Orthohepadnavirus infection in a neotropical bat (Platyrrhinus lineatus).

Hepatitis B virus (HBV) is the prototype of the Orthohepadnavirus genus and represents an important cause of chronic hepatitis, liver cirrhosis, and hepatic cancer in humans worldwide. To verify the occurrence and genetic variability of orthohepadnavirus among neotropical bats, we tested 81 liver samples of New World bats from São Paulo State, Southeastern Brazil, collected during 2012. PCR, sequencing, and phylogenetic analysis of Surface/Polymerase and Core viral genes confirmed the occurrence of the first isolate of bat orthohepadnavirus detected in South America. These results may contribute to subsequent studies of the origin, variability, host species, and evolution of bat orthohepadnaviruses in South America.

A Retrospective Survey of Rodent-borne Viruses in Rural Populations of Brazilian Amazon.

INTRODUCTION: The Amazon tropical rainforest has the most dense and diverse ecosystem worldwide. A few studies have addressed rodent-borne diseases as potential hazards to humans in this region. METHODS: A retrospective survey was conducted using enzyme-linked immunosorbent assay for detecting mammarenavirus and orthohantavirus antibodies in 206 samples collected from rural settlers of the Brazilian Western Amazonian region. RESULTS: Six (2.91%) individuals in the age group of 16 to 36 years were found to possess antibodies against mammarenavirus. CONCLUSION: Evidence of previous exposure to mammarenavirus in the rural population points to its silent circulation in this region.

Orthohantavirus pulmonary syndrome in Santa Cruz and Tarija, Bolivia, 2018.

INTRODUCTION: Orthohantaviruses are still a significant public health threat in endemic countries, with high case fatality rates (CFR). In Bolivia, the reporting of small outbreaks occurred until 2012. The findings of 40 laboratory-confirmed cases diagnosed in two departments are reported herein. METHODS: This was an observational, retrospective and cross-sectional study. Data on laboratory-confirmed cases in 2018 were collected from the hospitals and departmental health services (SEDES) of Santa Cruz and Tarija. An ELISA was used for the detection of IgM antibody to hantavirus in the patient blood samples. RESULTS: Forty patients were IgM-positive. The median age of the patients was 24 years (interquartile range 19-41 years) and 72.5% were male. All patients were hospitalized; 57.5% were admitted to the intensive care unit and had cardiopulmonary compromise, with 83% of these presenting acute respiratory distress syndrome and 89.5% of these requiring mechanical ventilation. Six patients died (CFR 15%). Patients <15 or >60 years old were more prone to die (odds ratio 10.33, 95% confidence interval 1.411-75.694), as were those with comorbidities (odds ratio 16.5, 95% confidence interval 1.207-225.540). CONCLUSIONS: Orthohantavirus infections were associated with a high CFR. These cases occurred in areas with eco-epidemiological conditions facilitating viral transmission, including the presence of rodents, as well as the risk of spillover to humans due to social, environmental, and occupational factors.

A Retrospective Survey of Rodent-borne Viruses in Rural Populations of Brazilian Amazon

Abstract INTRODUCTION: The Amazon tropical rainforest has the most dense and diverse ecosystem worldwide. A few studies have addressed rodent-borne diseases as potential hazards to humans in this region. METHODS: A retrospective survey was conducted using enzyme-linked immunosorbent assay for detecting mammarenavirus and orthohantavirus antibodies in 206 samples collected from rural settlers of the Brazilian Western Amazonian region. RESULTS: Six (2.91%) individuals in the age group of 16 to 36 years were found to possess antibodies against mammarenavirus. CONCLUSION: Evidence of previous exposure to mammarenavirus in the rural population points to its silent circulation in this region.

Development of a serosurveillance assay for detection of Necoclí virus exposure.

Hantavirus cardiopulmonary syndrome (HPS) has gained importance in Latin America as an emerging disease, with reports of about 4000 HPS cases; however, this is probably an underestimate because of limited surveillance programs and diagnostic tools to confirm HPS. In order to address this issue and develop better serosurveillance capability, we evaluated three recombinant peptides from the Necoclí virus (NECV) nucleocapsid in antibody-capture ELISA. We cloned and expressed antigens representing the whole NECV nucleocapsid protein (NECV-rN), the immunodominant domain (NECV-rN100), and a serospecific domain (NECV-rN428), and then we compared these antigens in ELISA to detect IgG antibodies to NECV in human sera. We evaluated human sera collected during two epidemiological studies from the area where NECV was discovered. The first group included 609 sera from healthy individuals, and the second one included 89 samples from patients with undifferentiated febrile illness. In these two groups, hantavirus infection had previously been determined by the presence of IgG to Maciel virus (MCLV), a hantavirus closely related to NECV. The number of IgG-positive sera was higher using the Necoclí ELISA with the rN100 protein, which detected antibodies in a higher percentage of healthy individuals, 129/609 (21.2%), as well as in febrile patients, 11/89 (12.3%). In contrast, using MCLV ELISA, 8 of 609 (1.3%) and 4 of 89 (4.5%) samples from healthy and febrile patients, respectively, were seropositive. The agreement between the NECV and MCLV ELISA assays was ≥ 82.3%; however, the kappa indices were weak but statistically significant for rN (0.251 CI; 0.138-0.365) and rN100rN (0.153 CI; 0.084-0.223). The weak kappa indices were attributed to decreased MCLV ELISA assay sensitivity. These results suggest that NECV rN and rN100 have increased specificity and could be further validated for improved diagnosis of hantavirus infections.

Silent Orthohantavirus Circulation Among Humans and Small Mammals from Central Minas Gerais, Brazil.

New World orthohantaviruses are emerging RNA viruses that cause hantavirus cardiopulmonary syndrome (HCPS). These viruses are a burden to public health around the world with a lethality rate of around 60%. In South America, rodents of Sigmodontinae subfamily are the main reservoirs of orthohantaviruses. We described a serosurvey for orthohantaviruses circulation in an apparently healthy human population and small mammals from rural areas in Central Minas Gerais State, Brazil. A total of 240 individuals and 50 small mammals (26 rodents belonging to 10 different species and 24 marsupials from 4 different species) were sampled during 2012-2013. The seroprevalence rates of IgG/IgM antibodies in humans were 7.1 and 1.6%, respectively. Only one rodent, an Oligoryzomys nigripes captured in peridomestic area, tested positive for IgG antibodies and viral RNA. Our findings suggest a silent circulation of orthohantaviruses in a region of intensive agriculture production. The detection of seropositive humans in an area with a lack of previous HCPS reports highlights potential oligosymptomatic cases and the need for surveillance strategies that could reduce the risk of future outbreaks.

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.

Padronização da PCR em Tempo Real para o Diagnóstico da Hepatite B / Standardization of real-time PCR for the hepatitis B diagnosis

A hepatite B é uma doença de interesse em saúde pública, acometendo cerca de 2 bilhões de pessoas no mundo, das quais aproximadamente 350 milhões tornam-se portadoras crônicas. Os ensaios quantitativos de detecção do DNA viral são empregados para avaliar a infectividade do vírus e para o monitoramento do tratamento, sendo importante para avaliar a progressão da doença. O objetivo deste estudo foi padronizar a técnica de amplificação quantitativa em tempo real (PCR em tempo real) do DNA do vírus da Hepatite B e comparar a técnica com métodos comerciais (COBAS AmpliPrep/COBAS TaqMan® e COBAS AmpliPrep/COBAS TaqMan®). A amostragem foi constituída por 397 amostras de soro ou plasma recebidas e estocadas no laboratório de hepatites virais do Centro de Virologia do Instituto Adolfo Lutz, no período de 2009 a 2011. Do total de amostras, 345 foram utilizadas para a comparação com os testes comerciais disponíveis e 52 amostras foram utilizadas para o teste de especificidade, sendo 27 amostras HIV positivas e 25 amostras HCV positivas, todas com AgHBS e Anti-HBc total não reagentes. Estas amostras foram inicialmente analisadas por testes comerciais e posteriormente pela técnica padronizada. Para o controle da quantificação, uma curva padrão foi construída em todas as reações, utilizando-se padrões internacionais produzidos pela Organização Mundial de Saúde, em diluições seriadas. Para a extração do DNA das amostras e padrão, foi utilizado kit comercial (QIAGEN®), seguindo os procedimentos descritos pelo fabricante. Os testes foram realizados empregando o método do TaqMan – PCR, em um equipamento ABI 7300 (Applied Biosystems, Foster City, CA). Os “primers” e sondas utilizadas para a padronização foram selecionados a partir da literatura e os que apresentaram melhores resultados nos testes iniciais foram escolhidos para o estudo. A técnica de sequenciamento foi realizada nas amostras com carga viral quantificável...

Hepatitis B is a disease of public health concern. Worldwide, it affects approximately 2 billion people, of whom approximately 350 million become chronic carriers. Quantitative assays for the detection of viral DNA are used to evaluate the infectivity of the virus and to monitor treatment, which is crucial to evaluate the progression of the disease. The aim of this study was to standardize real-time (real-time PCR) quantitative amplification of the DNA of hepatitis B and to compare this technique with commercial methods (COBAS AmpliPrep / COBAS ® TaqMan and COBAS AmpliPrep / COBAS TaqMan ®). The sample consisted of 397 serum or plasma samples received and stored at the laboratory of viral hepatitis from the Center of Virology, Instituto Adolfo Lutz, from 2009 to 2011. Of the total samples, 345 were used for comparison with the available commercial tests and 52 samples were used for the specificity test. Of these, 27 were HIV positive and 25, HCV positive, and all were HBsAg and anti- HBc nonreactive. These samples were initially analyzed by commercial tests and later by the standardized technique. To control the quantification, a standard curve was constructed in all reactions using international standards established by the World Health Organization, in serial dilutions. For the extraction of DNA samples and standard, a commercial kit used was (QIAGEN ®) according to the manufacturer’s instructions. The tests were performed using the TaqMan method - ABI 7300 PCR system (Applied Biosystems, Foster City, CA). Primers and probes used for standardization were selected from the literature and those presenting better results in initial tests were chosen for the study. Sequencing technique was performed on samples with measurable viral load. The standardized real-time PCR technique yielded satisfactory results...

Artículo de revisión: hepatitis viral B y su manejo / Review article: viral hepatitis B and its handling

La Hepatitis B es causada por el virus del mismo nombre (VHB). El paciente presenta ictericia, acolia, coluria, astenia, adinamia, anorexia, distensión abdominal, lienteria, náuseas y vómitos sin fiebre. Blumberg y colaboradores detectaron y caracterizaron al nuevo Virus. Además de hepatitis postranfusional puede causar epidemias por trasnmisión sexual, alimentos y en las zonas endémicas, perinatal. Trabajos en Asia, América y África demostraron relación del VHB con cirrosis macro y micronodular, hepatocarcinoma. Se presenta una parte de nuestra modesta contribución en Colombia. Varias vacunas están en uso clínico y se describen en el artículo. El tratamiento con Inmunomoduladores como interferones y antivirales inespecíficos como rivabirina, isoprinosine, lamivudina, adefovir, entecavir, tenofovir, emtricitabina, clevudina, alamifovir se comenta en el artículo. La respuesta a antivirales depende del subtipo y genotipo del VHB y mutaciones inducidas por el tratamiento. Esto y progresos en métodos diagnósticos en los cinco últimos años renovaron el interés por estudiar las mutaciones naturales y espontáneas del VHB, sus genotipos, subtipos y respuesta inmune que se revisan.

Hepatitis B is caused by the virus with the same name (HBV). The patient has symptoms of jaundice, acolia, dark urine, fatigue, weakness, anorexia, abdominal distension, lienteria, nausea and vomiting without fever. Blumberg and his colleagues identified and characterized the new virus. Besides postranfusional hepatitis, it can cause epidemics by sexual transmission, food, and in endemic areas, perinatal. Studies in Asia, America and Africa showed a relationship between the HBV and macro and micronodular cirrhosis, hepatocellular carcinoma. We present a part of our modest contribution in Colombia. Several vaccines are being used clinically and they are described in the article. In the article, we discuss the treatment with immunomodulators such as interferons and non-specific antiviral drugs such as isoprinosine, lamivudina, adefovir, entecavir, tenofovir, emtricitabina, clevudina, alamifovir. The response to antiviral drugs depends on the HBV subtype, genotype and mutations induced by the treatment. This and advances in diagnostic methods in the last five years renewed the interest in studying the natural and spontaneous mutations of HBV, its genotypes, subtypes, and the immune response that is analyzed here.