All specimens were collected between 1994 and 2020 by Dr. Hendrik Freitag and his team (Ateneo de Manila University) and preserved in 70%–96% ethanol.
The dissection of larvae was done in Cellosolve (2-Ethoxyethanol) with subsequent mounting on slides with Euparal liquid, using an Olympus SZX7 stereomicroscope.
The DNA of part of the specimens was extracted using non-destructive methods allowing subsequent morphological analysis (see Vuataz et al. 2011 (link) for details). We amplified a 658 bp fragment of the mitochondrial gene cytochrome oxidase subunit 1 (COI ) using the primers LCO 1490 and HCO 2198 (Folmer et al. 1994 (link); see Kaltenbach and Gattolliat 2020 (link) for details). Sequencing was done with Sanger’s method (Sanger et al. 1977 (link)). The genetic variability between specimens was estimated using Kimura-2-parameter distances (K2P , Kimura 1980 (link)), calculated with the program MEGA 7 (Kumar et al. 2016 (link), http://www.megasoftware.net ).
The GenBank accession numbers are given in Table1 , nomenclature of gene sequences follows Chakrabarty et al. (2013) (link).
The nomenclature used for Molecular Operational Taxonomic Units (MOTUs ) is somewhat different as the one proposed by Morard et al. (2016) (link).
Drawings were made using an Olympus BX43 microscope. To facilitate the determination of species and the comparison of important structures, we partly used a combination of dorsal and ventral aspects in one drawing. Explanations are given in Fig.1 .
Photographs of larvae were taken using a Canon EOS 6D camera and the Visionary Digital Passport imaging system (http://www.duninc.com ) and processed with Adobe Photoshop Lightroom (http://www.adobe.com ) and Helicon Focus version 5.3 (http://www.heliconsoft.com ). Photographs were subsequently enhanced with Adobe Photoshop Elements 13.
The distribution maps were generated with SimpleMappr (https://simplemappr.net , Shorthouse 2010 ). Google Earth (http://www.google.com/earth/download/ge/ ) was used to attribute approximate GPS coordinates to sample locations of Müller-Liebenau (1982) .
The taxonomic descriptions were generated with a DELTA (Dallwitz 1980 (link); Dallwitz et al. 1999 ; Coleman et al. 2010 (link)) database containing the morphological states of characters of theLabiobaetis species of the Philippines.
The dichotomous key was elaborated with the support of DKey version 1.3.0 (http://drawwing.org/dkey , Tofilski 2018 (link)).
The terminology follows Hubbard (1995) and Kluge (2004) (link). The character states of some of the characters are depicted in Fig.2 .
Abbreviations:
AdMU Ateneo de Manila University, Quezon City (Philippines)
MZL Musée de Zoologie Lausanne (Switzerland)
PCSD Palawan Council for Sustainable Development, Puerto Princesa, Palawan (Philippines)
PNM Museum of Natural History of the Philippine National Museum, Manila (Philippines)
ZSM Zoologische Staatssammlung München (Germany).
The dissection of larvae was done in Cellosolve (2-Ethoxyethanol) with subsequent mounting on slides with Euparal liquid, using an Olympus SZX7 stereomicroscope.
The DNA of part of the specimens was extracted using non-destructive methods allowing subsequent morphological analysis (see Vuataz et al. 2011 (link) for details). We amplified a 658 bp fragment of the mitochondrial gene cytochrome oxidase subunit 1 (
The GenBank accession numbers are given in Table
The nomenclature used for Molecular Operational Taxonomic Units (
Drawings were made using an Olympus BX43 microscope. To facilitate the determination of species and the comparison of important structures, we partly used a combination of dorsal and ventral aspects in one drawing. Explanations are given in Fig.
Photographs of larvae were taken using a Canon EOS 6D camera and the Visionary Digital Passport imaging system (
The distribution maps were generated with SimpleMappr (
The taxonomic descriptions were generated with a DELTA (Dallwitz 1980 (link); Dallwitz et al. 1999 ; Coleman et al. 2010 (link)) database containing the morphological states of characters of the
The dichotomous key was elaborated with the support of DKey version 1.3.0 (
The terminology follows Hubbard (1995) and Kluge (2004) (link). The character states of some of the characters are depicted in Fig.
Abbreviations:
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