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.

Detection of Toxoplasma gondii antibodies in captive non-human primates in the Amazon region, Brazil.

BACKGROUND: Toxoplasma gondii infections in captive non-human primates are of interest because often they die due to severe toxoplasmosis. Thus, we aimed to evaluate samples from a serum bank to T. gondii antibodies. METHODS: Serum samples of 179 monkeys from the National Center of Primates, Brazil, were retrospective analyzed for T. gondii IgG antibodies by modified agglutination test using formalin-fixed whole parasites as antigen (cutoff 1:25). RESULTS: Among the 179 samples tested, 88 (49.2%) were positive. Twelve of the 18 evaluated species presented at least one positive animal. A higher occurrence of positive animals was found in New World (52.2%) than Old World (22.2%) monkeys (P = .023) and in the animals from domestic donation origin, which had lived in human homes as pets (P = .004). CONCLUSIONS: We confirm the widespread presence of T. gondii in captive monkeys and contribute to the range of species that can be infected by this parasite.

Association of killer cell immunoglobulin-like receptor polymorphisms with chronic hepatitis C and responses to therapy in Brazil.

Soroprevalence for Hepatitis C virus is reported as 2.12% in Northern Brazil, with about 50% of the patients exhibiting a sustained virological response (SVR). Aiming to associate polymorphisms in Killer Cell Immunoglobulin-like Receptors (KIR) with chronic hepatitis C and therapy responses we investigated 125 chronic patients and 345 controls. Additionally, 48 ancestry markers were genotyped to control for population stratification. The frequency of the KIR2DL2 and KIR2DL2+HLA-C(Asp80) gene and ligand was higher in chronic infected patients than in controls (p < 0.0009, OR = 3.4; p = 0.001, OR = 3.45). In fact, KIR2DL3 is a weaker inhibitor of NK activity than KIR2DL2, which could explain the association of KIR2DL2 with chronic infection. Moreover, KIR2DS2 and KIR2DS2+HLA-C(Asp80) (p < 0.0001, OR = 2.51; p = 0.0084, OR = 2.62) and KIR2DS3 (p < 0.0001; OR = 2.57) were associated with chronic infection, independently from KIR2DL2. No differences in ancestry composition were observed between control and patients, even with respect to therapy response groups. The allelic profile KIR2DL2/KIR2DS2/KIR2DS3 was associated with the chronic hepatitis C (p < 0.0001; OR = 3). Furthermore, the patients also showed a higher mean number of activating genes and a lower frequency of the homozygous AA profile, which is likely secondary to the association with non-AA and/or activating genes. In addition, the KIR2DS5 allele was associated with SVR (p = 0.0261; OR = 0.184).The ancestry analysis of samples ruled out any effects of population substructuring and did not evidence interethnic differences in therapy response, as suggested in previous studies.

Serological and molecular retrospective analysis of hepatitis E suspected cases from the Eastern Brazilian Amazon 1993-2014.

INTRODUCTION: We evaluated the anti-hepatitis E virus (HEV) antibody prevalence and HEV-RNA in archived serum samples of non-A-C hepatitis, or suspected cases of HEV infection from the Eastern Brazilian Amazon from 1993 to 2014. METHODS: Serum samples (n = 318) were tested using ELISA and immunoblotting, and screened for HEV-RNA by RT-qPCR. RESULTS: Anti-HEV IgM and IgG were detected in 3.4% (11/318) and 5.9% (19/318) of the samples, respectively. All samples were HEV-RNA negative. CONCLUSIONS: HEV was detected at a low prevalence. Broader serological and molecular evaluation of HEV infection in the Amazon region should be carried out.

A paruterinid metacestode in the liver of a Neotropical bat (Molossus molossus).

We describe a case of microscopic liver lesion caused by larval cestode in a frugivorous bat (Molossus molossus) from the Western Brazilian Amazon. Histopathological analysis of liver indicated the occurrence of metacestode associated with multifocal histiocytic response and the phylogenetic analysis of Cox-I and 18S rDNA genes indicated that the parasite belonged to the family Paruterinidae. This is the first identification of cestodes of the family Paruterinidae in bats and may suggest a broad range of paruterinid tapeworm hosts to be investigated in the Brazilian Amazon.

Hepatitis E virus infection among rural Afro-descendant communities from the eastern Brazilian Amazon.

INTRODUCTION: The prevalence, public health impact, and epidemiological characteristics of hepatitis E virus (HEV) are poorly understood in Brazil. METHODS: Serum samples from 535 individuals from three rural Afro-descendant communities located in eastern Brazilian Amazon were collected in October 2015 and tested for presence of anti-HEV IgM and IgG antibodies. Serologically positive samples were also tested for HEV-RNA. RESULTS: Two cases were confirmed for anti-HEV IgM (0.3 %) and two cases for anti-HEV IgG (0.3 %). No sample was positive for HEV-RNA. CONCLUSIONS: Results indicated low prevalence of HEV infection in Afro-descendant rural communities from the eastern Brazilian Amazon.

[Prevalence of hepatitis B and D serological markers in the Parakanã, Apyterewa Indian Reservation, Pará State, Brazil]. / Prevalência dos marcadores sorológicos dos vírus das hepatites B e D na área indígena Apyterewa, do grupo Parakanã, Pará, Brasil.

In order to study the prevalence of hepatitis B (HBV) and D (HDV) viruses in the Parakanã Indians and to evaluate the impact of hepatitis B vaccination beginning there in 1995, 258 serum samples were analyzed in the year 2004 for hepatitis B and D serological markers using immunoenzymatic techniques; the results showed a moderate endemic pattern, with a total prevalence of HBV infection of 55.7% and 5.4% of virus carriers in the Apyterewa village and 49.5% with 1.1% of HBV carriers in the Xingu village; 31.4% of anti-HBs+ as an isolated marker in both villages and no detection of positive serological tests for HDV among HBV carriers. The laboratory analysis thus showed the presence of chronic HBV carriers, absence of HDV carriers, and an emerging vaccine profile among susceptibles, confirming the effectiveness and need to maintain vaccination, especially in the first year of life, and the need to implement effective epidemiological surveillance for early detection of HDV infection among HBV carriers.

SEROLOGICAL DETECTION OF HEPATITIS A VIRUS IN FREE-RANGING NEOTROPICAL PRIMATES (Sapajus spp., Alouatta caraya) FROM THE PARANÁ RIVER BASIN, BRAZIL.

Nonhuman primates are considered as the natural hosts of Hepatitis A virus (HAV), as well as other pathogens, and can serve as natural sentinels to investigate epizootics and endemic diseases that are of public health importance. During this study, blood samples were collected from 112 Neotropical primates (NTPs) (Sapajus nigritus and S. cay, n = 75; Alouatta caraya, n = 37) trap-captured at the Paraná River basin, Brazil, located between the States of Paraná and Mato Grosso do Sul. Anti-HAV IgG antibodies were detected in 4.5% (5/112) of NTPs, specifically in 6.7% (5/75) of Sapajus spp. and 0% (0/37) of A. caraya. In addition, all samples were negative for the presence of IgM anti-HAV antibodies. These results suggest that free-ranging NTPs were exposed to HAV within the geographical regions evaluated.

Contágio intradomiciliar e status vacinal entre comunicantes de portatores do vírus da hepatite B / House hold contagion and vaccination status among communicants of hepatitis B virus carriers

Objetivo: Avaliar a cobertura da vacina contra hepatite B e a frequência da infecção em comunicantes de portadores do Vírus da Hepatite B vinte anos após a implementação da vacina na cidade de Belém, Pará, Brasil. Método: Estudo descritivo quantitativo, desenvolvido entre 2016 e 2018. Foram realizadas visitas domiciliares aos portadores do vírus, notificados por meio da busca de comunicantes, com orientações sobre a pesquisa, preenchimento e assinatura de formulários, coleta de amostras para testes sorológicos de hepatite B (HBsAg, anti-HBc total, anti-HBs) por técnica imunoenzimática. Resultados: Na população analisada 53,5% são do sexo masculino, com média de idade de 9,4 anos (entre 4 meses a 19 anos com mediana de 10 anos). Não foram detectados portadores de HBsAg reagente; anti-HBc isolado total foi detectado em 2,1%; 4,2% eram anti-HBc total / anti-HBs reagente e 42% anti-HBs isolado, atribuídos à vacinação; 41,7% receberam esquema vacinal completo e 58,3% eram suscetíveis. Conclusões: Os resultados encontrados podem ser utilizados como norteadores de ações estratégicas visando ampliar a cobertura vacinal e reduzir a prevalência de infectados na população de comunicantes intradomiciliares. (AU)

Objective: To evaluate the coverage of the hepatitis B vaccine and the frequency of infection in communicants of Hepatitis B virus carriers twenty years after the vaccine was implemented in the city of Belém, Pará, Brazil. Method: Quantitative descriptive study developed between 2016 and 2018. Home visits were carried out to the carriers of the virus notified through the search for communicants, with guiding on research, filling and signing forms, collecting samples for serological tests for Hepatitis B (HBsAg, total anti - HBc, anti - HBs) by immunoenzymatic technique. Results: In the population analyzed 53.5% are male, with a mean age of 9.4 years (ranging from 4 months to 19 years and median of 10 years). Carriers HBsAg reagent were not detected; total anti - HBc isolated was detected in 2.1%; 4.2% were total anti - HBc / anti - HBs reagent and 42% anti - HBs alone, attributed to vaccination; 41.7% received a complete vaccination schedule and 58.3% were susceptible. Conclusions:The results found can be used as guidelines for strategic actions aimed at expanding vaccine coverage and reducing the prevalence of infected people in the population of household communicators. (AU)

Objetivo: Evaluarla cobertura de la vacuna contra lahepatitis B y lafrecuencia de infecciónen portadores delVirus de laHepatitis Bveinteañosdespués de laimplementación de lavacuna enlaciudad de Belém, Pará, Brasil. Método: Estudiodescriptivocuantitativo realizado entre 2016 y 2018. Fueron realizadas visitas domiciliarias a los portadores del vírus, notificados a través de labúsqueda de domiciliares, conorientaciones sobre lainvestigación, registro y asignatura de formularios, toma de muestras de sangre para pruebas serológicas de Hepatitis B (HBsAg, anti-HBc total, anti-HBs) por la técnica inmunoenzimática. Resultados: Enlapoblaciónanalizada 53,5% del sexo masculino, conedadpromedio de 9,4 años (entre 4 meses a 19 añoscon mediana de 10 años) no fueron detectados portadores de HBsAgreactivo; el total de anti-HBcfué detectado en 2,1%; 4,2% anti-HBc total/anti-HBs reactivo, y, 42% anti-HBs aislado, atribuido a lavacunación; 41,7% recibieronel esquema completo de vacunación y 58,3% fueronsusceptibles. Conclusiones: Los resultados encontrados puedenusarse como pautas para acciones estratégicas dirigidas a ampliar la cobertura de vacunación y reducirlaprevalencia de personas infectadas enlapoblación de comunicadores domésticos. (AU)

First report of equine Pegivirus in South America, Brazil.

The human Pegivirus (HPgV, also known as GBV-C virus or hepatitis G virus) is a lymphotropic RNA-virus phylogenetically related to the Hepatitis C virus, which infects approximately 5% of the world's human population. Recently, two novel, presumably hepatotropic, pegiviruses, designated as equine Pegivirus (EPgV) and Theiler's Disease Associated Virus (TDAV), were discovered in horses with clinical and laboratory evidence of hepatic disease. To verify the occurrence of pegiviruses infection in horses from Pará State, northern Brazil, serum samples from 114 horses located in four cities (Acará, Belém, Dom Eliseu and Ananindeua) were submitted for the molecular analysis of EPgV by nested RT-PCR. The results of nucleotide sequencing and phylogenetic analysis of EPgV NS3 and NS5B genomic regions confirmed one positive sample among 114 tested samples (1/114; 0.8%). No evidence of TDAV infection was found, but despite the low prevalence and unknown clinical significance among the studied population, these results represent the first molecular detection of EPgV in horses in South America.

Would Sacaca, Croton cajucara Benth (Euphorbiaceae) be an hepatotoxic plant like Germander, Teucrium chamaedrys L. (Labiatae)?

Clinical and experimental studies have consistently incriminated the medicinal plant germander (Teucrium chamaedrys L.) in epidemic and sporadic cases of liver diseases. The sacaca (Croton cajucara Benth), a common plant in Brazilian Amazon region also comes being incriminated in similar clinical cases. Of both plants were isolated diterpenoid compounds with similar chemical structures.

[The evolution of knowledge about viral hepatitis in Amazon region: from epidemiology and etiology to the prophilaxy]. / A evolução do conhecimento sobre as hepatites virais na região amazônica: da epidemiologia e etiologia à prevenção.

Since the 1950 years a disease similar to yellow fever but thought to be a new disease with unknown etiology has been described to health and researcher authorities. This disease occurs in Jurua, Purus and Madeira Rivers valleys. It is feared by local people by its high lethality. It is clinically a hepato-encephalopathy (Average survival time of 5-6 days) About 90% of sick people with typical symptoms go to death. The disease is popularly known as black fever of Lábrea and by pathologist as Lábrea hepatites after the city where the first cases were observed. The specific histopatologic picture of vesicular degeneration of hepatocytes like spider cells motivate the local pathologist to think as a new disease: Lábrea hepatitis. The finding of HBsAg and marker of hepatites D virus (HDV) in the serum motivate the researchers to think the disease as a superinfection of HDV in chronic carriers of HBV. In absence of a specific vaccine against HDV, the vaccine against HBV, must be given soon after the birth is the recommended prevention.

[Hepatotoxicity of the microcystin cyanotoxin]. / Hepatotoxicidade da cianotoxina microcistina.

At public health, there is increasingly interest on evaluating the possibility of human intoxication by biotoxins from blue-green algae, mainly the hepatotoxins from the microcystin group. Microcystin, a monocyclic heptapeptide, is mainly produced by a cyanobacteria called Microcistis aeruginosa. It is characterized by a few variable amino acids, from which two of them have an unusual structure and play an important role in the hepatotoxicity of the microcystin. Although human illnesses include gastroenteritis, allergic or irritative reactions, and neurotoxicity, the main target of this toxin is the liver. Inside the hepatocytes, microcystins are carried by the transportation system of the bile acid, inhibiting the activity of the protein phosphatase in the cytoplasm. This inhibition causes a morphologic change in the plasmatic membrane because of the hyperphosphorylation of cytokeratins, and also the tumoral promotion by the hyperphosphorylated proteins. The techniques used in the detection and quantification of the microcystins in the environment include liquid chromatography, bioanalysis of mice, and immunoenzymatic tests using mono and polyclonal antibodies against those toxins. The latter has been remarked because of its practicality and its high sensibility.

[Hepatitis A virus infection in Amerindian area in the east Brazilian Amazon]. / Infecção pelo vírus da hepatite A em área indigena da Amazônia oriental brasileira.

The hepatitis A virus infection represents an important problem of public health all over the world, being related to the socioeconomic and hygienic conditions of the population. In Brazilian Amazon, seroepidemiological studies in amerindians populations have been demonstrating high endemicity related to the infection. With the objective of evaluate the prevalence of the hepatitis virus A infection in xicrin village, in the municipality district of Altamira-Pará-Brazil, whose investigation was unchained by indigenous child's obit, that clinical developed in nine days with a picture icterus-hemorrhagic, without confirmation by serologic exams, 352 samples of blood were analyzed by serologic tests of the markers of the hepatitis A, B, C and D virus, for immunoenzymatic technic, that indicated a prevalence of 98% of antibodies against the hepatitis A virus, which 30.5% with recent infection, characterizing in laboratorial basis, the outbreak of infection for the virus of the hepatitis A and raising the possibility to be associated with the obit happened in the village.

[The Liver Diseases Program of the Hospital Santa Casa de Misericórdia do Pará--infrastructure and clinical epidemiology in 10 years of attendance]. / Programa de Hepatopatias do Hospital da Fundação Santa Casa de Misericórdia do Pará--infra-estrutura e epidemiologia clinica em 10 anos de atendimento.

The Liver Diseases Program of the Hospital Santa Casa de Misericordia do Pará was create because of the need to attend patients with liver diseases of the Amazônia area, taking as priority to attend with quality, diagnosis of aetiologies, clinical following and specific treatment. This study aim to describe dates related to epidemiology, aetiologics agents and histopathologic analysis. One thousand sixty nine patients were evaluated through medical, laboratory, endoscopic, ultrasound or computerized tomography and histopathologic examination. Nine hundred thirty five (63.6%) patients within 1469 patients were diagnose as chronic liver disease. The average age was 50 year, 666 (71.2%) were male, and the most patients lived in Belem, State of Pará. The aetiologic agents most prevail were alcoholism (53.7%) and viral hepatitis (39.1%). Hepatic biopsy were done in 403 (43.1%) within the 935 patients and the results showed chronic hepatitis (34%) and chirrosis (34%). In summary the chronic liver disease in the amazon region is more prevail in male than female, the alcoholism is the principal aetiologie, and the most of these cases were diagnose in the severe phase.

[Polycystic echinococcosis in the Eastern Brazilian Amazon: an update]. / Equinococose policística na Amazônia oriental brasileira: atualização da casuística.

By means of epidemiological and clinical-laboratorial approaches was consolidated an update of polycystic echinococcosis in the Eastern Brazilian Amazon, period from 1962 to 2003, including unpublished cases and those already published. In that way, they were identified 40 cases of the disease in referred period, understanding cases coming from the States of Pará and Amapá, Brazil. The width of the ages went from 10 to 72 years and 47.5% belonged to the masculine sex. The liver was the attacked organ (82.5% of the cases). The Echinococcus vogeli (Rausch and Bernstein, 1972), comes as the main agent involved. Starting from the recognition of the importance and of the implications of the handling of the echinococcosis for the tropical area, it is believed that should happen an improvement of the diagnosis, appropriate treatment and of a better registration of the disease.

[Prevalence of hepatitis B virus genotypes and the occurrence of precore mutation A-1896 and to correlate them with the clinical presentation of chronic hepatitis, in a population group of the Eastern Amazon region]. / Prevalência de genótipos e de mutantes pré-core A-1896 do vírus da hepatite B e suas implicações na hepatite crônica, em uma população da Amazônia oriental.

Hepatitis B virus (HBV) infection presents itself with a variety of clinical manifestations. The present work aims to describe the prevalence of HBV genotypes and the occurrence of precore mutation A-1896 in a population group of the Eastern Amazon region of Brazil and to correlate them with the clinical presentation of chronic HBV infection. 51 HBsAg carriers (HBV-DNA positive) were selected and divided into three groups: A (14 asymptomatic subjects), B (20 HBeAg positive symptomatic patients) and C (17 HBeAg negative symptomatic patients). Using an automa ed DNA sequencer ABI model 377 by sequencing for determined of genotypes and precore mutation. The results showed that the genotype A was the most commonly found (81.1%, 89.5% and 93.7% in groups A, B and C, respectively) and precore mutation A-1896 was described in 11.5% (3/26) of group A subjects. Genotype A of HBV was the most prevalent (89.1%) and low occurrence of precore mutation A-1896, both not associate with the worst outcome of the chronic infection of HBV.

[Hepatitis B and C virus infection and the hepatocellular carcinoma in the East Amazon, Brazil]. / Infecções pelos vírus das hepatites B e C e o carcinoma hepatocelular na Amazônia oriental.

In order to contribute to a better understanding of the possible role of hepatits B and C in the etiopathogenis of HCC in the East Amazon, there were studied 36 patients in Belém/PA. Serological hepatitis markers were evaluated and polymerase chain reaction assays were used to detect HBV-DNA and HCV-RNA. Alcohol abuse was observed in 33.3% and cirrhosis in 83.3%. In 88.9% of the sample, one or more hepatitis B markers were positive. Also, 8.3% those patients had anti-HCV simultaneously positive. The HBsAg serological test was positive in 58.3%; anti-HBc in 86%; anti-HBe in 85.7%; anti-HBe in 9.5%; IgM anti-HBc in 57.1%. The HBV DNA was found in 37.7% and in 65% of the HBsAg positive. The HCV RNA was detected in 8.5% and in 100% of the patients positive to anti-HCV. The AFP was above the normal value in 88.9% of patients, with levels up to 400ng/ml in 75% of them. In conclusion, hepatitis B virus infection seems to be important in the etiology of HCC and improving measures such immunization and screening in the risk population should be emphasyzed.

Serological and molecular retrospective analysis of hepatitis E suspected cases from the Eastern Brazilian Amazon 1993-2014

Abstract INTRODUCTION We evaluated the anti-hepatitis E virus (HEV) antibody prevalence and HEV-RNA in archived serum samples of non-A-C hepatitis, or suspected cases of HEV infection from the Eastern Brazilian Amazon from 1993 to 2014. METHODS Serum samples (n = 318) were tested using ELISA and immunoblotting, and screened for HEV-RNA by RT-qPCR. RESULTS Anti-HEV IgM and IgG were detected in 3.4% (11/318) and 5.9% (19/318) of the samples, respectively. All samples were HEV-RNA negative. CONCLUSIONS HEV was detected at a low prevalence. Broader serological and molecular evaluation of HEV infection in the Amazon region should be carried out.

Anatomo-clinical and molecular description of liver neotropical echinococcosis caused by Echinococcus oligarthrus in human host.

Since humans rarely play the role as Echinococcus oligarthrus host, there is lack of knowledge about the complex infectious process. Only three cases have been reported to occur in humans in the neotropics until now. We present the anatomo-clinical and molecular findings describing a new case of infection by E. oligarthrus in a man. The muscular or subcutaneous tissues tropism described for this species in the previously reported cases was not present, but a liver tropism was observed. Additionally, the larval stage rostellar hooks morphometry differed from E. oligarthrus in the other human cases.