Naslov
The role of ABC-mediated transport in biocidal activity of natural and synthetic compounds on Anisakis pegreffii larvae

Autori
Trumbić, Željka ; Bušelić, Ivana ; Hrabar, Jerko ; Vrbatović, Anamarija ; Ujević, Ivana ; Roje Busatto, Romana ; Babić, Ivana ; Messina, Concetta Maria ; Mladineo, Ivona

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Abstract Book of the 18th International Conference on Diseases of Fish and Shellfish / Mladineo, I. - , 2017, 39-39

Skup
18th EAFP International Conference on Diseases of Fish and Shellfish

Mjesto i datum
Belfast, Ujedinjeno Kraljevstvo, 04.09.2017. - 08.09.2017

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
ABC transmembrane transporters, nerolidol, farnesol, abamectin, levamisol

Sažetak
Anisakiasis, caused by accidental ingestion of live L3 stage larvae of Anisakis spp. through undercooked seafood, is a widespread, often underdiagnosed illness. One of the mechanisms commonly implicated in the development of drug resistance in nematodes is associated with the cellular efflux activity of ABC (ATP binding cassette) transmembrane transporters. Nematodes possess numerous homologues of ABCBs (Pgps and half-transporters) and ABCCs (MRPs) transporters with important physiological roles, aside from their role in multixenobiotic resistance. The scope of this study was to compare biocidal activity of two natural monoterpens (farnesol, nerolidol) and two synthetic widely-used anthelminthics (abamectin, levamisol) on Anisakis spp. larvae and the degree of involvement of ABC efflux mechanism in their pharmacokinetics. Ten Anisakis pegreffii larvae were exposed to growing concentrations of target chemicals ; experiment was conducted in six replicates with or without simultaneous administration of two inhibitors ; Valspodar (Pg-p inhibitor) and MK571 (MRP inhibitor). The concentration of target compound at which only 50% of larvae were alive (LC50) was determined: 135.9 μM for nerolidol, 134.4 μM farnesol, 0.3 μM abamectin and 0.84 for levamisol, significantly influenced by presence of one or both inhibitors for all chemicals except for abamectin. Toxicity curves were used to extrapolate doses where highest difference in larvicidal activity between the compound and one or both inhibitors was observed, suggesting it as point where ABC transporters elicit highest effect. Anisakis larvae were exposed to this concentration and sampled at 2, 6, 12 and 18 h after treatment for further analyses: quantification of Pg-p and MRP-like transcripts by real-time PCR, production of reactive oxygen species (ROS), intake of target compounds by HPLC, activity of transporters by Calcein-AM assay and effect on larval cytoskeleton by IHC and confocal microscopy. The results are discussed and provide integral view and understanding of the processes involved in larvicidal activity and protection mechanisms conferred by Anisakis pegreffii against tested compounds.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Veterinarska medicina, Biotehnologija

POVEZANOST RADA