Inaktivacija virusa Y krompirja v vodi s hidrodinamsko kavitacijo

Autor: Ješelnik, Meta
Přispěvatelé: Dermastia, Marina
Jazyk: slovinština
Rok vydání: 2023
Předmět:
Popis: Z vodo se poleg človeških in živalskih patogenih virusov prenašajo tudi rastlinski. Eden od virusov, za katerega je bila možnost prenosa z vodo potrjena, je virus Y krompirja (PVY). PVY je eden od najnevarnejših virusov krompirja, saj uničuje pridelek ene najpomembnejših poljščin. Poleg krompirja napada tudi druge rastline v družini razhudnikovk (Solanaceae) npr. paradižnik. Zaradi vse večjega pomanjkanje čiste vode in varčevanja z njo, se v kmetijstvu, ki porablja za namakanje ogromne količine vode, vse bolj uveljavljajo zaprti namakalni sistemi z reciklirano vodo. Pred ponovno uporabo moramo iz take vode odstraniti patogene organizme. Za njihovo odstranjevanje razvijamo nove, okolju prijaznejše metode, kot je hidrodinamska kavitacija za inaktivacijo virusov v vodi. V raziskavi smo vodovodni vodi dodali PVY in jo nato obdelali s hidrodinamsko kavitacijo. Po različnih časih obdelave smo testirali virusno infektivnost. Pokazali smo, da po 500 kavitacijskih prehodih pod tlačno razliko 700 000 Pa, virus ne more več okužiti rastlin tobaka, v nekaterih poskusih pa smo virusno inaktivacijo dosegli že po krajših časih obdelave. Virusno infektivnost smo testirali na testnih rastlinah, ki smo jih okuževali z odvzetimi alikvoti obdelanega vzorca v različnih časovnih točkah. Poleg virusne infektivnosti smo z metodo RT-PCR preverjali tudi, če je prišlo do razgradnje RNA. Po vizualizaciji z agarozno gelsko elektroforezo smo v nekaterih primerih opazili le manjši vpliv na genomsko RNA in tako sklepali, da hidrodinamska kavitacija vpliva na različne virusne strukture. Inaktivacija virusa naj bi bila posledica poškodb virusne kapside, kar smo opazili s transmisijsko elektronsko mikroskopijo. Da je inaktivacija virusa v glavnem posledica mehanskih učinkov kavitacije, smo dokazali z dodatkom lovilcev prostih radikalov, ki niso imeli vpliva na virusno infektivnost. Human, animal, and even plant pathogens can be transmitted through water. One of the plant pathogenic viruses, for which the possibility of transmission through water has been confirmed, is potato virus Y (PVY). PVY is one of the most important potato viruses because it can destroy a large proportion of crops, causing major agricultural losses. PVY can also destroy some other plants in the Solanaceae family, such as tomato. Lack of clean water is a major global problem, and since agriculture consumes large amounts of it, closed irrigation systems that recycle water are being used. In order to prevent the spread of viruses with water, we need to remove or inactivate them. Therefore, new environmentally friendly methods are being developed to serve this purpose. One of these new methods is hydrodynamic cavitation. In this study, we treated tap water contaminated with PVY using hydrodynamic cavitation and observed the ability of the virus to infect plants after different treatment times. We showed that after 500 cavitation passes below a pressure difference of 700 000 Pa, the virus was no longer able to infect tobacco plants. In some experiments, the virus lost this ability after shorter treatment times. Viral infectivity was tested with test plants that were infected with aliquots of the treated sample. In addition to viral infectivity, we also tested RNA degradation using RT-PCR and its visualization using agarose gel electrophoresis. In some cases, only minor effect of cavitation on viral RNA was observed, suggesting that cavitation affects various viral structures to different extents, but viral inactivation is due to damage to the viral capsid, which was also observed by transmission electron microscopy. By adding radical scavengers, we showed that the radicals known to be generated during cavitation did not affect viral infectivity, suggesting that viral inactivation is likely caused by the mechanical effects of cavitation.
Databáze: OpenAIRE
Externí odkaz: