Sequence analysis of Schmallenberg virus genomes detected in Hungary.

Since its emergence near the German-Dutch border in 2011, Schmallenberg virus (SBV) has been identified in many European countries. In this study, we determined the complete coding sequence of seven Hungarian SBV genomes to expand our knowledge about the genetic diversity of circulating field strains. The samples originated from the first case, an aborted cattle fetus without malformation collected in 2012, and from the blood samples of six adult cattle in 2014. The Hungarian SBV sequences shared ≥99.3% nucleotide (nt) and ≥97.8% amino acid (aa) identity with each other, and ≥98.9 nt and ≥96.7% aa identity with reference strains. Although phylogenetic analyses showed low resolution in general, the M sequences of cattle and sheep origin SBV strains seemed to cluster on different branches. Both common and unique mutation sites were observed in different groups of sequences that might help understanding the evolution of emerging SBV strains.

Emergence and characterisation of pandemic H1N1 influenza viruses in Hungarian swine herds.

In 2010, two novel porcine H1N1 influenza viruses were isolated from pigs with influenza-like illness in Hungarian swine herds. Sequence and phylogenetic analysis of these strains revealed that they shared molecular features with the pandemic H1N1 influenza virus strains, which emerged globally during 2009. The PB2, HA and NA genes contained unique amino acid changes compared to the available new H1N1 influenza virus sequences of pig origin. Furthermore, the investigated strains could be separated with respect to parallel amino acid substitutions affecting the polymerase genes (PB2, PB1 and PA) and the nucleoprotein (NP) gene, supporting the proposed complementarities between these proteins, all required for the viral fitness. Molecular characterisation of two Hungarian human pandemic H1N1 isolates was also performed, so that we could compare contemporaneous strains of different host species origins. Shared molecular motifs in various genes of animal and human influenza strains suggested that the Hungarian porcine strains could have originated from humans through direct interspecies transmission. This study is among the few that support the natural human-to-pig transmission of the pandemic H1N1 influenza virus.

Genome sequence of a monoreassortant H1N1 swine influenza virus isolated from a pig in Hungary.

The genome of a porcine H1N1 influenza A strain is reported in this study. The strain proved to be a monoreassortant strain with a typical porcine N1 gene on the genetic backbone of the pandemic H1N1 influenza A virus strain. Monitoring of descendants of the pandemic 2009 H1N1 strain is needed because of concerns that more-virulent strains may emerge in forthcoming epidemic seasons.

Putative novel genotype of avian hepatitis E virus, Hungary, 2010.

To explore the genetic diversity of avian hepatitis E virus strains, we characterized the near-complete genome of a strain detected in 2010 in Hungary, uncovering moderate genome sequence similarity with reference strains. Public health implications related to consumption of eggs or meat contaminated by avian hepatitis E virus, or to poultry handling, require thorough investigation.

Novel circovirus in European catfish (Silurus glanis).

Circular single-stranded DNA viral genomes had been identified worldwide in different species and in environmental samples. Among them, viruses belonging to the genus Circovirus of the family Circoviridae are present in birds and pigs, and recently, they were detected in barbels. The present study reports the identification of a new circovirus in fish. PCR amplification and sequencing were used to identify the novel circular DNA virus in European catfish (Silurus glanis). Full genome characterization and phylogenetic analysis showed that the virus belonged to the family Circoviridae and that it was distantly related to the previously described barbel circovirus.

Detection of Borrelia burgdorferi Sensu Lato and Anaplasma phagocytophilum in small mammals and ectoparasites in Hungary.

The aim of our study was to investigate the presence of Borrelia burgdorferi sensu lato (s.l.) and Anaplasma phagocytophilum in small mammals and ticks using polymerase chain reaction and to gain information about the prevalence and possible coexistence of these pathogens at a selected site in Hungary. Two hundred seventy-seven small mammals were trapped in South-Eastern Hungary during 2009. Tissue samples and a total of 831 ectoparasites (Ixodes ricinus, Ixodes acuminatus, Haemaphysalis concinna, Ctenophtalmus assimilis, and Nosopsyllus fasciatus) were collected from small mammals. One thousand one hundred and six I. ricinus and 476 H. concinna were collected from the vegetation during the investigation. Neither A. phagocytophilum nor B. burgdorferi s.l. was detected in any of the mammal tissue samples. A. phagocytophilum was not found in ticks collected from small mammals. Very low minimum prevalence was found for all pathogens (0.62% for Borrelia afzelii in ticks collected from small mammals, and 0.57%, 0.06%, and 0.19% for A. phagoctyophilum, B. afzelii, and Borrelia garinii, respectively, in questing ticks). The present study is the first report of borreliae from I. acuminatus and H. concinna from Hungary.

Molecular epidemiology and antimicrobial susceptibility of Campylobacter jejuni and Campylobacter coli isolates of poultry, swine, and cattle origin collected from slaughterhouses in Hungary.

Campylobacter spp. are the most common cause of bacterial enteritis in Hungary, and the aim of this study was to identify the distribution, genotypes, and antimicrobial susceptibility of Campylobacter species in the most important food-producing animals at the time of slaughter during 2008 and 2009. Of 1,110 samples, 266 were identified as Campylobacter coli (23.9%) and 143 as C. jejuni (12.9%) by real-time PCR. Resistance to enrofloxacin-ciprofloxacin and nalidixic acid was significant, especially in C. jejuni (73.3%) and C. coli (77.2%) from broilers. Higher erythromycin (P = 0.043) and tetracycline (P = 1.865e-14) resistance rates were found among C. coli isolates (9.7 and 74.1%, respectively) than among C. jejuni isolates (3.1 and 36.6%, respectively). A total of 47 fla short variable region sequences were identified among 73 selected C. coli and C. jejuni isolates, with 35 fla types detected only once. At the nucleotide level, fla types A66 and A21 were the most common. Using the pulsed-field gel electrophoresis method, 66% of strains exhibited unique profiles after Sma I digestion. Forty-two isolates assigned to 18 Sma I clusters were further typed by Kpn I, and of these, 24 were assigned to 10 Kpn I clusters. For isolates in five Kpn I clusters, epidemiological links were observed. Stable C. jejuni and C. coli clones were detected, indicating that further studies involving broiler and human isolates need to be conducted to elucidate the importance of these stable clones in human infections.

Molecular characterization of Campylobacter lanienae strains isolated from food-producing animals.

During 2008 and 2009, within the framework of the Hungarian monitoring program of antibiotic resistance of zoonotic agents from food-producing animals, a significant number (43 strains) of Campylobacter lanienae were detected for the first time in Hungary. The isolates were genotyped using partial 16S rRNA gene sequencing and pulsed-field gel electrophoresis using three different restriction enzymes. The antimicrobial resistance of the isolates was determined by microtiter broth dilution. C. lanienae isolation was successful only from swine but not from other animal species. According to phylogenetic analysis, clustering of the isolates shows the same extensive genetic diversity as other Campylobacter species. Sequence analysis of the partial 16S rRNA gene showed that additional variations exist in variable regions Vc2 and Vc6. SmaI restriction enzyme proved to be the most efficient for pulsed-field gel electrophoresis analysis of C. lanienae. A significant tetracycline resistance (60.9%) and the presence of erythromycin-, enrofloxacin-, and multiresistant C. lanienae strains were found. Although the pathogenic potential of C. lanienae in humans is currently unknown, this study demonstrates that C. lanieanae is common in pigs in the country, provides further details on the genotypic and phenotypic properties of C. lanienae, and offers a genotyping method for use in source tracing.