Lena Maria Barros

Hepatitis Delta virus genotype 8 infection in Northeast Brazil: Inheritance from African slaves?

Hepatitis Delta virus (HDV) is endemic worldwide, but its prevalence varies in different geographical areas. While in the Brazilian Amazon, HDV is known to be endemic and to represent a significant public health problem, few studies have assessed its prevalence in other regions in the country. This study evaluated the seroprevalence of HDV among HBsAg chronic carriers from Maranhão state, a region located in the Northeast of Brazil. Among 133 patients, 5 had anti-HD, of whom 3 had HDV RNA. HDV genotypes were characterized by Bayesian phylogenetic analysis of nucleotide sequences from the HDAg coding region. HDV-3 was identified in one patient who lives in Maranhão, but was born in Amazonas state (Western Amazon basin). Phylogenetic analysis shows that this HDV-3 sequence grouped with other HDV-3 sequences isolated in this state, which suggests that the patient probably contracted HDV infection there. Surprisingly, the other two patients were infected with HDV-8, an African genotype. These patients were born and have always lived in Urbano Santos, a rural county of Maranhão state, moreover they had never been to Africa and denied any contact with people from that continent. This is the first description of the HDV-8 in non-native African populations. This genotype may have been introduced to Brazil through the slaves brought to the country from the West Africa regions during the 16-18th centuries. Our results indicate that the need of clinical and epidemiological studies to investigate the presence of this infection in other areas in Brazil.

HBV carrying drug-resistance mutations in chronically infected treatment-naive patients.

BACKGROUND Nucleoside/nucleotide analogue (NA) treatment causes selection pressure for HBV strains carrying mutations conferring NA resistance. Drug-resistance mutations occur in the reverse transcriptase (RT) region of the HBV polymerase gene and spontaneously arise during viral replication. These mutations can also alter the hepatitis B surface (HBs) protein and in some cases reduce binding to HBs antibodies. The spread of NA-resistant HBV may impact the efficacy of antiviral treatment and hepatitis B immunization programmes. In this study, we used direct sequencing to assess the occurrence of HBV carrying known mutations that confer NA resistance in the largest cohort of treatment-naive patients with chronic hepatitis B (CHB) to date. METHODS HBV DNA samples isolated from 702 patients were sequenced and the RT region subjected to mutational analysis. RESULTS There was high genetic variability among the HBV samples analysed: A1 (63.7%), D3 (14.5%), A2 (3.3%), A3 (0.1%), B1 (0.1%), B2 (0.1%), C2 (0.9%), D1 (0.9%), D2 (4.6%), D4 (5.1%), D unclassified subgenotype (0.7%), E (0.6%), F2a (4.6%), F4 (0.4%) and G (0.4%). HBV strains harbouring mutations conferring NA resistance alone or combined with compensatory mutations were identified in 1.6% (11/702) of the patients. CONCLUSIONS HBV strains harbouring resistance mutations can comprise the major population of HBV quasispecies in treatment-naive patients. In Brazil, there is a very low frequency of untreated patients who are infected with these strains. These findings suggest that the spread and natural selection of drug-resistant HBV is an uncommon event and/or most of these strains remain unstable in the absence of NA selective pressure.

High prevalence of hepatitis B virus subgenotypes A1 and D4 in Maranhão state, Northeast Brazil

In this study, we determined the prevalence of HBV subgenotypes in Maranhão state, located in northeastern Brazil, where the population is heterogeneous, with a high proportion of African descendants. HBV was detected in 119 of 133 (89.5%) chronic hepatitis B patients, including 103 (86.5%) who were HBeAg-negative. Using phylogenetic analysis of the S/Polymerase region of HBV DNA, subgenotype A1 was found to be the most prevalent (67%), followed by genotype D (28%; subgenotype D4 was detected in 24%, D3 in 3%, and D2 in 1%). Genotype F, clustering with subgenotype F2a, was found in six (5%) patients. The topology of the phylogenetic tree showed that HBV/A1 sequences did not cluster together, suggesting that more than one strain was introduced into Maranhão. On the other hand, HBV/D4 sequences formed a monophyletic cluster, suggesting a single entry of this strain in this population. This study showed that HBV/A1 was the only subgenotype of HBV/A present in the population from Maranhão and indicated that in this region HBV/A1 was not restricted to an Afro-descendant community where it was previously reported, but is widely distributed among general population of HBV chronic carriers. Unexpectedly, we found a high frequency of HBV subgenotype D4. Together with previously reported data on the distribution of HBV/D4 in the world, these findings suggest that this subgenotype was more prevalent in the African continent in the past and may have been introduced in Maranhão by means of the slave trade during the late XVIII century, when the largest number of African slaves arrived to this region.

The hepatitis delta genotype 8 in Northeast Brazil: The North Atlantic slave trade as the potential route for infection

Hepatitis Delta virus (HDV) is not well known, even though HDV and Hepatitis B virus (HBV) co-infection leads to severe forms of acute and chronic liver diseases. HDV is endemic in the Western Amazon region. Recently, the HDV genotype 8 was found in chronic patients followed at the center for liver studies in the Northeast Brazil, Maranhão. Previous studies suggested that this genotype was introduced in Maranhão during the slave trade. The presence of HDV in that study, which was done outside the Amazon region, led us to investigate whether the virus is found infecting individuals in other regions of Maranhão as well. Thus, we screened ninety-two HBsAg positive individuals from five Municipalities of Maranhão for anti-HD antibody and eight were found positive (8.7%). These eight positive individuals were submitted to polymerase chain reaction (PCR) to investigate active HDV infection. Half of them were positive for a fragment sequence of the delta antigen; their sequence samples were submitted to genotype characterization by phylogenetic analysis. All sequences clustered in a unique branch of the tree separated from the other branch described in Africa. Our study confirmed the presence of HDV-8 in Maranhão. These infected individuals had no evidence of contact with African people. Furthermore, we found individuals infected with HDV-8 in two more different municipalities. More studies like ours are urgent because the co-infection HBV/HDV is more difficult to treat. Identification of the endemic regions and implementation of healthy policies for preventing this infection are urgent in this region.