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The spread of lineage 2 West Nile virus (WNV) from sub-Saharan regions to Europe and the unpredictable change in pathogenicity indicate a potential public and veterinary health threat and requires scientific awareness. To describe the results of clinical and virological investigations of the 1st outbreak of a genetic lineage 2 WNV encephalomyelitis in horses. Seventeen horses with neurologic signs. Information regarding signalment, clinical signs, and outcome was obtained for each animal. Serology was performed in 15 cases, clinicopathological examination in 7 cases, and cerebrospinal fluid was collected from 2 horses. Histopathology was carried out in 4 horses, 2 of which were assessed for the presence of WNV in their nervous system. WNV neutralizing antibody titers were between 10 and 270 (median, 90) and the results of other serological assays were in agreement with those of the plaque reduction neutralization test. Common signs included ataxia, weakness, asymmetric gait, muscle tremors, hypersensitivity, cranial nerve deficits, and recumbency. Twelve animals survived. Amplicons derived from the infection-positive specimens allowed molecular characterization of the viral strain. From our results, we conclude that this outbreak was caused by a lineage 2 WNV strain, even though such strains often are considered nonpathogenic. Neurological signs and survival rates were similar to those reported for lineage 1 virus infections. The disease occurrence was not geographically limited as had been the typical case during European outbreaks; this report describes a substantial northwestern spread of the pathogen. [ABSTRACT FROM AUTHOR]

The article describes the inclusion body rhinitis in pigs in Hungary. Inclusion body rhinitis in pigs is caused by porcine cytomegalovirus (PCMV) occurs worldwide and the virus belongs to the Betaherpesvirinae family. It indicates that PCMV infection is widespread in Hungarian pig herds, with occasional outbreaks of inclusion body rhinitis.

Porcine circovirus type 3 (PCV3) is an emerging pathogen that has been reported worldwide in all ages of healthy and clinically ill pigs. The presence of this virus in Hungary has been confirmed in a commercial farm experiencing reproductive failures, but there were no data on the circulation of PCV3 in the country. Here we report the prevalence and the genetic diversity of PCV3 in Hungarian herds. To estimate the prevalence, 1855 serum samples, 176 oral fluid and 97 processing fluid samples were collected in a systematic, cross-sectional method from 20 large scale swineherds and tested by real-time qPCR. PCV3 was present in at least one type of diagnostic matrix in 19 out of the 20 (95%) pig farms. The highest detection rates were observed in the processing fluid samples (61%), but 41% of the oral fluid and 23% of the serum samples were positive. The virus was found in all age groups, and slightly more adult animals were infected than growing pigs, but the viral burden was lower amongst them. Phylogenetic analysis of nine complete genomes, obtained from either the sampled herds or organ samples of PCV3-positive carcasses, showed high nucleotide identity between the detected sequences, which all belonged to the PCV3a genotype. Our results indicate that PCV3 is widespread in Hungary, but in most cases, the virus seems to circulate subclinically, infecting all age groups and production phases without the presence of apparent clinical disease.

The genomes of two Tick-borne encephalitis virus (TBEV) strains were fully sequenced and compared to those of known Hungarian strains. One was a laboratory strain (KEM-1) isolated in 1952, which had gone through hundreds of passages both on Vero cell cultures and in laboratory mice, while the other was a recent isolate (2019) from questing female ticks. The laboratory strain formed a monophyletic group with the already published 4 Hungarian strains on the evolutionary tree, located relatively close to Finnish (Kumlinge) and Russian (Absettarov) strains. This KEM-1 strain was phylogenetically distantly related both to the geographically close reference strain Neudörfl and the chronologically close Czech isolates from 1953. The 2019 isolate, KEM-195 was related to TBEV isolates from Southern Slovakia and Styria, and had the longest (328 nucleotides) deletion in its 3'-non-coding region among published sequences of strains of European subtype. Our results show that decades of laboratory passage have not altered the viral genome too much and that at least two distinct branches of TBEV strains circulate in Hungary., (Copyright © 2021. Published by Elsevier GmbH.)

The last surveys on methicillin-resistant Staphylococcus aureus (MRSA) isolated from bovine milk in Hungary took place in the 2000s. To elucidate the genetic variability and to estimate the burden of the pathogen, MRSA from our strain collection and prospectively collected Staphylococcus aureus (SA) isolates originating from two milk hygiene laboratories were investigated. Between 2003 and 2018, 27 MRSA strains originating from 10 dairy farms were deposited and characterised. Most strains (n = 20) belonged to ST1-t127-SCCmecIV and were recovered from three unrelated farms. From other farms, variable genotypes were identified sporadically: ST22-t032-SCCmecIV from three farms; a newly described double locus variant of ST97, ST5982-t458-SCCmecIV from two farms; and ST398-t011-SCCmecIV and ST398-t011-SCCmecV from two respective farms. The prospective screening of 626 individual SA isolates originating from 42 dairy farms resulted in four (0.48 %) MRSA strains from three (7.14 %) farms. All MRSA isolates belonged to the clonal complex 398 and a novel spa-type t19251 was also identified. Most isolates were resistant to three or more antimicrobial classes. The occurrence and significance of MRSA of dairy origin seems to be unchanged in the past decade in Hungary. However, the low host specificity and multiresistance of the identified genotypes calls for periodic revision on the role and distribution of the pathogen in the Hungarian dairy sector.

Between July 2011 and May 2016, a total of 40 Staphylococcus aureus strains originating from 36 horses were confirmed as methicillin resistant (methicillin-resistant Staphylococcus aureus [MRSA]) in a university equine clinic. An additional 10 MRSA strains from 36 samples of clinic workers were obtained in October 2017. The first equine isolate represented the sequence type ST398, spa -type t011, and SCC mec IV. This isolate was resistant to a wide spectrum of antimicrobial agents. MRSA strains with the same genotype and with very similar resistance profiles were isolated on 21 more occasions from September 2013 to September 2014. A second outbreak occurred from May 2015 until May 2016. The first isolate in this second outbreak shared the same genotype, but was additionally resistant to chloramphenicol. The second isolate from August 2015 also showed resistance to rifampicin. The clone was isolated 18 times. Most of the human isolates shared the same genotype as the isolates from horses and their resistance patterns showed only slight differences. We can conclude that the MRSA-related cases at the Department and Clinic of Equine Medicine were all nosocomial infections caused by the same clonal lineage belonging to the clonal complex 398. The clonal complex 398 of equine origin is reported for the first time in Hungary. In addition, our observation of the emergence of new resistance to antimicrobial agents within the clonal lineage after treatment with antibiotics is of concern. Strict hygiene regulations have been introduced to lower the incidence of MRSA isolation and the related clinical disease.

Porcine epidemic diarrhoea virus (PEDV) is an emerging enteropathogen, causing great economic losses in the pig industry. After many years of quiescence, PEDV was detected in Hungary in 2016 with a recombination in its S gene. In order to determine the extent of this change, an attempt was made to isolate the recombinant PEDV. This study was extended with a variety of samples collected from three separate farms with newly identified PEDV in 2018. The recombinant PEDV from 2016 was isolated successfully along with three viruses from 2018, and one isolate from the new cases was used for whole genome determination. Whole genome sequence alignment revealed the highest identity with recombinant Hungarian and Slovenian PEDV within the low-pathogenic European viruses. This suggests that these recombinant PEDV are circulating in this area and may spread to other parts of the continent.

Mycoplasma hyorhinis is a common pathogen of swine causing mainly polyserositis and arthritis, but it has also been implicated as a cause of pneumonia. The economic losses due to M. hyorhinis infection could be reduced by antibiotic treatment. The aim of this study was to determine minimal inhibitory concentrations (MIC) of antibiotics potentially used to combat M. hyorhinis in swine production. Thirty-eight Hungarian M. hyorhinis strains isolated between 2014 and 2017 were examined by microbroth dilution tests for fifteen antimicrobial agents. Low MIC values of tetracyclines (MIC 50 0.078 μg/ml for doxycycline, ≤0.25 μg/ml for oxytetracycline) and pleuromutilins (MIC 50 0.156 μg/ml for tiamulin, ≤0.039 μg/ml for valnemulin) were detected against all strains. Fluoroquinolones (MIC 50 0.625 μg/ml), gentamicin (MIC 50 1 μg/ml) and florfenicol (MIC 50 2 μg/ml) inhibited the growth of Hungarian isolates at moderate MIC values. Most of the strains were inhibited by spectinomycin with low or moderate MIC values (MIC 50 4 μg/ml) except one strain (>64 μg/ml). Numerous isolates showed decreased susceptibility to macrolides and lincomycin (MIC 90 >64 for tylosin, tilmicosin, tulathromycin, gamithromycin, lincomycin, 8 μg/ml for tylvalosin). This study serves as evidence for the increasing resistance to macrolides and lincomycin in mycoplasmas, and also reports the occurrence of strains with extremely high MIC values to spectinomycin thus emphasizes the importance of the prudent use of antibiotics. Based on our results, tetracyclines and pleuromutilins are the most active compounds in vitro against the Hungarian M. hyorhinis strains., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Atypical porcine pestivirus (APPV) is a recently identified RNA virus within the Flaviviridae family, causing congenital tremor (CT) in the piglets of infected sows. We have investigated 25 cases of CT from 2005, 2007, 2010 and 2016-2018, originating from six different farms. RT-PCR has been performed on these samples and all of the affected piglets were positive to APPV. Our phylogenetic analysis showed that Hungarian strains show a high degree of variability and are clustered into five distinct lineages. Four strains originating from one farm have shown exceptional similarity (99.9%) to an Austrian sequence, whereas another one from a different herd was grouped close to a Chinese strain (96.4% similarity). Our results suggest multiple events of introduction of the virus from various sources into Hungary. This is the first report of the presence and clinical relevance of APPV in the Hungarian pig population., (© 2018 Blackwell Verlag GmbH.)

The aim of this study was to determine whether measurements of certain metabolic (non-esterified fatty acid, β-hydroxybutyrate, glucose, total protein, albumin, urea-nitrogen, aspartate aminotransferase, total calcium, inorganic phosphate and magnesium) and endocrine (cortisol, thyroxine, triiodothyronine, insulin and insulin-like growth factor) parameters in the peripartal period (2 months and 3 weeks before expected calving and within 1 hr after calving) were related to the prevalence of stillbirth in a Holstein-Friesian farm in Hungary. All together 155 dairy cattle (n = 22 primiparous, n = 133 multiparous cows) were monitored in two separate years selected randomly on the same farm. Overall, the prevalence of stillbirth was 11% (n = 17). Significantly higher stillbirth rate was detected in case of heifer calvings (OR = 8.5), and when ≥3 assistants (severe dystocia; OR = 8.9) were needed to assist at calving while the body condition score of the dams, the bodyweight and gender of the newborn calves, the percentage of posterior presentations had no significant effect on stillbirth rate. There were no significant differences between cows without and with stillbirth in case of any measured metabolic and endocrine parameters during the examined time periods. At the same time, some of the metabolic parameters (TP, AST and inorg.P) showed some significant differences among the stillbirth groups, but stillbirth could not be predicted by the measured parameters and therefore the role of metabolic and/or endocrine changes on the prevalence of stillbirth needs further elucidation., (© 2018 Blackwell Verlag GmbH.)

Background: Reports on Sarcocystis-infection of cattle are outdated or lacking in many European countries, including those in the Central-Eastern part of the continent. Therefore, to assess the prevalence of Sarcocystis spp. among bovids in Hungary, a countrywide survey was initiated. In addition, fulminant deaths of four cattle, that showed clinical signs and post mortem lesions resembling acute sarcocystiosis ("Dalmeny disease"), were investigated., Methods: During the countrywide survey individual heart and oesophagus samples were collected at slaughterhouses from 151 beef cattle and from 15 buffalo, kept in 31 places of Hungary. Analysis for Sarcocystis spp. was carried out with conventional PCRs for the 18S rDNA gene and gel electrophoresis, followed by sequencing of 36 strongly positive samples. Mortality cases were evaluated by histological, molecular, bacteriological and virological analyses of samples from various organs., Results: Among slaughtered cattle the rate of Sarcocystis-infection was 66%. S. cruzi was identified as the most prevalent species in aurochs-like breed, and the zoonotic S. hominis in Hungarian grey cattle. Concerning the sudden deaths of cattle, Sarcocystis-infection could not be demonstrated in organs showing haemorrhages, but S. cruzi cysts were present in the muscles. In one case "S. sinensis" was molecularly identified in the blood (indicating sarcocystaemia). Results of analyses for bacterial/viral pathogens were negative., Conclusions: S. cruzi appears to be the most prevalent Sarcocystis sp. in cattle in Hungary, followed by the zoonotic S. hominis. However, the rate of infection with both species was shown to differ between cattle breeds. The suspected role of Sarcocystis spp. as causative agents of the fatal cases could not be confirmed.

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.

Hungary is traditionally regarded as a leishmaniasis-free country, and human or canine cases diagnosed locally have been recorded as imported. However, recent entomological surveys have verified the presence in Hungary of Phlebotomus neglectus and Phlebotomus perfiliewi perfiliewi, which have been incriminated as competent vectors of Leishmania infantum elsewhere in Europe. Following the occurrence in October 2007 of an undisputable clinical case of L. infantum canine leishmaniasis (CanL) in a 4-year-old female pug in a kennel of 20 dogs in Tolna province, an investigation was performed to assess the infection status in that canine population and to search for putative phlebotomine vectors. Another female pug became sick during the study period (May-November 2008) and L. infantum was confirmed as the causative agent. The other animals appeared clinically healthy; however, 4 additional dogs were found positive by indirect fluorescent antibody test (2 dogs), or by buffy-coat PCR (1 dog), or by both methods (1 dog). Hence the overall Leishmania infection prevalence in the kennel was 30% (6/20). All dogs were born in the same place and had been always kept outdoors. They had neither been abroad nor received a blood transfusion. No sand flies were collected with CDC Standard Miniature Light traps, Mosquito Magnet(®) X (MMX) dry ice-baited traps, or sticky traps placed either in or around the kennel and at nearby chicken yards during July and August of 2008 and 2009. Considering the dogs' historical background and the failure to trap any sand fly vectors in the kennel area, the origin of CanL in this site remains unexplained.

A number of newly identified porcine parvoviruses had been described during the last decade, but the presence and prevalence of these viruses are unknown in Hungary and only partly known for Europe. The present study was conducted to detect and measure the prevalence of these viruses, namely porcine parvovirus (PPV) 2, PPV3, PPV4, porcine bocavirus (PBoV) 1, PBoV2, PBo-likeV and the 6V and 7V parvoviruses. The prevalence of PPV1 and porcine circovirus type 2 (PCV2) was also investigated. Faecal samples, blood serum samples, organ tissues, foetuses and semen were collected from different swine herds in Hungary and tested by polymerase chain reaction methods specific for the different viruses. The results indicated that all of the examined parvoviruses were present in Hungary, hence in Europe. The prevalence was 18.1% for PCV2, 0.5 % for PPV1, 6.4% for PPV2, 9.7% for PPV3, 6.4% for PPV4, 1.5% for PBo-likeV, 4.8% for PBoV1 and PBoV2 and 1.8% for 6V and 7V. Based on the analysis of partial PPV4 and PBo-likeV sequences, these viruses showed a high degree of sequence conservation, whereas PPV3 and the majority of PPV2, PBoV1, PBoV2, 6V and 7V sequences showed higher variability. Possible sites of recombination were also identified between PBoV1 and PBoV2 genomes.

The aim of this study was to obtain prevalence estimates about the most important enteropathogenic bacteria: Lawsonia intracellularis, Brachyspira hyodysenteriae, Brachyspira pilosicoli, Salmonella enterica and Clostridium perfringens A and C in Hungarian farrow-to-finish pig herds. A total of 31 herds were selected, from where six pooled faecal samples, each containing three individual rectal faecal samples were collected from fattening pigs of 5-6 months of age. All 186 samples were examined by polymerase chain reaction (PCR) for the presence of the pathogens mentioned above. Lawsonia intracellularis was found in 29 herds (93.55%) and in 108 samples (58.06%); B. hyodysenteriae in 14 herds (45.16%) and in 23 samples (12.37%); B. pilosicoli in 19 herds (61.29%) and in 53 samples (28.49%); S. enterica in 17 herds (54.83%) and in 40 samples (21.50%). We detected the presence of C. perfringens A in 19 herds (61.29%) and in 46 samples (24.73%), while C. perfringens C was found in 8 herds (25.81%) and in 11 samples (5.91%). All examined herds were infected with one or more of these agents. Herds with diarrhoea in the mid- to late finishing phase had almost 10 times higher prevalence of B. hyodysenteriae than herds without such a history.

The authors report the data of the first survey on the incidence of post-weaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS) in Hungary. A PCR method specific for the detection of porcine circovirus 2 (PCV-2) was developed, which proved to be suitable for diagnostic purposes. PCR screening of organ samples from pigs suspected to be affected with PMWS or PDNS revealed the presence of PCV-2 in 80% of the cases. Six PCV-2 genomes from Hungarian isolates were completely sequenced. Phylogenetic comparison with all the available PCV-2 sequences showed that porcine circoviruses circulating in Hungary are more variable than in several other European countries. Two Hungarian strains clustered together with the Spanish strains forming a distinct group; two others fell in a common group with the French, UK, and Dutch strains, whereas another two strains showed the closest relationship to two of the three known German PCV-2 sequences.

Slaughterhouse sampling and examination of urogenital tracts of 499 sows and gilts culled for reproductive reasons from 21 Hungarian herds were performed over a 6-year period. The aim was to estimate the prevalence of different urogenital tract lesions, and to provide sensitivity and specificity estimates for macroscopic and bacteriological examinations in the diagnosis of urocystitis and endometritis. Furthermore, the association between endometritis and urocystitis was assessed. The prevalence of main lesions of the urogenital tract was similar to that reported in other studies. The 'sensitivity' of macroscopic and bacteriological methods was determined statistically by taking histopathology as the 'Gold Standard'. As a result, the 'sensitivity' of macroscopic methods for the diagnosis of endometritis and urocystitis was < or = 18.1% and 47.9%, respectively, while the 'sensitivity' of bacteriology for the diagnosis of the same conditions was < or = 31.8% and 63.0%, respectively. The presumed positive association between urocystitis and endometritis was confirmed; it was not confounded by parity. Animals affected by urocystitis had a 3.5 times higher odds to simultaneously have endometritis than animals without urocystitis.

Pooled faecal samples and/or intestinal contents from pig carcasses were collected from 11 Hungarian swine farms and subjected to DNA extraction/ purification and subsequent polymerase chain reaction (PCR) in order to detect Lawsonia intracellularis, the aetiological agent of the porcine proliferative enteropathy complex. Specific PCR positivity was detected in 6 individual and 22 pooled samples out of 46, originating from eight herds. The PCR products of collected faecal samples hybridised in Southern blot hybridisation with the DNA of the type strain L. intracellularis NCTC 12657. This is the first confirmed detection of L. intracellularis by PCR in Hungary.

Eubacterium suis was isolated from the preputium of seven out of 16 mature boars on two farms and from the urinary bladder of one out of five sows originating from a third herd. The morphological and biochemical characteristics of the isolated strains were identical to that of the reference strain of E. suis ATTC 33144. Three out of four strains isolated from Farm A were successfully subcultured aerobically, and then anaerobically again. E. suis together with Proteus mirabilis was isolated from cystitis of a sow 4 days after artificial insemination. These are the first strains of E. suis isolated in Hungary.