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      Madariaga and Venezuelan equine encephalitis virus seroprevalence in rodent enzootic hosts in Eastern and Western Panama

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          Abstract

          While rodents are primary reservoirs of Venezuelan equine encephalitis virus (VEEV), their role in Madariaga virus (MADV) transmission remains uncertain, particularly given their overlapping geographic distribution. This study explores the interplay of alphavirus prevalence, rodent diversity, and land use within Darien and Western Panama provinces. A total of three locations were selected for rodent sampling in Darien province: Los Pavitos, El Real de Santa Maria and Santa Librada. Two sites were selected in Western Panama province: El Cacao and Cirí Grande. We used plaque reduction neutralization tests to assess MADV and VEEV seroprevalences in 599 rodents of 16 species across five study sites. MADV seroprevalence was observed at higher rates in Los Pavitos (Darien province), 9.0%, 95% CI: 3.6–17.6, while VEEV seroprevalence was elevated in El Cacao (Western Panama province), 27.3%, 95% CI: 16.1–40.9, and El Real de Santa María (Darien province), 20.4%, 95% CI: 12.6–29.7. Species like Oryzomys coesi, 23.1%, 95% CI: 5.0–53.8, and Transandinomys bolivaris, 20.0%, 95% CI: 0.5–71.6 displayed higher MADV seroprevalences than other species, whereas Transandinomys bolivaris, 80.0%, 95% CI: 28.3–99.4, and Proechimys semispinosus, 27.3%, 95% CI: 17.0–39.6, exhibited higher VEEV seroprevalences. Our findings provide support to the notion that rodents are vertebrate reservoirs of MADV and reveal spatial variations in alphavirus seropositivity among rodent species, with different provinces exhibiting distinct rates for MADV and VEEV. Moreover, specific rodent species are linked to unique seroprevalence patterns for these viruses, suggesting that rodent diversity and environmental conditions might play a significant role in shaping alphavirus distribution.

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          Most cited references41

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          Eastern equine encephalitis in Latin America.

          The eastern equine encephalitis (EEE) and Venezuelan equine encephalitis (VEE) viruses are pathogens that infect humans and horses in the Americas. Outbreaks of neurologic disease in humans and horses were reported in Panama from May through early August 2010. We performed antibody assays and tests to detect viral RNA and isolate the viruses in serum samples from hospitalized patients. Additional cases were identified with enhanced surveillance. A total of 19 patients were hospitalized for encephalitis. Among them, 7 had confirmed EEE, 3 had VEE, and 1 was infected with both viruses; 3 patients died, 1 of whom had confirmed VEE. The clinical findings for patients with EEE included brain lesions, seizures that evolved to status epilepticus, and neurologic sequelae. An additional 99 suspected or probable cases of alphavirus infection were detected during active surveillance. In total, 13 cases were confirmed as EEE, along with 11 cases of VEE and 1 case of dual infection. A total of 50 cases in horses were confirmed as EEE and 8 as VEE; mixed etiologic factors were associated with 11 cases in horses. Phylogenetic analyses of isolates from 2 cases of equine infection with the EEE virus and 1 case of human infection with the VEE virus indicated that the viruses were of enzootic lineages previously identified in Panama rather than new introductions. Cases of EEE in humans in Latin America may be the result of ecologic changes that increased human contact with enzootic transmission cycles, genetic changes in EEE viral strains that resulted in increased human virulence, or an altered host range. (Funded by the National Institutes of Health and the Secretaría Nacional de Ciencia, Tecnología e Innovación, Panama.).
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            A generic nested-RT-PCR followed by sequencing for detection and identification of members of the alphavirus genus.

            A specific and sensitive nested RT-PCR method was developed for the detection of members of the alphavirus genus. Based on available sequences, degenerated primers were selected in the nsP4 gene. Reaction components and thermal cycling parameters were investigated and standardised, and optimal ones were selected. As few as 25 pfu/tube could be detected. The identities of the amplified fragments were confirmed by sequencing, and phylogenetic analysis was carried out. The resulting phylogenetic tree could be applied to classify every alphavirus according to its serogroup. This technique is suitable for rapid, sensitive and reliable detection of these viruses and may be very valuable for diagnostic applications and surveillance.
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              Genetic and antigenic diversity among eastern equine encephalitis viruses from North, Central, and South America.

              Eastern equine encephalitis virus (EEEV), the sole species in the EEE antigenic complex, is divided into North and South American antigenic varieties based on hemagglutination inhibition tests. Here we describe serologic and phylogenetic analyses of representatives of these varieties, spanning the entire temporal and geographic range available. Nucleotide sequencing and phylogenetic analyses revealed additional genetic diversity within the South American variety; 3 major South/Central American lineages were identified including one represented by a single isolate from eastern Brazil, and 2 lineages with more widespread distributions in Central and South America. All North American isolates comprised a single, highly conserved lineage with strains grouped by the time of isolation and to some extent by location. An EEEV strain isolated during a 1996 equine outbreak in Tamaulipas State, Mexico was closely related to recent Texas isolates, suggesting southward EEEV transportation beyond the presumed enzootic range. Plaque reduction neutralization tests with representatives from the 4 major lineages indicated that each represents a distinct antigenic subtype. A taxonomic revision of the EEE complex is proposed.
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                Author and article information

                Journal
                bioRxiv
                BIORXIV
                bioRxiv
                Cold Spring Harbor Laboratory
                29 August 2023
                : 2023.08.28.555226
                Affiliations
                [1 ]Department of Biology, University of Oxford, Oxford, United Kingdom
                [2 ]Pandemic Sciences Institute, University of Oxford, Oxford United Kingdom
                [3 ]Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies, Panama City, Panama
                [4 ]Viral Emerging Disease Dynamics Group, Gorgas Memorial Institute of Health Studies, Panama City, Panama
                [5 ]Carson Centre for Research in Environment and Emerging Infectious Diseases, La Peñita, Darien, Panama
                [6 ]Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas, USA
                [7 ]Department of Microbiology, Immunology, and Molecular Genetics, University of Kentucky, College of Medicine, Lexington, Kentucky, USA
                [8 ]Institute for Human infection and Immunity, University of Texas Medical Branch, Texas, USA
                [9 ]Department of Medical Entomology, Gorgas Memorial Institute of Health Studies, Panama City, Panama
                [10 ]Geographic System Information Unit, Gorgas Memorial Institute of Health Studies, Panama City, Panama
                [11 ]Department of Pathology, University of Texas Medical Branch, Galveston Texas, USA
                [12 ]Center for Vector-Borne and Zoonotic Diseases, University of Texas Medical Branch, Galveston, Texas, USA
                [13 ]Center for Biodefense and Emerging Infectious Diseases, University of Texas Medical Branch, Galveston, Texas, USA
                [14 ]Center for Tropical Diseases, University of Texas Medical Branch, Galveston, Texas, USA
                [15 ]World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, Texas, USA
                [16 ]Department of Medicine, Division of Infectious Disease and Global Medicine, University of Florida, Gainesville, Florida, USA
                [17 ]Sociedad Mastozoológica de Panamá, Panama City, Panama
                [18 ]Clinical Research Unit, Gorgas Memorial Institute of health Studies, Panama City, Panama
                [19 ]Department of Statistics, University of Oxford, Oxford United Kingdom
                [20 ]Department of Infectious Disease Epidemiology, MRC Centre for Global Infectious Disease Analysis, J-IDEA, Imperial College London, London, UK.
                Author notes
                [±]

                Contributed equally

                [+]

                These are joint senior authors in this study.

                Article
                10.1101/2023.08.28.555226
                10491141
                37693579
                e094bff4-7e04-4fc6-8bcd-3b698a469e35

                This work is licensed under a Creative Commons Attribution-NoDerivatives 4.0 International License, which allows reusers to copy and distribute the material in any medium or format in unadapted form only, and only so long as attribution is given to the creator. The license allows for commercial use.

                History
                Funding
                JPC is funded by the Clarendon Scholarship from University of Oxford and Lincoln-Kingsgate Scholarship from Lincoln College, University of Oxford [grant number SFF1920_CB2_MPLS_1293647]. JFG is a masters student studying Epidemiological Research at Universidad Peruana Cayetano Heredia supported by training grant D43 TW007393 awarded by the Fogarty International Center of the US National Institutes of Health. This work was supported by SENACYT [grant number FID- 16-201] grant to JPC and AV. Proyecto: Estudio de las Enfermedades Emergentes y Síndromes Febriles en la Población Migrante, Ministerio de Economia y Finanzas de Panamá (Código: 019911.013) The Centers for Research in Emerging Infectious Diseases (CREID) Coordinating Research on Emerging Arboviral Threats Encompassing the NEOtropics (CREATE-NEO) 1U01AI151807 grant awarded to NV. WMS is supported by the Global Virus Network fellowship and the NIH (AI12094). CAD acknowledges funding the National Institute of Health Research for support of the Health Protection Research Unit in Emerging and Zoonotic Infections. WMS is supported by the Global Virus Network fellowship and the NIH (AI12094) Global Virus Network fellowship, Burroughs Wellcome fund (#1022448) and Wellcome Trust-Digital Technology Development award (Climate Sensitive Infectious Disease Modelling; (226075/Z/22Z). NRF acknowledges support from Wellcome Trust and Royal Society Sir Henry Dale Fellowship (204311/Z/16/Z), Bill and Melinda Gates Foundation (INV034540) and Medical Research Council-Sao Paulo Research Foundation (FAPESP) CADDE partnership award (MR/S0195/1 and FAPESP 18/14389-0).
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