The Northeast region is the location of most cases of acute hepatitis A virus (HAV) in Brazil. In the present study, the genotypes of HAV strains from Pernambuco State, one of most populous states in the Northeast region, were characterized. Blood samples positive for anti-HAV IgM from 145 individuals (mean age = 29.1 years), collected during 2002 and 2003, were submitted to nested RT-PCR for amplification of the 5'non-translated region (5'NTR) and VP1/2A regions of the HAV genome. The VP1/2A and 5'NTR regions were amplified in 39 and 21% of the samples, respectively. Nucleotide sequencing was carried out in 46% of VP1/2A and in 53% of 5'NTR isolates. The identity in nucleotide sequence of the VP1/2A region ranged from 93.6 to 100.0%. Phylogenetic analysis of the VP1/2A sequences showed that 65% belong to sub-genotype IA and 35% to sub-genotype IB. Co-circulation of both sub-genotypes was observed in the two years studied. Distinct clusters of highly related sequences were observed in both sub-genotypes, suggesting endemic circulation of HAV strains in this area. In the 5'NTR isolates, 92.7-99.2% identity was observed and two isolates presented one deletion at position 413. Phylogenetic analysis showed that genotype IA strains cluster in the tree in the same way as genotype IB strains, but one IIIA isolate from Spain clusters with genotype IB strains. These results do not allow us to state that 5'NTR could be used to genotype HAV sequences. This is the first report of co-circulation of sub-genotypes IA and IB in this region, providing additional information about the molecular epidemiology of HAV strains in Brazil.

The epidemiology of hepatitis A virus (HAV) infection is shifting from high to intermediate endemicity in Brazil, resulting in increased numbers of susceptible individuals and a greater potential for the emergence of outbreaks. Universal vaccination against HAV has been recommended for children, but updated sero-epidemiological data are necessary to analyze the level of natural immunity and to identify candidates for preventive measures. In addition, more molecular studies are necessary to characterize the genotypes involved in HAV infections and outbreaks. Sera from 299 school children (5-15 years old) and 25 school staff members, collected during an outbreak of HAV at a rural public school in June 2000, were tested for IgM and total anti-HAV antibodies (ELISA). Viral RNA was amplified by RT-PCR from anti-HAV IgM-positive sera and from 19 fecal samples. Direct nucleotide sequencing of the VP1/2A region was carried out on 18 PCR-positive samples. Acute HAV infection was detected by anti-HAV IgM in 93/299 children and in 3/25 adult staff members. The prevalence of total anti-HAV antibodies in IgM-negative children under 5 years of age was only 10.5%. HAV-RNA was detected in 46% IgM-positive serum samples and in 16% stool samples. Sequence analysis showed that half the isolates belonged to subgenotype IA and the other half to IB. On the basis of these data, mass vaccination against HAV is recommended without prevaccination screening, especially for children before they enter school, since nearly 90% of the children under 5 years were susceptible. Molecular characterization indicated the endemic circulation of specific HAV strains belonging to subgenotypes IA and IB.

Hepatitis A virus (HAV) replicates relatively slowly in cell culture without a cytopathic effect, a fact that limits the use of tissue culture assays. The radioimmunofocus assay is the standard method for HAV titration, although it is labor intensive and requires the use of radioisotopes. A simple, rapid and objective infectivity assay based on an in situ enzyme immunoassay (EIA) is described here for a Brazilian cell culture-adapted HAV strain (HAF-203). The assay uses a peroxidase-labeled polyclonal antibody to fixed monolayers as an indicator of infection. EIA may be completed within 7 days using serial 5-fold dilutions of the virus, yielding a titer of 5.024 log 50% tissue culture infective dose (TCID50)/ml for HAF-203. This technique had a detection limit of 1.1 log TCID50/ml and the specificity was demonstrated by detecting no reaction on the columns of uninfected wells. The reproducibility (with intra- and inter-assay coefficients of variation ranging from 1.9 to 3.8% and from 3.5 to 9.9%, respectively) and quantitation of the assay were demonstrated by close agreement in virus infectivity titers among different assays of the same amount of virus and between assays of different amounts of virus. Furthermore, this assay does not require the use of radiolabeled antibodies. We describe here an efficient EIA that is highly reproducible and that could be used to monitor HAV growth in cell culture and to determine the quantity of HAV antigen needed for diagnostic assays. This is the first report of the infectious titer of the Brazilian cell culture-adapted HAV strain (HAF-203).

In the region of Precordillera of Jagüe, in La Rioja province, the Palaeozoic volcanic rocks, known as the Punta del Agua, Cerro Chuscho and Las Casitas formations, have been grouped into one unit, the "Andesita Cerro Chuscho". These rocks have been considered to be comagmatic and to form a subvolcanic complex of autoclastic breccias and calc-alkaline lava flows of the precordilleran orogenic cycle. In our study, the Cerro Chuscho Formation is reinterpreted as a mafic volcanic complex. Of Ordovician, it includes dykes, sills and pillow-lavas of basaltic composition, representing an ophiolitic facies interbedded in the sedimentary pile. The Cerro Chuscho Formation belongs to the "Famatinian Ophiolitic Belt", which is related to the opening of a marginal basin during Chilenia accretion in Cambro-Ordovician times. The geochemical signature of the basaltic rocks changes along the rift, from N-MORB in the Cordillera Frontal to E-MORB in the Precordillera of San Juan and La Rioja. This variation probably reflects the changing altitude of the rift relative to the mantle. By contrast, the "Andesitas Punta del Agua", of Late Carboniferous to Early Permian age, are considered to be subduction-related andesites according to their geochemical characteristics.

En la Precordillera de Jagüé (provincia de la Rioja), fueron agrupados bajo la denominación de Andesita Cerro Chuscho los afloramientos de volcanitas de: Punta del Agua, Cerro Chuscho y Las Casitas. Se consideraba a estas rocas comagmáticas, constituyendo el cerro Chuscho un cono subvolcánico heterogéneo compuesto por una autobrecha de flujo, integrada por rocas calcoalcalinas correspondientes al ciclo precordillerano. En este trabajo se asigna a los afloramientos del cerro Chuscho (Formación Chuscho) como un complejo de volcanitas máficas intraordovícicas, representado por diques y filones de diabasas y por basaltos de estructura almohadillada. Conforman una facies ofiolítica asociada con las sedimentitas ordovícicas y, por lo tanto, quedarían incluidas en la Faja Ofiolítica Famatiniana, asociada a la apertura de un rift oceánico de cuenca marginal formado durante la acreción de Chilenia en el Cambro-Ordovícico. Sus características parecen cambiar a lo largo del rift, pasando de N-Morb en la Cordillera Frontal a E-Morb (P-MORB) en la Precordillera de San Juan y La Rioja; estas diferentes características se deben muchas veces al cambio de elevación del rift respecto del manto. Por su parte, las Andesitas Punta del Agua (Formación Punta del Agua), son andesitas orogénicas de arco volcánico, de acuerdo a sus características geoquímicas, con edades que irían desde el Carbonífero superior al Pérmico inferior.