Outbreak In Brazil Research Articles

Inter- and Intra-Host Viral Diversity in a Large Seasonal DENV2 Outbreak

BackgroundHigh genetic diversity at both inter- and intra-host level are hallmarks of RNA viruses due to the error-prone nature of their genome replication. Several groups have evaluated the extent of viral variability using different RNA virus deep sequencing methods. Although much of this effort has been dedicated to pathogens that cause chronic infections in humans, few studies investigated arthropod-borne, acute viral infections.Methods and Principal FindingsWe deep sequenced the complete genome of ten DENV2 isolates from representative classical and severe cases sampled in a large outbreak in Brazil using two different approaches. Analysis of the consensus genomes confirmed the larger extent of the 2010 epidemic in comparison to a previous epidemic caused by the same viruses in another city two years before (genetic distance = 0.002 and 0.0008 respectively). Analysis of viral populations within the host revealed a high level of conservation. After excluding homopolymer regions of 454/Roche generated sequences, we found 10 to 44 variable sites per genome population at a frequency of >1%, resulting in very low intra-host genetic diversity. While up to 60% of all variable sites at intra-host level were non-synonymous changes, only 10% of inter-host variability resulted from non-synonymous mutations, indicative of purifying selection at the population level.Conclusions and SignificanceDespite the error-prone nature of RNA-dependent RNA-polymerase, dengue viruses maintain low levels of intra-host variability.

Serological study of vaccinia virus reservoirs in areas with and without official reports of outbreaks in cattle and humans in São Paulo, Brazil

Vaccinia virus (VACV), the etiological agent of an exanthematic disease, has been associated with several bovine outbreaks in Brazil since the end of the global vaccination campaign against smallpox. It was previously believed that the vaccine virus used for the WHO global campaign had adapted to an unknown wild reservoir and was sporadically re-emerging in outbreaks in cattle and milkers. At present, it is known that Brazilian VACV is phylogenetically different from the vaccinia virus vaccinal strain, but its origin remains unknown. This study assessed the seroprevalence of orthopoxviruses in domestic and wild animals and farmers from 47 farms in three cities in the southwest region of the state of São Paulo with or without official reports of outbreaks in cattle or humans. Our data indicate a low seroprevalence of antibodies in wild animals and raise interesting questions about the real potential of wild rodents and marsupials as VACV reservoirs, suggesting other routes through which VACV can be spread.

Ultrastructural Study of Bovine Papillomavirus During Outbreaks in Brazil

Los virus del papiloma que pertenecen a la familia Papillomaviridae son pequenos virus oncogenicos que causan papilomas y fibropapilomas en epitelio cutaneo y mucoso de distintas especies de animales. En el ganado vacuno, trece tipos (BPV- 1-13) se caracterizaron hasta el momento. En este estudio, se documenta la ocurrencia de cuatro brotes de papilomatosis en los rebanos de ganado, procedentes de estados brasilenos. Las lesiones papilomatosas se localizaron en los pezones, la ubre, la cabeza y el cuello. Al microscopio electronico de transmision, en la tecnica de tincion negativa fueran visualizadas particulas del virus del papiloma redondeadas, con simetria icosaedrica, caracterizadas como llenas y vacias, midiendo unos 60 nm de diametro en todas las 40 muestras de fragmentos de lesion de piel estudiado. Los cortes ultra finos mostraron la presencia de grupos de particulas virales, intranucleares redondeadas con 35 nm de diametro y tubulares 35-39 nm de diametro. En la tecnica de microscopia inmunoelectronica, la positividad obtenida se caracterizo por la presencia de agregados de particulas virales formadas por la interaccion antigeno-anticuerpo. En la aplicacion de la tecnica de inmunocitoquimica, la reaccion antigeno-anticuerpo mostro particulas de oro coloidal distribuidos de manera uniforme sobre la superficie del virus. Estos resultados muestran la importancia de las tecnicas de microscopia electronica de transmision en el diagnostico de papilomatosis bovina, que pueden ser utilizados en los procedimientos de rutina para la identificacion del agente viral causante de esta importante enfermedad.

Mortality associated with Weissellosis (Weissella sp.) in USA farmed rainbow trout: Potential for control by vaccination

Recent reports indicate that novel Weissella sp. bacteria have been associated with disease outbreaks in cultured rainbow trout (Oncorhynchus mykiss) at commercial farms in China and Brazil. In the summer of 2011, a severe disease outbreak displaying similar clinical signs occurred at a commercial rainbow trout farm in western North Carolina. Observed signs included dark skin coloration, lethargic swimming, bilateral exophthalmia, corneal opacity, ocular hemorrhage, occasional corneal rupture, and in some cases cerebral hemorrhage. Mortality was most severe in larger fish approaching market size. Bacteria isolated from moribund fish were identified to the genus level as Weissella sp. by 16S rRNA gene sequence analysis and were 99% identical to the sequences of isolates collected from the Chinese and Brazilian outbreaks. Laboratory-based disease challenge experiments using the isolated pathogen replicated both the disease signs and induction of mortality in exposed healthy rainbow trout. Aqueous vaccine formulations containing formalin-inactivated whole-cell Weissella sp. antigens conferred significant protection against experimental infection when both the vaccine and the pathogen were delivered by injection (relative percent survival [RPS] of 87.5% and 85% at 38 and 72days after vaccination, respectively). The Weissella sp. vaccine was equally effective when combined with a commercially available Yersinia ruckeri vaccine, and this bivalent formulation did not alter the efficacy of the Y. ruckeri component of the vaccine. This is the first identification of this emerging rainbow trout disease, which we have termed Weissellosis, in the United States, and the spread of this pathogen might pose a significant threat to the domestic rainbow trout aquaculture industry. Our results also suggest that a bivalent Weissella/Y. ruckeri vaccine could be used as an effective and economical means for controlling this pathogen.

Molecular characterization of Aeromonas spp. and Vibrio cholerae O1 isolated during a diarrhea outbreak

This work aimed to assess pathogenic potential and clonal relatedness of Aeromonas sp. and Vibrio cholerae isolates recovered during a diarrhea outbreak in Brazil. Clinical and environmental isolates were investigated for the presence of known pathogenic genes and clonal relatedness was assessed by intergenic spacer region (ISR) 16S-23S amplification. Four Aeromonas genes (lip, exu, gcat, flaA/B) were found at high overall frequency in both clinical and environmental isolates although the lip gene was specifically absent from selected species. A fifth gene, aerA, was rarely found in A. caviae, the most abundant species. The ISR profile revealed high heterogeneity among the Aeromonas isolates and no correlation with species identification. In contrast, in all the V. cholerae isolates the four genes investigated (ctxA, tcpA, zot and ace) were amplified and revealed homogeneous ISR and RAPD profiles. Although Aeromonas isolates were the major enteric pathogen recovered, their ISR profiles are not compatible with a unique cause for the diarrhea events, while the clonal relationship clearly implicates V. cholerae in those cases from which it was isolated. These results reinforce the need for a better definition of the role of aeromonads in diarrhea and whether they benefit from co-infection with V. cholerae.

Characterization of a New Vaccinia virus Isolate Reveals the C23L Gene as a Putative Genetic Marker for Autochthonous Group 1 Brazilian Vaccinia virus

Since 1999, several Vaccinia virus (VACV) isolates, the etiological agents of bovine vaccinia (BV), have been frequently isolated and characterized with various biological and molecular methods. The results from these approaches have grouped these VACV isolates into two different clusters. This dichotomy has elicited debates surrounding the origin of the Brazilian VACV and its epidemiological significance. To ascertain vital information to settle these debates, we and other research groups have made efforts to identify molecular markers to discriminate VACV from other viruses of the genus Orthopoxvirus (OPV) and other VACV-BR groups. In this way, some genes have been identified as useful markers to discriminate between the VACV-BR groups. However, new markers are needed to infer ancestry and to correlate each sample or group with its unique epidemiological and biological features. The aims of this work were to characterize a new VACV isolate (VACV DMTV-2005) molecularly and biologically using conserved and non-conserved gene analyses for phylogenetic inference and to search for new genes that would elucidate the VACV-BR dichotomy. The VACV DMTV-2005 isolate reported in this study is biologically and phylogenetically clustered with other strains of Group 1 VACV-BR, the most prevalent VACV group that was isolated during the bovine vaccinia outbreaks in Brazil. Sequence analysis of C23L, the gene that encodes for the CC-chemokine-binding protein, revealed a ten-nucleotide deletion, which is a new Group 1 Brazilian VACV genetic marker. This deletion in the C23L open reading frame produces a premature stop-codon that is shared by all Group 1 VACV-BR strains and may also reflect the VACV-BR dichotomy; the deletion can also be considered to be a putative genetic marker for non-virulent Brazilian VACV isolates and may be used for the detection and molecular characterization of new isolates.

Evaluation of a generic RT-nested-PCR for detection of flaviviruses in suspected fatal cases of dengue infection, Rio de Janeiro, Brazil

Flaviviruses are significant causes of disease worldwide and can be classified serologically into several antigenic complexes. The purpose of the present study was to evaluate the effectiveness of a generic RT-nested-PCR for detection of flavivirus during a dengue outbreak in Brazil in 2008. A total of 105 serum samples were collected from patients with fatal outcome and examined by generic RT-PCR, conventional RT-PCR, and IgM serology. The generic RT-PCR confirmed 19 of 105 (18%) cases. Conventional RT-PCR performed on 105 serum samples detected 45 (42.8%) dengue virus infections. The IgM serology confirmed 44 of 102 (43.1%) cases. The infecting serotype was identified by generic RT-PCR in 19 cases (18 DENV-2 and 1 DENV-3) and by conventional RT-PCR in 45 cases (40 DENV-2 and 5 DENV-3). In addition, we analyzed the performance of the generic and conventional RT-PCRs and IgM serology on serum samples stratified by the day of onset of symptoms. Our results indicate that different methods should be included in flavivirus surveillance programs, including virological and serological approaches.

Detection of dengue virus in sera of Brazilian blood donors

Dengue is the most important arboviral disease in the world. Dengue viruses (DENVs) have produced huge outbreaks in Brazil in the past 25 years with more than 5 million reported cases. During these epidemics, asymptomatic individuals infected with DENV could donate blood and serve as a source of virus dissemination in the community. Here, we studied the circulation of DENV in healthy individuals during an epidemic outbreak. The study included 500 serum samples from healthy blood donors collected at the Hemotherapy Center of Ribeirão Preto, Brazil, during a dengue outbreak. The presence of DENV RNA in the serum samples was screened by real-time reverse transcription-polymerase chain reaction (PCR). The virus serotype was determined by a heminested PCR procedure. A partial fragment of the NS5 gene sequence was used for phylogenetic analysis. DENV RNA was detected in the serum sample of 2 of 500 (0.4%) individuals. Both of them were infected with DENV-3 Genotype III, a virus that has been circulating in Brazil in the past decade. Individuals with asymptomatic DENV infection can be blood donors and serve as a source of virus dissemination in the community. Further studies are needed to determine the risk of recipient infection by DENV as a result of transfusion in Brazil, especially during epidemic periods.

Molecular Characterization and Phylogenetic Study of Coxsackievirus A24v Causing Outbreaks of Acute Hemorrhagic Conjunctivitis (AHC) in Brazil

BackgroundCoxsackievirus A24 variant (CA24v) is the most prevalent viral pathogen associated with acute hemorrhagic conjunctivitis (AHC) outbreaks. Sixteen years after its first outbreak in Brazil, this agent reemerged in 2003 in Brazil, spread to nearly all states and caused outbreaks until 2005. In 2009, a new outbreak occurred in the northeast region of the country. In this study, we performed a viral isolation in cell culture and characterized clinical samples collected from patients presenting symptoms during the outbreak of 2005 in Vitória, Espírito Santo State (ES) and the outbreak of 2009 in Recife, Pernambuco State (PE). We also performed a phylogenetic analysis of worldwide strains and all meaningful Brazilian isolates since 2003.Methods and FindingsSterile cotton swabs were used to collect eye discharges, and all 210 clinical samples were used to inoculate cell cultures. Cytopathic effects in HEp-2 cells were seen in 58 of 180 (32%) samples from Vitória and 3 of 30 (10%) samples from Recife. Phylogenetic analysis based on a fragment of the VP1 and 3C gene revealed that the CA24v causing outbreaks in Brazil during the years 2003, 2004 and 2005 evolved from Asian isolates that had caused the South Korean outbreak of AHC during the summer of 2002. However, the 2009 outbreak of AHC in Pernambuco was originated from the reintroduction of a new CA24v strain that was circulating during 2007 in Asia, where CA24v outbreaks has been continuously reported since 1970.ConclusionsThis study is the first phylogenetic analysis of AHC outbreaks caused by CA24v in Brazil. The results showed that Asian strains of CA24v were responsible for the outbreaks since 1987 and were independently introduced to Brazil in 2003 and 2009. Phylogenetic analysis of complete VP1 gene is a useful tool for studying the epidemiology of enteroviruses associated with outbreaks.

An Experimental Model of Meningoencephalomyelitis by Rocio Flavivirus in Balb/C Mice: Inflammatory Response, Cytokine Production, and Histopathology

Rocio virus (ROCV) is a flavivirus, probably transmitted by Culex mosquitoes and maintained in nature as a zoonosis of wild birds. Rocio virus caused a human epidemic of severe encephalitis that lasted from 1973 to 1980 in the Ribeira valley, in the southeastern coast of Brazil. After this outbreak, serologic evidence of ROCV circulation has been reported and public health authorities are concerned about a return of ROCV outbreaks in Brazil. We show here a study on the pathogenesis and the physiopathology of ROCV disease in the central nervous system of a Balb/C young adult mice experimental model. The animals were intraperitoneally infected by ROCV and followed from 0 to 9 days after infection, when all of them died. Nervous tissue samples were collected from infected animals for immunohistochemistry and molecular biology analysis. We observed the virus in the central nervous system, the inflammatory changes induced by Th1 and Th2 cytokines, and the final irreversible damage of nervous tissues by neuronal degeneration and apoptosis. These findings can help to better understand the pathogenesis and physiopathology of the human meningoencephalomyelitis by ROCV and other flaviviruses.

Changes in vancomycin-resistant Enterococcus faecium causing outbreaks in Brazil

Enterococci have been implicated in severe human infections as a consequence of associated determinants of virulence and antimicrobial resistance. The majority of vancomycin-resistant Enterococcus faecium (VRE(fm)) connected to outbreaks worldwide pertains to the clonal complex 17 (CC17). In Brazil, the majority of VRE(fm) involved in outbreaks reported so far are not related to CC17. VRE(fm) strains responsible for an outbreak and sporadic cases in hospitals located in the city of Campinas, Brazil, were compared to other VRE(fm) strains in the country. Twenty-two out of 23 E.faecium were vancomycin-resistant and harboured the vanA gene. One vancomycin-susceptible E.faecium (VSE(fm)) strain was included in this study because it was isolated from a patient who one week later harboured a VRE(fm). All strains, except VSE, showed the same alteration in the VanA element characterised by deletion of the left extremity of the transposon and insertion of IS1251 between the vanS and vanH genes. Genes codifying virulence factors such as collagen-adhesin protein, enterococcal surface protein and hyaluronidase were detected in the VRE(fm) and VSE(fm) studied. Both pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) revealed that VRE(fm) and VSE(fm) strains have a clonal relationship. New sequence types (STs) were identified by MLST as ST447, ST448, ST478 and ST412 but all belonged to the CC17. The present study revealed that VRE(fm) outbreaks in Brazil were caused by strains that did not share a common evolutionary history, and that VRE(fm) strains belonging to CC17 could be predominant in Brazil as in other countries.

National evaluation of rapidly growing mycobacteria outbreaks in Brazil

This study describes the epidemiologic characteristics of HAI outbreaks caused by RGM in Brazil.

Zoonotic Vaccinia Virus Outbreaks in Brazil

The vaccinia virus (VACV) was used as a live vaccine during the WHO-led smallpox eradication campaign in the second half of the 20th century. The program culminated with the obliteration of the disease, one of the most important achievements in modern medicine. Interestingly, one of the key factors in the successful vaccination campaign – the VACV itself – is poorly understood in relation to its natural reservoirs, evolutionary history and origins, being frequently considered extinct as a naturally occurring virus. Nevertheless, orthopoxviruses other than variola virus have been known to circulate in Brazil since the early 1960s. More specifically, VACV has been associated with naturally acquired infections in humans, cattle and possibly other reservoirs since 1999, when bovine vaccinia outbreaks started to be consistently described year after year. In this article, we list and discuss the most important VACV outbreaks that have occurred in Brazil in the last 20 years. Phylogenetic issues are considered, as the latest studies point to large genetic variance among isolates. Clinical and epidemiological data, both published and new, are presented.

Evaluation of two molecular methods for the detection of Yellow fever virus genome

Yellow fever virus (YFV), a member of the family Flaviviridae, genus Flavivirus is endemic to tropical areas of Africa and South America and is among the arboviruses that pose a threat to public health. Recent outbreaks in Brazil, Bolivia, and Paraguay and the observation that vectors capable of transmitting YFV are presenting in urban areas underscore the urgency of improving surveillance and diagnostic methods. Two novel methods (RT-hemi-nested-PCR and SYBR® Green qRT-PCR) for efficient detection of YFV strains circulating in South America have been developed. The methods were validated using samples obtained from golden hamsters infected experimentally with wild-type YFV strains as well as human serum and tissue samples with acute disease.

Vaccinia Virus Zoonotic Infection, São Paulo State, Brazil

Univ Estadual Paulista, Fac Med Vet & Zootecnia, Dept Higiene Vet & Saude Publ, BR-18610250 Sao Paulo, Brazil

An analysis of the 1978 African swine fever outbreak in Brazil and its eradication

The aim of this paper is to evaluate the African swine fever (ASF) outbreak that began in 1978 in Paracambi municipality, Rio de Janeiro State, Brazil, and the steps taken by the Brazilian authorities to eradicate the disease. The presence of ASF in the country was confirmed by isolating the virus, and its pathogenicity was certified by the laboratory of the Plum Island Disease Center, New York, United States. Even before the laboratory results became available, the Brazilian Agriculture Ministry declared an Animal Health Emergency, in which the official Veterinary Services adopted rapid control measures to restrain and eradicate the disease. These control measures contributed to the reorganisation of the national swine industry and stimulated the use of high-technology production techniques, as well as an improvement in herd health consciousness. All this contributed to Brazil becoming the largest meat exporter in the world.

Identification of an Atypical Strain ofToxoplasma gondiias the Cause of a Waterborne Outbreak of Toxoplasmosis in Santa Isabel do Ivai, Brazil

Multilocus DNA sequencing has identified a nonarchetypal strain of Toxoplasma gondii as the causal agent of a waterborne outbreak in Brazil in 2001. The strain, isolated from a water supply epidemiologically linked to the outbreak, was virulent to mice, and it has previously been identified as BrI. Using a serologic assay that detects strain-specific antibodies, we found that 13 (65%) of 20 individuals who were immunoglobulin (Ig) M positive during the outbreak possessed the same serotype as mice infected with the purported epidemic strain. The remaining 7 individuals, plus additional IgM-negative, IgG-positive individuals, possessed 1 of 4 novel serotypes, the most common of which matched the serotype of mice infected with strains isolated from chickens foraging near the outbreak site. The latter strains likely reflect the genetic diversity of T. gondii circulating in highly endemic regions of Brazil. The serotyping assay proved a useful tool for identification of specific individuals infected with the outbreak agent.

Prokaryotic Taxonomy Rules and Nomenclature Changes in the Mycobacterium Chelonae–Abscessus Group: Author Reply

Future MicrobiologyVol. 5, No. 10 Letter to the EditorFree AccessProkaryotic taxonomy rules and nomenclature changes in the Mycobacterium chelonae–abscessus group: author replySylvia Cardoso Leão, Rafael Silva Duarte & Erica ChimaraSylvia Cardoso Leão† Author for correspondenceDepartamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Rua Botucatu 862 3° andar, São Paulo-SP, Brazil. Search for more papers by this authorEmail the corresponding author at sylvia.leao@unifesp.br, Rafael Silva DuarteInstituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, BrazilSearch for more papers by this author & Erica ChimaraSetor de Micobactérias, Instituto Adolfo Lutz, São Paulo, SP, BrazilSearch for more papers by this authorPublished Online:12 Nov 2010https://doi.org/10.2217/fmb.10.121AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInReddit We thank Jorge Sampaio for his interest in our paper and appreciate his suggestions regarding the taxonomy of rapidly growing mycobacteria. His concern about the nomenclature of members of the Mycobacterium chelonae–abscessus group is justifiable. After an extensive phenotypic and genotypic characterization of members of the M. chelonae–abscessus group, Leao et al. proposed the union of M. abscessus, M. bolletii and M. massiliense and the recognition of two subspecies within M. abscessus: M. abscessus subsp. abscessus and M. abscessus subsp. massiliense [1]. The name ‘massiliense’ was chosen as it appeared first in the literature. A note has been submitted and is under evaluation in the International Journal of Systematic and Evolutionary Microbiology (ITSEM) to provide a complete and strict systematic description of M. abscessus and its subspecies in accordance with the Rules of the Bacteriological Code (1990 Revision) [2,3]. In this note, the name M. abscessus subsp. massiliense is corrected to M. abscessus subsp. bolletii because according to Rule 24b (2) the name M. bolletii has priority over the name M. massiliense; these taxa are heterotypic synonyms and, according to Rule 38, must be united under the name M. bolletii; and finally, according to Rule 50b, the name of the subspecies must be M. abscessus subsp. bolletii. However, we disagree with publishing an erratum because at the time our paper was published in Future Microbiology [4] the nomenclature of the proposed M. abscessus subspecies was not validated and there was only one reference available for citation, that of Leao et al. [1]. Moreover, a detailed description of the cited species and subspecies is required and a possible change in the nomenclature of these taxa in the future is still under consideration by the IJSEM.Financial & competing interests disclosureThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.Bibliography1 Leao SC, Tortoli E, Viana-Niero C et al.: Characterization of mycobacteria from a major Brazilian outbreak suggests that revision of the taxonomic status of members of the Mycobacterium chelonae–M. abscessus group is needed. J. Clin. Microbiol.47(9),2691–2698 (2009).Crossref, Medline, CAS, Google Scholar2 De Vos P, Trüper HG: Judicial commission of the International Committee on Systematic Bacteriology. IXth International (IUMS) Congress of Bacteriology and Applied Microbiology. Minutes of the meetings, 14, 15 and 18 August 1999, Sydney, Australia. Int. J. Syst. Evol. Microbiol.50,2239–2244 (2000).Crossref, Google Scholar3 Lapage SP, Sneath PHA, Lessel EF, Skerman VBD, Seeliger HPR, Clark WA: International Code of Nomenclature of Bacteria (1990 Revision). American Society for Microbiology, Washington, DC, USA (1992).Google Scholar4 Leao SC, Viana-Niero C, Matsumoto CK et al.: Epidemic of surgical-site infections by a single clone of rapidly growing mycobacteria in Brazil. Future Microbiol.5(6),971–980 (2010).Link, Google ScholarFiguresReferencesRelatedDetails Vol. 5, No. 10 Follow us on social media for the latest updates Metrics History Published online 12 November 2010 Published in print October 2010 Information© Future Medicine LtdFinancial & competing interests disclosureThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.PDF download

Characterization of Dengue Virus Type 2: New Insights on the 2010 Brazilian Epidemic

Dengue viruses (DENV) serotypes 1, 2, and 3 have been causing yearly outbreaks in Brazil. In this study, we report the re-introduction of DENV2 in the coast of São Paulo State. Partial envelope viral genes were sequenced from eighteen patients with dengue fever during the 2010 epidemic. Phylogenetic analysis showed this strain belongs to the American/Asian genotype and was closely related to the virus that circulated in Rio de Janeiro in 2007 and 2008. The phylogeny also showed no clustering by clinical presentation, suggesting that the disease severity could not be explained by distinct variants or genotypes. The time of the most recent common ancestor of American/Asian genotype and the São Paulo and Rio de Janeiro (SP/RJ) monophyletic cluster was estimated to be around 40 and 10 years, respectively. Since this virus was first identified in Brazil in 2007, we suggest that it was already circulating in the country before causing the first documented outbreak. This is the first description of the 2010 outbreak in the State of São Paulo, Brazil, and should contribute to efforts to control and monitor the spread of DENVs in endemic areas.

Vaccinia Virus Infection in Monkeys, Brazilian Amazon

To detect orthopoxvirus in the Brazilian Amazon, we conducted a serosurvey of 344 wild animals. Neutralizing antibodies against orthopoxvirus were detected by plaque-reduction neutralizing tests in 84 serum samples. Amplicons from 6 monkey samples were sequenced. These amplicons identified vaccinia virus genetically similar to strains from bovine vaccinia outbreaks in Brazil.