Author/Editor		Strajn, Breda; Kotnik, Vladimir; Vengušt, Andrej; Pestevšek, Uroš; Malovrh, Tadej
Title		The effects of mycotoxin deoxynivalenol on the immune system in pigs
Translated title		Učinek mikotoksina deoksinivalenola na imunski odziv prašičev
Type		članek
Source		Med Razgl
Vol. and No.		Letnik 43, št. Suppl 5
Publication year		2004
Volume		str. 167-71
Language		eng
Abstract		Deoxynivalenol (DON, vomitoxin), one of the world's most common mycotoxin contaminants, is a type B trichothecene, which occurs predominantly in grains. DON has been implicated in incidents of mycotoxicoses in both humans and animals. Pigs are more sensitive to trichothecenes than other farm animals. The research was performed to establish the effect of DON on the immune system in gilts. In a controlled field experiment, twenty pregnant gilts were randomly divided into control and experimental group. The composition and nutritive value of complete feeding stuffs were the same for both groups. Maize in experimental diet was naturally contaminated with DON. The experimental diet contained 5.08 mg/kg DON. During the 56-day feeding trial, blood samples were collected on day 0 (3 weeks before parturitions),17, 42 and at the end of experiment (day 56). Lymphocyte stimulation was measured by BrdU incorporation test using two different concentrations of PHA (10 and 20 mg/ml), three different concentrations of DON (5000 ng/ml, 1000 ng/ml or 100 ng/ml) and combination of both, mitogen and toxin. Using the photometric enzyme immunoassay for quantitative determination of apoptosis, the apoptosis induced by DON was studied in non-stimulated 72/h lymphocyte culture or 72/h culture stimulated with 1000 ng/ml of DON. Furthermore, the lymphocyte count in control and experimental group was evaluated by flow cytometry. Freshly drawn blood, anticoagulated with Lithium heparin, was labelled using monoclonal antibodies directed against CD3+CD4+ and CD3+CD8+ cells. Completely depressed lymphocyte response was observed in cultures deriving from control and experimental group stimulated with toxin or combination of both, toxin and mitogen. PHA reconstituted depressed lymphocyte response when cells were stimulated with a lower dose of toxin. The only identical response in lymphocytes from both groups was noted after PHA stimulation. (Abstract truncated at 2000 characters).
Summary		Mikotoksin deoksinivalenol (DON, vomitoksin) spada v skupino trihotecenov B in je eden od najbolj razširjenih mikotoksinov po vsem svetu. Najdemo ga predvsem v žitaricah in povzroča mikotoksikoze tako pri ljudeh kot pri živalih. Od farmskih živali so prašiči na DON najbolj občutljivi. Z raziskavo smo želeli ugotoviti učinek DON na imunski odgovor mladic, zato smo dvajset brejih mladic (prvič osemenjenih svinj) naključno razdelili v kontrolno in poskusno skupino. Za obe skupini je bila količina posameznih surovin in vsebnost hranilnih snovi v krmni mešanici enaka. Koruza v krmi za poskusno skupino pa je bila naravno kontaminirana z DON. V krmi je bilo 5,08 mg/kg DON. Poskus je trajal 56 dni. V tem času smo krvne vzorce odvzeli na dan začetka poskusa (dan 0, približno 3 tedne pred porodi),17. dan ter 42. dan po porodu in na koncu poskusa (56. dan). Stimulacijo limfocitov smo izmerili s testom BrDU, pri čemer smo uporabili dve različni koncentraciji PHA (10 in 20 mg/ml), tri različne koncentracije DON (5000 ng/ml,1000 ng/ml ali 100ng/ml) ter kombinacijo obeh, toksina in mitogena. Apoptozo smo preučevali na 72 ur starih limfocitnih kulturah, ki so bile brez dodatka DON ali pa stimulirane s 1000 ng/ml DON. Za kvantitativno determinacijo apoptoze smo uporabili fotometrično encimskoimunsko reakcijo, s katero ugotavljamo fragmentacijo DNK. Število limfocitov v kontrolni in poskusni skupini smo ugotavljali s pretočno citometrijo. Svežo odvzeto kri z dodatkom litijevega heparina smo označili z monoklonskimi protitelesi proti celicam CD3+CD4+ in CD3+CD8+. Ko smo za stimulacijo uporabili toksin ali kombinacijo toksina in mitogena, je bil v kulturah limfocitov kontrolne in poskusne skupine limfocitni odgovor popolnoma zatrt. Pri nižjih dozah toksina je bila stimulacija limfocitov boljša. Odgovor limfocitov kontrolne in poskusne skupine je bil identičen samo po stimulaciji s PHA. (Izvleček skrajšan pri 2000 znakih).
Descriptors		MYCOTOXINS T-LYMPHOCYTES APOPTOSIS SWINE FLOW CYTOMETRY LYMPHOCYTE TRANSFORMATION