Genitalia are rich source of characters in insect taxonomy. Usually, they are examined after dissection and cleaning with potassium hydroxide (KOH), procedure that may damage both genital morphological structures and intracellular molecular contents. Enzymatic procedure with Proteinase K has been used to clean muscle off the genitalia while extract DNA, but its damage to the genital structures has not been evaluated. Herein, we qualitatively compare the use of KOH and Proteinase K to prepare genital structures in minute insects (Hymenoptera: Bethylidae). We show that Proteinase K is better to preserve the genital structure and provides quality DNA.

The muriqui (Brachyteles) is one of the most endangered primates in the world, however little is known about the viability of the remaining populations. We evaluated the technique of extracting DNA from wild muriqui feces for PCR applications. In order to determine the effect of the DNA in subsequent amplifications, we analyzed five different extracts. The importance of the recommended BSA and the HotStarTaq DNA polymerase was tested. The minimal conditions to successfully amplify highly degraded fecal DNA were determined, showing that the recommended reagents are not required. We envision that this method may be useful in further conservation management studies

(1) Background: Trichomycterinae represent 60% of the species in the family and, while seven genera comprise 1–3 species each, Trichomycterus and Cambeva have over 180 known species between them. Although integrative studies aimed to clarify the relationships within the subfamily, the diversity of species of Trichomycterus remains an open question. Herein, we explored an unprecedented sample to investigate the divergence in the lineages of Trichomycterus. (2) Methods: we recovered the phylogenetic relationships of the subfamily using 566 sequences (999 bp) of the mitochondrial gene cytochrome b, calculated intra- and intergroup distance percentages, and estimated divergence times. (3) Results: we recovered 13 highly supported and geographically structured lineages; intergenus divergence was 11–20%, while interspecies divergence was 3–11%; Trichomycterus, Cambeva, Scleronema, Hatcheria, Eremophilus, and Ituglanis were recovered as monophyletic, with three other highly divergent clades: Guiana Shield, Magdalena basin, and Tapajós basin. (4) Conclusions: We propose that the trans-Andean austral clades be allocated into Hatcheria, and the Guiana clade supports a new genus. We also observed that the headwaters nearest the Magdalena and Orinoco basins showed a high diversity and endemism of Trichomycterinae lineages. We discussed the role of geomorphological events and the climatic features which may explain cladogenesis events in Trichomycterinae.

DNA sequences have proved valuable for associating males and females of the same dimorphic species in Hymenoptera. These molecular associations, however, depend on the used DNA sequences and the analysis methods. In the present study, we evaluated the molecular associations on 23 species of Dissomphalus (Hymenoptera, Bethylidae) using the three most common DNA sequences in molecular studies of Hymenoptera (28S rRNA, COI and ITS2). Our results indicated the male-female association with these three markers. COI, however, showed to be more suitable than the other markers because there was no overlap of genetic variation among species. In contrast, the 28S rRNA showed overlapping of genetic distance limits, indicating that the distance matrix is not enough to assign a sexual conspecificity. We were able to perform female association to the described species D. verus Mugrabi & Azevedo, demonstrating that phylogenetic analysis is a valuable tool for associating the sexes of dimorphic species.