J. Abreu-Silva

Seroprevalence of Hepatitis E Virus Antibodies in Portuguese Children

Background: Hepatitis E virus (HEV) has become a growing public health concern in industrialized countries. Most of the HEV seroprevalence studies have focused on adult populations, and reports regarding HEV seroepidemiology among children are scarce in these countries. The aims of this work were to perform a nationwide seroprevalence study on HEV infection in Portuguese children and to compare the HEV seropositivity in this 2015 children cohort with results in sera performed 20 years earlier. Methods: Sera (N = 352) from children collected in 2015 from all regions of Portugal were screened for anti-HEV IgG and IgM using the commercial enzyme-linked immunosorbent assay recomWell HEV IgG/IgM (2015 version; Mikrogen, Neuried, Germany), and positive samples were retested by immunodot assay recomLine HEV IgG/IgM (2015 version; Mikrogen, Neuried, Germany). For the comparative assessment of HEV seropositivity of the 2 children cohorts, children’s sera (N = 71) archived since 1995 were screened for anti-HEV IgG and results were compared with that of the 2015 cohort, matched by sex, age and region. Results: Anti-HEV antibodies were detected in 4 children giving an overall HEV seroprevalence in the 2015 cohort of 1.1%. A healthy 10–15-year-old female was found positive for anti-HEV IgM indicating a current or recent HEV infection. No statistically significant difference was observed in HEV seroprevalence regarding gender, age group and region of residence. Comparison of the HEV seropositivity of the 2 children cohorts showed a statistical significant decrease with time (P = 0.024). Conclusions: This is the first national study of HEV seroprevalence in Portuguese children and the first to demonstrate a decrease of anti-HEV antibodies in this age group over time.

Evidence of autochthonous hepatitis E in a Portuguese pediatric cohort, 1992-1995.

Despite the growing importance of hepatitis E virus (HEV) in industrialized countries, minimal attention has been given to autochthonous HEV infection in children. The present study screened archived sera (N = 71) from a Portuguese pediatric cohort collected in 1992–1995 for the presence of antibodies against HEV, using enzyme immunoassays and immunodot. Anti‐HEV IgG was detected in two children and anti‐HEV IgM in one, suggesting an acute HEV infection. This shows that HEV was circulating in the paediatric population of Portugal in the early 1990s. J. Med. Virol. 88:919–921, 2016.

Short communication: detection and molecular characterization of hepatitis E virus in domestic animals of São Tomé and Príncipe

As in most of the African continent, the status of hepatitis E virus (HEV) infection in domestic animals in São Tomé and Príncipe, an archipelago off the western equatorial coast of Central Africa, is also completely unknown. In the present study, we investigated the presence of HEV among domestic animals in São Tomé and Príncipe. A total of 93 stool samples from different animal species (goat, cow, pig, chicken, duck, and monkey) were tested for HEV RNA using two real-time RT-PCR assays, followed by a nested RT-PCR assay for sequencing and phylogenetic analysis. A total of six samples (1 cow stool and 5 pig stools) were found to be positive for HEV RNA of which one pig stool was positive by broad spectrum nested RT-PCR. Phylogenetic analysis showed that the retrieved sequence clustered within HEV subgenotype 3f, similar to zoonotic strains of European countries and posing interesting questions on past introduction of European HEV into São Tomé and Príncipe archipelago. This is the first report describing the presence and molecular characterization of HEV in São Tomé and Príncipe.

A nationwide serosurvey of hepatitis E virus antibodies in the general population of Portugal

Background Evidence has shown that Hepatitis E virus (HEV) genotype 3 is autochthonous in industrialized countries due to zoonotic transmission through direct contact or consumption of raw or undercooked meat from domestic swine or wild boar. As there is lack of data on seroprevalence of HEV in the general Portuguese population, a wide survey was conducted as part of the HEPeCONTROL project (60DT2), under EEA grants funding. Methods Sera from a representative sample of the Portuguese population (n = 1656) at different geographic locations (30 territorial units), and age (0-99 years) were collected between July 2015 and February 2016. The sera were tested for the presence of anti-HEV IgG and IgM by EIA using one of the two most commonly used commercial immunoassays in Europe. Results The overall HEV IgG seroprevalence was found to be 16.3% increasing with age (P < 0.05) from 0.6% in the 0-9 years group to 30.1% in people older than 70 years. The seroprevalence also varied geographically with generally higher seropositivities (25-30%) in the most rural areas of Portugal. However, the geographical differences were not statistically significant (P > 0.05). Out of 1656 samples, 8 were positive for anti-HEV IgM indicating current of recent HEV infection but no significant differences were found concerning age groups, regions and sex. Conclusions The present nation-wide survey provides insight in the epidemiology of HEV in Portugal and confirms that HEV is endemic in the Portuguese population.

Hepatitis E Virus Genotype 1 Cases Imported to Portugal from India, 2016

Hepatitis E in industrialized countries is mainly associated with genotype 3 hepatitis E virus (HEV) and normally causes a sporadic self-limiting disease in immunocompetent individuals. Unlike genotype 3, genotypes 1 and 2 circulate in developing countries, produce severe disease and occur in the epidemic form. Hepatitis E occurring in travellers returning from endemic areas in developing countries is not a novel epidemiological occurrence, however the vast majority of cases remain to be genetically studied. The present study describes two cases of severe acute hepatitis E that required hospitalization for 6 and 9 days in two individuals of Indian nationality that had recently migrated to Portugal to work. The retrieved HEV sequences both belonged to genotype 1 and had a high degree of nucleotide sequence identity, clustering with strains isolated in India and Nepal, in 2013 and 2014. Confirmed HEV genotypes of increased pathogenicity like genotype 1 are being introduced into otherwise naïve populations of industrialized countries such as European countries with consequences difficult to predict. As far as we know the present study is the first in Portugal to describe and genetically characterize imported cases of hepatitis E infection caused by HEV genotype 1.Hepatitis E in industrialized countries is mainly associated with genotype 3 hepatitis E virus (HEV) and normally causes a sporadic self-limiting disease in immunocompetent individuals. Unlike genotype 3, genotypes 1 and 2 circulate in developing countries, produce severe disease and occur in the epidemic form. Hepatitis E occurring in travellers returning from endemic areas in developing countries is not a novel epidemiological occurrence, however the vast majority of cases remain to be genetically studied. The present study describes two cases of severe acute hepatitis E that required hospitalization for 6 and 9 days in two individuals of Indian nationality that had recently migrated to Portugal to work. The retrieved HEV sequences both belonged to genotype 1 and had a high degree of nucleotide sequence identity, clustering with strains isolated in India and Nepal, in 2013 and 2014. Confirmed HEV genotypes of increased pathogenicity like genotype 1 are being introduced into otherwise naïve populations of industrialized countries such as European countries with consequences difficult to predict. As far as we know the present study is the first in Portugal to describe and genetically characterize imported cases of hepatitis E infection caused by HEV genotype 1.

First detection and molecular characterization of hepatitis E virus in water from wastewater treatment plants in Portugal

Hepatitis E virus (HEV) genotype 3 is widespread in industrialized countries and widely recognized as a serious public health issue. HEV genotype 3 is excreted in both human and animal stools, having the potential to be introduced in aquatic environments through wastewater discharge. For this reason, waterborne transmission of the HEV genotype 3 is drawing increasing attention from the scientific community. Although studies in Portugal have shown that HEV circulates in both humans and swine, no study has yet focused on the presence of HEV in wastewater. Hence, a total of 60 water samples (influent and effluent) from 15 Wastewater Treatment Plants (WWTP) of different regions of Portugal were tested for the presence of HEV using a broad spectrum real-time RT-PCR with amplification within the ORF2 region of HEV genome. Positive samples were retested by nested RT-PCR with amplification within the ORF1 region, sequenced followed by phylogenetic analysis. HEV sequences were retrieved from two influent samples collected in December 2013 from WWTPs of North and Central of Portugal and classified in subgenotype 3i and 3f. This is the first study showing the presence of HEV in aquatic environments of Portugal.