN560970 and MN560971). Putative genes were analyzed and compared with previously described genes, showing conservation in gag and pol genes and high variations in LTR and env sequences. Amino acid changes were identified in the p26 protein, one of the most common targets used for diagnosis, and p26 molecular modelling showed surface amino acid alterations in some epitopes. Brazilian genome sequences presented 88.6% nucleotide identity with one another and 75.8 to 77.3% with main field strains, such as EIAV Liaoning, Wyoming, Ireland, and Italy isolates. Furthermore, phylogenetic analysis suggested that this Brazilian strain comprises a separate monophyletic group. These results may help to better characterize EIAV and to overcome the challenges of diagnosing and controlling EIA in endemic regions.
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