2.1 Taxon sampling and data collection
Specimens were collected between 2014 and 2017 from 34 localities, in a
diverse array of habitats including small streams and ponds to lakes,
rivers and bays in Argentina, Brazil, Chile and Dominican Republic. The
main emphasis was on adult sampling, collected with a sweep net near
aquatic systems. A 20 cm diameter D-frame kick net (mesh size 250 μm)
was also used to collect immature stages at some localities. All sampled
adults were preserved in 75%-85% ethanol and larvae in 96-100%
ethanol and stored at 4°C in the dark prior to the extraction. Specimens
were identified using the classification proposed by Townes (1945),
Bidawid & Fittkau (1995), Bidawid-Kafka (1996), Shimabakuro et al.
(2019), Pinho & Silva (2020), and eventual examination of type
material. Voucher specimens are deposited in the Museum of Comparative
Zoology (MCZ) at Harvard University and in the National Institute of
Amazonian Research (INPA).
In addition to data generated for this publication, we also searched for
public COI barcodes in the Barcode of Life Data Systems (BOLD,www.boldsystems.org) belonging
to the genus Polypedilum that were longer than 300 base pairs and
without stop codons. Searches were performed on 25 January 2022 in BOLD.
In total, 9,540 COI barcodes were included in our dataset, of which 149
barcodes of 54 identified species were not previously used in any
molecular analysis. A reduced data set, containing 1,492 sequences, was
generated based on the manual deletion of the highly similar sequences
based on an UPGMA tree. Duplicate sequences occurring at different
sampling localities were retained in our dataset. The detailed specimen
records and sequence information, including trace files, are available
in BOLD through the dataset ‘DS-RPPPOL - Reduced personal and publicly
available records of Polypedilum (Diptera: Chironomidae)” with
DOI: https://doi.org/XXXX.