Criação in vitro da abelha sem ferrão Frieseomelitta varia Lepeletier 1836 (Apidae, Meliponini) e efeitos de doses subletais de glifosato na morfologia

Abstract

Pollination is one of the essential ecological services for maintaining natural and agricultural environments. Among pollinators, such as Meliponini bees represent a diverse group that plays a fundamental ecological role and are exposed to various pesticides in both natural and agricultural environments, including glyphosate. This compound has been designed and is used extensively as a broad spectrum herbicide. However, recent publications show that glyphosate has changed in the cognitive capacity of Apis mellifera bees. Here we hypothesize that glyphosate can promote morphophysiological changes in developing insects and thus be responsible for reducing bee populations, including how the native species Frieseomelitta varia. As an initial approach to test our hypothesis, we defined two fundamental objectives: (i) Establish an in vitro breeding protocol for F. varia workers, which would make it possible to carry out tests to evaluate the pesticide effects during post-embryonic development; (ii) Determine external morphological changes in newly emerged bees surviving larval exposure. The larvae were formed in acrylic ELISA plates (96 wells) under controlled conditions of humidity and temperature and fed with natural stored larval food. Our results show that (L1) larvae fed with 27 μL of food were the most successful when considering larval emergence rates (87%), enabling the emergence of a significant number of workers (93%) over the course of 33.3 ± 0.57 days of

development. The morphological analyzes of emerged bees indicated that there are no statistical differences between bees in vitro and in vivo. Thus, these results show the feasibility of using this species of stingless bees in ecotoxicological studies and as a model for other species that have an ecology similar to F. varia (Chapter 1). To achieve our second objective, L1 larvae received food containing glyphosate to determine morphological changes after ingesting the herbicide. Our results hindered that, in the tested options, glyphosate is not lethal to F. varia larvae. Although it was not lethal, we recorded newly emerged bees treated 1,000 ng / 27 μL of food with malformations in the wings. These results indicate that the herbicide glyphosate is a stressor and affects the post- embryonic development of F. varia bees (Chapter 2). Finally, during the handling of F. varia colonies, we first registered the predators of the mite Tyrophagus putrescentiae with parasitic behavior, causing weakening and consequently the mortality of all colonies in which the mite was registered (Chapter 3). Our results show that F. varia is sensitive to glyphosate and mite infestation and obtains the need to carry out experiments that show details of the ecological interactions between bees and the mite and the effects of exposure to glyphosate throughout the development period of the Bee. Together, the body of knowledge found will contribute to the planning of conservation actions for bee populations and, consequently, the maintenance of pollination services.