PhD Thesis Demeter Zoltan

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Az FPV és CPV2 együttes kimutatására alkalmas klasszikus PCR-alapú eljárás segítségével a vizsgált 230 minta közül 72 (31,3 %) pozitívnak bizonyult (17 macska, 1 oroszlán, 1 pálmasodró és 53 kutya eredető minta esetében). Egy korábban leközölt MboII alapú RFLP vizsgálat alkalmazását követıen 20 magyarországi CPV2 törzsbıl 15 esetében CPV2c genotípusú törzseknek megfelelı eredményt kaptunk. A nuklein- és aminosav szekvenciák vizsgálata alapján az összes vizsgált magyarországi törzs tulajdonképpen 2a típusú CPV-nak bizonyult. A félrevezetı eredmény magyarázata a PCR során keletkezett amplikonok másik végén létrejött pontmutációban rejlik. Ennek következtében a 20 magyarországi törzsbıl az RFLP alapú eljárást követıen 15 esetében félrevezetı genetipizálási eredményt kaptunk. Ez a tény a gyors genotipizálási technikák folyamatos felülvizsgálatának, illetve az eredmények megbízhatóbb eljárások segítségével történı alátámasztásának szükségességét emeli ki. Vizsgálataink során elsıként sikerült direkt eljárásokkal kimutatni FPV fertızést egy cibetmacskafélében (Viverridae). A megvizsgált ázsiai pálmasodróban (Paradoxurus hermaphroditus) keletkezett kórbonctani és kórszövettani elváltozások megegyeztek az FPV által más állatfajokban elıidézett elváltozásokkal. Ugyanakkor a fertızés sikeres kimutatása fényt derített arra is, hogy a kórokozó gazdaspektruma szélesebb az eddig ismertnél, és hogy az FPV iránt a cibetmacskafélék is fogékonyak. FPV fertızést egy Érd mellıl származó oroszlánban (Panthera leo) is sikerült kimutatni. Az ázsiai pálmasodróban, az oroszlánban és két macskában kimutatott vírustörzsek genetikai vizsgálatának eredménye arra enged következtetni, hogy a jelenleg Magyarországon cirkuláló FPV törzsek viszonylag heterogén csoportot alkotnak. 10
1. Introduction Based on modern scientific results an enormous variety of viral pathogens have been concurrently evolving with species belonging to all taxonomical classes, presumably ever since the early days of life on Earth. The life of all living creatures, starting with the simplest forms of life to the highest, most evolved forms of existence, was, is and most likely will be considerably influenced by these infinitesimal pathogens. Companion animals, such as the dog, cat and many other species are no exceptions. Virus infections, such as rabies, distemper, parvovirus infections, just to name a few of the most relevant ones, have shaped and decisively influenced receptive populations throughout the early, recent and present history. As “with great power comes great responsibility” the research, better understanding, treatment and most importantly the prevention of these diseases that threaten the well-being and survival of all receptive species fall under the responsibility of human beings, who did and will benefit of these species throughout the centuries. Viral diseases such as distemper and parvovirus infection constantly threaten susceptible animal populations. The understanding of these diseases, including the molecular characteristics of the pathogens, represents the first line of defense and constitutes the basis of the better prevention of infections and protection of animals. The hypothesis of the present study was that presently there is more than one genotype of CDV and CPV2 present in Hungary. The hypothesis was tested by detecting the pathogens in naturally infected animals and by analyzing relevant segments of the viral genomes. The infections were demonstrated using clinical and necropsy samples collected between 2004 and 2008 from several different species, such as dogs, foxes, ferrets, raccoons and an Asian palm civet. The genetic characterization of the pathogens was performed by determining the nucleic acid sequence of key segments of their genome and by using these data to determine the phylogenetic relationships between the Hungarian and strains previously isolated in other parts of the world. 11
Page 1 and 2: Szent István University Postgradua
Page 3 and 4: Contents Abbreviations ............
Page 5 and 6: aa amino acid Asn asparagine Asp as
Page 7 and 8: Summary Samples taken from various
Page 9: Összefoglaló (Summary in Hungaria
Page 13 and 14: 3.1.1 General informations 3. Liter
Page 15 and 16: stomach, and mononuclear cells of t
Page 17 and 18: (noninflammatory demyelination), ho
Page 19 and 20: according to the area of CNS involv
Page 21 and 22: Edward Jenner was probably the firs
Page 23 and 24: comparison, a study conducted by Bo
Page 25 and 26: 3.2.2.1 Neonatal animals Infection
Page 27 and 28: 3.2.4 Pathology Gross pathologic ch
Page 29 and 30: 3.3.1 General informations 3.3 Type
Page 31 and 32: ecovered from the intestinal-associ
Page 33 and 34: 3.3.5 Diagnosis, treatment and prev
Page 35 and 36: een overtaken by CPV2b and/or CPV2c
Page 37 and 38: Table 1: Description of the analyze
Page 39 and 40: purposes by a circus lion tamer fro
Page 41 and 42: Table 4: Primers used for the diagn
Page 43 and 44: 4.5.3.2 Classical PCR assays The 50
Page 45 and 46: 5.1.1 CDV infections 5. Results 5.1
Page 47 and 48: 5.1.2 FPV infections During necrops
Page 49 and 50: 5.2.1 CDV infection Most relevant c
Page 51 and 52: 5.3 Electron microscopy In the urin
Page 53 and 54: amplicons obtained from field virus
Page 55 and 56: 5.4.3 Sequence analysis and phyloge
Page 57 and 58: Fig. 19: Phylogenetic tree construc
Page 59 and 60: P gene Vanguard A75/17 Ferret Onder
Page 61 and 62:
(98.90 %) isolated from a naturally
Page 63 and 64:
Fig. 22: Phylogenetic tree construc
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5.4.3.2 FPV strains PCR positive sa
Page 67 and 68:
The alignment of the Hungarian CPV2
Page 69 and 70:
America. The similarity of the Hung
Page 71 and 72:
Only one strain from each subgroup
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Vanguard (Pfizer Animal Health, USA
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performed in order to reveal whethe
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a relatively quick evolutionary pat
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7. New Scientific Results Most impo
Page 81 and 82:
APPEL, M.J.G.: Canine distemper vir
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DE YBANEZ R.R., VELA C., CORTES E.,
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HAAS L., MARTENS W., GREISER-WILKE
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KIMURA, M.: A simple method for est
Page 89 and 90:
MEURS K.M., FOX P.R., MAGNON A.M.,
Page 91 and 92:
PARRISH C.R.: Pathogenesis of felin
Page 93 and 94:
VANDEVELDE M., ZURBRIGGEN A., HIGGI
Page 95 and 96:
10. Other Publications in Peer Revi
Page 97 and 98:
Academy of Sciences and the Doctora
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PhD Thesis Demeter Zoltan

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