2021 SIMMONS ET AL,: NEW SPECIES OF MYOTIS 5 
MATERIALS AND METHODS 
We surveyed subterranean habitats (abandoned mine adits and natural caves) by placing harp 
traps (a two-bank 4.2 m? harp trap [manufactured by Austbat] or a “cave catcher” 0.9 m* harp 
trap [Bat Conservation and Management]) at entrances at least 30 minutes before sunset (fig. 2). 
Harp traps were checked a maximum of every 10-15 minutes for the presence of bats for two to 
three hours after sunset and removed after bat emergence activity had subsided. Captured bats 
were placed in individual cloth bags and processed to identify species, sex, age, and reproductive 
status prior to release at place of capture following standard methods as described in Kunz and 
Parsons (2009). We collected standard field measurements (forearm length, tibia length, hind-foot 
length, tail length, ear length, tragus length, body length, and mass) and consulted Mammals of 
Africa, volume 4 (Hedgehogs, Shrews and Bats; Happold and Happold, 2013) to identify species. 
We collected tissue samples for DNA analysis using a 3 mm biopsy punch from the wing mem- 
brane and stored in desiccant. Echolocation calls were recorded upon release for echolocating bat 
species using a Pettersson M500 full-spectrum bat detector (Pettersson Elektronik) at a sampling 
rate of 500 kHz, Files were saved as uncompressed “wav” format and analyzed using BatSound 
v.4.1 (Pettersson Elektronik: FFT size 1024, Hanning window) to determine the following param- 
eters for each pulse: duration (D), maximum frequency (FMAX), minimum frequency (FMIN), 
peak frequency (PF), and interpulse interval (IPI). D, FMAX, FMIN, and IPI were measured from 
spectrograms, and PF from power spectrum. 
A male bat captured and released on 26 January 2018 at Kaiser Adit 3 could not be identi- 
fied using keys for the bats of Africa, Due to the likelihood that this individual represented an 
unknown species of Myotis, we resurveyed two adjacent adits (Kaiser Adit 1 and Kaiser Adit 
3) on 2 February 2018 and collected two individuals (one male, one female) emerging from 
Kaiser Adit 1 to confirm and describe the species, The collected specimens included the male 
bat originally captured and released on 26 January 2018, identifiable by the 3 mm hole in its 
wing membrane from tissue sampling. 
We employed analyses of mitochondrial cytochrome b gene sequences as well as standard 
morphological comparisons in our evaluation of the new taxon. The specimens examined and 
tissues used for this study (appendices 1 and 2) belong to the following collections: 
AMNH American Museum of Natural History, New York 
BMNH Natural History Museum, London 
FMNH Field Museum of Natural History, Chicago 
USNM National Museum of Natural History, Smithsonian Institution, 
Washington, DC 
MOLECULAR ANALYSES 
Our analysis was designed to cover all the major lineages previously recognized in 
the subgenus Chrysopteron to allow placement of our new species in phylogenetic context. 
We downloaded GenBank sequences of Myotis compiled or originally published by Csorba 
