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Group A rotaviruses are the main cause of acute dehydrating diarrhea in children, responsible for high mortality in developing countries and a significant socio-economic burden associated with treating the disease in developed countries. Two rotavirus vaccine candidates predicated on either homotypic or heterotypic protection have undergone clinical trials recently and await licensure for routine use. In anticipation of a future vaccination campaign in Hungary, the diversity of rotaviruses collected from Budapest between 2000 and 2003 were analyzed by polyacrylamide gel electrophoresis (PAGE) of the viral genome and by serotyping and genotyping of the outer capsid genes, VP7 and VP4. Among 2,763 rotavirus positive specimens available for analysis, we were able to determine the electropherotype of 2,227, and, of these, 1,517 (68.1%) were subjected to G typing and 1,173 (52.7%) were subjected to P typing. We successfully G typed 1,481 (97.6%) and P typed 1,130 (96.3%) strains, respectively. A total of six G types (G1, 50.2%; G2, 2.2%; G3, 1.7%; G4, 5.8%; G6, 0.6%; and G9, 34.4%) and four P types (P[4], 3.0%; P[6], 0.7%; P[8], 89.9%; and P[9], 1.7%) were identified in nine individual combinations (P[8],G1; P[4],G2; P[8],G3; P[8],G4; P[8],G9; P[6],G4; P[4],G1; P[9],G3; and P[9],G6). The prevalence of VP7 and VP4 specificities varied from year to year. In this regard, a shift in serotype predominance from G1 in 2000-2001 (61.8%) and 2001-2002 (69.7%) to G9 in 2002-2003 (51.3%) was an intriguing observation that has been reported recently in some other countries, as well. The emergence of serotype G9 rotaviruses in Hungary and other parts of the world may have implications for future vaccine development and use, particularly, if current vaccine candidates cannot confer adequate homotypic or heterotypic protection against these 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.

Human picobirnaviruses characterised in this study were serendipitously detected in a non-bacterial gastroenteritis outbreak when specimens were examined for the presence of human rotaviruses using polyacrylamide gel electrophoresis. Of ten stool samples sent for virological examination, two, three, and one specimens were positive for human caliciviruses, picobirnaviruses, and both viruses, respectively. Partial sequences of the RNA-dependent RNA polymerase gene were determined for three picobirnavirus-positive samples. The sequence identity among these three strains was 60% to 65% for the nucleic acid and 64% to 70% for the deduced amino acid sequences. Phylogenetic analysis revealed that each of the three strains clustered with strains identified in geographically separate areas. In contrast, human calicivirus strains co-incidentally identified, showed complete nucleotide sequence identity. These findings demonstrate a lack of common exposure to or point of source for picobirnavirus infection, suggesting that the outbreak was caused by human caliciviruses. Further studies are needed to determine the etiologic role and to establish the taxonomic basis of picobirnaviruses.

1Hepatitis E Virus (HEV) is an enterically transmitted human pathogen. HEV infections 2 are mainly associated with acute, self-limited, icteric hepatitis with an average mortality rate 3 of 1%. Animal reservoirs are considered to play an important role in the maintenance of the 4 virus and in the spread of HEV to humans. HEV-induced seroconversion was described in 5 several species, however clinical hepatitis in animals has not been observed to date. HEV 6 strains from animals are genetically closely related to human HEV isolates, which supports 7 the opinions on the zoonotic transmission of the virus. 8In this expansive study the occurrence of HEV was investigated in Hungarian wild and 9 domesticated animal samples. HEV RNA was detected by reverse transcription-polymerase 10 chain reaction in liver samples of wild boars, roe deer, and deer. The investigations of 11 domestic swine samples detected HEV in 39% of the investigated Hungarian pig farms. 12Simultaneous investigation revealed no definite difference between liver and faeces samples 13 of domestic pigs in the frequency of HEV positivity. The highest (36%) incidence of HEV 14 infection was found among the eleven to sixteen week old pigs. Samples from domestic cattle 15 and rodents collected in pig farms, forests and meadows were tested negative for HEV RNA.

Between January 2001 and December 2003, stool specimens from 262 (45%) of 581 reported outbreaks of gastroenteritis were investigated for noroviruses in Hungary. Specimens collected from outbreaks of non-bacterial gastroenteritis were examined by reverse-transcription polymerase chain reaction and enzyme immunoassay. In 253 (97%) of 262 outbreaks, norovirus was detected and confirmed by sequencing in 211 (81%). Hospitals (35%), day care centers (30%), and elderly homes (27%) were the most common settings. Diversity and frequency of the genotypes changed over time but with predominance (95%) of genogroup (GG) II strains. Strains grouped into 11 genotypes including an epidemic spread of new-variant GGII4 (Lordsdale virus) and a recently emerged group of natural recombinant strains (GGIIb/Hilversum polymerase) with four capsid types (Hawaii, Mexico, Snow Mountain, and Lordsdale). Clusters of epidemics including food-borne outbreaks were detected. According to this study, noroviruses are the predominant etiological agents causing gastroenteritis outbreaks in Hungary.