HTLV-2 infection in Manaus, Brazil: first description of HTLV-2c subtype in an urban area of the Western Amazon region

Abstract INTRODUCTION: We investigated the prevalence of human T-cell lymphotropic virus types 1 and 2 (HTLV-1/2) infection in patients with hematological diseases from the western Amazon region of Brazil. METHODS: Samples from 306 patients were submitted for the molecular diagnosis of HTLV-1/2 infection by real time PCR (qPCR), with amplification, sequencing, and phylogenetic analysis of the long terminal repeat (LTR) region. RESULTS: A 29-year-old male carrier of sickle cell anemia with a history of multiple blood transfusions was diagnosed with the HTLV-2c subtype. CONCLUSIONS: This study describes the first known occurrence of HTLV-2c in the urban area of Brazil's western Amazon region.

HTLV-2 infection in Manaus, Brazil: first description of HTLV-2c subtype in an urban area of the Western Amazon region.

INTRODUCTION: We investigated the prevalence of human T-cell lymphotropic virus types 1 and 2 (HTLV-1/2) infection in patients with hematological diseases from the western Amazon region of Brazil. METHODS: Samples from 306 patients were submitted for the molecular diagnosis of HTLV-1/2 infection by real time PCR (qPCR), with amplification, sequencing, and phylogenetic analysis of the long terminal repeat (LTR) region. RESULTS: A 29-year-old male carrier of sickle cell anemia with a history of multiple blood transfusions was diagnosed with the HTLV-2c subtype. CONCLUSIONS: This study describes the first known occurrence of HTLV-2c in the urban area of Brazil's western Amazon region.

Cacipacore virus as an emergent mosquito-borne Flavivirus.

INTRODUCTION:: Cacipacore virus (CPCV), a possible bird-associated flavivirus, has yet to be detected in mosquitoes. Our purpose is examining CPCV in mosquitoes from the Amazon region of Brazil. METHODS:: Approximately 3,253 Culicidae (grouped into 264 pools) were collected from the Amazon region during 2002-2006 and analyzed using a Flavivirus genus-specific reverse transcription- polymerase chain reaction followed by nested polymerase chain reaction assay and by nucleotide sequencing of amplicons. RESULTS:: Nucleotide sequences from five mosquito samples showed high similarity to the those of CPCV originally isolated in the Amazon region. CONCLUSIONS:: This is the first report of CPCV-infected mosquitoes which has implications on the arbovirus maintenance in nature and transmission to man.

Cacipacore virus as an emergent mosquito-borne Flavivirus

Abstract INTRODUCTION: Cacipacore virus (CPCV), a possible bird-associated flavivirus, has yet to be detected in mosquitoes. Our purpose is examining CPCV in mosquitoes from the Amazon region of Brazil. METHODS: Approximately 3,253 Culicidae (grouped into 264 pools) were collected from the Amazon region during 2002-2006 and analyzed using a Flavivirus genus-specific reverse transcription- polymerase chain reaction followed by nested polymerase chain reaction assay and by nucleotide sequencing of amplicons. RESULTS: Nucleotide sequences from five mosquito samples showed high similarity to the those of CPCV originally isolated in the Amazon region. CONCLUSIONS: This is the first report of CPCV-infected mosquitoes which has implications on the arbovirus maintenance in nature and transmission to man.

Detection of DENV-4 genotype I from mosquitoes collected in the city of Manaus, Brazil.

BACKGROUND: Dengue epidemics have been reported in Brazil since 1981. In Manaus, a large city in the Amazon region, dengue is endemic with all four-virus serotypes (DENV-1, -2, -3, and -4) simultaneously causing human disease. In 2008, during a surveillance of dengue virus in mosquitoes in the district of Tancredo Neves in Manaus, 260 mosquitoes of Aedes genus were captured, identified and grouped into pools of 10 mosquitoes. FINDINGS: RNA extracts of mosquito pools were tested by a RT-Hemi-Nested-PCR for detection of flaviviruses. One amplicon of 222 bp, compatible with dengue virus serotype 4, was obtained from a pool of Aedes aegypti. The nucleotide sequence of the amplicon indicated that the mosquitoes were infected with DENV-4 of genotype I. This virus of Asian origin has been described in Manaus in 2008 infecting acute febrile illness patients. CONCLUSION: This is the first report of dengue virus serotype 4 genotype I infecting Aedes aegypti in the Americas.

Diversidade molecular de rotavírus do grupo A na cidade de Manaus, Estado do Amazonas, Brasil, 2004 a 2006 / Group A rotavirus molecular diversity in the city of Manaus, Amazonas state, Brazil 2004-2006

Objetivo: descrever a ocorrência de genótipos de rotavírus A em crianças na idade de 1 a 36 meses no município de Manaus, Estado do Amazonas, Brasil. Métodos: estudo descritivo, com resultados de análises laboratoriais de fezes diarréicas de crianças de 1 a 36 meses, atendidas em prontos-socorros infantis de Manaus-AM no período de junho/2004 a dezembro/2006; foram empregados os métodos de eletroforese de ácido ribonucleico em gel de poliacrilamida, transcriptase reversa e reação em cadeia da polimerase. Resultados: constatou-se 25 por cento de positividade para rotavírus A – 30,7 por cento e 44,7 por cento para genótipos G e P, respectivamente –, com predominância dos genótipos G2 (17 por cento) e P[8] (66,7 por cento); das amostras positivas para os genótipos G e P, 38,3 por cento e 11,7 por cento, respectivamente, não apresentaram características para qualquer dos genótipos G e P pesquisados. Conclusão: a elevada proporção de ausência de características dos genótipos G e P pesquisados sugere a circulação de genótipos diferentes desses.

Objective: verify rotavirus A genotype occurrence in children aged 1 to 36 months in Manaus-AM. Methods: descriptive and retrospective study conducted from June/2004 to December/2006, through laboratory analysis of diarrheal stools of children aged 1 to 36 months cared for at a Child Emergency service in anaus/Amazonas. Results: 25 per cent rotavirus A positivity was found using Polyacrylamide Gel Electrophoresis of Ribonucleic Acid. Reverse Transcriptase followed by Polymerase Chain Reaction were applied to positive samples and revealed 30.7 per cent G genotypes and 44.7 per cent P genotypes, with predominance of G2 (17 per cent) and P[8] (66.7 per cent) genotypes. 38.3 per cent and 11.7 per cent of the G and P genotype positive samples, respectively, showed no features of any of the G and P genotypes surveyed. Conclusion: 38.3 per cent and 11.7 per cent G and P genotypes, respectively, showed no characteristics of the genotypes studied, suggesting that genotypes different to these are in circulation.

Molecular epidemiology of hepatitis B virus among the indigenous population of the Curuçá and Itaquaí Rivers, Javari Valley, State of Amazonas, Brazil / Epidemiologia molecular do vírus da hepatite B em população indígena em torno dos Rios Curuçá e Itaquaí, Vale do Javari, Estado do Amazonas, Brasil

INTRODUCTION: Hepatitis B virus (HBV) infection is one of the most serious public health problems in the world. In Brazil, HBV endemicity is heterogeneous, with the highest disease prevalence in the North region. METHODS: A total of 180 samples were analyzed and subjected to polymerase chain reaction (PCR) and semi-nested PCR of the HBV S-gene, with the aim of determining the prevalence of HBV-DNA (deoxyribonucleic acid) in indigenous groups inhabiting the areas near the Curuçá and Itaquaí Rivers in the Javari Valley, State of Amazonas, Brazil. RESULTS: The prevalence of the HBV-DNA S-gene was 51.1% (92/180). The analysis found 18 of 49 (36.7%) samples from the Marubo tribe, 68 of 125 (54.4%) from the Kanamary, and 6 of 6 (100%) from other ethnic groups to be PCR positive. There was no statistically significant difference in gender at 5% (p=0.889). Indigenous people with positive PCR for HBV-DNA had a lower median age (p<0.001) of 23 years. There was no statistical difference found in relation to sources of contamination or clinical aspects with the PCR results, except for fever (p<0.001). The high prevalence of HBV-DNA of 75% (15/20) in pregnant women (p=0.009) demonstrates an association with vertical transmission. CONCLUSIONS: The results confirm the high prevalence of HBV-DNA in the Javari Valley, making it important to devise strategies for control and more effective prevention in combating the spread of HBV.

INTRODUÇÃO: A infecção pelo vírus da hepatite B (VHB) é um dos mais sérios problemas de saúde pública do mundo. No Brasil, a endemicidade do VHB é heterogênea, sendo a doença mais prevalente na região norte do país. MÉTODOS: Neste estudo, foram investigadas 180 amostras de sangue por meio da técnica da reação em cadeia da polimerase (PCR) e PCR semi-nested para o vírus da hepatite B, gene S, com o intuito de determinar a prevalência do DNA (ácido desoxirribonucléico) do vírus da hepatite B em povos de etnias indígenas habitantes dos Rios Curuçá e Itaquaí no Vale do Javari, Estado do Amazonas, Brasil. RESULTADOS: A prevalência encontrada para o DNA-VHB gene S foi de 51,1% (92/180). Entre as amostras positivas 18/49 (36,7%) pertenciam à etnia Marubo, 68/125 (54,4%) à Kanamary e 6/6 (100%) a outras etnias. Não houve diferença significante ao nível de 5% em relação ao gênero (p=0,889). Os indígenas com PCR positiva para DNA-VHB apresentaram mediana de idade menor de 23 anos (p<0,001). Não foi constatado nenhuma diferença estatística em relação às fontes de contágio e o resultado da PCR, como também aos aspectos clínicos, com exceção da febre (p<0,001). A alta prevalência do DNA-VHB de 75% (15/20) em gestantes (p=0,009) demonstra associação com a transmissão vertical. CONCLUSÕES: Os resultados comprovam a alta prevalência do DNA-VHB no Vale do Javari, tornando-se importante traçar estratégias de controle e prevenção mais eficazes no combate à disseminação do VHB.

[Dengue virus serotypes in children of Manaus, State of Amazonas, 2008]. / Sorotipos virais de dengue identificados em crianças de Manaus, Estado do Amazonas, 2008.

INTRODUCTION: Dengue is an arbovirus that continues to cause serious public health problems in tropical and sub-tropical regions of the planet. METHODS: In this study, blood samples from children were investigated by RT-PCR, aimed at identifying dengue virus serotypes in Manaus, AM, in 2008 in this infant population. RESULTS: DENV-3 was the only serotype identified. CONCLUSIONS: In this study, 83% of children examined were negative for dengue by RT-PCR, suggesting the occurrence of other febrile illnesses that need to be determined.

Sorotipos virais de dengue identificados em crianças de Manaus, Estado do Amazonas, 2008 / Dengue virus serotypes in children of Manaus, State of Amazonas, 2008

INTRODUCÃO: A dengue é uma arbovirose que vem causando sérios problemas de saúde pública, em regiões tropicais e subtropicais do planeta. MÉTODOS: Neste estudo, foram investigadas amostras de sangue de crianças, através da RT-PCR, com o intuito de se identificar sorotipos do vírus dengue nessa população infantil, em Manaus/AM, durante o ano de 2008. RESULTADOS: O DENV-3 foi o único sorotipo viral identificado. CONCLUSÕES: No presente estudo, 83 por cento das crianças analisadas apresentaram resultado negativo para dengue através do RT-PCR sugerindo a ocorrência de outras doenças febris que necessitam ser esclarecidas.

INTRODUCTION: Dengue is an arbovirus that continues to cause serious public health problems in tropical and sub-tropical regions of the planet. METHODS: In this study, blood samples from children were investigated by RT-PCR, aimed at identifying dengue virus serotypes in Manaus, AM, in 2008 in this infant population. RESULTS: DENV-3 was the only serotype identified. CONCLUSIONS: In this study, 83 percent of children examined were negative for dengue by RT-PCR, suggesting the occurrence of other febrile illnesses that need to be determined.

Detecção de rotavírus em um cão doméstico na cidade de Manaus-AM / Rotavirus detection in a domestic dog in the City of Manaus-AM

Os rotavírus do grupo A, são frequentemente associados com gastroenterites em mamíferos e aves. O objetivo deste trabalho foi detectar a presença de rotavírus em fezes de cães sintomáticos e assintomáticos para diarréia aguda. Foram coletadas 32 amostras de fezes de cães. Todas as amostras foram submetidas à extração do RNA viral seguida da Eletroforese em Gel de Poliacrilamida (EGPA), onde se identificou apenas 1 (um) caso de infecção por rotavírus, em amostra assintomática. A análise do eletroferótipo mostrou um perfil 4:2:3:2 longo, e a homologia dos eletroferótipos de rotavírus humano e canino foi muito alta, sugerindo uma possível infecção interespécie.

The group A rotaviruses are frequently associated with gastroenteritis in mammals and birds. The objective of this study was to detect the presence of rotavirus in feces of symptomatic and asymptomatic dogs for acute diarrhea. 32 samples of dog feces were collected. All the samples were submitted to viral RNA extraction followed by electrophoresis in poliacrylamide gels (PAGE), where only one case of rotavirus infection in one asymptomatic sample was found. The electrophoretic analysis showed 4:2:3:2 long profile, and the homology between human and dog rotaviruses was very high. This suggests a possible interspecies infection.

Detecção e tipagem de vírus dengue em Aedes aegypti (Diptera: Culicidae) na Cidade de Manaus, Estado do Amazonas / Detection and typing of dengue viruses in Aedes aegypti (Diptera: Culicidae) in the City of Manaus, State of Amazonas

O estudo teve por objetivo a detecção e tipagem do vírus dengue, nos vetores Aedes aegypti. Durante o período de dezembro de 2005 a dezembro de 2006, foram coletados 8.984 mosquitos, em 46 bairros da Cidade de Manaus abrangendo todas as zonas geográficas da cidade. Destes, 819 eram Aedes aegypti (414 fêmeas e 405 machos). As fêmeas de Aedes aegypti foram agrupadas em pools de 1 a 10 mosquitos totalizando 138 pools, sendo que 111 pools foram positivos para DENV 3. Porém, um pool mostrou-se positivo para dois sorotipos, DENV 1 e DENV 3. A prevalência de Aedes aegypti infectados com DENV 3, na Cidade de Manaus foi de 53 por cento. Entretanto, a prevalência por zona foi de 70 por cento no Centro-oeste, 60 por cento no Sul, 53 por cento no Oeste, 47 por cento no Centro-Sul, 30 por cento no Norte e 23 por cento na zona Leste. O monitoramento da circulação viral em mosquitos com o uso da técnica da transcrição reversa-reação da polimerase em cadeia que permite o conhecimento prévio dos níveis de disseminação viral em determinadas áreas contribuindo para determinar os locais para aplicar as medidas de prevenção e controle.

The aim of this study was to detect and type dengue viruses in the vector Aedes aegypti. Between December 2005 and December 2006, 8,984 mosquitoes were collected in 46 districts of the city of Manaus, covering all of the geographical zones of the city. Of these, 819 were Aedes aegypti (414 females and 405 males). The females of Aedes aegypti were grouped in pools of 1 to 10 mosquitoes, thus totaling 138 pools, of which 111 pools were positive for DENV 3 and a single pool was positive for two serotypes (DENV 1 and DENV 3). The prevalence of Aedes aegypti infected with DENV 3 in the city of Manaus was 53 percent. The zonal prevalence was 70 percent in the western central zone, 60 percent in the southern zone, 53 percent in the western zone, 47 percent in the southern central zone, 30 percent in the northern zone and 23 percent in the eastern zone. Monitoring of virus circulation among mosquitoes by means of the reverse transcription polymerase chain reaction technique enables prior knowledge of the levels of virus spread in given areas, thus contributing towards determining the localities where prevention and control measures should be applied.

Duplex-PCR assay for the detection of adenovirus and respiratory syncytial virus in nasopharyngeal samples.

Human adenovirus (HAdV) and human respiratory syncytial virus (HRSV) are important etiologic agents of acute respiratory infections. In this study, a duplex polymerase chain reaction (PCR) assay was developed for the simultaneous detection of HAdV and HRSV in clinical samples. Sixty previously screened nasopharyngeal aspirates were used: 20 HAdV-positive, 20 HRSV-positive and 20 double-negative controls. Eight samples were positive for both viruses. The duplex PCR assay proved to be as sensitive and specific as single-target assays and also detected the mixed infections with certainty. The identification of both viruses in a single reaction offers a reduction in both cost and laboratory diagnostic time.

Duplex-PCR assay for the detection of adenovirus and respiratory syncytial virus in nasopharyngeal samples

Human adenovirus (HAdV) and human respiratory syncytial virus (HRSV) are important etiologic agents of acute respiratory infections. In this study, a duplex polymerase chain reaction (PCR) assay was developed for the simultaneous detection of HAdV and HRSV in clinical samples. Sixty previously screened nasopharyngeal aspirates were used: 20 HAdV-positive, 20 HRSV-positive and 20 double-negative controls. Eight samples were positive for both viruses. The duplex PCR assay proved to be as sensitive and specific as single-target assays and also detected the mixed infections with certainty. The identification of both viruses in a single reaction offers a reduction in both cost and laboratory diagnostic time.

[Detection and typing of dengue viruses in Aedes aegypti (Diptera: Culicidae) in the City of Manaus, State of Amazonas]. / Detecção e tipagem de vírus dengue em Aedes aegypti (Diptera: Culicidae) na Cidade de Manaus, Estado do Amazonas.

The aim of this study was to detect and type dengue viruses in the vector Aedes aegypti. Between December 2005 and December 2006, 8,984 mosquitoes were collected in 46 districts of the city of Manaus, covering all of the geographical zones of the city. Of these, 819 were Aedes aegypti (414 females and 405 males). The females of Aedes aegypti were grouped in pools of 1 to 10 mosquitoes, thus totaling 138 pools, of which 111 pools were positive for DENV 3 and a single pool was positive for two serotypes (DENV 1 and DENV 3). The prevalence of Aedes aegypti infected with DENV 3 in the city of Manaus was 53%. The zonal prevalence was 70% in the western central zone, 60% in the southern zone, 53% in the western zone, 47% in the southern central zone, 30% in the northern zone and 23% in the eastern zone. Monitoring of virus circulation among mosquitoes by means of the reverse transcription polymerase chain reaction technique enables prior knowledge of the levels of virus spread in given areas, thus contributing towards determining the localities where prevention and control measures should be applied.

Ocorrência de Salmonella spp. em carcaças de frangos recém abatidos em feiras e mercados da cidade de Manaus-AM / Occurrence of Salmonella spp. in chicken carcasses commercialized in open markets in the city of Manaus - Amazonas

A presente pesquisa avaliou a contaminação em carcaças de frango por salmonelas. O estudo foi realizado em feiras e mercados nas diferentes zonas da Cidade de Manaus-AM, por um período de 11 semanas (março a maio de 1998). Sessenta amostras foram examinadas; destas, trinta (50 por cento) resultaram positivas para Salmonella spp., das quais foram isoladas 67 cepas, incluídas em 11 sorotipos diversos. Entre os sorotipos identificados, a S. panama, S. mbandaka, S. schwarzengrund, S. typhimurium, S. albany e S. agona, foram as predominantes, representando 85 por cento do total isolado. A contaminação de Salmonella é dependente de vários fatores, e a sua ocorrência pode estar relacionada com as condições de higiene da granja. A metodologia empregada para a detecção microbiológica foi a recomendada pela "Food and Drug Administration", dos Estados Unidos da América.

This research evaluates the levels of salmonella that are present in chicken carcasses commercialized in shops and public markets in different zones in the city of Manaus, Amazonas Brazil. Sixty samples were randomly collected in a period of eleven weeks and examined specifically for Salmonella presence. Fifty percent of the samples were positive for Salmonella, and 67 serovars were isolated and distributed mainly among 11 different sorotypes. Those identified,S. panama, S. mbandaka, S. schwarzengrund, S. typhimurium, S. albany, S. agona, corresponded to 85 percent. The contamination of Salmonella is dependent on various factors, and its occurrence may berelated to the hygeine conditions of the poultry farm. The methodology used for the microbiological detection was that recommende by the "Food and Drugs Administration" of the United States of America.

Ocorrência de Listeria monocytogenes em queijo artesanal tipo coalho comercializado na cidade de Manaus-AM, Brasil / Occurrence of Listeria monocytogenes in handmade type cheese coagulum marketed in the city of Manaus-Amazonas, Brazil

A ocorrência de Listeria sp. em queijo artesanal tipo coalho comercializado na cidade de Manaus foi estudada. No período de março a maio de 1998 foram coletadas 58 amostras do produto em feiras e mercados localizados em seis zonas da cidade de Manaus. Em duas amostras de queijo foram identificadas duas espécies de Listeria: L. monocytogenes e L. innocua. A primeira espécie é reconhecidamente patogênica para o ser humano. Os resultados apresentados demonstraram haver risco de transmissão de enfermidades pelo consumo de queijos processados sem os devidos cuidados de higiene.

The occurrence of Listeria sp. in cheese craft type coagulum marketed in the city of Manaus was studied. In the period of March to May of 1998 58 samples of the product were collected at fairs and located markets in six areas of the city. In two cheese samples we identified two species of Listeria: L. monocytogenes and L. innocua. The first species was found fobe pathogenic for the human being. The results demonstrated there to be risk of transmission of illnesses from the consumption of cheeses processed without the due hygiene care.