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Razširjenost okužbe z virusom hepatitisa E pri prašičih ob zakolu
ID Raspor Lainšček, Petra (Avtor), ID Kirbiš, Andrej (Mentor) Več o mentorju... Povezava se odpre v novem oknu, ID Toplak, Ivan (Komentor)

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Izvleček
Naša raziskava sovpada s porastom zanimanja za »novo porajajočo se« bolezen virusa hepatitisa E (HEV) v svetu in prinaša nove izsledke o razširjenosti okužbe v slovenskih rejah prašičev. Ker gre za zoonozo, smo z raziskavo želeli ugotoviti, kakšno je stanje v slovenskih klavnicah in ali hepatitis E virus ogroža zdravje ljudi. Vzorčenje prašičev je potekalo v štirih klavnicah po Sloveniji med junijem in septembrom 2014. Vzorce blata, jeter in žolča smo odvzeli 811 klinično zdravim prašičem različnih starostnih kategorij. Ob vzorčenju v klavnici smo jemali tudi brise površin na klavni liniji, vzorčili mleto meso in pečenice. Z metodo RT-PCR v realnem času smo dokazovali prisotnost nukleinske kisline HEV v odvzetih vzorcih. Istim prašičem smo odvzeli tudi vzorec krvi in določali prisotnosti protiteles IgG proti HEV. Največ HEV pozitivnih živali smo ugotovili med 322 odojki, in sicer 17 %. Med 400 pitanci smo ugotovili en pozitiven vzorec jeter in en pozitiven vzorec žolča, kar je bistveno manj, kot smo pričakovali. Med 89 plemenskimi svinjami nismo ugotovili pozitivne živali. Med 63 brisi površin smo odkrili dva pozitivna vzorca. V vzorcih pečenic in mletega mesa prisotnosti HEV nismo ugotovili. Z genetsko tipizacijo 45 HEV pozitivnih vzorcev in filogenetsko primerjavo nukleotidnih zaporedij slovenskih sevov skupaj z zaporedji sevov HEV, ki so dostopni v podatkovni zbirki GenBank, smo ugotovili pojav velike genetske raznolikosti med vzorci ter prisotnost štirih različnih podtipov genotipa 3 (3a, 3b, 3e in 3f) HEV v Sloveniji. V izvedeni študiji genotipa 4 nismo ugotovili. V določanje nukleotidnega zaporedja smo vključili tudi pet humanih vzorcev, pozitivnih na prisotnost HEV. Zaradi premajhne količine virusne RNA v HEV pozitivnih humanih vzorcih nam pri nobenem ni uspelo določiti nukleotidnega zaporedja. Z dvema komercialnima encimskoimunskima testoma (ID Screen® Hepatitis E Indirect Multi-species proizvajalca IDvet in HEV-IgG ELISA porcine proizvajalca Axiom) smo pregledali vzorce krvi domačih prašičev na prisotnost protiteles IgG proti HEV. Pregledali smo 351 vzorcev, naključno izbranih iz vzorcev odvzetih na celotnem območju Slovenije v okviru rednega monitoringa kužnih bolezni pri prašičih v letih 2013 in 2014, ter 359 vzorcev, odvzetih v klavnici ob zakolu leta 2014. Primerjava rezultatov obeh testov je pokazala, da je encimskoimunski test proizvajalca Axiom ustreznejši za presejalna testiranja. Za razjasnitev 38 rezultatov, ki se pri vzorcih iz klavnice pri obeh encimskoimunskih testih niso ujemali, smo uporabili modificiran humani imunoblot test recomLine HEV IgG/IgM (Mikrogen diagnostik). Izvedena raziskava je prva tovrstna študija, ki dokazuje prisotnost HEV pri prašičih na klavni liniji v Sloveniji, prav tako je prva izvedena sistematična raziskava v tako širokem obsegu v Sloveniji. Rezultati raziskave prispevajo k boljšemu razumevanju stanja okužb prašičev s HEV ob vstopu na klavno linijo, in podajajo natančnejšo sliko o stanju prekuženosti prašičev v rejah. Glede na številčnost pregledanih vzorcev in razpršenost rej po državi, iz katerih so izhajali vzorčeni prašiči, lahko z gotovostjo trdimo, da rezultati podajajo natančen podatek o razširjenosti okužb s HEV med domačimi prašiči v Sloveniji.

Jezik:Slovenski jezik
Ključne besede:bolezni prašičev, virologija, zoonoze, hepatitis E, filogenija, blato, jetra, mesni izdelki, verižna reakcija s polimerazo, encimsko vezani imunosorbentni test, analiza zaporedja DNA, RNA, prašiči
Vrsta gradiva:Doktorsko delo/naloga
Tipologija:2.08 - Doktorska disertacija
Organizacija:VF - Veterinarska fakulteta
Leto izida:2022
Št. strani:126 f.
PID:20.500.12556/RUL-135555 Povezava se odpre v novem oknu
COBISS.SI-ID:102455811 Povezava se odpre v novem oknu
Datum objave v RUL:19.03.2022
Število ogledov:1398
Število prenosov:100
Metapodatki:XML DC-XML DC-RDF
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Sekundarni jezik

Jezik:Angleški jezik
Naslov:Prevalence of hepatitis E virus infection in pigs at slaughter
Izvleček:
Our research coincides with increasing interest in the newly emerging zoonosis, the hepatitis E virus, and provides new information regarding infection distribution in Slovenian pig herds. Since HEV is a zoonosis, our goal was to investigate the existing condition in Slovenian slaughterhouses and to assess associated health risks posed to consumers and personnel. Sampling of pigs was conducted in four Slovenian slaughterhouses, between June and September 2014. Feces, liver and bile samples were collected from 811 healthy pigs (different age categories). At the time of slaughter, swab samples of different surfaces and samples of minced meat and sausages were also taken. All samples were analyzed with real time PCR (RT-PCR) for the presence of HEV. Blood samples taken from the same pigs were analyzed for the presence of IgG HEV antibodies. The highest number of HEV positive animals was found in the 3 month old age group of pigs (322 animals), where 17% of animals were HEV positive. In the age group of 6 month old pigs (400 animals), only one liver sample and one bile sample was positive, which was significantly less than we would have expected. Out of 89 pigs that were older than 1 year, none was HEV positive. Out of 63 surface swabs, two were positive for the presence of HEV. None of the minced meat and sausage samples was positive. A phylogenetic tree, generated combining sequences obtained from Slovenian strains (45 positive samples) and nucleotide sequences of HEV strains derived from GeneBank, revealed high genetic variability between samples, allowing classification of the identified Slovenian strains into four different subtypes of genotype 3 (3a, 3b, 3e and 3f). Genotype 4 was not found. Additionally, five HEV positive human samples were sequenced, although sequencing was unsuccessful due to an insufficient amount of viral RNA. With two commercial ELISA (ID Screen® Hepatitis E Indirect Multi-species (IDvet) and HEV-IgG ELISA porcine (Axiom)) we analyzed blood samples from domestic pigs for the presence of IgG HEV antibodies. We analyzed 351 randomly selected serum samples collected from pigs located in different parts of Slovenia (these samples were collected within the scope of annual monitoring of infectious diseases in 2013 and 2014) and 359 samples collected from pigs at slaughter in 2014. Comparison of results from both ELISA showed that Axiom ELISA is more suitable as a screening method. A human immunoblot test was optimized and standardized to test 38 serum samples that showed discrepancies in results obtained with Axion and IDvet ELISA kits. The work presented herein represents the first systematic, wide scope analysis on the presence of HEV in pigs at slaughter in Slovenia. Results of the research have contributed to a better understanding of pig HEV infection status at slaughter and have shown a more detailed picture of seroprevalence status in pig farming. Since our research included a great number of samples and dispersion of different farms around the country, we can safely say that the obtained results show accurate information about pig HEV infections in Slovenia.

Ključne besede:swine diseases, virology, zoonoses, hepatitis E, phylogeny, feces, liver, meat products, polymerase chain reaction, encyme-linked immunosorbent assy test, sequence analysis DNA, RNA, swine

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