Domenica Donia

Presence of hepatitis E RNA in mussels used as bio-monitors of viral marine pollution

Mussels (Mytilus galloprovincialis), collected from a harvesting area approved by European Community Regulation, were transplanted to four polluted sites located in the Northwestern Mediterranean area (Tuscany). They were used as bio-monitors to test the quality of the marine water pollution. At different times after the transplantation, mussels were withdrawn and tested for presence of phages and enteric viruses by molecular tests. 52.4% of the transplanted mussel samples were positive for at least one enteric virus. Hepatitis A virus (HAV) was identified in each site (17/37; 45.9%). Three samples were positive for hepatitis E virus (HEV) (8.1%) and two (5.4%) for norovirus (NoV) genogroup I. Coliphages and RYC 2056 phages were detected in all sites, while HSP 40 phages were detected in three sites. Results demonstrate the ability of transplanted mussels in accumulating and retaining different species of enteric microorganisms. Their utility as bio-monitor organisms enables testing for viral marine pollution.

Detection and molecular characterization of human rotaviruses isolated in Italy and Albania

Rotaviruses are one of the most important causes of gastroenteritis in children under 5 years old. Analysis of G and P rotavirus genotypes in circulation is crucial in evaluating the appropriacy of mass vaccination of children worldwide. Overall, 592 stool samples were collected in Tirana (Albania), the Salento peninsula (South Italy), and three different hospitals in Rome (Central Italy). Of the total samples, 31.3% were rotavirus positive in Albania, 78.3% in the Salento, and 40.3% in Rome. The samples collected in Tirana and Rome were G–P typed, whereas the samples collected in the Salento were only G typed. Overall, in Italy the most frequent combinations were G4 P[8] (54.5%), G1 P[8] (27.3%), and G2 P[4] (18.2%); in Albania they were G9 P[8] (72.1%), G4 P[8] (8.8%), G1 P[8] (5.9%), and G2 P[4] (2.9%). The prevalence in Albania of atypical combinations was 7.4% for G4 P[4] and 2.9% for G9 P[4]. Phylogenetic analysis was also performed to assess the genetic relatedness of the strains. J. Med. Virol. 82:510–518, 2010.

Genetic Diversity of Human Adenovirus in Children with Acute Gastroenteritis, Albania, 2013–2015

The objectives of the present study were to assess the occurrence of human adenoviruses (HAdVs) in paediatric patients with gastroenteritis in Albania and to characterize HAdV strains. Faecal specimens from children admitted with acute gastroenteritis to the Paediatric Hospital in Tirana were screened for HAdV, using broad-range primers targeting the hexon gene, in combination with species-specific primers targeting the fiber gene. Phylogenetic analysis was then performed to assess the genetic relationships among the different sequences and between the sequences of the samples and those of the prototype strains. Adenovirus DNA was detected in 33/142 samples (23.2%); 14 belonged to species F (13 HAdV-41 and 1 HAdV-40), 13 to species C (1 HAdV-1, 8 HAdV-2, and 4 HAdV-5), 5 to species B (HAdV-3), and 1 to species A (HAdV-12). Rotavirus coinfection was present in 9/33 (27.2%) positive samples. In the remaining 24 positive samples (12 enteric—F species; 12 nonenteric—A, B, or C species), HAdVs were detected as unique viral pathogens, suggesting that HAdV may be an important cause of diarrhoea in children requiring hospitalization. This is the first study investigating the presence of human adenoviruses (species A–G) as etiologic agents of viral gastroenteritis in children in Albania.

Evaluation of the wastewater treatment plant of Rome airport

The wastewater plant of Rome airport, which receives all the sewage from the airport as well as the cess from aeroplanes, was analysed for microbiological parameters. From the bacteriological point of view, in the water and sludge samples the densities of the faecal indicator of pollution and the presence of Salmonella spp and Vibrio cholerae as bacteriological pathogens were determined. At the same time, samples were analysed for the presence of enteric viruses and phages. Overall, the mean reduction of the faecal coliforms was 96%, E. coli 92% and faecal streptococci 99%. Salmonella spp was identified in all but one of the final effluents and V. cholerae in 2/10. Enteric viruses were identified in all but one of the raw waters and in three samples of final effluent. Bacteriophages (somatic coliphage, F-plus phage and B40-8), were found in all the samples but irregularly. Phages and enteric viruses were also found in the prefilter membranes used for prefiltering the raw water samples.

Human bocavirus in children with acute gastroenteritis in Albania

Human Bocavirus (HBoV) has been recently identified in association with acute viral gastroenteritis (AGE). The objective of this work was to investigate the prevalence of HBoV in children with AGE in Albania. Stool specimens collected from 142 children were analyzed by amplification of partial NP1 and Vp1/Vp2 genes. HBoV was detected in 13 samples (9.1%), 12 HBoV‐1 and one HBoV‐2. All HBoV‐positive patients were co‐infected with rotavirus and/or adenovirus, a finding which might indicate that there is no clear causal association of this agent with diarrhea. Further investigation is needed to assess the pathogenic role of HBoV in childhood diarrhea. J. Med. Virol. 88:906–910, 2016.

Interaction between single wall carbon nanotubes and a human enteric virus.

Activated single wall carbon nanotubes have been used for biomedical purposes as carriers for drugs, peptides, proteins and nucleic acids. A large volume of data speaks to their suitability to act as a carrier. The ability of two differently activated SWNTs (with carboxyl groups or with carboxyl groups and polyethylenimine (PEI)) to form a complex with the hepatitis A virus was evaluated. Both types of activations permitted the formation of a virus-SWNT complex. However, their patterns were different. The carboxyl-activated nanotubes had a somewhat low adsorptive capacity that was related inversely to the concentrations of the SWNTs and viruses. Statistical analysis, using the chi(2)-test, showed no significant differences between the SWNT-PEI ratios of 1:2.5, 1:1 and 1:0.5. The addiction of PEI improved the adsorption, probably because of the electropositive charge of the molecule. Adsorption was optimal between 100 microg and 10 ng with a SWNTs-PEI weight ratio of 1:0.2 up to an inoculum of 10(5) genome equivalents of hepatitis A virus. Reducing or increasing this weight ratio reduced the adsorptive capacity of the PEI, and this adsorption activity was time and contact-dependent. Thus, SWNTs coated with PEI are able to complex with viruses, and they might be used in the future to transfect non-permissive cell lines.

Are PEI-coated SWCNTs conjugated with hepatitis A virus? A chemical study with SEM, Z-potential, EDXD and RT-PCR

The conjugation between nanotubes, coated with different doses of polyethylene imine (PEI) and hepatitis A virus (HAV) was investigated by scanning electron microscopy, Z-potential, thermogravimetric and differential thermal analysis, transmission electron microscopy, energy dispersive x-ray diffraction (EDXD) and reverse transcript polymerase chain reaction (RT-PCR). For the first time, to our knowledge, evidence is obtained that conjugation between the nanotubes and the HAV occurs and that it has an (at least a partial) electrostatic character. Since all components of the conjugated systems, nanotubes, coating material and virus are characterized by different peak shapes in the selected q range, it was possible to infer that conjugation occurred. RT-PCR measurements confirmed that the conjugation of the coated nanotubes and HAV occurred and the result was stable. This opens up the prospect of probing the coated nanotubes as intra-cellular carriers in transfection processes of the virus. Further biological applications will concern a possible vaccine especially for non-replicative viruses.

Concomitant poliovirus infection during an outbreak of hepatitis A

AIM OF THE STUDY The present study was designed to evaluate the possible co-infection, with other enteric viruses, during an outbreak of hepatitis A (HA). MATERIAL AND METHODS Forty-two stool samples and sera were collected during an outbreak of hepatitis A. Sera were analysed by the Abbott test for IgG-IgM anti-HAV antibodies. Stool samples were used to identify the presence of enteric viruses. HAV genome was identified by a RT-PCR test, other enteric viruses were identified, after cell passage and seroneutralization test on BGM cells, by RT-PCR and RFLP assay. RESULTS The samples were obtained from 27 employees of an industrial plant, nine household contacts and six non-employee controls. The attack rate was 12.5%, whereas the overall prevalence was 63%. In the employee group, 12 out of 27 stool samples were positive for the presence of HAV by reverse transcriptase polymerase chair reaction (RT-PCR). All the other samples (30) were negative. Five samples from employees, three from household contacts and one from non-employees were also found positive for enteroviruses. These viruses were classified by seroneutralization as poliovirus and RFLP assay as Sabin poliovirus type 1. Four samples were positive both for HAV and poliovirus. CONCLUSIONS This study confirms co-infection with different enteric viruses may occur and also emphasizes the wide circulation of HAV and the existence of silent infection with poliovirus.

Molecular characterization of norovirus GII strains identified in Albania.

Noroviruses (NoVs) are considered as the leading cause of diarrheal diseases in all groups of age. In the last decade the number of NoV outbreaks worldwide is increasing. Data published by the systems of NoV surveillance show the GII.4 strain as the human predominant genotype circulating worldwide and new genetic variants of GII.4 were associated with epidemic events. In Albania the economy transformation has damaged significantly the environment and a large circulation of enteric viruses was reported in the past with the presence of NoV among the genotyped strains. This study aimed to characterize, by molecular analysis, the NoV GII strains detected in Albania during two time periods: in 2010 from the outbreak occurred in Ballsh and in 2002–2003 from sporadic cases of diarrhoea. A total of 21 Nov GII strains were characterized. The NoV GII.4 was genotyped more frequently and it was related closely to the pandemic variants recorded in GenBank. During 2002–2003, six NoV GII recombinant strains have been characterized. J. Med. Virol. 85:731–736, 2013.