Ana Maria Coimbra Gaspar

Identification of a new hepatitis B virus (HBV) genotype from Brazil that expresses HBV surface antigen subtype adw4

The complete genome of a hepatitis B virus (HBV) from Brazil that expressed the subtype adw4 of HBV surface antigen (HBsAg) was cloned and sequenced. The genome, termed w4B, consists of 3215 bp. The overall genetic organization of typical hepadnaviruses with four open reading frames including the preC region was found to be conserved. When comparing the w4B sequence with 19 complete HBV genomes it was, however, found to be more divergent (15%) than any other HBV sequence thus far reported. Until now, no more than 11% divergence has been reported. Distinct from the five known HBV genotypes A to E, w4B made up a new, sixth genotype. The importance of the conserved third start codon in the HBV X gene became apparent in isolate w4B. By mutation, this ATG was out of frame, and by what appears to have been a linked mutation, a new start site two codons downstream was re-established. The significance of several other mutations is discussed.

Serological evidence of hepatitis E virus infection in different animal species from the Southeast of Brazil

Serological evidence of hepatitis E virus infection (HEV) has been observed in both humans and different animal species living in non-endemic areas, suggesting that animals could be important reservoir for virus transmission to man. Antibodies to HEV have been detected in some Brazilian population groups. Nevertheless, sporadic cases of acute HEV infection have never been reported. We collected 271 serum samples from several domestic animals and also from pig handlers from Southeast of Brazil in order to investigate the seroprevalence of HEV infection. Anti-HEV IgG was detected in cows (1.42%), dogs (6.97%), chickens (20%), swines (24.3%), and rodents (50%), as well as in pig handlers (6.3%). The recognition of swine HEV infections in pigs in many countries of the world led us to investigate a larger sample of pigs (n = 357) from the same Brazilian region with ages ranging from 1 to > 25 weeks. IgG anti-HEV was detected in 100% of 7-day old pigs. Following a gradual decline between weeks 2 and 8 (probably due to loss of maternal IgG), the prevalence then steady increased until it reached 97.3% of animals older than 25 weeks. Besides the detection of anti-HEV antibodies in different animal species, the results showed that swine HEV infection seems to be almost universal within this Brazilian pig population. This is the first report that shows evidences of HEV circulation in Brazilian animal species and pig handlers.

Clinical experience with miglustat therapy in pediatric patients with Niemann-Pick disease type C: A case series

Niemann-Pick disease type C (NP-C) is an inherited neurovisceral lysosomal lipid storage disease characterized by progressive neurological deterioration. Different clinical forms have been defined based on patient age at onset: perinatal, early-infantile (EI), late-infantile (Li), juvenile and adult. We evaluated the efficacy and tolerability of miglustat in 16 symptomatic NP-C patients, with comparative reference to one neurologically asymptomatic, untreated patient. All patients were categorized according to age at neurological disease onset, and were assessed using a standardized clinical assessment protocol: disability and cognitive function scales, positron emission tomography (PET), and biochemical markers. PET and disability scale evaluations indicated that cerebral hypometabolism and neurological symptoms were stabilized during treatment in juvenile-onset NP-C patients. EI and Li NP-C patients, who had higher disease severity at baseline (treatment start), showed increased disability scores and progressive cerebral hypometabolism during follow up. Similarly, while cognitive scale scores remained relatively stable in patients with juvenile NP-C, cognition deteriorated in EI and Li patients. Plasma chitotriosidase (ChT) activity was lower in the juvenile NP-C subgroup than in EI and Li patients, and generally increased in patients who discontinued treatment. Plasma CCL18/PARC and ChT activities indicated greater macrophagic activity in EI and Li patients versus juveniles. Miglustat was generally well tolerated; frequent adverse events included diarrhea and flatulence, which were managed effectively by dietary modification and loperamide. Overall, miglustat appeared to stabilize neurological status in juvenile-onset NP-C patients, but therapeutic benefits appeared smaller among younger patients who were at a more advanced stage of disease at baseline.

Seroprevalence of viral hepatitis in riverine communities from the Western Region of the Brazilian Amazon Basin

The western region of the Brazilian Amazon Basin has long been shown to be a highly endemic area for hepatitis B and hepatitis D viruses. Data concerning the prevalence of hepatitis C and E viruses in this region are still scarce. In this study we investigated the presence of hepatitis A, B, C, D and E viruses infection in communities that live along the Purus and Acre rivers in the states of Acre and Amazonas within the Amazon Basin. A total of 349 blood samples were collected and tested for hepatitis A-E serological markers (antibodies and/or antigens) using commercial enzyme linked immunosorbent assays. Anti-HCV positive sera were further assayed by an immunoblot. HBsAg positive sera were subtyped by immunodifusion. The overall prevalence for hepatitis A, B, C, and E were 93.7%, 66.1%, 1.7%, and 4%, respectively. A very high prevalence of delta hepatitis (66.6%) was found among HBsAg positive subjects. Hepatitis A, B and D viruses were shown to be largely disseminated in this population, while hepatitis C and E viruses infection presented low prevalence rates in this region. The analysis of risk factors for HBV infection demonstrated that transmission was closely associated with sexual activity.

Age-specific Prevalence of Antibodies to Hepatitis A in Children and Adolescents from Rio de Janeiro, Brazil, 1978 and 1995: Relationship of Prevalence to Environmental Factors

The age-specific prevalence of antibodies to hepatitis A virus (anti-HAV) was determined in two different population groups with low socio-economic status from Rio de Janeiro city, Brazil, whose serum samples were collected 17 years apart (Population 1, 1978; Population 2, 1995). In Population 2, analysis of the anti-HAV prevalence was also carried out with respect to environmental factors. Population 1 was composed of 520 stored sera collected from the umbilical cord of term neonates and children aged 1 month to 6 years. In population 2, 720 serum samples were collected from children and adolescents with ages ranging from 1 to 23 years. The overall prevalence rate of anti-HAV in Population 1 and Population 2 was 65.6% and 32.1%, respectively. In Population 1, the anti-HAV prevalence reached 88% at the age of 3, while in Population 2, it increased from 4.5% in children under the age of 3 to 66% in the group of adolescents over the age of 14. The low exposure to HAV infection in younger children from Population 2 could be a result of improved environmental hygiene and sanitation, as demonstrated by the presence of piped water, waste and sewage disposal systems in most houses from this population group. These findings indicate a possible change in the prevalence of hepatitis A in Rio de Janeiro.

First report of a human autochthonous hepatitis E virus infection in Brazil

BACKGROUND Sporadic acute hepatitis E cases occurring in non-endemic areas have been associated to genotypes 3 and 4 of hepatitis E virus. Several studies have demonstrated the relationship among human and animals strains, mostly pigs and deers, from respective areas characterizing zoonosis. Circulation of genotype 3 of HEV in Brazilian swine herds have already been demonstrated. Nevertheless, no confirmed human cases have been reported to date in Brazil. OBJECTIVES A study was developed to attempt the identification of hepatitis E acute cases in Brazil. STUDY DESIGN A retrospective study carried out with 64 serum samples from patients with acute non-A-C hepatitis was performed to identify human cases of acute hepatitis E. RESULTS We could identify a confirmed case of acute hepatitis E. The patient seroconverted to hepatitis E virus-specific IgM and IgG antibody, HEV-RNA was amplified from serum, and the analysis of the sequence of a 242 nucleotide fragment from the ORF1 genome region classified the strain within genotype 3 and subgenotype 3b. Investigation of risk factors and results from phylogenetic analysis suggested a likely zoonotic origin for the infection. CONCLUSIONS The first report of a human autochthonous in Brazil contributes with new information for hepatitis E epidemiology in Latin America and to considerate further broadly epidemiological studies.

Serological and molecular evidence of hepatitis E virus in swine in Brazil

Active hepatitis E virus (HEV) infections in two Brazilian swine herds were investigated. In study 1, 26 piglets born to five anti-HEV positive sows were monitored from birth to post-partum week 22. Serum samples were screened for the detection of anti-HEV antibodies and a nested RT-PCR used to examine the HEV genome. Passive transfer of immunity was confirmed. At week 22, 23/26 (88.4%) of the piglets had seroconverted. Genome amplification was achieved in a feces pool from one holding pen and in one serum sample, both from 13-week-old animals. Histology was suggestive of a potential HEV infection. In the second study, 47 piglets born to six anti-HEV-positive sows were monitored after weaning. Seroconversion was determined in eight animals at 6-8 weeks of age. HEV RNA was detected in two pools from a holding pen for 12-16-week-old animals. Brazilian isolates were classified as genotype 3. This is the first molecular evidence of HEV infection in Brazilian pig herds.

Epidemiological pattern and mortality rates for hepatitis A in Brazil, 1980-2002: a review

The prevalence of hepatitis A virus (HAV) infection is high in developing countries, in which low standards of sanitation promote the transmission of the virus. In Latin America, which is considered an area of high HAV endemicity, most HAV-positive individuals are infected in early childhood However recent studies have shown that prevalence rates are decreasing. Herein, we review the data on HAV prevalence and outbreaks available in scientific databases. We also use official government data in order to evaluate mortality rates in Brazil over the last two decades. Studies conducted in the northernmost regions of Brazil have indicated that, although improved hygiene has led to a reduction in childhood exposure to HAV, the greatest exposure still occurs early in life. In the Southeastern region, the persistence of circulating HAV has generated outbreaks among individuals of low socioeconomic status, despite adequate sanitation. Nationwide, hepatitis A mortality rates declined progressively from 1980 to 2002. During that period, mortality rates in the Northern region consistently exceeded the mean national rate and those for other regions. Excluding the North, the rates in all regions were comparable. Nevertheless, the trend toward decline observed in the South was paralleled by a similar trend in the North.

Seroepidemiological markers of enterically transmitted viral hepatitis A and E in individuals living in a community located in the North Area of Rio de Janeiro, RJ, Brazil

We investigated the seroprevalence of hepatitis A virus (HAV) and hepatitis E virus (HEV) infection in subjects living in the community of Manguinhos, Rio de Janeiro, Brazil, and assisted at the Health Unit of Escola Nacional de Saúde Pública, Fundação Oswaldo Cruz. After formal consent, individuals were submitted to an interview using a standardized questionnaire. Anti-HAV and anti-HEV antibodies were detected by ELISA. Statistical analysis was carried out using the Epi-Info 6.04b software, to investigate possible associations between serological markers and risk factors. Results were regarded as significant when p value < 0.05. Although a high prevalence of anti-HAV was observed (87%), almost 50% of subjects under the age of 10 were susceptible to HAV infection, an unexpected rate in endemic areas. This fact could be attributed to improvements in environmental sanitation, occurring in this area in the last years. The increasing proportion of susceptible people may result in outbreaks of HAV infection, since the virus still circulates in this area, as verified by the detection of anti-HAV IgM in some individuals. No statistical association was met between HAV infection and the risk factors here assessed. The anti-HEV IgG prevalence found in this population was 2.4%, consistent with the one found in non-endemic areas.

Detection of hepatitis A, B, and C virus-specific antibodies using oral fluid for epidemiological studies

In this report, we examine the adaptability of commercially available serological kits to detect antibodies markers for viral hepatitis in oral fluid samples. We also assessed the prevalence of hepatitis A, B, and C virus-specific antibodies, and related risk factors for these infectious diseases through sensitivity of the tests in saliva samples to evaluate if oralfluid can be an alternative tool to substitute serum in diagnosis of acute viral hepatitis and in epidemiological studies. One hundred and ten paired serum and saliva specimens from suspect patients of having acute hepatitis were collected to detect antibodies to hepatitis A (total and IgM), hepatitis B (anti-HBs, total anti-HBc and IgM anti-HBc), and hepatitis C (anti-HCV) using commercially available enzyme-linked immunossorbent assay (EIA). In relation to serum samples, oral fluid assay sensitivity and specificity were as follows: 87 and 100% for total anti-HAV, 79 and 100% for anti-HAVIgM, 6 and 95% for anti-HBs, 13 and 100%for total anti-HBc, 100 and 100% for anti-HBc IgM, and 75 and 100% for anti-HCV The consistency observed between antibodies tests in saliva and expected risk factors for hepatitis A and C suggests that the saliva method could replace serum in epidemiological studies for hepatitis A and C.