Edina Meleg

Genogroup I picobirnaviruses in pigs: Evidence for genetic diversity and relatedness to human strains

Picobirnaviruses (PBVs) are small, non-enveloped viruses with a bisegmented double-stranded RNA genome. Their pathogenic potential, ecology, and evolutionary features are largely unexplored. Here, we describe the molecular analysis of porcine PBVs identified in the intestinal content of dead pigs. Six of 13 positive samples were cloned and then subjected to single-strand conformation polymorphism analysis and nucleotide sequencing. All clones belonged to genogroup I PBVs and almost all clones clustered on separate branches from human strains. A single strain shared a notably close genetic relationship with a Hungarian human PBV strain (89.9 nt and 96.4% aa identity). Genetic diversity was also observed among strains identified in mixed infections. Single point mutations and deleterious mutations within highly related strains suggested that PBVs exist as quasispecies in the swine alimentary tract. Clones with complete sequence identities originating from different animals suggested effective animal-to-animal transmission of the virus. Our findings indicate that infection with genogroup I PBVs is common in pigs.

Detection and quantification of group C rotaviruses in communal sewage.

ABSTRACT Group C rotaviruses have been recognized as a cause of acute gastroenteritis in humans, cattle, and swine, although the true epidemiologic and clinical importance of this virus in these hosts has not yet been fully established. A real-time PCR assay based on a broadly reactive primer pair was developed and used to quantitatively determine the viral load of group C rotaviruses in environmental samples. A total of 35 raw and 35 treated sewage samples collected at the same sampling time in four Hungarian sewage treatment plants during a survey in 2005 were tested for the presence of group C rotaviruses. The overall detection rates were 91% (32 of 35) for the influent and 57% (20 of 35) for the effluent samples. Molecular characterization of the amplified partial VP6 gene revealed the cocirculation of human and animal (i.e., bovine and porcine) strains that were easily distinguishable by melting curve analysis. Human strains yielded relatively high viral loads (mean, 1.2 × 107; median, 6.9 × 105 genome equivalents per liter influent sewage) and appeared to display seasonal activity over the study period, whereas animal strains appeared to circulate throughout the year at much lower average titers (bovine strains mean, 9.9 × 104; median, 3.0 × 104; porcine strains mean, 3.9 × 104; median, 3.1 × 104 genome equivalents per liter influent sewage). Our findings suggest that monitoring of communal sewage may provide a good surrogate for investigating the epidemiology and ecology of group C rotaviruses in humans and animals.

Human astroviruses in raw sewage samples in Hungary

Aims:  Routine procedures for monitoring viruses in water samples have not been drawn up for the water‐microbiology screening panel. Enteric viruses, including astroviruses, are able to persist under environmental conditions and may cause public health problems by contaminating natural and drinking water resources. The aim of this study was to detect human astroviruses (HAstVs) from raw wastewater samples.

Human astrovirus infection associated with childhood intussusception

Human astroviruses (HAstV) are small (27 – 34 nm in diameter), round particles with a fi veor six-pointed star-like surface structure. They were fi rst identifi ed by electronmicroscopy in stool samples from children with gastro-enteritis in 1975. The positive-sense, single-stranded, polyadenylated RNA genome is 6.8 – 7.2 kb in length and consists of three open reading frames. Based on the genome organization eight genotypes of HAstV (HAstV-1 – HAstV-8) have been identifi ed. HAstV are one of the major causes of gastro-enteritis worldwide among infants, children and occasionally among adults and immunosuppressed patients. The prevalence of astroviruses has been reported to be 2 – 16% among children hospitalized with diarrhea and 5 – 17% in community studies. 1 The main clinical manifestations of HAstV infection are diarrhea, vomiting, and fever. 2

Comparison of clinical characteristics between astrovirus and rotavirus infections diagnosed in 1997 to 2002 in Hungary

AIM To determine the severity and clinical characteristics of human astrovirus (HAstV) infections among hospitalized children and compare them with children infected by rotavirus. METHODS Retrospective, case-control study of astrovirus-infected and rotavirus-infected children. Astroviruses were detected in stool samples by enzyme immunoassay and/or reverse transcriptase-polymerase chain reaction. All stool samples were tested for rotavirus and bacterial pathogens, and all negative samples were further tested for human astrovirus. Children with astrovirus-positive stool samples and complete clinical data were included in this study. RESULTS Astrovirus was detected in 29 (1.8%) children, and 63 rotavirus-infected children were included as controls. Astrovirus-infected children had shorter duration of diarrhea than rotavirus-infected children (median 4 and 6 d, respectively; p<0.05), and 79% of the astrovirus infections were associated with a short duration of vomiting (median 1 and 4 d, respectively; p<0.0001). Rotavirus-infected children had longer hospitalization (p<0.050) than astrovirus-infected children. CONCLUSION HAstV-infected children had similar symptoms to those occurring in rotavirus infection. However, astrovirus-infected patients had a significantly shorter duration of diarrhea and vomiting, and they required a shorter hospitalization. On the basis of the clinical data and severity scores, children with rotavirus infection had more severe illness.

Adenovírus 8-as típusa okozta járványos keratoconjunctivitis molekuláris diagnosztikája

INTRODUCTION Both infectious and non-infectious forms of acute conjunctivitis are known. Viruses, especially different types of adenoviruses are the etiological agents of infectious epidemic conjunctivitis (conjunctivitis epidemica). AIMS The authors aims were to describe an outbreak of keratoconjunctivitis and to detect the viral agent by molecular methods in Hungary. MATERIALS AND METHODS Classical epidemiological methods were used for investigation. Polymerase chain reaction (PCR) followed by sequencing were used for the detection of adenoviral hexon region from freshly collected conjunctival swabs. RESULTS Between 9 October and 18 December 2006, a total of 60 patients became ill with keratoconjunctivitis in 7 settlements in Southwest Hungary. Mean age was 51,2 years. Conjunctivitis (100%), lacrimation (94%), foreign body sensation (83%), and dim vision (76%) were the main clinical symptoms. Both eyes were affected in half of the cases. Direct contact was the main transmission route including nosocomial spread associated with ophthalmology practices. Five (62.5%) of 8 conjunctival swabs were PCR-positive for adenovirus type 8 (HAdV8/Baranya/2006/HUN; EF210714) which was genetically identical to adenovirus strain detected in Austria in 2004 (DQ149614). CONCLUSIONS The outbreak of keratoconjunctivitis was partially associated with nosocomial infection caused by type 8 adenovirus. Both the recognition of the clinical illness, laboratory diagnosis and public health measures are necessary for the prevention of keratoconjunctivitis infection and epidemic.

Adenovírus 8-as típusa okozta járványos keratoconjunctivitis molekuláris diagnosztikája@@@Molecular detection of adenovirus type 8 epidemic keratoconjunctivitis in Hungary

INTRODUCTION Both infectious and non-infectious forms of acute conjunctivitis are known. Viruses, especially different types of adenoviruses are the etiological agents of infectious epidemic conjunctivitis (conjunctivitis epidemica). AIMS The authors aims were to describe an outbreak of keratoconjunctivitis and to detect the viral agent by molecular methods in Hungary. MATERIALS AND METHODS Classical epidemiological methods were used for investigation. Polymerase chain reaction (PCR) followed by sequencing were used for the detection of adenoviral hexon region from freshly collected conjunctival swabs. RESULTS Between 9 October and 18 December 2006, a total of 60 patients became ill with keratoconjunctivitis in 7 settlements in Southwest Hungary. Mean age was 51,2 years. Conjunctivitis (100%), lacrimation (94%), foreign body sensation (83%), and dim vision (76%) were the main clinical symptoms. Both eyes were affected in half of the cases. Direct contact was the main transmission route including nosocomial spread associated with ophthalmology practices. Five (62.5%) of 8 conjunctival swabs were PCR-positive for adenovirus type 8 (HAdV8/Baranya/2006/HUN; EF210714) which was genetically identical to adenovirus strain detected in Austria in 2004 (DQ149614). CONCLUSIONS The outbreak of keratoconjunctivitis was partially associated with nosocomial infection caused by type 8 adenovirus. Both the recognition of the clinical illness, laboratory diagnosis and public health measures are necessary for the prevention of keratoconjunctivitis infection and epidemic.

Molecular detection of adenovirus type 8 epidemic keratoconjunctivitis in Hungary

INTRODUCTION Both infectious and non-infectious forms of acute conjunctivitis are known. Viruses, especially different types of adenoviruses are the etiological agents of infectious epidemic conjunctivitis (conjunctivitis epidemica). AIMS The authors aims were to describe an outbreak of keratoconjunctivitis and to detect the viral agent by molecular methods in Hungary. MATERIALS AND METHODS Classical epidemiological methods were used for investigation. Polymerase chain reaction (PCR) followed by sequencing were used for the detection of adenoviral hexon region from freshly collected conjunctival swabs. RESULTS Between 9 October and 18 December 2006, a total of 60 patients became ill with keratoconjunctivitis in 7 settlements in Southwest Hungary. Mean age was 51,2 years. Conjunctivitis (100%), lacrimation (94%), foreign body sensation (83%), and dim vision (76%) were the main clinical symptoms. Both eyes were affected in half of the cases. Direct contact was the main transmission route including nosocomial spread associated with ophthalmology practices. Five (62.5%) of 8 conjunctival swabs were PCR-positive for adenovirus type 8 (HAdV8/Baranya/2006/HUN; EF210714) which was genetically identical to adenovirus strain detected in Austria in 2004 (DQ149614). CONCLUSIONS The outbreak of keratoconjunctivitis was partially associated with nosocomial infection caused by type 8 adenovirus. Both the recognition of the clinical illness, laboratory diagnosis and public health measures are necessary for the prevention of keratoconjunctivitis infection and epidemic.

Comparison of clinical characteristics between astrovirus and rotavirus infections diagnosed in 1997 to 2002 in Hungary: Astrovirus and rotavirus gastroenteritis

Aim: To determine the severity and clinical characteristics of human astrovirus (HAstV) infections among hospitalized children and compare them with children infected by rotavirus. Methods: Retrospective, case‐control study of astrovirus‐infected and rotavirus‐infected children. Astroviruses were detected in stool samples by enzyme immunoassay and/or reverse transcriptase‐polymerase chain reaction. All stool samples were tested for rotavirus and bacterial pathogens, and all negative samples were further tested for human astrovirus. Children with astrovirus‐positive stool samples and complete clinical data were included in this study. Results: Astrovirus was detected in 29 (1.8%) children, and 63 rotavirus‐infected children were included as controls. Astrovirus‐infected children had shorter duration of diarrhea than rotavirus‐infected children (median 4 and 6 d, respectively; p<0.05), and 79% of the astrovirus infections were associated with a short duration of vomiting (median 1 and 4 d, respectively; p<0.0001). Rotavirus‐infected children had longer hospitalization (p<0.050) than astrovirus‐infected children.