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Equine herpesvirus-1 (EHV-1) can be classified into distinct groups by single nucleotide polymorphisms (SNPs) in their genomes. Only a few of these can be associated with a special attribute of the virus. Differences in the ORF30 region can determine the neuropathogenic potential, while by substitutions in the ORF68 region several strain groups can be made. In previous studies no connection was found between the neuropathogenic potential and the SNPs in ORF68, but the occurrence of members of distinct groups in different outbreaks can facilitate epidemiological investigations because the geographical distribution of a particular group is very often specific. The present study aimed at the molecular examination and grouping of 35 EHV-1 strains isolated from aborted equine fetuses in Hungary between 1977 and 2008. Genotyping was based on the comparison of nucleotide sequences of a polymorphic segment located in the ORF68 region, which had previously been found to be a useful tool for classification. After sequencing this region, the Hungarian EHV-1 isolates could be classified into seven groups. Only 23 of the 35 isolates belonged to the formerly described groups, while the SNPs of 12 isolates diverged, and four new groups could be set up. In addition, phylogenetic analysis was performed to compare the ORF68 sequences of the Hungarian strains with the sequences of isolates from Europe, America and Australia. The number of newly formed groups suggests that the further analysis of unknown EHV-1 isolates would involve the emergence of extended numbers of new groups, which can impair the usability of this grouping method.

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Absztrakt

Bevezetés: A megfelelő emelt szintű újraélesztési ismeretek elengedhetetlenek a sürgősségi ellátásban dolgozó szakemberek számára. Célkitűzés: A szerzők célja a magyarországi mentőtiszthallgatók felnőtt emelt szintű újraélesztési ismereteinek felmérése volt. Módszer: Felmérésüket a Pécsi Tudományegyetem Egészségtudományi Karának, a Semmelweis Egyetem Egészségtudományi Karának és a Debreceni Egyetem Egészségügyi Karának III. és IV. évfolyamos hallgatói körében végezték, saját szerkesztésű kérdőív segítségével. A mintába 97 fő hallgató került be (n = 97). Eredmények: A hallgatók átlagosan 67,79%-os eredménnyel teljesítettek a felmérésen. A férfiak és a nők összpontszámai között nem volt szignifikáns különbség (p = 0,725). Az alacsony életkor szignifikánsan javította az elért összpontszámokat (p = 0,003). A nappali tagozatosok szignifikánsan magasabb összpontszámot értek el, mint a levelezősök (p = 0,004). Az egyes képzőintézmények hallgatóinak összpontszámai között szignifikáns különbség nem volt a felmérés során (p = 0,254). Következtetések: Célszerű lenne a mentőtisztképzésben a témával foglalkozó tantárgyak számát növelni, a számonkérések kritériumrendszerét szigorítani. A szerzők a jövőben indokoltnak tartanák a gyakorlati ismereteket vizsgáló felmérés elvégzését. Orv. Hetil., 2016, 157(37), 1476–1482.

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Acta Veterinaria Hungarica
Authors: Ádám Bálint, István Kiss, Krisztián Bányai, Imre Biksi, Katalin Szentpáli-Gavallér, Tibor Magyar, István Jankovics, Mónika Rózsa, Bálint Szalai, Mária Takács, Ádám Tóth and Ádám Dán

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.

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Acta Veterinaria Hungarica
Authors: István Szabó, Tamás Molnár, Imre Nemes, Tamás Abonyi, Zsolt Terjék and Ádám Bálint

Eradication of porcine reproductive and respiratory syndrome virus (PRRSV) from the pig population of Hungary started in 2014 on the basis of the territorial principle. In order to reach this goal it was crucial to render each fattening unit free of this disease, since fattening units play a significant role in spreading the virus all over the country. In 2015, 188 out of 307 large-scale fattening farms (61.2%) kept PRRS-positive animals. The main source of infection of these farms was the import of PRRS-infected fattening pigs. The following methods were used during the eradication from 2017: (1) Only pigs coming from PRRS-free farms were allowed to be used for fattening in Hungary; (2) Quarantine of all herds for 60 days; (3) PCR test for PRRS 48 hours after the arrival of the prefattening animals; (4) Serological test for PRRS at the end of the quarantine period. If any diagnostic test gave even one positive result and the result was confirmed by another test, the stock had to be sold for slaughter within 15 days or placed outside Hungary, so that the infected stock would not compromise the PRRS status of that area. PRRSV eradication on large-scale fattening units applying all-in/all-out operation was relatively simple, using the depopulation-repopulation method. On permanently operating farms, the infected herd was sold from time to time, without having to be repopulated until the last delivery. After cleaning, disinfection and restocking, the repopulation was done with PRRS-free animals. As the eradication progressed over the years, a ban on the import of infected fattening pigs was imposed. As a consequence of these measures, by the end of 2018, Hungarian large-scale fattening farms became free of PRRS. Maintaining the national-level PRRS-free status of large-scale pig fattening units contributes to eliminating a significant cost factor from the Hungarian pork production industry, and opens the way for a significant reduction in antibiotic consumption as well.

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Acta Veterinaria Hungarica
Authors: Imre Nemes, Tamás Molnár, Tamás Abonyi, Zsolt Terjék, Ádám Bálint and István Szabó

In the EU Member States with a traditionally significant pig industry, the prevalence of PRRS infections is high. Therefore, the Pig Strategy of the Government of Hungary prioritises eradication of PRRSV in Hungary. For the first time among the EU Member States, a National PRRS Eradication Programme was introduced in order to reach a more efficient, economical and competitive international market position. Although its significance has decreased in recent decades, 20% of the Hungarian pig population is still kept on small-scale (backyard) farms (< 100 animals). The prevalence of PRRSV in backyard farms was 3.9% at the beginning of the programme. The present paper details the measures applied during the different phases of the programme in backyard farms. During all the phases, serological testing of the breeding animals of the registered small-scale herds was performed, including the highest number of individual animals. Seropositive individuals were tested by PCR and were removed from the backyard farm within the framework of official measures. By sequencing the identified PRRS strains, the possible epidemic relationships between small-scale and large-scale farms were continuously monitored. As a result of the programme, PRRS-free status of the small-scale herds was achieved by the end of 2015, and this status was maintained in 2016-2018.

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Acta Veterinaria Hungarica
Authors: István Mészáros, Ferenc Olasz, Enikő Kádár-Hürkecz, Ádám Bálint, Ákos Hornyák, Sándor Belák and Zoltán Zádori

Feline enteric coronaviruses have three open reading frames (ORFs) in region 3 (3a, 3b, and 3c). All three ORFs were expressed with C-terminal eGFP and 3xFLAG tags in different cell lines and their localisation was determined. ORF 3a is predicted to contain DNA-binding and transcription activator domains, and it is localised in the nucleus and in the cytoplasm. ORF 3b is also predicted to contain DNA-binding and activator domains, and was found to localise in the mitochondrion. Besides that, in some of the non-infected and FIPV-infected cells nucleolar, perinuclear or nuclear membrane accumulation of the eGFP-tagged 3b was observed. The exact compartmental localisation of ORF 3c is yet to be determined. However, based on our co-localisation studies 3c does not seem to be localised in the ER-Golgi network, ERGIC or peroxisomes. The expression of 3c-eGFP is clearly cell type dependent, it is more stable in MARC 145 cells than in Fcwf-4 or CrFK cells, which might reflect in vivo stability differences of 3c in natural target cells (enterocytes vs. monocytes/macrophages).

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Acta Veterinaria Hungarica
Authors: Attila Farsang, Ildikó Bódi, Orsolya Fölker, Krisztina Minkó, Zsófia Benyeda, Ádám Bálint and Imre Oláh

The aim of this immunocytochemical study was to compare mannose-binding lectin (MBL) production induced by avian coronavirus in the spleen and caecal tonsil (CT). One-day-old specific-pathogen-free (SPF) chickens were experimentally infected with six QX field isolates and the H120 vaccine strain. In the negative control birds, the spleen was MBL negative, while the CT showed scattered MBL-positive cells in close proximity and within the surface epithelium and germinal centre (GC)-like cell clusters. MBL was detectable in the ellipsoid-associated cells (EACs) and cell clusters in the periarterial lymphoid sheath (PALS) by 7 days post infection (dpi). In both organs, the MBL-positive cells occupy antigen-exposed areas, indicating that GC formation depends on resident precursors of dendritic cells. The majority of MBL-positive EACs express the CD83 antigen, providing evidence that coronavirus infection facilitated the maturation of dendritic cell precursors. Surprisingly, co-localisation of MBL and CD83 was not detectable in the CT. In the spleen (associated with circulation), the EACs producing MBL and expressing CD83 are a common precursor of both follicular (FDC) and interdigitating dendritic cells (IDC). In the CT (gut-associated lymphoid tissue, GALT) the precursors of FDC and IDC are MBL-producing cells and CD83-positive cells, respectively. In the CT the two separate precursors of lymphoid dendritic cells provide some ‘autonomy’ for the GALT.

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Acta Veterinaria Hungarica
Authors: István Kiss, Tamás Mató, Zalán G. Homonnay, Judit Kojer, Attila Farsang, Ádám Bálint and Vilmos Palya

Understanding the epidemiology and improving vaccinal protection against the highly variable chicken infectious bronchitis virus (IBV) requires the knowledge of circulating IBV serotypes/genotypes in defined geographic areas. Accordingly, the authors initiated a survey among the major poultry producers in Hungary in order to reveal the prevailing IBV serotypes in the country. Tracheal swabs and organ samples (caecal tonsils, kidneys, and trachea) were collected from broiler, layer, and meat-type breeder flocks, and were subjected to IBV detection by virus isolation and polymerase chain reaction (PCR). The IBV-positive samples were further characterised by nucleotide sequencing and phylogenetic analysis of a portion of the S1 IBV gene. Seventeen out of the 26 submitted samples proved to be positive for IBV. Sequence analyses revealed ten 4/91 and six QX serotypes, and a single D274 type IB virus. One sample contained a mixture of QX and Massachusetts serotype viruses. Presumably most of the 4/91 and D274 type viruses were vaccine strains. The proportion of QX type viruses and their observed variation are in good agreement with the situation in a few other European countries. The detected viruses clustered largely according to their geographic origin, with a few exceptions. If updated regularly, the preliminary ‘virus map’ will be useful for the adjustment of vaccination protocols.

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The 2006 epidemic due to highly pathogenic avian influenza virus (HPAIV) subtype H5N1 in Hungary caused the most severe losses in waterfowl which were, according to the literature at the time, supposed to be the most resistant to this pathogen. The presence of pathological lesions and the amount of viral antigen were quantified by gross pathology, histopathology and immunohistochemistry (IHC) in the organs of four waterfowl species [mute swans (n = 10), domestic geese (n = 6), mulard ducks (n = 6) and Pekin ducks (n = 5)] collected during the epidemic. H5N1 subtype HPAIV was isolated from all birds examined. Quantitative real-time reverse transcriptase-polymerase chain reaction (qRRT-PCR) was also applied on a subset of samples [domestic geese (n = 3), mulard (n = 4) and Pekin duck (n = 4)] in order to compare its sensitivity with IHC. Viral antigen was detected by IHC in all cases. However, the overall presence of viral antigen in tissue samples was quite variable: virus antigen was present in 56/81 (69%) swan, 22/38 (58%) goose, 28/46 (61%) mulard duck and 5/43 (12%) Pekin duck tissue samples. HPAIV subtype H5N1 was detected by qRRT-PCR in all birds examined, in 19/19 (100%) goose, 7/28 (25%) mulard duck and 12/28 (43%) Pekin duck tissue samples. As compared to qRRTPCR, the IHC was less sensitive in geese and Pekin ducks but more sensitive in mulard ducks. The IHC was consistently positive above 4.31 log10 copies/reaction but it gave very variable results below that level. Neurotropism of the isolated virus strains was demonstrated by finding the largest amount of viral antigen and the highest average RNA load in the brain in all four waterfowl species examined.

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Acta Veterinaria Hungarica
Authors: Attila Farsang, Ildikó Bódi, Orsolya Fölker, Krisztina Minkó, Zsófia Benyeda, Ádám Bálint, Anna L. Kiss and Imre Oláh

Coronavirus infection delays the development of the cortico-medullary (CM) capillary network which results in retarded development of bursal follicles. The smaller size of the medulla in the coronavirus-infected birds may lead to a lower number of B lymphocytes and bursal secretory dendritic cells, which negatively affects the reactivity and efficacy of the immune system. Contrary to the wild-type infectious bronchitis virus (IBV) strain, infection induced by H120 vaccine virus exerts only a moderate influence on caveolin-1 expression of the CM capillary web and on follicular development compared to the untreated controls.

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