Ispitivanje imunofenotipa mononuklearnog ćelijskog infiltrata u jetri svinja prirodno inficiranih virusom hepatitisa E
Immunophenotyping of mononuclear cell infiltrate in swine liver naturally infected with hepatitis E virus
Докторанд
Kureljušić, Branislav I.Ментор
Knežević, MilijanaЧланови комисије
Knežević, MilijanaAleksić-Kovačević, Sanja
Delić, Dragan
Ivetić, Vojin
Метаподаци
Приказ свих података о дисертацијиСажетак
U ovoj disertaciji ispitano je 233 rektalna brisa od klinički zdravih svinja,
različitih uzrasnih kategorija, poreklom sa četiri farme, na prisustvo nukleinske
kiseline virusa hepatitisa E metodom RT-PCR. Pored toga, RT-PCR, bakteriološkom i
PCR analizom su ispitani uzorci jetre od 12 svinja kod kojih je RT-PCR nalaz iz
uzorka rektalnog brisa pokazao prisustvo HEV RNK. Kod 12 svinja kod kojih je
metodom RT-PCR u uzorku rektalnog brisa utvrđena nukleinska kiselina virusa
hepatitisa E, izvršena je punkcija vena cava cranialis i uzeta je krv za biohemijska
ispitivanja. Nakon koagulisanja krvi i izdvajanja seruma uz pomoć poluautomatskog
biohemijskog spektrofotometra određeni su sledeći parametri: ukupni proteini,
albumini, aspartat-aminotransferaza (AST), alanin-aminotransferaza (ALT), γ-
glutamil-transpeptidaza (GGT) i ukupni bilirubin.
Histopatološka i imunohistohemijska analiza urađena je na uzorcima jetre koji
su fiksirani u 10% neutralnom formalinu, obrađeni u automatskom tkivnom pro...cesoru
i uklopljeni u parafin. Parafinski isečci debljine oko 3-5μm bojeni su hematoksilineozin
i Masson trihrom metodom. Imunofenotipizacija ćelija mononuklearnog
infiltrata izvršena je streptavidin-biotin (LSAB2) metodom i tehnikom dvostrukog
imunohistohemijskog bojenja, uz upotrebu antitela za CD3 i CD79. Ekspresija TGFβ1
molekula postignuta je streptavidin-biotin (LSAB2) metodom. Za procenu gustine i
distribucije limfocitno – plazmaćelijskog infiltrata u jetri korišćena je
semikvantitativna analiza. Broj CD3 i CD79 pozitivnih ćelija u jetri određen je
morfometrijskom analizom. Pored deskriptivnih statističkih parametara upotrebljeni
su ANOVA i Tukey test.
Ukupno je ispitano 233 svinje, od kojih je kod dvanaest u uzorku rektalnog
brisa dokazana HEV RNK (5,15%). Istovremeno prisustvo HEV RNK u uzorku
rektalnog brisa i jetre je ustanovljeno kod pet svinja (41,67%). U dvanaest uzoraka
jetre nije dokazano prisustvo aerobnih i anaerobnih bakterija i genoma svinjskog
cirkovirusa tip 2...
In this thesis from clinically healthy pigs of different ages, originating from 4
farms, a total of 233 rectal swabs were tested for the presence of HEV RNA by RTPCR.
In addition, liver samples of 12 pigs which were RT-PCR positive to HEV RNA
were tested by RT-PCR, bacteriological and PCR analysis. A total of 12 pigs which
rectal swabs tested positive by RT-PCR analysis were further on sampled by puncture
of vena cava cranialis, as well. After coagulation blood sera were analyzed on a
biochemical spectrophotometer and the following parameters were determined: total
protein, albumin, aspartate aminotransferase (AST), alanine aminotransferase (ALT),
γ – glutamil transpeptidase (GGT) and total bilirubin.
Histopathological and immunohistochemical analyses were performed on liver
samples fixed in 10% neutral formalin, processed in an automated tissue processor
and embedded in paraffin. Paraffin sections about 3-5μm thick were stained with
hematoxilin-eosin and by Masson trichrome method. Im...munophenotypisation of
mononuclear infiltrate cells was carried out by the streptavidin – biotin method
(LSAB2) and by the double immunohistochemical staining technique with antibodies
for CD3 and CD79. Expression of TGF-β1 molecule was obtained by the streptavidin
– biotin method (LSAB2). Estimation of the density and distribution of lymphocyticplasmacytic
infiltrate in the liver was done by semiquantitative analysis. The number
of CD3 and CD79 positive cells in the liver was determined by morphometric
analysis. In addition to descriptive statistical parameters ANOVA and Tukey test were
used, as well.
A total of 233 pigs were tested. Out of this number in 12 rectal swabs HEV
RNA (5.15%) was confirmed. Concurrent presence of HEV RNA in both rectal and
liver samples was determined in 5 pigs (41.67%). In 12 liver samples the presence of
aerobic and anaerobic bacteria, as well as circovirus type 2 genome were not
confirmed.
Results of blood serum biochemical testing did not show statistical
significance between values for total proteins, albumins, and AST between the
infected pigs and the control group. A statistically very significant difference (p<0.01) between the infected and the control group was determined for ALT, and a significant
difference (p<0.05) was recorded for GGT and total bilirubin...
Факултет:
Универзитет у Београду, Факултет ветеринарске медицинеДатум одбране:
12-06-2012Пројекти:
- Унапређење здравља и добробити високопродуктивних крава идентификацијом и отклањањем стресогених фактора (RS-MESTD-Technological Development (TD or TR)-31062)