Browsing by Author "Vitrio, Nathalia"
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- Comparative Analysis of DNA Extraction Methods for Individual Varroa destructorPublication . Costa, Maíra; Lopes, Ana; Yadró Garcia, Carlos A.; Vitrio, Nathalia; Gonçalves, Telma; Pinto, M. Alice; Henriques, DoraThe ectoparasitic mite Varroa destructor is one of the major honey bee threats and it is associated to population worldwide decline. Genetic analyses using the mtDNA of V. destructor are fundamental for establishing the taxonomy and distribution of the mites. Consequently, low-quality DNA can lead to inaccurate or inconsistent data, making genetic interpretation more challenging. In this study, we compared the concentration and quality of DNA extracted from individual female V. destructor using two different commercial kits, aiming to identify the optimal method for obtaining high-quality DNA. Total DNA was extracted from mites using both an automated and manual extracted method. In addition, manual kit extraction tested three incubation procedure (1h, 5h, and overnight). DNA concentration was quantified using three different instruments: the SpectroStarVR Nano LVis Plate spectrophotometer, the Quantus™ Fluorometer apparatus, and NanoDrop™. The manual extraction DNA concentration did not vary across incubation times and the concentration values varied between 0.240-0.545 ng/μl (Quantus), 0.72-4.49 ng/μl (spectrophotometer), and 0.0-1.47 ng/μl (NanoDrop). While extraction automatic approach yielded higher respectively 0.483-0.631 ng/μl, 20.33-9.0 ng/μl, and 5.3-6.8 ng/μl. In conclusion, the automated kit extraction seems to be the best extraction method since it produced the higher-concentration DNA using only one individual mite.
- Wing geometric morphometrics of european honey bee populationsPublication . Costa, Maíra; Gonçalves, Telma; Lopes, Ana; Vitrio, Nathalia; Albo, Alexandre; Blažytė-Čereškienė, Laima; Brodschneider, Robert; Brusbardis, Valters; Carreck, Norman L.; Charistos, Leonidas; Chlebo, Robert; Coffey, Mary F.; Danneels, Ellen; Graaf, Dirk C.; Dobrescu, Constantin; Filipi, Janja; Gajda, Anna; Gratzer, Kristina; Hatjina, Fani; Johannesen, Jes; Körmendy-Rácz, János; Kovačić, Marin; Kristiansen, Preben; Martikkala, Maritta; Martín-Hernández, Raquel; Pietropaoli, Marco; Poirot, Benjamin; Radev, Zheko; Raudmets, Aivar; Douarre, Vincent; Roessink, Ivo; Škerl, Maja Ivana Smodiš; Titera, Dalibor; Steen, Jozef van der; Varnava, Andri; Vejsnæs, Flemming; Webster, Matthew T.; Rodrigues, Pedro João; Henriques, Dora; Pinto, M. AliceWing venation patterns have long been used to identify honey bee subspecies, singly or in combination with other morphological traits, using different approaches. Beekeepers have traditionally used identification methods that only require estimations of the Cubital Index, Hantel Index, and/or Discoidal Shift Angle. However, these measures do not consider all the information carried by wing patterns, and a more accurate approach is geometric morphometrics, which requires the annotation of 19 landmarks in the forewing vein junctions. While manual wing annotation is a time-consuming and error-prone endeavour, the recently developed software DeepWings© performs this task automatically, allowing a fast and relatively accurate identification of European honey bee subspecies, and is appropriate for large-scale projects. In this study, we analyse the patterns of forewings extracted from over 850 colonies sampled across 29 European countries. To that end, the right forewings of five workers per colony are photographed with a digital camera attached to a stereomicroscope. The images are then analysed using DeepWings©. Our main objective is to assess (i) how closely the colonies identified by DeepWings© match the endemic evolutionary lineages and subspecies; and (ii) the association between the identification produced by DeepWings© and that inferred from mitochondrial DNA. While geometric morphometrics does not replace molecular tools for the identification of colonies, it can provide a preliminary estimation of their genetic integrity.