Search

Noroviruses (NoV) are a major cause of gastroenteritis worldwide. Recently, a novel variant of NoV GII.17 (GII.P17_GII.17 NoV), termed Kawasaki 2014, has been increasingly reported in NoV outbreaks in Asia, and has also been described in Europe and North America. In this study, sewage samples were investigated to study the occurrence and genetic diversity of NoV genogroup II (GII) along a 6-year period. Moreover, the spread of GII.17 strains (first appearance and occurrence along time) was specifically assessed. A total of 122 sewage samples collected from 2011 to 2016 from four wastewater treatment plants in Rome (Italy) were initially tested using real-time RT-(q)PCR for GII NoV. Positive samples were subsequently subjected to genotypic characterization by RT-nested PCRs using broad-range primes targeting the region C of the capsid gene of GII NoV, and specific primers targeting the same region of GII.17 NoV. In total, eight different genotypes were detected with the broad-range assay: GII.1 (n = 6), GII.2 (n = 8), GII.3 (n = 3), GII.4 (n = 13), GII.6 (n = 3), GII.7 (n = 2), GII.13 (n = 2), and GII.17 (n = 3), with the latter two genotypes detected only in 2016. Specific amplification of GII.17 NoV was successful in 14 out of 110 positive samples, spanned over the years 2013-2016. The amplicons of the broad-range PCR, pooled per year, were further analyzed by next-generation sequencing (NGS) for a deeper analysis of the genotypes circulating in the study period. NGS confirmed the circulation of GII.17 NoV since 2013 and detected, beyond the eight genotypes identified by Sanger sequencing, three additional genotypes regarded as globally uncommon: GII.5, GII.16, and GII.21. This study provides evidence that GII.17 NoV Kawasaki has been circulating in the Italian population before its appearance and identification in clinical cases, and has become a major genotype in 2016. Our results confirm the usefulness of wastewater surveillance coupled with NGS to study the molecular epidemiology of NoV and to monitor the emergence of NoV strains.

The original version of this article unfortunately contained a mistake. The presentation of Table 1 was incorrect. The corrected table is given below. The original article has been corrected.

Worldwide, 12.5% of the more than 170 million people infected by hepatitis C virus (HCV), live in Eastern Mediterranean countries. In Turkey, the prevalence of HCV infection ranges from 0.3% to 0.4% of the general population. We investigated the distribution of HCV subtype 1b in Turkey by analysing the NS5b viral genomic region, using a Bayesian coalescent-based framework and phylogeographical analysis to estimate the origin of the HCV 1b subtype epidemic and the genetic diversification of the virus in Turkey. The dataset consisted of 24 NS5b sequences obtained from patients chronically infected with HCV subtype 1b admitted to the different health districts of Ankara hospital plus the reference sequences for phylogenetic analysis. An independent dataset including the same 342-nt NS5b fragment from all over the world (203 sequences) was used to calibrate the evolutionary rate. Using the relaxed clock model, we estimated a mean evolutionary rate of 0.84 × 10 −3 sub/site/year (95% highest posterior density interval HPD 0.16–1.5 × 10 −3 ). The results of the phylogeographical analysis suggested that the HCV epidemic probably originated in Greece during the first decade of 1900 and, a few years later (in the 1920s or 1930s), successfully spread to neighboring countries such as Turkey and Cyprus. The clustering of the majority of the Turkish strains in a single monophyletic group suggests the subsequent segregated circulation of the virus in the country during the years 1940–1999, which was probably due to unsafe medical parenteral procedures, with drug addiction playing a relatively negligible role. The Bayesian skyline plot (BSP) showed a growth in the number of effective infections between the 1940s and the 1990s, when the curve reached a plateau that still remains today, suggesting a partial success of improved transfusional policies. A coalescent-based approach to population dynamics can improve our understanding of the origin and spread of epidemics in a limited geographical area.

Background Autochthonous (locally acquired) cases of acute hepatitis E virus have been recently reported in several developed countries. Aim To evidence cases, if any, and characteristics of acute hepatitis E virus infections in North-East of Italy several years ago. Methods In 2014, stored sera of 165 nonA–nonB acute hepatitis referred to the hospital of Padua during the period 1978–1991 were tested for hepatitis C virus antibodies by EIA III and for anti-hepatitis E virus IgM by Wantai HEV IgM ELISA. Anti-hepatitis E virus IgM positive sera were tested by Real Star HEV RT-PCR kit (Altona Diagnostics, Hamburg, Germany). Results Ninety-six (58.1%) sera resulted anti-HCV positive, and thus classified as acute C hepatitis. None of these subjects was anti-HEV IgM positive. Out of the 69 anti-HCV negative cases, 4 (5.8%) resulted anti-HEV IgM positive (one case hepatitis E virus-RNA positive), with an increasing trend from 2.8% during the years 1978–1984 to 9.1% during the years 1985–1991. All cases occurred in Italian patients with no travel abroad history. Conclusions There is evidence for the presence of autochthonous cases of acute hepatitis E virus infections in Italy since 80s.

In early May 2013, Germany reported through the Epidemic Intelligence Information Systems (EPIS) and the Early Warning and Response Systems (EWRS) seven Hepatitis A (HAV) cases associated with a travel history in Northern Italy. Following the alert, the Italian Integrated Epidemiological System of Acute Viral Hepatitis (SEIEVA) reported also an increase of HAV cases in the same area in 2013. Cases identified were infected with HAV genotype IA with an identical “outbreak” sequence (KF182323). To monitor HAV cases …

Introduction: Infection with hepatitis C virus (HCV) represents the major cause of liver disease, affecting more than 170 million individuals worldwide. Several data indicate that HCV replication can influence the progression and the severity of liver diseases by a modulation of cellular gene expression. Methods: To evaluate HCV-induced modification in the global expression profile, we compare gene expression of the human hepatoma cell line 2 1-5 carrying the full-length HCV replicon with the parental cell line Huh7 and the cured replicon cells in which the HCV replicon was removed by IFN-a treatment. Gene expression was examined using Applied Biosystems Human Genome Survey Arrays containing 3 1,700, 60-mer oligonucleotides probes representing a set of27,868 individual human genes and more than 1,000 control probes. The study was designed to analyze gene expression for three biological replicates for each cell line and two technical replicates for each biological replicate, for a total of 18 microarrays. The fold change of HCV replicon vs. parental or cured cell lines was analyzed by filtering the dataset of normalized signals (using p value 10.01 and 3) and performing ANOVA statistical analysis. PANTHER Classification system (Celera Genomics) was used to interpret gene expression data obtained comparing HCV replicon vs parental or cured cells. Alteration of specific genes was confirmed by Real-time PCR. Results and Discussion: Data obtained from this study indicated that HCV replication results in a specific and significant alteration of genes involved in cellular process such as endoplasmic reticulum (ER) and oxidative stress, apoptosis, lipid metabolism, immune defense and cell cycle which are relevant for HCV pathogenesis. Virul Hepatitis linit

Frequent occurrence of woodchuck hepatitis virus DNA (WHV DNA) integration into or in proximity tomyconcogenes and in thewinlocus of cellular genome in woodchuck hepatocellular carcinomas (HCC) has been described by several authors. We report a further cellular locus as a recurrent target for WHV integration in woodchuck HCCs. A WHV DNA integration and its cellular flanking regions were cloned from a HCC developed in a chronically WHV-infected woodchuck. Sequence analysis showed integration of rearranged C, PreS1, and 5′ truncated X regions of the WHV genome, located in a cellular locus previously described for WHV integration in another woodchuck HCC. The two integration sites are only about 0.5 kb apart. In addition toAlu-like repeats and agag-like coding region, previously described, we found several features of MAR (matrix attachment region) chromosomal sequences in the normal cellular locus, leading us to predict that part of it might be a previously unrecognized MAR.