Magyarországi hüllő és kétéltű gyűjtemények virológiai vizsgálata, különös tekintettel a járványos megbetegedést okozó vírusokra. = Virological analysis of the Hungarian reptile and amphibian collections with respect of viruses causing epizootics.

Az OTKA-téma keretében hagyományos és modern diagnosztikai módszereket vezettünk be Magyarországon a hüllő virológiai kutatásokba. A projekt keretében végzett kutatásaink során elsőként számoltunk be hazai mintában vipera adenovírus- és parvovírus-fertőzöttségről. Egy felnőtt dobozteknősben az adenovírus fertőzöttségre jellemző zsíros májelfajulást, sejtzárványos hepatitist állapítottunk meg, általános PCR-rel egy különleges adenovírus jelenlétét sikerült kimutatni, mely az eddig leírt hüllő és teknős adenovírusokkal ellentétben egyik ismert nemzetségbe sem volt besorolható az Adenoviridae családon belül. Coronavírus-szerű részecskéket mutattunk ki transzmissziós elektronmikroszkópos vizsgálattal amursikló savós-fibrines légcsőgyulladásával összefüggésben. Együttműködő partnereinkkel a világon elsőként sikerült hazai halakból circovírusokat kimutatni, a vírusok teljes genom szekvenciáját meghatározni és elemezni. Circovírusokat később gazdasági szempontból jelentős indiai halfajokból is dokumentáltunk. Munkánk során egy új hüllő paramyxovírust sikerült jellemeznünk egy homalopsid kígyó mucopurulens tüdőgyulladással járó elhullásával kapcsolatban. Németországi együttműködésünk keretében eddig nem ismert picornavírus izolátumok szekvenciáját részlegesen meghatároztuk. Hüllő reovírusokból mindeddig csak részleges szekvencia volt elérhető a génbankban, azonban az Ion-Torrent új generációs szekvenáló berendezés segítségével sikerült több hüllő reovírus izolátumot szekvenálni. | Within the OTKA project traditional and modern diagnostic methods have been introduced into the reptilian virological research in Hungary. During our studies we could document a viper adenovirus and parvovirus infection for the first time in a Hungarian sample. Fatty liver, inclusion body hepatitis characteristic for adenoviral infection was described in an adult ornate box turtle. General PCR also revealed the presence of a novel adenovirus distinct from the previously described chelonian and reptilian adenoviruses, not belonging to any of the recognized genera of the family Adenoviridae. Coronavirus-like particles were detected by transmission electron microscopic examination in connection with a serofibrinous tracheitis of an Amur ratsnake. In a collaborative study we were the first to describe a circovirus in a Hungarian fish species; the complete sequence of the virus was determined and analyzed. Circoviruses were later documented in economically important Indian fish species. During the course of our research we could describe a novel reptilian paramyxovirus related to the death of a homalopsid snake associated with mucopurulent pneumonia. Partial genome sequences of reptilian picornavirus isolates were determined with our German cooperating partners. So far only partial sequences of reptilian reoviruses were available in Genbank, however with the Ion-Torrent new generation sequencer we could determine the whole genome sequence of reptilian reovirus isolates

Állati eredetű adenovírusok genetikai jellemzése az evolúciót hűen tükröző rendszertan kialakítása céljából = Genetic characterization of adenoviruses of different animal hosts to achieve an improved taxonomy, which reflects the viral evolution

A pályázat által támogatott kutatások során rendszertani szempontból különösen jelentősnek ítélt adenovírusokat vizsgáltunk. Két óvilági majom-adenovírus teljes DNS-ének bázissorrendjét meghatároztuk és analizáltuk, mert ilyen vírusokból még nem volt ismert teljes genom. Befejezés előtt áll egy liba- és egy pulyka-adenovírus teljes szekvenciájának meghatározása is. Ezzel célunk az Aviadenovirus genusba tartozó ismert vírusok számának növelése volt. Filogenetikai számításokkal igazoljuk a madár adenovírusok gazdáikkal való koevolúciójára utaló jeleket. Egy kígyó- és egy hal-adenovírus genom elemzése az evolúciós viszonyok nagy vonalainak tisztázásához szolgáltat majd hasznos adatokat. Egy újnak bizonyult siadenovírus teljes genomanalízise folyik. A vírus, mely elhullást okozott Angliában ragadozómadár tenyészetekben a Siadenovirus nemzetség harmadik tagja lehet. Valamennyi vizsgált vírus új vírusfajt is képvisel, így elemzésük eredménye várhatóan tovább erősíti vagy cáfolja az általunk kialakított kategóriák helyességét. Az adenovírusok természetben megfigyelhető diverzitásának vizsgálatára érzékeny PCR módszert dolgoztunk ki. A PCR szűrés során pozitív, új típusnak látszó adenovírusok további vizsgálatát tervezzük. | In the framework of the project, we have studied adenoviruses that seemed to have great importance from the viewpoint of taxonomy. We determined and analyzed the nucleotide sequence of the full genome of two adenovirus isolates originating from Old World monkeys, because such viruses have not been examined before. The full genome sequencing of a goose and a turkey adenovirus, both isolated in Hungary, is almost finished. The purpose of the analysis of these avian adenoviruses was to increase the number of well-characterized members of the genus Aviadenoviruses. With phylogenetic calculations, we have confirmed that aviadenoviruses have a long co-evolutionary history with their avian hosts. Full genome analysis of a snake and a fish adenovirus will help to understand the large-scale evolutionary relations within the family Adenoviridae. Full genome sequencing of a novel siadenovirus is also in progress. The virus, that has caused mortality among captive birds of prey in England, will be only the third member in the genus Siadenovirus. Every adenovirus being studied represents a novel adenovirus species, thus criteria set for the demarcation of this taxon can also be evaluated. For the detection of biodiversity of adenoviruses circulating in the nature, a very sensitive PCR method was elaborated. We plan to further characterize those novel adenovirus candidates that are being detected during screening with PCR

Ubiquiter circovirus sequences raise challenges in laboratory diagnosis: The case of honey bee and bee mite, reptiles, and free living amoebae

Circoviruses of pigs and birds are established pathogens, however, the exact role of other, recently described circoviruses and circovirus-like viruses remains to be elucidated. The aim of this study was the detection of circoviruses in neglected host species, including honey bees, exotic reptiles and free-living amoebae by widely used broad-spectrum polymerase chain reaction (PCR) assays specific for the replication initiation protein coding gene of these viruses. The majority of sequences obtained from honey bees were highly similar to canine and porcine circoviruses, or, were distantly related to dragonfly cycloviruses. Other rep sequences detected in some honey bees, reptiles and amoebae showed similarities to various rep sequences deposited in the GenBank. Back-to-back PCR primers designed for the amplification of whole viral genomes failed to work that suggested the existence of integrated rep-like elements in many samples. Rolling circle amplification and exonuclease treatment confirmed the absence of small circular DNA genomes in the specimens analysed. In case of honey bees Varroa mite DNA contamination might be a source of the identified endogenous rep-like elements. The reptile and amoebae rep-like sequences were nearly identical with each other and with sequences detected in chimpanzee feces raising the possibility that detection of novel or unusual rep-like elements in some host species might originate from the microbial community of the host. Our results indicate that attention is needed when broad-spectrum rep gene specific polymerase chain reaction is chosen for laboratory diagnosis of circovirus infections

Nimodipine augments cerebrovascular reactivity in aging but runs the risk of local perfusion reduction in acute cerebral ischemia

IntroductionThe efficacy of cerebrovascular reactivity (CVR) is taken as an indicator of cerebrovascular health.Methods and ResultsWe found that CVR tested with the inhalation of 10 % CO2 declined in the parietal cortex of 18-20-month-old rats. The CVR deficit in old rats was coincident with cerebrovascular smooth muscle cell and astrocyte senescence, revealed by the immuno-labeling of the cellular senescence marker p16 in these cells. In a next series of experiments, CVR was severely impaired in the acute phase of incomplete global forebrain ischemia produced by the bilateral occlusion of the common carotid arteries in young adult rats. In acute ischemia, CVR impairment often manifested as a perfusion drop rather than blood flow elevation in response to hypercapnia. Next, nimodipine, an L-type voltage-gated calcium channel antagonist was administered topically to rescue CVR in both aging, and cerebra ischemia. Nimodipine augmented CVR in the aged brain, but worsened CVR impairment in acute cerebral ischemia.DiscussionA careful evaluation of benefits and side effects of nimodipine is recommended, especially in acute ischemic stroke

Candidate New Rotavirus Species in Sheltered Dogs, Hungary

We identified unusual rotavirus strains in fecal specimens from sheltered dogs in Hungary by viral metagenomics. The novel rotavirus species displayed limited genome sequence homology to representatives of the 8 rotavirus species, A–H, and qualifies as a candidate new rotavirus species that we tentatively named Rotavirus I

Genetic Parameters and Inbreeding Depression of Litter Weight in Pannon White Rabbits

Genetic parameters and inbreeding depression were estimated for litter weight at day 21 of a synthetic Pannon White rabbit population selected as a closed population since 1992. The data file consisted of 15652 kindling records of 3711 rabbit does (mated to 933 bucks) collected between 1992 and 2009. The total number of animals in the pedigree file was 4804. REML and BLUP procedures were applied using repeatability animal model (VCE and PEST soft ware). The estimated narrow sense heritability for litter weight was low (0.08±0.01). The estimated permanent environmental effects showed higher relative importance compared to the additive genetic effects (0.13±0.01). Analyzing the inbreeding depression simultaneously for all parities, increasing the inbreeding coefficient by 10% had small positive effect on litter weight (0.028 kg) but the estimate was not significant (p=0.30). When the inbreeding depression was estimated by parities small depressions (-0.023, -0.045 and -0.021 kg) were found for the 2-4th parity groups but the estimates were not significant (p=0.59, 0.23 and 0.61, respectively). On the contrary a significant positive effect was observed for the 1st parity (0.198 kg; p=0.01). The small positive estimate estimated at the 5th parity (0.033 kg) was not significant (p=0.39)