20.500.12556/RUL-127031 Optimizacija metod za spremljanje procesov, povezanih z imunostjo v hemolimfi kopenskih enakonožcev Optimisation of methods to evaluate the immune-related processes in the haemolymph of terrestrial isopods Imunski sistem rakov gradijo imunske celice in molekule, katerih glavna funkcija je razlikovanje med lastnim in tujim ter oblikovanje ustreznega odgovora na potencialne grožnje za organizem. V hemolimfi kontrolnih oziroma asimptomatskih kopenskih rakov vrste Porcellio scaber (navadni prašiček) smo optimizirali in določili referenčne vrednosti celičnih in humoralnih parametrov, ki so posredno ali pa neposredno vpleteni v delovanje prirojenega imunskega sistema rakov. S kombinacijo tehnik svetlobne in elektronske mikroskopije smo v hemolimfi asimptomatskih P. scaber opisali tri glavne tipe hemocitov (semigranulociti, granulociti in hialinociti) in podali informacije o njihovi morfologiji ter ultrastrukturi. Diferencialno interferenčno kontrastno mikroskopsko tehniko smo prepoznali kot hiter in zanesljiv pristop za merjenje gostote različnih tipov hemocitov v svežih vzorcih hemolimfe asimptomatskih živali, pri čemer so semigranulociti najpogosteje zastopan tip (povprečno 65 %), sledijo hialinociti (18 %) in granulociti (17 %). Uspešnost optimizacije metod za merjenje celičnih in humoralnih parametrov v hemolimfi P. scaber smo preverili še z ovrednotenjem vrednosti izbranih parametrov v hemolimfi živali, ki so imele izražene simptome bakterijske (Rhabdochlamydia porcellionis) in virusne okužbe (Iridovirus IIV-31). Vrednosti merjenih parametrov v hemolimfi simptomatskih P. scaber se značilno razlikujejo od asimptomatskih P. scaber, hkrati pa se enakonožec na bakterijsko oziroma virusno okužbo odzove z aktivacijo različnih procesov prirojene imunosti. V primeru bakterijske okužbe P. scaber smo zaznali izrazitejšo modulacijo humoralnih parametrov, medtem ko je virusna okužba izzvala občutno spremembo celičnih parametrov v hemolimfi P. scaber. The immune system of crustaceans consists of immune cells and molecules whose main function is to distinguish between self and non-self and to maintain an adequate response to a potential threat to the organism. In the haemolymph of control or asimptomatic terrestrial isopods Porcellio scaber, we optimized and evaluated reference values of cellular and humoral parameters indirectly or directly involved in the function of the innate immune system. Using a combination of light and electron microscopic techniques, we have described three main haemocyte types (semigranulocytes, granulocytes, hyalinocytes) in the haemolymph of asimptomatic P. scaber and provided information on their morphology and ultrastructure. Differential interference contrast microscopic tehnique was recognised as a rapid and reliable method for measuring the density of different haemocyte types in fresh haemolymph samples from asimptomatic animals, with semigranulocytes being the most common type (65% on average), followed by hyalinocytes (18%) and granulocytes (17%). The performance of the method optimization for measuring cellular and humoral parameters in the haemolymph of P. scaber was validated with the evaluation of the values of selected parameters in the haemolymph of animals with evident symptoms of bacterial (Rhabdochlamydia porcellionis) and viral (Iridovirus IIV-31) infection. The values of measured parameters in the haemolymph of symptomatic P. scaber differ significantly from asymptomatic P. scaber. Isopods respond differently to bacterial or viral infection, i. e. different set of innate immunity processes are induced in each of the infection. While bacterial infection triggers significant changes in humoral parameters, cellular parameters were significantly altered upon viral infection. porcellio scaber optimizacija metod procesi povezani z imunostjo hemolimfa bakterijska in virusna okužba porcellio scaber optimisation of methods immune-related processes haemolymph bacterial and viral infection true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2021-05-14 07:15:14 2021-05-14 07:15:30 2022-09-06 03:49:26 0000-00-00 00:00:00 2021 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 191179 71802627 12858.pdf 12858.pdf 1 31F9E9FF4A37F7DF1933B9981C55B452 2c064b82cde4df35125614e3675906110a64216b74ef3dd260ff6c37637a3610 67bdf225-b473-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=143233 Biotehniška fakulteta 0 0 0