Unexpected rabies variant identified in kinkajou (Potos flavus), Mato Grosso, Brazil.

A second case of a novel rabies variant described once in a capuchin monkey from Mato Grosso, Brazil, was discovered in a rabid wild kinkajou from the same region, indicating a public health risk following exposure to either of the two animals.

Yellow Fever Virus Reemergence and Spread in Southeast Brazil, 2016-2019.

The recent reemergence of yellow fever virus (YFV) in Brazil has raised serious concerns due to the rapid dissemination of the virus in the southeastern region. To better understand YFV genetic diversity and dynamics during the recent outbreak in southeastern Brazil, we generated 18 complete and nearly complete genomes from the peak of the epidemic curve from nonhuman primates (NHPs) and human infected cases across the Espírito Santo and Rio de Janeiro states. Genomic sequencing of 18 YFV genomes revealed the estimated timing, source, and likely routes of yellow fever virus transmission and dispersion during one of the largest outbreaks ever registered in Brazil. We showed that during the recent epidemic, YFV was reintroduced from Minas Gerais to the Espírito Santo and Rio de Janeiro states multiple times between 2016 and 2019. The analysis of data from portable sequencing could identify the corridor of spread of YFV. These findings reinforce the idea that continued genomic surveillance strategies can provide information on virus genetic diversity and transmission dynamics that might assist in understanding arbovirus epidemics.IMPORTANCE Arbovirus infections in Brazil, including yellow fever, dengue, zika, and chikungunya, result in considerable morbidity and mortality and are pressing public health concerns. However, our understanding of these outbreaks is hampered by the limited availability of genomic data. In this study, we investigated the genetic diversity and spatial distribution of YFV during the current outbreak by analyzing genomic data from areas in southeastern Brazil not covered by other previous studies. To gain insights into the routes of YFV introduction and dispersion, we tracked the virus by sequencing YFV genomes sampled from nonhuman primates and infected patients from the southeastern region. Our study provides an understanding of how YFV initiates transmission in new Brazilian regions and illustrates that genomics in the field can augment traditional approaches to infectious disease surveillance and control.

Seroprevalence of selected flaviviruses in free-living and captive capuchin monkeys in the state of Pernambuco, Brazil.

Mosquito-borne diseases such as dengue, yellow fever and, more recently, Chikungunya virus (CHIKV) and Zika virus (ZIKV) have a great impact in the public health. In addition, the presence of such viruses might have an impact on wild animal conservation as well as their possible role as animal reservoir. Here, we performed a serological survey searching for antibodies against a panel of flaviviruses [ZIKV, Dengue virus (DENV), Yellow Fever virus (YFV), West Nile virus (WNV), Saint Louis Encephalitis virus (SLEV), Ilheus virus (ILHV) and Rocio virus (ROCV)] using plaque reduction neutralization test (PRNT90 ) in both free-ranging and captive capuchin monkeys (Sapajus flavius and Sapajus libidinosus). Captive and free-living monkeys were sampled between June 2015 and January 2016 in the state of Pernambuco, including in the border with State of Paraíba, the epicentre of the ZIKV epidemics in Brazil. We have found neutralizing antibodies for ZIKV, DENV-1, DENV-2, DENV-3, DENV-4, YFV, ILHV and SLEV in both S. flavius and S. libidinosus samples. No positives samples were found for ROCV and WNV. Our results suggest that these flaviviruses might be circulating in capuchin monkey in the studied region. The possible presence of these viruses represents a risk for public health, as well as for animal conservation, especially for S. flavius which is a critically endangered species, facing high risk of extinction.

A novel hepatitis B virus species discovered in capuchin monkeys sheds new light on the evolution of primate hepadnaviruses.

BACKGROUND & AIMS: All known hepatitis B virus (HBV) genotypes occur in humans and hominoid Old World non-human primates (NHPs). The divergent woolly monkey HBV (WMHBV) forms another orthohepadnavirus species. The evolutionary origins of HBV are unclear. METHODS: We analysed sera from 124 Brazilian monkeys collected during 2012-2016 for hepadnaviruses using molecular and serological tools, and conducted evolutionary analyses. RESULTS: We identified a novel orthohepadnavirus species in capuchin monkeys (capuchin monkey hepatitis B virus [CMHBV]). We found CMHBV-specific antibodies in five animals and high CMHBV concentrations in one animal. Non-inflammatory, probably chronic infection was consistent with an intact preCore domain, low genetic variability, core deletions in deep sequencing, and no elevated liver enzymes. Cross-reactivity of antisera against surface antigens suggested antigenic relatedness of HBV, CMHBV, and WMHBV. Infection-determining CMHBV surface peptides bound to the human HBV receptor (human sodium taurocholate co-transporting polypeptide), but preferentially interacted with the capuchin monkey receptor homologue. CMHBV and WMHBV pseudotypes infected human hepatoma cells via the human sodium taurocholate co-transporting polypeptide, and were poorly neutralised by HBV vaccine-derived antibodies, suggesting that cross-species infections may be possible. Ancestral state reconstructions and sequence distance comparisons associated HBV with humans, whereas primate hepadnaviruses as a whole were projected to NHP ancestors. Co-phylogenetic analyses yielded evidence for co-speciation of hepadnaviruses and New World NHP. Bayesian hypothesis testing yielded strong support for an association of the HBV stem lineage with hominoid ancestors. Neither CMHBV nor WMHBV was likely the ancestor of the divergent human HBV genotypes F/H found in American natives. CONCLUSIONS: Our data suggest ancestral co-speciation of hepadnaviruses and NHP, and an Old World origin of the divergent HBV genotypes F/H. The identification of a novel primate hepadnavirus offers new perspectives for urgently needed animal models of chronic hepatitis B. LAY SUMMARY: The origins of HBV are unclear. The new orthohepadnavirus species from Brazilian capuchin monkeys resembled HBV in elicited infection patterns and could infect human liver cells using the same receptor as HBV. Evolutionary analyses suggested that primate HBV-related viruses might have emerged in African ancestors of New World monkeys millions of years ago. HBV was associated with hominoid primates, including humans and apes, suggesting evolutionary origins of HBV before the formation of modern humans. HBV genotypes found in American natives were divergent from those found in American monkeys, and likely introduced along prehistoric human migration. Our results elucidate the evolutionary origins and dispersal of primate HBV, identify a new orthohepadnavirus reservoir, and enable new perspectives for animal models of hepatitis B.

Chapparvoviruses occur in at least three vertebrate classes and have a broad biogeographic distribution.

Chapparvoviruses are a highly divergent group of parvoviruses (family Parvoviridae) that have recently been identified via metagenomic sampling of animal faeces. Here, we report the sequences of six novel chapparvoviruses identified through both metagenomic sampling of bat tissues and in silico screening of published vertebrate genome assemblies. The novel chapparvoviruses share several distinctive genomic features and group together as a robustly supported monophyletic clade in phylogenetic trees. Our data indicate that chapparvoviruses have a broad host range in vertebrates and a global distribution.

Evidence of leptospiral exposure in neotropical primates rescued from illegal trade and a Zoo in Bahia, Brazil / Indícios de exposição a leptospiras em primatas neotropicais resgatados do comércio ilegal e de um Zoológico da Bahia

Few studies have compared the seroprevalence of antileptospiral agglutinins with the demonstration of urinary shedding of leptospires or evidence of active infection in the bloodstreams of non-human primates. The study population consists of 58 animals, including d 42 monkeys from the Zoological Park of Salvador (Parque Zoobotânico Getúlio Vargas), Bahia, Brazil. The study also evaluated 16 primates (Cebus sp.) rescued from illegal trade that were housed in the Wildlife Rehabilitation Center of Salvador (CETAS), Bahia, Brazil. The seroprevalence of antileptospiral antibodies was low (2%) in the animals from the Zoo. A higher rate (31%) was observed among the animals that were rescued from illegal trade in the state of Bahia. Even if all the blood and urine samples were negative for leptospiral DNA fragments, the high frequency of serological evidence of exposure suggests a potential risk of leptospirosis transmission when keeping these animals as pets.(AU)

Poucos estudos compararam a soroprevalência de aglutininas antileptospira com a demonstração de excreção urinária de leptospiras ou evidência de infecção ativa em primatas não humanos. A população estudada consistiu em 58 animais, sendo 42 primatas do Parque Zoobotânico Getúlio Vargas, Bahia, Brasil. O estudo avaliou ainda 16 primatas (Cebus sp.) resgatados do tráfico ilegal e abrigados no Centro de Triagem de Animais Silvestres Chico Mendes, Salvador, Bahia, Brasil. A soroprevalência de anticorpos antileptospira foi baixa (2%) nos animais do Zoológico. Uma taxa mais elevada (31%) foi observada nos animais resgatados do tráfico ilegal. Mesmo que todas as amostras de sangue e urina tenham sido negativas para DNA de leptospiras, a alta frequência de evidência de exposição nos animais de origem selvagem indicam o risco potencial da adoção de primatas como animais de estimação.(AU)

Detection of antibodies to Oropouche virus in non-human primates in Goiânia City, Goiás.

INTRODUCTION: Arboviruses are associated with human disease, and non-human primates (NHPs) are important primary hosts. This study shows the detection of antibodies to Oropouche virus (OROV) in NHPs either living in urban parks or acclimatized at the Wild Animal Screening Center, Goiânia city. METHODS: Fifty blood samples were analyzed by hemagglutination-inhibition and neutralization assays. RESULTS: Two monkeys (Alouatta caraya) had antibodies to OROV by both techniques. CONCLUSIONS: This is the first report demonstrating the detection of OROV antibodies in Goiás State and may represent the introduction/circulation of OROV in the region and a potential risk to the human population.

Isolation of Madre de Dios Virus (Orthobunyavirus; Bunyaviridae), an Oropouche Virus Species Reassortant, from a Monkey in Venezuela.

Oropouche virus (OROV), genus Orthobunyavirus, family Bunyaviridae, is an important cause of human illness in tropical South America. Herein, we report the isolation, complete genome sequence, genetic characterization, and phylogenetic analysis of an OROV species reassortant, Madre de Dios virus (MDDV), obtained from a sick monkey (Cebus olivaceus Schomburgk) collected in a forest near Atapirire, a small rural village located in Anzoategui State, Venezuela. MDDV is one of a growing number of naturally occurring OROV species reassortants isolated in South America and was known previously only from southern Peru.

Characterization and comparative analysis of a simian foamy virus complete genome isolated from Brazilian capuchin monkeys.

Foamy viruses infect a wide range of placental mammals, including primates. However, despite of great diversity of New World primates, only three strains of neotropical simian foamy viruses (SFV) have been described. Only after 40 years since serological characterization, the complete sequence of an SFVcap strain infecting a family of six capuchin monkeys (Sapajus xanthosternos) was obtained. Co-culture of primate peripheral blood mononuclear cells with Cf2Th canine cells was established and monitored for the appearance of cytopathic effects, PCR amplification of integrated SFV proviral genome and viral reverse transcriptase activity. The novel SFVcap was fully sequenced through a next-generation sequencing protocol. Phylogenetic analysis of the complete genome grouped SFVcap and SFVmar, both infecting primate species of the Cebidae family with a genetic similarity of approximately 85%. Similar ORF sizes were observed among SFV from neotropical primates, and env and pol genes were the most conserved. Neotropical SFV presented the smallest LTRs among exogenous mammalians. The novel SFVcap strain provides a valuable research tool for the FV community.

Evaluation of arboviruses of public health interest in free-living non-human primates (Alouatta spp., Callithrix spp., Sapajus spp.) in Brazil.

INTRODUCTION: The aim of the present study was to evaluate the presence of arboviruses from the Flavivirus genus in asymptomatic free-living non-human primates (NHPs) living in close contact with humans and vectors in the States of Paraná and Mato Grosso do Sul, Brazil. METHODS: NHP sera samples (total n = 80, Alouatta spp. n = 07, Callithrix spp. n = 29 and Sapajus spp. n = 44) were screened for the presence of viral genomes using reverse transcription polymerase chain reaction and 10% polyacrylamide gel electrophoresis techniques. RESULTS: All of the samples were negative for the Flavivirus genome following the 10% polyacrylamide gel electrophoresis analysis. CONCLUSIONS: These negative results indicate that the analyzed animals were not infected with arboviruses from the Flavivirus genus and did not represent a risk for viral transmission through vectors during the period in which the samples were collected.

Evaluation of arboviruses of public health interest in free-living non-human primates (Alouatta spp., Callithrix spp., Sapajus spp.) in Brazil

INTRODUCTION: The aim of the present study was to evaluate the presence of arboviruses from the Flavivirus genus in asymptomatic free-living non-human primates (NHPs) living in close contact with humans and vectors in the States of Paraná and Mato Grosso do Sul, Brazil. METHODS: NHP sera samples (total n = 80, Alouatta spp. n = 07, Callithrix spp. n = 29 and Sapajus spp. n = 44) were screened for the presence of viral genomes using reverse transcription polymerase chain reaction and 10% polyacrylamide gel electrophoresis techniques. RESULTS: All of the samples were negative for the Flavivirus genome following the 10% polyacrylamide gel electrophoresis analysis. CONCLUSIONS: These negative results indicate that the analyzed animals were not infected with arboviruses from the Flavivirus genus and did not represent a risk for viral transmission through vectors during the period in which the samples were collected. .

Levantamento soroepidemiológico para arbovírus em macaco-prego-galego (Cebus flavius) de vida livre no estado da Paraíba e em macaco-prego (Cebus libidinosus) de cativeiro do nordeste do Brasil / Epidemiologic survey for arbovirus in galician capuchin monkeys (Cebus flavius) free living in Paraíba and captive capuchin monkey (Cebus libidinosus) from northeast Brazil

Este estudo descreve a primeira investigação de anticorpos para arbovírus em primatas não humanos do Novo Mundo no nordeste brasileiro. No período de março de 2008 a setembro de 2010 foram colhidos soros sanguíneos de 31 macacos-prego-galegos (Cebus flavius) de vida livre na Paraíba e de 100 macacos-prego (Cebus libidinosus) em cativeiro nos estados de Alagoas, Paraíba, Pernambuco, Piauí e Rio Grande do Norte. Para a pesquisa de anticorpos utilizou-se o teste de inibição da hemaglutinação (IH), usando antígenos de 19 diferentes tipos de arbovírus, pertencentes aos gêneros Flavivirus,Alphavirus e Bunyavirus. As amostras de soro foram testadas nas diluições de 1:20 a 1:1280. Dentre as amostras examinadas, todas as de C. flavius foram negativas e 46 por cento das de C. libidinosus em cativeiro apresentaram anticorpos para arbovírus. Foram detectados anticorpos para nove (9/19) arbovírus. Foram observadas 17 reações heterotípicas, para dois ou mais vírus, do gênero Flavivirus, e 15 para o gênero Alphavirus, com títulos variando de 1:20 a 1:1280. Quinze amostras apresentaram reação monotípica para ILHV (n=4), MAYV (n=6), SLEV (n=1), ROCV (n=2), OROV (n=1) e MUCV (n=1). Estes resultados sugerem que houve intensa circulação de arbovírus na população estudada de macacos-prego em cativeiro.

This paper describes the first investigation of arbovirus antibodies on New World non-human primates from Northeast Brazil. From March 2008 to September 2010 blood serum samples were collected from 31 wild blond capuchin monkeys (Cebus flavius) from Paraíba and 100 captive capuchin monkeys from Alagoas, Paraíba, Pernambuco, Piauí and Rio Grande do Norte. The haemagglutination-inhibition test (HI) was employed for 19 arbovirus of the Flavivirus,Alphavirus and Bunyavirus genus. Serum samples were tested from 1:20 to 1:1280 dilutions. Among the primates tested all C. flavius were negative and 46 percent C. libidinosus presented antibodies to arbovirus. Antibodies were detected for nine arbovirus (9/19). Seventeen heterotypic reactions were observed for at least two Or Flavirus and 15 for Alphavirus, at titers varying between 1:20 to 1:1280. Fifteen samples presented monotypic reaction for ILHV (n=4), MAYV (n=6), SLEV (n=1), ROCV (n=2), OROV (n=1) and MUCV (n=1). These results suggest that there was an intense arbovirus circulation in the studied population of captive capuchin monkeys.

Isolation of a phylogenetically distinct rabies virus from a tufted capuchin monkey (Cebus apella) in Brazil.

A rabies virus isolate (BRmk1358 strain) was discovered from a rabid tufted capuchin monkey in Brazil. The present study determined the nucleotide sequence of the BRmk1358 strain and compared with the rabies viruses isolated from marmosets and other animals in the Americas. Phylogenetic analyses showed that the BRmk1358 strain formed a lineage distant from that of marmoset rabies virus within the Chiroptera-related rabies virus cluster. This result suggests that the source of rabies infection in the tufted capuchin monkey may have been bat, and that they have a risk to act as rabies reservoir in Brazil.

Seroepidemiological monitoring in sentinel animals and vectors as part of arbovirus surveillance in the state of Mato Grosso do Sul, Brazil.

INTRODUCTION: From February-September 2010, seroepidemiological surveys were conducted on non-human primates and transmitter vector capture was used to investigate the possible circulation of arboviruses in the municipalities of Bonito, Campo Grande, and Jardim, State of Mato Grosso do Sul, Brazil. METHODS: A total of 65 primates from the wild and captivity were used, and potential vectors were captured using Castro and dip nets. Serum samples were tested at the Instituto Evandro Chagas, State of Pará, using the hemagglutination inhibition test to detect total antibodies against 19 different arboviruses. Virus isolation was attempted from serum samples and arthropod suspensions using newborn mice and the C6/36 cell line clone. In addition, identification of the vector species was conducted. RESULTS: From the 19 serum samples from Campo Grande, 1 sample had a 1:20 titer for Flavivirus. From the 35 samples collected in Bonito, 17 samples had antibodies to arboviruses, 4 (11.4%) were positive for Alphavirus, and 5 (14.2%) were positive for Flavivirus. Monotypic reactions were observed for the Mayaro (n = 10) and Oropouche (n = 5) viruses, and 6 (17.1%) samples had titers for >1 virus. We captured 120 Culicidae individuals that were potential arbovirus transmitters in Jardim; however, all the samples were negative for the viruses. CONCLUSIONS: Mato Grosso do Sul has a variety of vertebrate hosts and transmission vectors, thereby providing ideal conditions for the emergence or reemergence of arboviruses, including some pathogenic to human beings.

Seroepidemiological monitoring in sentinel animals and vectors as part of arbovirus surveillance in the state of Mato Grosso do Sul, Brazil / Monitoramento soroepidemiológico em animais-sentinela e vetores como parte da vigilância de arbovírus, no estado de Mato Grosso do Sul, Brasil

INTRODUCTION: From February-September 2010, seroepidemiological surveys were conducted on non-human primates and transmitter vector capture was used to investigate the possible circulation of arboviruses in the municipalities of Bonito, Campo Grande, and Jardim, State of Mato Grosso do Sul, Brazil. METHODS: A total of 65 primates from the wild and captivity were used, and potential vectors were captured using Castro and dip nets. Serum samples were tested at the Instituto Evandro Chagas, State of Pará, using the hemagglutination inhibition test to detect total antibodies against 19 different arboviruses. Virus isolation was attempted from serum samples and arthropod suspensions using newborn mice and the C6/36 cell line clone. In addition, identification of the vector species was conducted. RESULTS: From the 19 serum samples from Campo Grande, 1 sample had a 1:20 titer for Flavivirus. From the 35 samples collected in Bonito, 17 samples had antibodies to arboviruses, 4 (11.4%) were positive for Alphavirus, and 5 (14.2%) were positive for Flavivirus. Monotypic reactions were observed for the Mayaro (n = 10) and Oropouche (n = 5) viruses, and 6 (17.1%) samples had titers for >1 virus. We captured 120 Culicidae individuals that were potential arbovirus transmitters in Jardim; however, all the samples were negative for the viruses. CONCLUSIONS: Mato Grosso do Sul has a variety of vertebrate hosts and transmission vectors, thereby providing ideal conditions for the emergence or reemergence of arboviruses, including some pathogenic to human beings.

INTRODUÇÃO: No período de fevereiro a setembro de 2010, foram realizados inquéritos soroepidemiológicos em primatas não humanos e captura de vetores transmissores, com o intuito de investigar a possível circulação de arbovírus nos municípios de Bonito, Campo Grande e Jardim, no Estado do Mato Grosso do Sul, Brasil. MÉTODOS: Foram utilizados 65 primatas de vida livre e de cativeiro, e potenciais vetores capturados por Castro e puçás. As amostras séricas foram testadas pelo teste de inibição da hemaglutinação para a detecção de anticorpos totais contra 19 diferentes arbovírus e a tentativa de isolamento viral (camundongo recém-nascido e linhagem celular-clone C6/36) nas amostras séricas e suspensões de artrópodes, bem como a identificação das espécies vetoriais foram realizadas no Instituto Evandro Chagas-IEC no Estado do Pará. RESULTADOS: Das 19 amostras séricas do município de Campo Grande, 1 apresentou titulo de 1:20 para Flavivirus. Das 35 amostras coletadas em Bonito, 17 apresentaram anticorpos para arbovírus, sendo 4 (11,4%) positivos para Alphavirus, e 5 (14,2%) positivos para Flavivirus. Reações monotipicas foram observados para o vírus Mayaro (n=10) e para o vírus Oropouche (n=5) e 6 (17,1%) amostras apresentaram títulos para mais de um dos vírus estudados. Foram capturados 120 Culicídeos potenciais transmissores de arbovírus no município de Jardim. Todas as amostras coletadas foram negativas para o isolamento viral. CONCLUSÕES: Por possuir uma variedade de hospedeiros vertebrados e vetores transmissores, o Mato Grosso do Sul apresenta condições propícias para a emergência ou reemergência de arbovírus, inclusive alguns patogênicos para os seres homem.

The basal ganglia communicate with the cerebellum.

The basal ganglia and cerebellum are major subcortical structures that influence not only movement, but putatively also cognition and affect. Both structures receive input from and send output to the cerebral cortex. Thus, the basal ganglia and cerebellum form multisynaptic loops with the cerebral cortex. Basal ganglia and cerebellar loops have been assumed to be anatomically separate and to perform distinct functional operations. We investigated whether there is any direct route for basal ganglia output to influence cerebellar function that is independent of the cerebral cortex. We injected rabies virus (RV) into selected regions of the cerebellar cortex in cebus monkeys and used retrograde transneuronal transport of the virus to determine the origin of multisynaptic inputs to the injection sites. We found that the subthalamic nucleus of the basal ganglia has a substantial disynaptic projection to the cerebellar cortex. This pathway provides a means for both normal and abnormal signals from the basal ganglia to influence cerebellar function. We previously showed that the dentate nucleus of the cerebellum has a disynaptic projection to an input stage of basal ganglia processing, the striatum. Taken together these results provide the anatomical substrate for substantial two-way communication between the basal ganglia and cerebellum. Thus, the two subcortical structures may be linked together to form an integrated functional network.

Basal-ganglia 'projections' to the prefrontal cortex of the primate.

We used retrograde transneuronal transport of the McIntyre-B strain of herpes simplex virus type 1 to examine the extent and organization of basal-ganglia-thalamocortical projections to five regions of prefrontal cortex in the cebus monkey (Cebus apella): medial and lateral area 9 (9m and 9l), dorsal and ventral area 46 (46d and 46v) and lateral area 12 (12l). All of these prefrontal areas were found to be targets of basal-ganglia output that originated in the internal segment of the globus pallidus (GPi) and/or the pars reticulata of the substantia nigra (SNpr). Approximately one-third of the total volume of these nuclei was directed toward prefrontal cortex, a volume comparable to that directed at the cortical motor areas. The origins of the outputs to different prefrontal areas were topographically organized. Different portions of SNpr (the rostral and caudal thirds) projected to areas 9m and 12l. Similarly, different output nuclei (GPi and SNpr) projected to adjacent portions of the same cytoarchitectonic field (46d and 46v). Furthermore, the outputs to prefrontal areas were segregated from those to motor areas of cortex. Thus, basal-ganglia outputs to prefrontal cortex are both extensive and topographically organized, forming a rich anatomical substrate for basal-ganglia influences on the cognitive operations of the frontal lobe.