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Accurate taxonomic delimitation in ecological research is absolutely critical as studies that seek to evaluate levels of biodiversity and qualify human effects on the environment are rapidly undertaken. Coloration is a widely used morphological character for species identification through dichotomous keys. However, taxonomic identification based upon coloration is often unreliable because this character can exhibit high degree of intraspecific variation. In this study, we use a DNA barcoding approach to investigate the interpretation of two color morphs (yellow or dark) in the eucerine bee Melissodes nigroaenea. Our hypothesis is that if significant genetic divergence exists between each morphotype of M. nigroaenea, coloration reflects two distinct evolutionary lineages within this species, which may require taxonomic revision. Our alternative hypothesis is that, if genetic divergence is low between each morphotype of M. nigroaenea, we can attribute this variation to color polymorphism. Our Bayesian phylogenetic reconstruction revealed that both yellow and black individuals clustered together in a highly supported phylogenetic group. Additionally, pairwise genetic distances between M. nigroaenea color morphotypes were lower than 3%. These results indicate that both mesosome color morphs correspond to intraspecific variability within the same evolutionary unit. Together, our results indicate that mesosome coloration is not a reliable character for taxonomic differentiation of these Melissodes species, and that the incorporation of DNA barcoding approaches to taxonomic classification can help resolve some of the problems that originate while relying on purely morphological taxonomy.

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The drastic decline of bees is associated with several factors, including the immune system suppression due to the increased exposure to pesticides. A widely used method to evaluate these effects on these insects' immune systems is the counting of circulating hemocytes in the hemolymph. However, the extraction of hemolymph from larvae is quite difficult, and the collected material is frequently contaminated with other tissues and gastrointestinal fluids, which complicates counting. Therefore, the present work established a high quality and easily reproducible method of extracting hemolymph from honeybee larvae (Apis mellifera), the extraction with ophthalmic scissors. Extraction methods with the following tools also were tested: 30G needle, fine-tipped forceps, hypodermic syringe, and capillaries tubes. The hemolymph was obtained via an incision on the larvae's right side for all methods, except for the extraction with ophthalmic scissors, in which the hemolymph was extracted from the head region. To assess the purity of the collected material, turbidity analyses of the samples using a turbidimeter were proposed, tested, and evaluated. The results showed that the use of ophthalmic scissors provided the clearest samples and was free from contamination. A reference range between 22,432.35 and 24,504.87 NTU (nephelometric turbidity units) was established, in which the collected samples may be considered of high quality and free from contamination.

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