Pimplinae

2.3. Pimplinae Collections We collected pimplines using 30 Malaise traps (a form a flight interception trap) distributed at 15 broad elevation sites throughout the transect, with two replicate traps at each site. Malaise traps are one of the most efficient methods for sampling Ichneumonidae and are widely used in ecological studies of insects [94]. Traps were set at ground level, with the collecting head 1.5 m above the ground. Traps were placed at least 50 m from the road and the 15 sites were roughly spaced at 100 m to 200 m elevation intervals. At each elevational site, the two traps were placed at least 50 m apart from each other to ensure that neither trap affected the catch of the other and to sample different environmental space [65]. Trap collecting bottles (1 L capacity) contained 98% ethanol for preservation of the sampled material and were replaced monthly (after 30 days of collecting). The samples were collected during both the rainy hot season, from December 2014 to February 2015, and the dry cooler season from June to August 2015, totaling 180 Malaise trap months. These months were chosen to represent the opposite environmental extremes throughout the year to capture seasonal variations in species composition, but also to encompass the warmest, wettest months when insect activity is expected to be highest (December–February). Insects were preserved in 98% ethanol and stored in plastic bottles. Sample sorting was performed in the laboratory using a stereoscopic microscope. Identification was initially carried out according to subfamily by D.R.R.F. and D.G.P. following [92]. Then, the Pimplinae were identified according to genus following [89]. Thereafter, morphospecies were identified by D.G.P. in conjunction with I.E.S., and where possible, species (using specific bibliography and large reference collections of neotropical Darwin wasps in the Biodiversity Unit, University of Turku, Finland). All the researchers involved in identification were experienced in neotropical ichneumonid taxonomy. The use of morphospecies (i.e., individuals sorted based on phenotypic characteristics) as surrogates for species is widely used in the estimation of species richness for comparisons over time and space [95, 96]. Although the designation of morphospecies can lead to the split of a single species into many different morphospecies (“splitting”) or aggregation of different species into a single species (“lumping”), it is often the only way to assess species diversity in groups that have not been fully described [95, 96]. Collections were performed under license number 21409-10 (Ministério do Meio Ambiente—MMA; Instituto Chico Mendes de Conservação da Biodiversidade—ICMBio; Sistema de Autorização e Informação em Biodiversidade—SISBIO) to Ricardo Ferreira Monteiro. The sampled material is deposited at the following Brazilian entomological collections: Invertebrate Collection of Instituto Nacional de Pesquisas da Amazônia, Manaus, Brazil (INPA), (curator: Marcio L. Oliveira); Museu de Zoologia da Universidade de São Paulo, São Paulo, Brazil (MZUSP), (curator: Gabriela P. Camacho); and Taxonomic Collection of the Departamento de Ecologia e Biologia Evolutiva from Universidade Federal de São Carlos, São Carlos, Brazil (DCBU), (curator: Angelica M. Penteado-Dias).

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