Rapid spread and emergence of heartworm resulting from climate and climate-driven ecological changes in Hungary.

In Europe, Dirofilaria immitis persists mainly in the southern countries with a Mediterranean climate. Because spreading of heartworms from these countries towards the northern ones could be observed in the past decades, necropsy records of 4076 Hungarian dogs were reviewed for heartworm infections. The first autochthonous canine D. immitis case was detected on the Great Hungarian Plain in 2007. Until 2011, the number of heartworm infection cases was low, and these cases were restricted to a small part of the Great Hungarian Plain. Since 2012, the number of cases has increased considerably, and the rapid expansion of the parasite's geographic range could also be observed. Our retrospective study has revealed that most of the Hungarian territory became a heartworm endemic region, and the prevalence of infection greatly multiplied over the past 12 years. The establishment, rapid spread, and emergence of D. immitis may be mainly explained by the warming climate in Hungary. However, the partly climate-driven spread of the most important reservoir host in wildlife, the golden jackal (Canis aureus) from the Mediterranean Balkan Peninsula might have also played a significant role. This study is an example of the rapid spread and emergence of pathogens resulting from climate and climate-driven ecological changes. Because a continuous increase in the temperature and further dispersal of golden jackals in Europe are projected, further spread and emergence of heartworm can be expected. Similar spread and emergence of D. immitis could be observed in North America. It cannot be excluded that similar reasons (global warming and rapid dispersal and population growth of the most important wild canine reservoir host) are in the background on both continents.

Establishment of Fascioloides magna in a new region of Hungary: case report.

During the monitoring of red deer (N = 124) and fallow deer (N = 13) populations in four neighbouring areas, the presence of Fascioloides magna was confirmed in southwestern Hungary. The prevalence and the mean intensity of the infection within the host populations ranged between 0 and 100% and 0-36.3, respectively. The determined prevalences are similar to that observed earlier in other European natural foci. The authors hypothesise that the appearance of F. magna in this region should have been a partly natural- and partly human-influenced process.

Data on the epidemiology and pathology of anatipestifer disease in Hungary (2010-2014).

Anatipestifer disease is a contagious disease caused by Riemerella anatipestifer, affecting primarily ducks, geese and turkeys, and characterised by listlessness, diarrhoea, sneezing, nasal discharge, and nervous signs. Sporadically, it occurs in a wide range of other domesticated and wild birds as well. The incidence and characteristics of the disease seen in the three main host species are summarised based on birds submitted for routine laboratory investigation in Hungary over the period 2010-2014. The infection was diagnosed in a higher percentage in geese (9.9%) and ducks (7.5%). It occurred in 5-day-old to 17-week-old geese and 3- to 6.5-week-old ducks, respectively. The pathological lesions were comparable in these two species: enlarged spleen, serofibrinous pericarditis, perihepatitis, airsacculitis, catarrhal enteritis, subcutaneous oedema and hyperaemia over the cranium, mucopurulent exudate in the nasal cavity and occasionally pneumonia, conjunctivitis, purulent arthritis and caseous salpingitis. In some cases, R. anatipestifer produced only secondary lesions, which complicated other diseases such as circovirus infection, mycotoxicosis, mycoplasmosis, or Derzsy's disease. In turkeys, the disease occurred rarely (0.5%) and at an older age (12 to 19 weeks). The lesions most frequently seen were purulent osteomyelitis of the cranium and seropurulent meningitis. Purulent osteomyelitis in the cranium caused by R. anatipestifer infection had not been reported in turkeys previously. To various extents, other local lesions such as serofibrinous pericarditis, airsacculitis, arthritis, and in one case septicaemia were also observed. The high incidence of the disease in waterfowl underlines the importance of appropriate treatment and prevention that should be based on accurate diagnosis and antimicrobial susceptibility testing, proper biosecurity and vaccination with regard to the serotype(s) present on the farm.

Molecular characterization of a novel picobirnavirus in a chicken.

Picobirnaviruses (PBVs) are bisegmented viruses with a wide geographical and host species distribution. The number of novel PBV sequences has been increasing with the help of the viral metagenomics. A novel picobirnavirus strain, pbv/CHK/M3841/HUN/2011, was identified by viral metagenomics; the complete segment 1 (MH327933) and 2 (MH327934) sequences were obtained by RT-PCR from a cloacal sample of a diseased broiler breeder pullet in Hungary. Although the conserved nucleotide (e.g., ribosome binding site) and amino acid motifs (e.g., ExxRxNxxxE, S-domain of the viral capsid and motifs in the RNA-dependent RNA polymerase) were identifiable in the chicken picobirnavirus genome, the putative segment 1 showed low (< 30%) amino acid sequence identity to the corresponding proteins of marmot and dromedary PBVs, while segment 2 showed higher (< 70%) amino acid sequence identity to a wolf PBV protein sequence. This is the first full-genome picobirnavirus sequence from a broiler breeder chicken, but the pathogenicity of this virus is still questionable.

Characterization of Ornithobacterium rhinotracheale field isolates from Hungary.

Ornithobacterium rhinotracheale is a widely distributed rod-shaped Gram-negative bacterium that infects several avian species including chickens and turkeys. It is associated with respiratory signs, growth retardation, mortality, and reduced egg production, thus causing severe economic losses to the poultry industries. In this study, 37 field isolates of O. rhinotracheale, collected from various locations in Hungary between 1997 and 2015, were identified and characterized by the analysis of partial 16S rRNA gene sequences, enterobacterial repetitive intergenic consensus (ERIC)-PCR, and random amplified polymorphic DNA (RAPD) PCR assays with the OPG11, OPH19, and M13 primers. Most of the field isolates were serotype A, one was serotype B, and four were serotype D. One isolate could not be typed with antisera against serotypes A-E. In a phylogenetic analysis of the 16S rRNA sequences, the isolates formed two clusters. Thirteen distinct patterns were identified with ERIC-PCR, and the RAPD assay with the M13 primer assigned the isolates to 10 different patterns. The other two RAPD assays were unsuitable for distinguishing and grouping the isolates. Neither ERIC type nor RAPD pattern correlated with the place or year of isolation. However, the strains isolated from chickens were more heterogeneous on ERIC-PCR than the isolates recovered from turkeys. In this study, ERIC-PCR was the most discriminatory method for investigating the genetic diversity of O. rhinotracheale isolates.

Lethal Angiostrongylus vasorum infection in a Hungarian dog.

During post-mortem examination of lungs and heart of a 7-month-old female French bulldog, 158 worms were collected from the lung vessels and they proved Angiostrongylus vasorum by their morphological and genetic identification by PCR. The histopathological investigation found a multifocal interstitial inflammation characterized by numerous lymphocytes and a smaller number of plasma cells and eosinophils whilst L1 stage larvae could be seen inside dilated alveoli. We suggested a lethal angiostrongylosis supposed to lead to a fatal effect. Our report attracts attention to the presence of the nematode A. vasorum as causative agent of canine cardiopulmonary disorder in the south-western region of Hungary.

Occurrence of Coxiella burnetii and Chlamydiales species in abortions of domestic ruminants and in wild ruminants in Hungary, Central Europe.

Coxiella burnetii and certain members of the Chlamydiales order are zoonotic, intracellular, Gram-negative bacteria, with abortigenic potential in ruminants. These pathogens have a broad host range and worldwide geographical distribution. The current study aimed to reveal the importance of C. burnetii and Chlamydiales spp. in abortions in domestic ruminants and their occurrence in wild ruminants with real-time polymerase chain reaction (qPCR) assays, histology, and immunohistochemical staining (IHC). From the 111 abortion cases of domestic ruminants examined, C. burnetii was detected in 33 placenta samples (cattle, n = 22; sheep, n = 10; goat, n = 1), and members of the Chlamydiales order were detected in 32 placenta samples (cattle, n = 14; sheep, n = 16; goat, n = 2) using qPCR. Coinfection with both C. burnetii and Chlamydiales spp. were identified in 12 cases (cattle, n = 3; sheep, n = 8; goat, n = 1) out of the qPCR-positive samples. The presence of the relevant antigen was confirmed by IHC in 20 cases (C. burnetii, n = 2, in sheep; Chlamydiaceae, n = 17, in sheep [n = 15] and goat [n = 2]; and both pathogens in 1 sheep). Coxiella burnetii was identified in 2.2% (2/91) of the wild ruminants, but the samples were negative by IHC. Uncultured Chlamydiales spp. were detected in 4.4% (4/91) of the placenta samples by qPCR. In conclusion, Q fever is widespread among domestic ruminants in Hungary, and, in several cases, C. burnetii was implicated as the primary cause of abortions. Waddlia chondrophila, Parachlamydia spp., and uncultured Chlamydiales spp. were present only sporadically in samples from cattle and wild ruminants.

Comparative complete genome analysis of chicken and Turkey megriviruses (family picornaviridae): long 3' untranslated regions with a potential second open reading frame and evidence for possible recombination.

UNLABELLED: Members of the family Picornaviridae consist of small positive-sense single-stranded RNA (+ssRNA) viruses capable of infecting various vertebrate species, including birds. One of the recently identified avian picornaviruses, with a remarkably long (>9,040-nucleotide) but still incompletely sequenced genome, is turkey hepatitis virus 1 (THV-1; species Melegrivirus A, genus Megrivirus), a virus associated with liver necrosis and enteritis in commercial turkeys (Meleagris gallopavo). This report presents the results of the genetic analysis of three complete genomes of megriviruses from fecal samples of chickens (chicken/B21-CHV/2012/HUN, GenBank accession no. KF961186, and chicken/CHK-IV-CHV/2013/HUN, GenBank accession no. KF961187) (Gallus gallus domesticus) and turkey (turkey/B407-THV/2011/HUN, GenBank accession no. KF961188) (Meleagris gallopavo) with the largest picornavirus genome (up to 9,739 nucleotides) so far described. The close phylogenetic relationship to THV-1 in the nonstructural protein-coding genome region and possession of the same internal ribosomal entry site type (IVB-like) suggest that the study strains belong to the genus Megrivirus. However, the genome comparisons revealed numerous unique variations (e.g., different numbers of potential 2A peptides, unusually long 3' genome parts with various lengths of a potential second open reading frame, and multiple repeating sequence motifs in the 3' untranslated region) and heterogeneous sequence relationships between the structural and nonstructural genome regions. These differences suggest the classification of chicken megrivirus-like viruses into a candidate novel species in the genus Megrivirus. Based on the different phylogenetic positions of chicken megrivirus-like viruses at the structural and nonstructural genome regions, the recombinant nature of these viruses is plausible. IMPORTANCE: The comparative genome analysis of turkey and novel chicken megriviruses revealed numerous unique genome features, e.g., up to four potential 2A peptides, unusually long 3' genome parts with various lengths containing a potential second open reading frame, multiple repeating sequence motifs, and heterogeneous sequence relationships (possibly due to a recombination event) between the structural and nonstructural genome regions. Our results could help us to better understand the evolution and diversity (in terms of sequence and genome layout) of picornaviruses.

Genetic characterization of a novel picornavirus in turkeys (Meleagris gallopavo) distinct from turkey galliviruses and megriviruses and distantly related to the members of the genus Avihepatovirus.

This study reports the metagenomic detection and complete genome characterization of a novel turkey picornavirus from faecal samples of healthy (1/3) and affected (6/8) commercial turkeys with enteric and/or stunting syndrome in Hungary. The virus was detected at seven of the eight farms examined. The turkey/M176-TuASV/2011/HUN genome (KC465954) was genetically different from the currently known picornaviruses of turkey origin (megriviruses and galliviruses), and showed distant phylogenetic relationship and common genomic features (e.g. uncleaved VP0 and three predicted and unrelated 2A polypeptides) to duck hepatitis A virus (DHAV) of the genus Avihepatovirus. The complete genome analysis revealed multiple distinct genome features like the presence of two in-tandem aphthovirus 2A-like sequence repeats with DxExNPG/P 'ribosome-skipping' sites (76 %, 23/30 amino acids identical), with the first aphthovirus 2A-like sequence being located at the end of the VP1 capsid protein (VP1/2A1 'ribosome-skipping' site). The phylogenetic analyses, low sequence identity (33, 32 and 36 % amino acid identity in P1, P2 and P3 regions) to DHAV, and the type II-like internal ribosome entry site suggests that this turkey picornavirus is related to, but distinct from the genus Avihepatovirus and it could be the founding member of a novel Avihepatovirus sister-clade genus. This is the third, taxonomically highly distinct picornavirus clade identified from turkeys exhibiting varied symptoms.

Porcine kobuvirus in wild boars (Sus scrofa).

Fecal samples (N = 10) from 6- to 8-week-old wild boar piglets (Sus scrofa), collected from an animal park in Hungary in April 2011, were analyzed using viral metagenomics and complete genome sequencing. Kobuvirus (genus Kobuvirus, family Picornaviridae) was detected in all (100 %) specimens, with the closest nucleotide (89 %) and amino acid (94 %) sequence identity of the strain wild boar/WB1-HUN/2011/HUN (JX177612) to the prototype porcine kobuvirus S-1-HUN (EU787450). This study suggests that genetically highly similar (practically the same geno-/serotype) porcine kobuvirus circulate in wild boars, the wildlife counterparts of domestic pigs. Wild boars could be an important host and reservoir for kobuvirus.

Identification and complete genome characterization of a novel picornavirus in turkey (Meleagris gallopavo).

Members of the family Picornaviridae are important pathogens of humans and animals, although compared with the thousands of known bird species (>10 000), only a few (n = 11) picornaviruses have been identified from avian sources. This study reports the metagenomic detection and complete genome characterization of a novel turkey picornavirus from faecal samples collected from eight turkey farms in Hungary. Using RT-PCR, both healthy (two of three) and affected (seven of eight) commercial turkeys with enteric and/or stunting syndrome were shown to be shedding viruses in seven (88 %) of the eight farms. The viral genome sequence (turkey/M176/2011/HUN; GenBank accession no. JQ691613) shows a high degree of amino acid sequence identity (96 %) to the partial P3 genome region of a picornavirus reported recently in turkey and chickens from the USA and probably belongs to the same species. In the P1 and P2 regions, turkey/M176/2011/HUN is related most closely to, but distinct from, the kobuviruses and turdivirus 1. Complete genome analysis revealed the presence of characteristic picornaviral amino acid motifs, a potential type II-like 5' UTR internal ribosome entry site (first identified among avian-origin picornaviruses) and a conserved, 48 nt long 'barbell-like' structure found at the 3' UTR of turkey/M176/2011/HUN and members of the picornavirus genera Avihepatovirus and Kobuvirus. The general presence of turkey picornavirus - a novel picornavirus species - in faecal samples from healthy and affected turkeys in Hungary and in the USA suggests the worldwide occurrence and endemic circulation of this virus in turkey farms. Further studies are needed to investigate the aetiological role and pathogenic potential of this picornavirus in food animals.

Porcine teschovirus in wild boars in Hungary.

The genus Teschovirus, family Picornaviridae, currently includes 12 serotypes (PTV 1 to 12) isolated from swine. PTVs have been well studied in domestic pigs, but knowledge about PTVs in wild boars is deficient. Here, we report the first complete PTV genome sequence from 7 (70 %) of 10 fecal samples of wild boar piglets (Sus scrofa) by RT-PCR and pyrosequencing. Analysis of the wild boar PTV strain WB2C-TV/2011/HUN (JQ429405) showed considerable difference, especially in VP1 (66-74 % amino acid identity) compared to the available PTVs. PTV is present in wild boars, and WB2C-TV/2011/HUN represents a novel PTV genotype, provisionally named PTV-13.

Astrovirus in wild boars (Sus scrofa) in Hungary.

The family Astroviridae consists of two genera, Avastrovirus and Mamastrovirus whose members are associated with gastroenteritis in avian and mammalian hosts, respectively. In this study, we report the first detection of astrovirus from fecal specimens of wild boars (Sus scrofa) using viral metagenomics and complete genome sequencing. The wild boar astrovirus (WBAstV-1/2011/HUN, JQ340310) genome is 6707 nucleotide long and had 76%, 95% and 56% amino acid (aa) identity in the ORF1a (852aa), ORF1b (522aa) and ORF2 (845aa) regions, respectively, to porcine astrovirus 4 (PAstV-4, JF713713), the closest match. This study indicates that wild boar could be a reservoir for astroviruses.

Characterization of a novel porcine enterovirus in wild boars in Hungary.

Porcine enteroviruses (PEVs) are members of the family Picornaviridae, genus Enterovirus. Until now, only three different PEV genotypes (PEV-9 and -10, and PEV-3H/PEV-14) have been detected in domestic pigs, and there is no information about the presence of PEVs in wild animals. Here, we identify and characterize the complete genomes of PEV originated from 5 of 10 (50%) of wild boar (Sus scrofa) piglets by RT-PCR and pyrosequencing. Wild boar/WBD/2011/HUN (JN807387) PEV showed only 67% amino acid identity in VP1 compared to the most closely related prototype PEV-3H/PEV-14. Wild boar enterovirus represents a novel PEV genotype, provisionally called PEV-15.

High prevalence of turkey parvovirus in turkey flocks from Hungary experiencing enteric disease syndromes.

Samples collected in 2008 and 2009, from 49 turkey flocks of 6 to 43 days in age and presenting clinical signs of enteric disease and high mortality, were tested by polymerase chain reaction and reverse transcription-polymerase chain reaction for the presence of viruses currently associated with enteric disease (ED) syndromes: astrovirus, reovirus, rotavirus, coronavirus, adenovirus, and parvovirus. Turkey astroviruses were found in 83.67% of the cases and turkey astrovirus 2 (TAst-2) in 26.53%. The investigations directly demonstrated the high prevalence of turkey parvovirus (TuPV) in 23 flocks (46.9%) experiencing signs of ED, making this pathogen the second most identified after astroviruses. Phylogenetic analysis on a 527 base pair-long region from the NS1 gene revealed two main clusters, a chicken parvovirus (ChPV) and a TuPV group, but also the presence of a divergent branch of tentatively named "TuPV-like ChPV" strains. The 23 Hungarian TuPV strains were separately positioned in two groups from the American origin sequences in the TuPV cluster. An Avail-based restriction fragment length polymorphism assay has also been developed for the quick differentiation of TuPV, ChPV, and divergent TuPV-like ChPV strains. As most detected enteric viruses have been directly demonstrated in healthy turkey flocks as well, the epidemiology of this disease complex remains unclear, suggesting that a certain combination of pathogens, environmental factors, or both are necessary for the development of clinical signs.

Association of Enterococcus cecorum with vertebral osteomyelitis and spondylolisthesis in broiler parent chicks.

Enterococcus cecorum is the most frequently occurring enterococcal species in the intestine of chickens of over 12 weeks of age, and there are few reports on its isolation from the skeleton of broiler parent chicks. In the present study, observations on vertebral osteomyelitis and spondylolisthesis ('kinky back syndrome') showing high incidence in 8 broiler parent flocks in different parts of Hungary are summarised. Clinical signs were seen only in roosters between 5 and 13 weeks of age. Diseased birds were alert and remained sitting on their hocks with their feet slightly raised off the ground. Incidence of the disease among male birds ranged from 8% to 30% depending on flocks. Enlargement and distortion of the body of the 6th vertebra were seen as the main pathological lesions. The cavity of the spinal canal was constricted by the distorted vertebral bodies. Resorption of bone tissue and sequestrum formation, signs of increased osteoclast activity, proliferation of fibrotic tissues, infiltration with heterophils and formation of sclerotic layers were detected in the vertebral bodies. From all 24 samples collected from the vertebral lesions, Enterococcus cecorum was isolated and identified using metabolic fingerprinting as well as 16S rRNA gene sequencing. Demonstration of E. cecorum from the vertebral lesions in all examined broiler breeder roosters showing the same clinical and pathological findings in different flocks suggested the pathogenic role of this microorganism for the first time in Hungary.

Harvey Cushing, a pioneer of neuroanesthesia.

Harvey Cushing's name is most frequently mentioned in conjunction with Cushing's syndrome, and Cushing's reflex following raised intracranial pressure. The aim of this review is to pay tribute to Cushing's contribution to anesthesia. Besides his own specialty, he used an anesthesia chart for the first time, he introduced blood pressure measurement and precordial auscultation to anesthesiological practice, he employed the first independent neurosurgical anesthetist, and he described the terminology of regional anesthesia.

[Adverse cognitive effects and ECT]. / Kognitive Störungen und Elektrokrampftherapie.

Electroconvulsive therapy (ECT) is a rapidly acting and highly effective treatment for severe and life threatening conditions seen in affective and schizophrenic diseases. Notwithstanding its therapeutic benefits, ECT remains controversial because of seizure induction, cognitive side effects, memory dysfunction and effects on cerebral physiology. These factors have raised the concern that ECT produces structural and functional brain damages. This issue continues to have a major impact on the acceptance of ECT as a therapeutic modality, both within the medical community and in public opinion. A close look at incidence, type, severity, neurofunctional and -anatomical correlates, aetiology and therapeutic approaches of the adverse cognitive effects attributed to ECT may contribute to rational and objective handling of this topic. The final chapter deals with the issue of whether ECT causes brain damage.

Phylogenetic diversity of avian nephritis virus in Hungarian chicken flocks.

Avian nephritis virus (ANV) infection was detected in 4-day-old to 22-day-old chickens collected on Hungarian farms between 2002 and 2005. The animals suffered from diarrhoea, growth retardation, runting-stunting syndrome, and 2 to 6% mortality was reported. Tubulonephrosis, interstitial nephritis and uricosis (gout) was diagnosed. The presence of ANV RNA was detected in chicken carcasses using reverse transcriptase-polymerase chain reaction. The virus was demonstrated in 69% of the investigated farms. The nucleotide sequence of the amplification products (corresponding to part of the genome that encodes the GP1 protein) was determined and phylogenetic analysis was performed. The nucleotide sequences showed 76 to 86% identity to the reference strain isolated in 1976 in Japan. The constructed phlyogenetic tree indicates high diversity of the Hungarian ANV sequences, regardless of their origin and year of sample collection. Analysis of the putative amino acid sequences encoded by the partial GP1 sequences also revealed high diversity of the virus. Even samples collected at the same farm, at the same time but from different flocks, differed in nucleotide and putative amino acid sequences. The possible effects of the sequence diversity on the pathogenicity, antigenicity and diagnostics of ANV are discussed.