TOKSIČNOST I KVANTIFIKACIJA ODREĐENIH SINTETIČKIH I PRIRODNIH ANTIHELMINTIKA U TKIVU INFEKTIVNE LIČINKE Anisakis pegreffii (Campana-Rouget & Biocca, 1955) PRI ANTIPARAZITSKOM TRETMANU

doctoral thesis

• Title (english): TOXICITY AND QUANTIFICATION OF CERTAIN SYNTHETIC AND NATURAL ANTHELMINTICS IN THE TISSUE OF INFECTIOUS Anisakis pegreffii (Campana-Rouget & Biocca, 1955) LARVAE DURING THE ANTIPARASITIC TREATMENT
• Author: Romana Roje Busatto
• Mentor: Ivana Ujević (mentor)
• Committee member: Romana Gračan (predsjednik povjerenstva)
• Committee member: Tvrtko Smital (član povjerenstva)
• Committee member: Damir Kapetanović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Geology); Zagreb
• Defense date and country: 2023-09-28, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Interdisciplinary Natural Sciences; Marine Science
• Universal decimal classification (UDC): 55 - Geology. Meteorology. Hydrology

Abstract

Uslijed sve češće pojave nametničkih infestacija i razvoja otpornosti nametnika na tretmane postojećim antihelminticima potrebno je ispitati učinkovitost i mehanizme istih, ali i okrenuti se alternativnim, prirodnim izvorima zaštite. Ovo istraživanje predstavlja nove informacije o učinkovitosti mehanizama samozaštite L3 ličinki nametničke vrste oblića Anisakis pegreffii (Campana-Rouget & Biocca, 1955) u interakciji sa sintetičkim antihelminticima, abamektinom i levamisolom, te prirodnim seskviterpenskim spojevima, farnesolom i nerolidolom. Provođenjem in vitro eksperimenata s nizom koncentracija ispitivanih spojeva uz prisustvo/odsustvo inhibitora ABC transportera, valspodara ili MK571, su metodom tekućinske kromatografije određene njihove letalne koncentracije i razlike u razini akumulacije u tkivima Anisakis ličinki što daje uvid u mehanizme samozaštite ličinki koji se očituju u staničnoj aktivaciji ABC transmembranskih transportnih proteina (P-gp i MRP). Toksičnost ispitivanih spojeva na stanice ličinki je dodatno potvrđena testom proizvodnje reaktivnih kisikovih čestica (engl. reactive oxygen species, ROS), dok je imunohistokemijski određen njihov štetni utjecaj na citoskelet ličinki. Navedena ispitivanja doprinose razumijevanju mehanizma procesa višestruke rezistencije na ksenobiotike (engl. multixenobiotic resistance, MXR) čime su se dobile smjernice za daljnja ispitivanja s ciljem odgode pojave rezistencije na korištene terapeutike.

Abstract (english)

Due to the widespread prevalence of parasitic infestations and numerous reports on parasites developing resistance to anthelmintic treatments, it is necessary to investigate the efficacy and mechanisms of action of the currently used therapeutics, but also to turn to alternative, natural sources. This research provides new information on the efficacy of the self-protective mechanism of the parasitic species Anisakis pegreffii (Campana-Rouget & Biocca, 1955) when interacting with frequently used anthelmintics, abamectin and levamisole, and natural sesquiterpene compounds, farnesol and nerolidol. After performing in vitro experiments with a range of concentrations of the tested compounds in the presence/absence of the ABC transporter inhibitors valspodar or MK571, the liquid chromatography technique was used to determine lethal concentrations and differences in toxicity and accumulation of the target compounds in Anisakis L3 larvae. Production of Reactive Oxygen Species (ROS) indicated the level of stress and immunohistochemistry was used to evaluate deleterious effects of the tested compounds on the cytoskeleton of Anisakis larvae. Insight into the defence mechanism of treated larvae manifested by cellular activation of ABC transporters (P-gp and MRP transmembrane proteins) contributes to the understanding of the Multixenobiotic Resistance (MXR) mechanisms and provides guidelines for further research to delay the onset of this type of anthelmintic resistance.

• Keywords: ABC transmembranski transporteri; Anisakis; antiparazitska sredstva; multiksenobiotička otpornost; nametnici; tekućinska kromatografija
• Keywords (english): ABC Transmembrane Transporters; Anisakis; Antiparasitic agents; Multixenobiotic Resistance; Parasite; Liquid Chromatography
• Language: croatian
• URN:NBN: urn:nbn:hr:217:551886
• Promotion: 2024
• Project: Number: IP-2013-11-5576; Title: Anisakis spp: genomska epidemiologija; Title: Anisakis spp: genomic epidemiology; Acronym: AnGEL; Leader: Ivona Mladineo; Jurisdiction: Croatia; Funder: Hrvatska zaklada za znanost; Funding stream: Research Projects
• Study programme: Title: DOCTORAL STUDY OF OCEANOLOGY; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti u području prirodnih znanosti (doktor/doktorica znanosti u području prirodnih znanosti)
• Type of resource: Text
• Extent: 143 str. ; 30 cm
• File origin: Born digital
• Access conditions: Open access
• Created on: 2025-01-30 14:45:34