Non-lethal method of DNA sampling in euglossine bees supported by mark–recapture experiments and microsatellite genotyping
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Non-lethal sampling methods are of great interest for conservation genetic studies to prevent the death of individuals in populations that are threatened or in decline. With this aim, we tested a non-lethal method of partial antennae removal for DNA sampling in two euglossine bee species: Euglossa cordata and Eulaema nigrita. We validated the survival of the individuals through mark–recapture experiments during 16 months. The quality and quantity of the tissue for DNA analysis was verified through amplification and genotyping of nine and eleven microsatellite loci, respectively. Our results from the mark–recapture experiments showed equal recapture rates of individuals with intact and removed antennae (E. cordata χ2 = 2.492, df = 1, p = 0.114; E. nigrita χ2 = 1.683, df = 1, p = 0.194). Microsatellite loci were successfully genotyped in 97.1 and 97.6 % of the E. cordata and E. nigrita individuals, respectively. Our results validate the feasibility of using antennae tissue for DNA genetic analysis without compromising the survival of individual bees.
KeywordsTissue sampling Antennae Orchid bees Euglossa cordata Eulaema nigrita
We acknowledge support from CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) for the master degree fellowship to C. A. Oi and from CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) and FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo, Processo 2004/15801-0) for the financial research support. We are thankful to N. Mortari for laboratory setting courtesy, and C. H. da Silva, J. C. Almeida and O. Lino e Silva for help in the field. We also would like to thank F. T. T. Hanashiro, K. Nagamati Jr., L. Packer, J. Gibbs and three anonymous reviewers for commentaries and suggestions that improved this manuscript.
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