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Ronai Z.,Veterinary Diagnostic Directorate | Csivincsik A.,University of Kaposvár | Dan A.,Veterinary Diagnostic Directorate | Gyuranecz M.,Hungarian Academy of Sciences
Infection, Genetics and Evolution | Year: 2016

Besides Mycobacterium avium subsp. paratuberculosis (MAP), M. avium subsp. avium (MAA), M. avium subsp. silvaticum (MAS), and 'M. avium subsp. hominissuis' (MAH) are equally important members of M. avium complex, with worldwide distribution and zoonotic potential. Genotypic discrimination is a prerequisite to epidemiological studies which can facilitate disease prevention through revealing infection sources and transmission routes. The primary aim of this study was to identify the genetic diversity within 135 MAA, 62 MAS, and 84 MAH strains isolated from wild and domestic mammals, reptiles and birds.Strains were tested for the presence of large sequence polymorphism LSPA17 and were submitted to Mycobacterial interspersed repetitive units-variable-number tandem repeat (MIRU-VNTR) analysis at 8 loci, including MIRU1, 2, 3, and 4, VNTR25, 32, and 259, and MATR9. In 12 strains hsp65 sequence code type was also determined.LSPA17 was present only in 19.9% of the strains. All LSPA17 positive strains belonged to subspecies MAH. The discriminatory power of the MIRU-VNTR loci set used reached 0.9228. Altogether 54 different genotypes were detected. Within MAH, MAA, and MAS strains 33, 16, and 5 different genotypes were observed. The described genotypes were not restricted to geographic regions or host species, but proved to be subspecies specific.Our knowledge about MAS is limited due to isolation and identification difficulties. This is the first study including a large number of MAS field strains. Our results demonstrate the high diversity of MAH and MAA strains and the relative uniformity of MAS strains. © 2016 Elsevier B.V.


Sulyok K.M.,Hungarian Academy of Sciences | Kreizinger Z.,Hungarian Academy of Sciences | Hornstra H.M.,Northern Arizona University | Pearson T.,Northern Arizona University | And 5 more authors.
BMC Veterinary Research | Year: 2014

Background: Information about the genotypic characteristic of Coxiella burnetii from Hungary is lacking. The aim of this study is to describe the genetic diversity of C. burnetii in Hungary and compare genotypes with those found elsewhere. A total of 12 samples: (cattle, n = 6, sheep, n = 5 and human, n = 1) collected from across Hungary were studied by a 10-loci multispacer sequence typing (MST) and 6-loci multiple-locus variable-number of tandem repeat analysis (MLVA). Phylogenetic relationships among MST genotypes show how these Hungarian samples are related to others collected around the world.Results: Three MST genotypes were identified: sequence type (ST) 20 has also been identified in ruminants from other European countries and the USA, ST28 was previously identified in Kazakhstan, and the proposed ST37 is novel. All MST genotypes yielded different MLVA genotypes and three different MLVA genotypes were identified within ST20 samples alone. Two novel MLVA types 0-9-5-5-6-2 (AG) and 0-8-4-5-6-2 (AF) (Ms23-Ms24-Ms27-Ms28-Ms33-Ms34) were defined in the ovine materials correlated with ST28 and ST37. Samples from different parts of the phylogenetic tree were associated with different hosts, suggesting host-specific adaptations.Conclusions: Even with the limited number of samples analysed, this study revealed high genetic diversity among C. burnetii in Hungary. Understanding the background genetic diversity will be essential in identifying and controlling outbreaks. © 2014 Sulyok et al.; licensee BioMed Central Ltd.


Erdelyi K.,Veterinary Diagnostic Directorate | Mezosi L.,Simba Veterinary Surgery | Vladov S.,Veterinary Diagnostic Directorate | Foldvari G.,Szent Istvan University
Ticks and Tick-borne Diseases | Year: 2014

Two adult male Eurasian grey wolves belonging to a group of 12 animals, kept in an open air 15,000-m2 enclosure at the Bear Farm facility near Veresegyháza, Hungary, were found dead in September 2002. Another 2 wolves died during the same period, but laboratory examination of their carcasses was not possible. During necropsy both animals were found to be in a good body condition. Oral mucosa, conjunctiva, sclera, and subcutaneous tissues revealed severe jaundice. The liver, gall bladder, and spleen were enlarged. The kidneys were paler than normal, and petechial haemorrhages were also seen under their fascia. Small, round Babesia-like organisms, 1.5-2μm in diameter, were demonstrated in large numbers in stained impression smears made from the spleens of both animals. PCR amplification and sequencing identified Babesia canis. There are very few reports on babesiosis in the grey wolf, and our findings draw attention to the potential threat posed by B. canis that will probably have to be taken into account in future ex situ and in situ wolf conservation efforts. © 2014 Elsevier GmbH.


Farkas R.,Szent Istvan University | Takacs N.,Szent Istvan University | Hornyak,Veterinary Diagnostic Directorate | Nachum-Biala Y.,Hebrew University of Jerusalem | And 2 more authors.
Parasites and Vectors | Year: 2015

Background: To date, only one report of a small Babesia infection based on microscopic observation which caused babesiosis in two dogs in Hungary has been published. Babesiosis due to Babesia canis - which is endemic in the local dogs - has only been detected in captive grey wolves. No information is available on babesial/theilerial infections in red foxes in Hungary. The aim of the study was to screen red foxes in Hungary for babesial parasites by PCR and to compare their partial 18S rRNA gene sequences to those parasites of domestic dogs and wild canids from other countries. Methods: Blood samples of 404 red foxes originating from 316 locations representing all 19 Hungarian counties were screened in Hungary for babesial parasites by PCR and the partial 18S rRNA gene sequences were compared to those parasites of domestic dogs and wild canids from other countries. Results: Altogether 81 red foxes out of 404 (20.0%; 95% CI: 16.4-24.2%) shot in 74 locations and in 17 of the 19 Hungarian counties were found to be infected with Babesia cf. microti by PCR. Conclusions: This is the first report to demonstrate the occurrence of Babesia cf. microti in Hungary, and its widespread presence in the fox population throughout the country. Further studies are needed to identify the tick species involved in its transmission, and whether other mechanisms of transmission are involved in its spread in fox populations. © 2015 Farkas et al.; licensee BioMed Central.


Gyuranecz M.,Hungarian Academy of Sciences | Rannals B.D.,Northern Arizona University | Allen C.A.,Northern Arizona University | Janosi S.,Veterinary Diagnostic Directorate | And 2 more authors.
BMC Veterinary Research | Year: 2013

Background: Little is currently known about Brucella evolution within the host during infection. The current study is the first to employ fine-scale genotyping on an isolate collection derived from a Brucella canis outbreak. Eight isolates of B. canis, cultured from different tissues of three dogs (female, stud dog, puppy of another female) from a single kennel over three months were genetically characterized with a 15-marker multi-locus, variable-number tandem repeat (VNTR) analysis (MLVA) to assess the genetic relatedness of isolates and potential rapid mutational changes.Results: MLVA discriminated among the otherwise indistinguishable isolates from different animals and from isolates collected at different time points within each host, with different VNTR alleles being detected at multiple dates and tissue sites. We suspect that all isolates cultured from the female, puppy, and stud dogs originated from the same strain, with subsequent rapid in vivo mutations. However, high mutation rates and apparent in several of the loci prevented making definitive epidemiological relationships among isolates.Conclusions: This investigation highlights the rapid in vivo genetic mutations of several VNTRs of B. canis over a short time period in the host and the emergence of alternate alleles. However, this work also suggests the challenges of using highly mutable VNTRs to infer epidemiological relationships of strains within a short duration outbreak. © 2013 Gyuranecz et al.; licensee BioMed Central Ltd.


Sulyok K.M.,Hungarian Academy of Sciences | Kreizinger Z.,Hungarian Academy of Sciences | Fekete L.,Hungarian Academy of Sciences | Janosi S.,Veterinary Diagnostic Directorate | And 5 more authors.
BMC Veterinary Research | Year: 2014

Background: Mycoplasma bovis is an important pathogen causing pneumonia, mastitis and arthritis in cattle worldwide. As this agent is primarily transmitted by direct contact and spread through animal movements, efficient genotyping systems are essential for the monitoring of the disease and for epidemiological investigations. The aim of this study was to compare and evaluate the multi locus sequence typing (MLST) and the multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) through the genetic characterization of M. bovis isolates from Hungary. Results: Thirty one Hungarian M. bovis isolates grouped into two clades by MLST. Two strains had the same sequence type (ST) as reference strain PG45, while the other twenty nine Hungarian isolates formed a novel clade comprising five subclades. Isolates originating from the same herds had the same STs except for one case. The same isolates formed two main clades and several subclades and branches by MLVA. One clade contained the reference strain PG45 and three isolates, while the other main clade comprised the rest of the strains. Within-herd strain divergence was also detected by MLVA. Little congruence was found between the results of the two typing systems. Conclusions: MLST is generally considered an intermediate scale typing method and it was found to be discriminatory among the Hungarian M. bovis isolates. MLVA proved to be an appropriate fine scale typing tool for M. bovis as this method was able to distinguish closely related strains isolated from the same farm. We recommend the combined use of the two methods for the genotyping of M. bovis isolates. Strains have to be characterized first by MLST followed by the fine scale typing of identical STs with MLVA. © 2014 Sulyok et al.; licensee BioMed Central Ltd.


Kreizinger Z.,Hungarian Academy of Sciences | Makrai L.,Szent Istvan University | Helyes G.,Szent Istvan University | Magyar T.,Hungarian Academy of Sciences | And 2 more authors.
Journal of Antimicrobial Chemotherapy | Year: 2013

Objectives: Determining the in vitro susceptibility to 11 antibiotics of Francisella tularensis subsp. holarctica strains belonging to the phylogenetic group B.13, from different areas of Hungary. Methods: Twenty-nine F. tularensis strains isolated between 2003 and 2010 from free-ranging European brown hares (Lepus europaeus) and a captive patas monkey (Erythrocebus patas) were collected from different parts of Hungary and examined for antibiotic susceptibility with commercially available MIC test strips on modified Francis agar plates; values were interpreted according to CLSI breakpoints. Results: The strains were susceptible to aminoglycosides (MIC90 values: gentamicin, 0.75mg/L; and streptomycin, 6.0mg/L), tetracyclines (MIC90 values: tetracycline, 0.5mg/L; and doxycycline, 1.0mg/L), quinolones (MIC90 values: ciprofloxacin, 0.047mg/L; and levofloxacin, 0.023mg/L) and chloramphenicol (MIC90 value: 1.5mg/L), i.e. antibiotics commonly used in therapy. Tigecycline (MIC90 value: 0.19mg/L) and rifampicin (MIC90 value: 1.0mg/L) were also active against F. tularensis strains, while resistance to erythromycin (MIC90 value:.256mg/L) and linezolid (MIC90 value: 32mg/L) was observed in all strains. Conclusions: Based on the results, quinolones are recommended as first choice therapy for F. tularensis infection. The in vitro susceptibility of the strains to tigecycline may encourage the application of this antibiotic as well. The similar antibiotic susceptibilities of the Hungarian strains belonging to different subclades of phylogenetic group B.13 indicates that strains from other Central and Eastern European countries belonging to this group might also have the same susceptibility profile. © The Author 2012. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.


Banyai K.,Hungarian Academy of Sciences | Toth A.G.,Veterinary Diagnostic Directorate | Ivanics E.,Veterinary Diagnostic Directorate | Glavits R.,Veterinary Diagnostic Directorate | And 2 more authors.
Emerging Infectious Diseases | Year: 2012

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.


Dandar E.,Hungarian Academy of Sciences | Balint A.,Veterinary Diagnostic Directorate | Kecskemeti S.,Veterinary Diagnostic Directorate | Szentpali-Gavaller K.,Veterinary Diagnostic Directorate | And 4 more authors.
Archives of Virology | Year: 2013

Avian orthoreoviruses have been associated with a variety of diseases in chickens, including tenosynovitis, runting-stunting syndrome, hepatitis, myocarditis, osteoporosis, respiratory diseases, and central nervous system disease. The primary objective of our study was the molecular characterization of an avian reovirus strain, T1781, which was isolated from a broiler chicken with a central nervous system disorder in Hungary during 2012. The complete genome sequence was determined using a traditional sequencing method after cell culture adaptation of the strain. Sequence and phylogenetic analyses showed that T1781 shared only moderate nucleic acid sequence identity in several genes to previously analyzed reovirus strains from chickens, and each gene formed separate branches in the corresponding phylogenetic trees. The maximum nucleotide sequence identities of strain T1781 genes to reference avian reovirus strains ranged from 79 % to 90 %. Collectively, our analyses indicated that T1781 is a divergent chicken reovirus strain. The genetic background of this and other avian reoviruses associated with various disease manifestations needs further investigation. © 2013 Springer-Verlag Wien.


Meszaros I.,Hungarian Academy of Sciences | Toth R.,Hungarian Academy of Sciences | Balint A.,Veterinary Diagnostic Directorate | Dan A.,Veterinary Diagnostic Directorate | And 2 more authors.
Avian Pathology | Year: 2014

Duck circovirus, duck hepatitis A virus 1, goose parvovirus and goose haemorrhagic polyomavirus are economically damaging pathogens of waterfowl, and replicate poorly or not at all in established cell lines. AGE1.CR, AGE1.CR.pIX and AGE1.CS cell lines, originating from the Muscovy duck, were tested for their suitability to isolate and identify these viruses. Immunofluorescence (IF) and quantitative polymerase chain reaction investigations verified that all cell lines are permissive for all four viruses; however, AGE1.CR.pIX proved to be the most productive and most sensitive for viral infection. IF experiments revealed that the time of one infectious cycle is approximately 12 to 14 h in the AGE1.CR.pIX cells in the case of the three DNA viruses, while it is 10 to 12 h for DHAV-1. Specific viral infectivity and the limit of detection by IF varied between 55 and 1484 copies, depending on the viruses and cell lines. Despite the high sensitivity of the cell lines for viruses, their viral productivity remained relatively low for the investigated field isolates. However, optimization of virus infection and/or the adaptation of the viruses to the cells can raise viral productivity and can make these cell lines suitable for vaccine development and production. © 2014 Houghton Trust Ltd.

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