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Rabies is a contagious viral disease that can be easily transmitted by the saliva and brain/nervous system tissues of the infected animals, causing severe and fatal encephalitis in both animals and humans. Vaccination campaigns are crucial to combat and prevent rabies's spread in dogs and humans. The Modified Fuenzalida & Palicios vaccines have been widely used since the 70s and have proven effective in producing a solid serological response. Since 2008, the Brazilian Ministry of Health has introduced a Cell Culture Rabies Vaccine (CCRV) for all dog mass vaccination campaigns in Brazil. However, to date, there is limited evidence on the immunologic response of dogs to this type of vaccine in field conditions. The present study evaluated the serological response in dogs vaccinated with CCRV from blood samples of 724 dogs using the Simplified Fluorescence Inhibition Microtest - SFIMT. Dogs with a titer equal to 0.5 IU/mL or above were considered seropositive. The results revealed that 59.12% (428/724) of all dogs tested and 48.49% (32/66) of primo-vaccinated animals were seropositive. The percentage of seronegative animals was higher than seropositive for animals that received a single dose during their life (p < 0.05). The opposite was observed in animals with five or more doses. The results of this study demonstrated that the CCRV vaccines elicit a satisfactory immunological response in field conditions and can constitute an essential population-level preventive strategy as part of annual canine rabies vaccination campaigns. Although its effectiveness has been studied, there is limited evidence of its immunological response in dogs under field conditions. This paper evaluates the serological response to CCRV in dogs vaccinated during mass vaccination campaigns from 2012 to 2017.
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Rabies virus is recognized as one of the most fatal zoonotic agents affecting all mammals. Wild boars (Sus scrofa), classified as a large-size exotic invasive species in Brazil with nationwide hunting permitted, may serve as an extra blood source for the common vampire bat (Desmodus rotundus). Our aim was to document wild boar exposure to vampire bats to determine the seroprevalence of rabies virus antibodies in wild boars and to determine the immune status of hunters in southern and central-western Brazilian regions. Serum samples were collected from 80 wild boars and 49 hunters from natural and degraded areas of the Atlantic Forest biome of southern Brazil and in degraded areas of the Cerrado biome of central-western Brazil. The rabies-modified rapid fluorescent focus inhibition test was performed to detect the presence of rabies virus neutralizing antibodies in wild boars and considered seropositive when ≥0.10 IU/mL. The simplified fluorescence inhibition microtest was used for samples from hunters with a titer of ≥0.50 IU/mL and considered indicative of seroconversion. While 11% (9/80) of wild boars had serum titers for rabies exposure (≥0.10 IU/mL), 88% (43/49) of corresponding hunters lacked immune protective titers (<0.50 IU/mL). Wild boars showed serum titers for rabies likely due to contact with contaminated saliva of vampire bats or from infected carcass consumption. Additionally, Brazilian wild boars can be exposed to rabies and may play an important role in the sylvatic rabies cycle by providing a blood supply for vampire bats, highlighting the possibility of direct transmission of rabies virus to hunting dogs and hunters. These findings suggested hunters are a potential risk group for contracting rabies, and the World Health Organization may consider adding this occupation to their recommendations of who should receive the pre-exposure rabies vaccination.
Assuntos
Quirópteros , Vírus da Raiva , Animais , Brasil/epidemiologia , Cães , Estudos Soroepidemiológicos , Sus scrofa , SuínosRESUMO
We evaluated the presence of antibodies for rabies virus in 177 serum samples from 125 wild lowland tapirs (Tapirus terrestris) from three different Brazilian biomes. The rapid fluorescent focus inhibition test was performed. No antibody titers suggesting the circulation of the rabies virus in tapir habitat were detected.
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Ecossistema , Perissodáctilos/sangue , Vírus da Raiva , Raiva/veterinária , Animais , Anticorpos Antivirais/sangue , Brasil/epidemiologia , Raiva/epidemiologia , Raiva/virologia , Estudos SoroepidemiológicosRESUMO
The direct rapid immunohistochemical test (dRIT) has been recommended for laboratorial diagnosis of rabies, especially in developing countries. The absence of commercial primary antibodies, however, still represents a major limitation to its wider use in testing. We describe here the development of a biotinylated polyclonal antibody against Rabies lyssavirus (RABV) ribonucleoprotein (RNP) and its use as a primary reagent in dRIT. Anti-RNP polyclonal horse IgG was purified by ionic exchange chromatography followed by immunoaffinity column chromatography, and its affinity, diagnostic sensitivity, and specificity were evaluated. CNS samples (120) of suspected rabies cases in different animal species were tested by dRIT, with the positive (n = 14) and negative (n = 106) results confirmed by direct fluorescence antibody testing (dFAT). Comparing the results of dRIT and dFAT, we found that the biotinylated anti-RNP IgG delivered 100% diagnostic specificity and sensibility for rabies diagnosis. Our findings show that the biotinylated anti-RNP polyclonal IgG can be produced with the quality required for application in dRIT. This work represents an important step in efforts to diagnose rabies in developing countries.
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Anticorpos Monoclonais/imunologia , Antígenos Virais/imunologia , Imunoglobulina G/imunologia , Vírus da Raiva/imunologia , Raiva/imunologia , Ribonucleoproteínas/imunologia , Animais , Anticorpos Monoclonais/metabolismo , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/metabolismo , Biotinilação , Encéfalo/imunologia , Encéfalo/virologia , Gatos , Bovinos , Quirópteros , Cães , Técnica Direta de Fluorescência para Anticorpo/métodos , Cavalos , Imunoglobulina G/metabolismo , Imuno-Histoquímica/métodos , Primatas , Raiva/diagnóstico , Raiva/virologia , Sensibilidade e Especificidade , Especificidade da Espécie , SuínosRESUMO
Rabies is a zoonotic viral disease that remains a serious threat to public health worldwide. The rabies lyssavirus (RABV) genome encodes five structural proteins, multifunctional and significant for pathogenicity. The large protein (L) presents well-conserved genomic regions, which may be a good alternative to generate informative datasets for development of new methods for rabies diagnosis. This paper describes the development of a technique for the identification of L protein in several RABV strains from different hosts, demonstrating that MS-based proteomics is a potential method for antigen identification and a good alternative for rabies diagnosis.(AU) i
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Animais , Raiva/diagnóstico , Proteínas Virais/genética , Vírus da Raiva/genética , Espectrometria de Massas , Proteômica/métodosRESUMO
Here, we compared the growth kinetics, cell-to-cell spread, and virus internalization kinetics in N2a cells of RABV variants isolated from vampire bats (V-3), domestic dogs (V-2) and marmosets (V-M) as well as the clinical symptoms and mortality caused by these variants. The replication rate of V-3 was significantly higher than those of V-2 and V-M. However, the uptake and spread of these RABV variants into N2a cells were inversely proportional. Nevertheless, V-3 had longer incubation and evolution periods. Our results provide evidence that the clinical manifestations of infection with bat RABV variant occur at a later time when compared to what was observed with canine and marmoset rabies virus variants.
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Quirópteros/virologia , Vírus da Raiva/fisiologia , Raiva/veterinária , Animais , Antígenos Virais , Callithrix/virologia , Linhagem Celular Tumoral , Cães/virologia , Camundongos , Raiva/patologia , Raiva/virologia , Vírus da Raiva/classificaçãoRESUMO
This study investigates the exposure of free-living jaguars from two federal protected areas in the Pantanal of Mato Grosso, Brazil, to a variety viral agents. These viral agents, particularly causing zoonotic diseases, were analyzed using serological and molecular methods. None of the jaguars was positive by RT-PCR for the molecular detection of avian influenza and West Nile Fever (WNF). Only one animal was serologically positive for Eastern Equine Encephalitis (EEE) by virus neutralization test in VERO cell cultures, representing the first reported case of jaguar exposure to EEE virus. However, all the animals were negative for Western Equine Encephalitis (WEE) virus and Venezuelan Equine Encephalitis (VEE) virus. Eleven jaguars were tested by two tests for the detection of antibodies against rabies virus (Simplified Fluorescent Inhibition Microtest SFIMT and Rapid Fluorescent Focus Inhibition Test RFFIT), resulting in five positive animals, two animals in each test and one in both serological tests. Furthermore, three out of 14 samples subjected to the neutralization test were positive for antibodies against canine distemper virus (CDV), and 15 out of 17 samples subjected to the hemagglutination-inhibition test (HI) were positive for antibodies against canine parvovirus (CPV). In view of the findings of this study, it is unlikely that the viruses examined here represent a threat to the jaguar populations in this region.(AU)
Este estudo investigou a exposição de onças-pintadas de vida livre a agentes virais selecionados em duas unidades de conservação federais no Pantanal de Mato Grosso, Brasil. Para a análise desses agentes virais, a maioria de caráter zoonótico, foram utilizados métodos sorológicos e moleculares. Nenhuma das onze onças-pintadas examinadas foi positiva na técnica de real-time RT-PCR para a detecção molecular dos agentes da Influenza aviária e Febre do Nilo Ocidental (WNF). Somente um animal foi positivo sorologicamente para a o vírus da Encefalite Equina do Leste (EEE) pela Microtécnica de vírus neutralização em culturas de células VERO, sendo este o primeiro relato da exposição de onças-pintadas. Todos os animais examinados s foram negativos para o vírus da Encefalite Equina do Oeste (WEE) e Venezuelana (VEE). Amostras de soro colhidas de 11 onças-pintadas foram submetidas a adois testes distintos para a detecção de anticorpos contra o vírus da raiva (Teste Rápido de Inibição de Foco de Fluorescência RFFIT e Microteste Simplificado de Inibição da Fluorescência - SFIMT), resultando em cinco animais positivos, dos quase dois positivos para cada teste e um positivo quando submetido aos dois testes sorológicos. Além disso, três das 14 amostras submetidas a técnica de soroneutralização foram positivas para a pesquisa de anticorpos contra o vírus da cinomose (CDV) e 15 amostras positivas das 17 analisadas para a pesquisa de anticorpos contra o parvovírus canino (CPV) foram identificadas pela técnica de Inibição da Hemaglutinação (HI). De acordo com os resultados deste estudo, é pouco provável que os agentes virais aqui analisados representem ameaça à população de onçaspintadas nesta região.(AU)
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Animais , Panthera/virologia , Pesquisa , Animais Selvagens/virologia , Técnicas de Diagnóstico Molecular/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Testes Sorológicos/veterináriaRESUMO
Background: Rabies cell culture infection test was developed for the isolation of Rabies lyssavirus and as an alternative for the mouse inoculation test. However, tissue culture for street rabies strains produces low viral titer. Here, we assessed the quantity of brain tissue for successful viral isolation toward increased virus titer in effective way.Methods: Brain tissue isolates from different reservoirs species of Brazil were harvested in different concentration and inoculated in mouse neuroblastoma cells (N2a). These isolates were measured infectious viral titer and cell viability followed by consecutive passages in N2a cells.Results: Inoculum containing were prominent Rabies lyssavirus due to higher viral titer and not significantly dead cell. After consecutive passages in N2a cells Rabies lyssavirus variant maintained by vampire bat had remarkable adaptation to the culture system, while isolates from marmoset presents distinct pattern of propagation in N2a cell when compared with other groups.Conclusion: Based on these results, the isolation followed by viral replication assay may be used in isolates from different reservoirs which enable an effective amplification of the wild type virus strains
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Callitrichinae/virologia , Cães/virologia , Quirópteros/virologia , Replicação Viral , Vírus da Raiva/isolamento & purificação , Raiva/diagnóstico , Técnicas de Cultura de CélulasRESUMO
BACKGROUND: Rabies is an incurable neglected zoonosis with worldwide distribution characterized as a lethal progressive acute encephalitis caused by a lyssavirus. Animal venoms and secretions have long been studied as new bioactive molecular sources, presenting a wide spectrum of biological effects, including new antiviral agents. Bufotenine, for instance, is an alkaloid isolated from the skin secretion of the anuran Rhinella jimi that inhibits cellular penetration by the rabies virus. Antimicrobial peptides, such as ocellatin-P1 and ocellatin-F1, are present in the skin secretion of anurans from the genus Leptodactylus and provide chemical defense against predators and microorganisms. METHODS: Skin secretion from captive Leptodactylus labyrinthicus was collected by mechanical stimulation, analyzed by liquid chromatography and mass spectrometry, and assayed for antiviral and cytotoxic activities. Synthetic peptides were obtained using solid phase peptide synthesis, purified by liquid chromatography and structurally characterized by mass spectrometry, and assayed in the same models. Cytotoxicity assays based on changes in cellular morphology were performed using baby hamster kidney (BHK-21) cells. Fixed Rabies virus (Pasteur Virus - PV) strain was used for virological assays based on rapid fluorescent focus inhibition test. RESULTS: Herein, we describe a synergic effect between ocellatin-F1 and bufotenine. This synergism was observed when screening the L. labyrinthicus skin secretion for antiviral activities. The active fraction major component was the antimicrobial peptide ocellatin-F1. Nevertheless, when the pure synthetic peptide was assayed, little antiviral activity was detectable. In-depth analyses of the active fraction revealed the presence of residual alkaloids together with ocellatin-F1. By adding sub-effective doses (e.g. < IC50) of pure bufotenine to synthetic ocellatin-F1, the antiviral effect was regained. Moreover, a tetrapetide derived from ocellatin-F1, based on alignment with the virus's glycoprotein region inferred as a possible cell ligand, was able to maintain the synergic antiviral activity displayed by the full peptide. CONCLUSIONS: This novel antiviral synergic effect between a peptide and an alkaloid may present an innovative lead for the study of new antiviral drugs.
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INTRODUCTION: In Brazil, various isolates of rabies virus (RABV) show antigenic profiles distinct from those established by the reduced panel of eight monoclonal antibodies (MAbs) determined by the Centers for Disease Control and Prevention (CDC), utilized for the antigenic characterization of RABV in the Americas. The objective of this study was to produce MAbs from RABV isolates from insectivorous bats with an antigenic profile incompatible with the pre-established one. METHODOLOGY: An isolate of RABV from the species Eptesicus furinalis that showed an antigenic profile incompatible with the panel utilized was selected. Hybridomas were produced utilizing the popliteal lymph nodes of mice immunized with ribonucleoproteins purified from the isolate. RESULTS: Two MAbs-producing clones were obtained, BR/IP1-3A7 and BR/IP2-4E10. Fifty-seven isolates of RABV from different species of animals and different regions of Brazil were analyzed utilizing the MAbs obtained. In the analysis of 23 RABV isolates from non-hematophagous bats, the MAbs cross-reacted with ten isolates, of which four were of the species Nyctinomops laticaudatus, one of the species Eptesicus furinalis, and five of the genus Artibeus. Of the nine isolates of non-hematophagous isolates that displayed an incompatible profile analyzed, characteristic of insectivorous bats, BR/IP1-3A7 reacted with five (55.55%) and BR/IP2-4E10 with four (44.44%). CONCLUSIONS: The MAbs obtained were able to recognize epitopes common between the three genera, Artibeus, Eptesicus, and Nyctinomops, thereby allowing the antigenic characterization of RABV isolates in Brazil.