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Systematic review of Myotis (Chiroptera, Vespertilionidae) from Chile based on molecular, morphological, and bioacoustic data

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Abstract

Myotis is the most diverse genus of bats in the world, with more than 30 species recognized in the Neotropics. However, many of these species represent cryptic complexes and are evidence of the existence of hidden diversity in several regions. Using an integrative approach based on molecular, morphological, and bioacoustic data, we performed a systematic review of Myotis species from Chile. Phylogenetic inference using cytochrome-b indicated the existence of three monophyletic lineages, and qualitative and quantitative morphological analyses supported these lineages as distinct and morphologically diagnosable taxa. Analysis of discriminant functions using parameters of echolocation calls also indicates the existence of three distinct bioacoustic clusters. Thus, all lines of evidence congruently indicate the existence of three distinct taxa. As a result, we recognize Myotis arescens as a valid and distinct species and define its taxonomic limits from the other species from Chile, Myotis atacamensis and Myotis chiloensis.
https://doi.org/10.11646/zootaxa.5188.5.2
http://zoobank.org/urn:lsid:zoobank.org:pub:8CE3B18D-44A3-4873-A759-CA372B21555E
430 Accepted by P. Velazco: 4 Sept. 2022; published: 21 Sept. 2022
Article ZOOTAXA
ISSN 1175-5326 (print edition)
ISSN 1175-5334 (online edition)
Zootaxa 5188 (5): 430–452
https://www.mapress.com/zt/
Copyright © 2022 Magnolia Press
Systematic review of Myotis (Chiroptera, Vespertilionidae) from Chile based on
molecular, morphological, and bioacoustic data
ROBERTO LEONAN M. NOVAES1*, ANNIA RODRÍGUEZ-SAN PEDRO2,3,4,
MÓNICA M. SALDARRIAGA-CÓRDOBA5, OMAYRA AGUILERA-ACUÑA3, DON E. WILSON6 &
RICARDO MORATELLI1
1Fundação Oswaldo Cruz, Fiocruz Mata Atlântica, 22713-375, Rio de Janeiro, RJ, Brazil
2Universidad Santo Tomás, Centro de Investigación e Innovación Para el Cambio Climático, 8370003, Santiago, Chile
3Bioecos E.I.R.L., Las Condes, Santiago, Chile
4Programa para la Conservación de los Murciélagos de Chile, Santiago, Chile
5Universidad Bernardo O’Higgins, Centro de Investigación en Recursos Naturales y Sustentabilidad, Santiago, Chile
6Smithsonian Institution, National Museum of Natural History, 20560, Washington, DC, USA
*Corresponding author.
robertoleonan@gmail.com
Abstract
Myotis is the most diverse genus of bats in the world, with more than 30 species recognized in the Neotropics. However,
many of these species represent cryptic complexes and are evidence of the existence of hidden diversity in several regions.
Using an integrative approach based on molecular, morphological, and bioacoustic data, we performed a systematic review
of Myotis species from Chile. Phylogenetic inference using cytochrome-b indicated the existence of three monophyletic
lineages, and qualitative and quantitative morphological analyses supported these lineages as distinct and morphologically
diagnosable taxa. Analysis of discriminant functions using parameters of echolocation calls also indicates the existence
of three distinct bioacoustic clusters. Thus, all lines of evidence congruently indicate the existence of three distinct taxa.
As a result, we recognize Myotis arescens as a valid and distinct species and define its taxonomic limits from the other
species from Chile, Myotis atacamensis and Myotis chiloensis.
Key words: Echolocation calls, mitochondrial gene, multivariate analysis, Myotinae, Myotis arescens, phylogenetics,
South America
Introduction
Myotis Kaup, 1829 is the most speciose genus of mammals, with more than 140 species distributed worldwide
(Moratelli et al. 2019a; MDD 2022). Thirty-three species are recognized in the Neotropics (Novaes et al. 2021a,
2022a), but this number is clearly low, considering that several studies have indicated the existence of complexes
of cryptic species (Larsen et al. 2012a; Carrión-Bonilla & Cook 2020; Novaes et al. 2021a, b). Recent systematic
studies have focused on solving taxonomic problems, including the description and revalidation of several species
in addition to defining the taxonomic and geographic limits of these taxa (e.g., Moratelli & Wilson 2011, 2014;
Moratelli et al. 2011, 2013, 2016, 2017, 2019b; Novaes et al. 2018, 2021a, b, c). However, few studies have focused
on the Myotis assemblage of the southern portion of South America.
In a review of Chilean mammals, Osgood (1943) recognized Myotis chiloensis (Waterhouse, 1840) as the only
species of the genus occurring in Chile. In addition, Osgood (op. cit.) considered Myotis atacamensis (Lataste,
1892) as a subspecies of M. chiloensis and described a new subspecies from Valparaíso, named Myotis chiloensis
arescens Osgood, 1943. However, after an extensive morphological review, LaVal (1973) raised M. atacamensis
to the species level and did not recognize M. c. arescens as valid, placing it under synonymy with M. chiloensis.
This arrangement has been maintained so far (Wilson 2008; Ossa & Rodríguez-San Pedro 2015; Moratelli et al.
2019a).
Using a set of molecular, morphological, morphometric, and bioacoustic data, we reassess the taxonomic status
of M. c. arescens and comment on the taxonomic and distributional limits of Myotis species from Chile.
SYSTEMATIC REVIEW OF CHILEAN MYOTIS Zootaxa 5188 (5) © 2022 Magnolia Press · 431
Material and methods
The present study was based on the analysis of datasets composed of molecular, morphological, and bioacoustic
characters, including qualitative and quantitative approaches. Taxonomic decisions were based on the integrative
taxonomy protocol by Padial et al. (2010), which uses congruence among different sources of evidence to guide
taxonomic decisions. We adopted the Phylogenetic Species Concept (q.v., Wheeler 1999; Zachos 2016), which
predicts that a species is a lineage evolutionarily independent of any other, where its populations form a monophyletic
group identifiable from character distribution patterns, whether phenotypic or genotypic.
Molecular analyses
Molecular analyses were based on 63 sequences of the mitochondrial gene cytochrome-b (cyt-b, ca. 1,140 bp),
from 26 species of Neotropical Myotis (Appendix 1). Most sequences were obtained from NCBI’s GenBank (54
out of 64, including outgroups), and the remaining nine sequences were generated in the present study (Appendix
1). Wing tissue samples were obtained from live bats captured during fieldwork in Chile using disposable biopsy
punches (4 mm, Derma-Punch, Dolphin Medical Spa, Santiago, Chile). Bat captures in Chile were carried out with
permission of Servicio Agrícola y Ganadero, División de Protección de Recursos Naturales Renovables (Diproren;
Res Ex 9895/2015, 7612/2017, 4572/2021, 4728/2021, and 130/2022). Specimens were identified based on
external morphology (fur color, forearm, and ear length), photographed and released at the same capture location.
DNA extraction was made using E.Z.N.A.® Omega Tissue DNA Kit and the cyt-b gene was amplified by PCR and
sequenced using the primers Bat 05A (sense: 5’-CGACTAATGACATGAAAAATCACCGTTG-3’, Tm: 63.2˚C)
designed by Martins et al. (2007) and Bat-Ep (antisense: 5’ -TAGTTTAABTAGAAYHYCAGCTTTGGG-3’, Tm:
61.6) designed by Caraballo et al. (2020). PCR-reaction mixtures consisted of 2 µL of DNA extracted directly
from the wing tissue samples, 1.5 mM MgCl2, 0.2 mM dNTPs, 1 µM of each primer, 1 mg/mL of BSA and 4
U/rx of GoTaq® G2 Flexi DNA Polymerase (Promega, Madison, WI, USA) in a final volume of 50 μL. The PCR
program consisted of an initial denaturation step at 94°C for 5 min, followed by 35 cycles with a denaturation step
at 94°C for 45 seconds, an annealing stage at 55°C for 45 seconds, an extension at 72°C for two minutes and a final
extension at 72°C for 10 minutes. Amplicons were separated by electrophoresis on 1.5% agarose gels in 0.5X TBE
buffer, dyed with GelRed™ Nucleic Acid Gel Stain (Biotium, Inc.) and visualized under UV light. We performed
Sanger sequencing in a capillary automated ABI3500 sequencer (Applied Biosystems®), at the AUSTRAL-omics
(Santiago, Chile). The DNA sequences were edited (Trim Ends and de Novo Assemble) and aligned in Geneious
Prime v2022.2 (Kearse et al. 2012). The nucleotide sequences were translated into proteins to evaluate the reading
frame and ensure the absence of premature stop codons or other nonsense mutations (Triant & Dewoody 2007).
Novel sequences were deposited in GeneBank (accession number OP270158–OP270166).
The cyt-b sequences were aligned using the UPGMB clustering method implemented in the MUSCLE algorithm
(Edgar 2004) in the MEGA X software (Kumar et al. 2018) with default settings. We chose the evolutionary model of
nucleotide substitution for phylogenetic analyses using the software JModelTest 2 (Darriba et al. 2012) employing
the Bayesian Information Criterion (BIC). The Hasegawa–Kishino–Yano model (Hasegawa et al. 1985) yielded
the best fit to our dataset regarding the substitution of nucleotides, corrected for rate heterogeneity with gamma
distribution and proportion of invariant sites parameters (i.e., HKY + Γ + I).
Phylogenetic reconstruction was performed using the Bayesian Inference (BI) probabilistic method (Huelsenbeck
et al. 2001) in the software MrBayes v. 3.4 (Ronquist & Huelsenbeck 2003) using the coupled Markov Chain
Monte Carlo (MCMC). Four simultaneous Markov chains were performed for 100 million generations with trees
sampled every 10,000 generations. The first 25,000 trees were discarded as burn-in. Posterior probabilities were
calculated from the consensus of the remaining trees. The confidence of the Bayesian sampling was verified for the
free parameters using the effective sample size statistic (ESS) implemented in the software Tracer v. 1.5 (Rambaut
& Drummond 2009). The analyses were also checked for convergence by plotting the value of the log-likelihood
against the generation time for each model. All parameters showed ESS greater than 300 and the analyses converged
asymptotically, indicating reliable performance.
Estimation of genetic distances was made using the Kimura two-parameter model (K2p) implemented in MEGA
X (Kumar et al. 2018). This method measures the distance between pairs of sequences, estimating the proportion of
different nucleotides between two or more sequences (Kimura 1980).
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Specimens examined
This research is part of an exhaustive review of Neotropical Myotis, and more than 7,500 specimens from different
localities in South and Central Americas have been examined, covering all species currently recognized and
their primary type-specimens. The analyses of this study were based on 222 specimens (Appendix 2) deposited
in Muséum d’Histoire Naturelle (MHNG, Geneva, Switzerland), American Museum of Natural History (AMNH,
New York, USA), Carnegie Museum of Natural History (CM, Pittsburgh, USA), Field Museum of Natural History
(FMNH, Chicago, USA), Museum of Natural History of the Kansas University (KU, Lawrence, USA), Museum of
Natural Science, Louisiana State University (LSUMZ, Baton Rouge, USA), Museum of Vertebrate Zoology (MVZ,
Berkeley, USA), Sam Noble Oklahoma Museum of Natural History (OMNH, Norman, USA), and Smithsonian
National Museum of Natural History (USNM, Washington D.C., USA). Morphological comparative analyses were
performed using samples from Chile, Argentina, and Peru, including all species occurring in the Western Andes
and those morphologically similar with occurrence in the Southern Cone, including Myotis albescens (É. Geoffroy,
1806), Myotis atacamensis (Lataste, 1892), Myotis bakeri Moratelli, Novaes, Carrión & Wilson, 2019, Myotis
chiloensis chiloensis (Waterhouse, 1840), Myotis chiloensis arescens Osgood, 1943, Myotis keaysi J.A. Allen, 1914,
and Myotis oxyotus (Peters, 1866).
Morphological and morphometric analyses
The specimens examined were allocated to Operational Taxonomic Units (OTU) formed by geographical groups
with phenotypic cohesion, and the classification of these specimens was made based on qualitative morphological
analyses, considering the fur color and cranial traits. Thus, specimens were classified into three OTUs (Figure
1). The intra and interspecific morphological variation was evaluated from qualitative and quantitative analyses
based on adults (classified from ossified epiphyses; see Brunet-Rossini & Wilkinson 2009). The total length (TL),
tail length (TL), hind foot length (HF), ear length (EL), and body mass (BM) were recorded from skin labels and
reported to the nearest millimeter and to the nearest gram. Skull and other external dimensions were taken using
digital calipers accurate to 0.01 mm, including: forearm length (FA), length of dorsal fur (LDF), length of ventral
fur (LVF), and 16 craniodental measurements (Table 1).
TABLE 1. Description of cranial, mandibular, and external dimensions (and their abbreviations). Lengths were measured
from the anteriormost point or surface of the 1st structure to the posteriormost point or surface of the 2nd structure, except
as specified.
Measurements Acronyms Descriptions
Forearm length FA From the elbow to the distal end of the forearm including carpals
Ear length EL From the base to the apex of the ear
Length of dorsal fur LDF Measured at the midpoint of the scapulae
Length of ventral fur LVF Measured at the midpoint of the sternum
Greatest length of skull GLS From the apex of the upper internal incisors, to the occiput
Condylo-canine length CCL From the anterior surface of the upper canines to a line connecting the occipital
condyles
Condylo-basal length CBL From the premaxillae to a line connecting the occipital condyles
Condylo-incisive length CIL From the apex of upper internal incisors to a line connecting the occipital condyles
Basal length BAL Least distance from the apex of upper internal incisors to the ventral margin of the
foramen magnum
Zygomatic breadth ZYG Greatest breadth across the outer margins of the zygomatic arches
Mastoid breadth MAB Greatest breadth across the mastoid region
Braincase breadth BCB Greatest breadth of the globular part of the braincase
Interorbital breadth IOB Least breadth between the orbits
Postorbital breadth POB Least breadth across frontals posterior to the postorbital bulges
Breadth across canines BAC Greatest breadth across outer edges of the crowns of upper canines, including cingulae
Breadth across molars BAM Greatest breadth across outer edges of the crowns of upper molars
Maxillary toothrow length MTL From the upper canine to M3
Molariform toothrow length M1–3 From M1 to M3
Mandibular length MAL From the mandibular symphysis to the condyloid process
Mandibular toothrow length MAN From the lower canine to m3
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FIGURE 1. Operational Taxonomic Units (OUT) for Myotis from Chile. Circles indicate geographical origin of specimens
analyzed morphologically; diamonds indicate the geographical origin of specimens analyzed genetically.
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To characterize and discriminate samples, Principal Component and Discriminant Function analyses were
performed using “MASS” and “Lattice” packages implemented in the R platform (Vanables & Ripley 2002; Sarkar
2008). To test whether OTUs have the same average for different cranial measurements, we used unifactorial
Multivariate Analysis of Variance (MANOVA) to calculate Wilks’ lambda statistic (λ-Wilks’) and the associated
value of F’s Rao (Tabachnick & Fidell 2014). Pairwise comparisons were made with the Hotelling-pairwise test with
Bonferroni-corrected p-value. MANOVA was performed using ‘Dplyr’ package implemented in the R platform, and
the differences were considered statistically significant when p-values were less than 0.05 (Wickham et al. 2017).
For these morphometric analyses, we selected a subset of skull dimensions (GLS, CCL, CBL, CIL, BAL, MAB,
BCB, IOB, POB, BAC, BAM, MTL, M1–3, MAL, MAN) representing different axes of length and width of skull,
rostrum, and mandible. As multivariate procedures require complete data sets, missing values (less than 5% of
the total dataset) were estimated from the existing raw data using the Amelia II package (Honaker et al. 2011)
implemented in R platform. Measurements were transformed to natural logs, and covariance matrices were computed
considering all variables. To avoid unbalanced samples, we randomly selected a maximum of 11 individuals per
OTU. All samples were previously tested to verify the existence of intraspecific secondary sexual dimorphism using
MANOVA applied on datasets from the same OTU. However, no statistically significant differences were detected
in cranial and external dimensions between males and females. Thus, all analyses were performed with grouped
males and females. Qualitative traits employed here to characterize and distinguish species follow Moratelli et al.
(2013) and Novaes et al. (2021a, b, c). Capitalized color nomenclature follows Ridgway (1912).
Bioacoustics analyses
Acoustic recordings used in the analyses came from the personal library of one of the authors (ARSP) containing
reference echolocation calls of M. atacamensis from the Arica and Parinacota, Antofagasta, and Atacama regions
in northern Chile, M. c. arescens from Coquimbo, Valparaíso, and Metropolitan regions in central Chile, and M. c.
chiloensis from Los Ríos and Los Lagos regions in southern Chile, following the same geographic organizations
from morphological OTUs (Figure 1). Reference calls were obtained from captured bats and recorded directly by
hand-release. Other recordings were obtained from free-flying bats, foraging within a radius of 50 m around known
roosts for each species, using an automatic recorder Song Meter SM4BAT FS with an external omnidirectional
SMM-U1 ultrasound microphone (Wildlife Acoustics, Inc., Maynard, MA, USA). Detectors were placed at an
approximate height of 4.5 m above ground level in uncluttered flyways, in both natural and anthropogenic habitats,
and ran for a minimum of one night at each site from sunset to sunrise. Recordings were conducted using a sampling
frequency of 256 kHz or higher.
For sound analysis, spectrograms were generated on BatSound 2.1 software (Pettersson Elektronic AB, Upsala,
Sweden), with a Hanning window type, frame overlap of 99%, and 512 FFT size. From each echolocation sequence
(defined as a recording file of 15 s maximum containing two or more pulses emitted by a bat), one set of search-
phase pulses with good signal-to-noise ratio (peak intensity with more than 20 dB above noise level measured in
the power spectrum) was chosen and described. Following this criterion, a total of 176 pulses of M. atacamensis,
162 pulses of M. c. arescens and 105 pulses of M. c. chiloensis, were selected for analysis. Interspecific variation in
echolocation calls was evaluated from quantitative analyses based on the following acoustic parameters: duration
(Dur; time between start and end of a pulse, measured in ms in the oscillogram), start and end frequencies (SF
and EF; measured in kHz in the spectrogram), bandwidth (BW; difference between start and end frequencies),
slope of frequency modulation (SFM; difference in kHz between the start and end frequencies of the call divided
by call duration), peak frequency (PF; frequency in kHz corresponding to the maximal intensity in the power
spectrum), maximum and minimum frequencies (MaxF and MinF; measured 20 dB below peak intensity in the
power spectrum). In each call sequence, we also measured the interpulse interval (IPI) from the beginning of a call
to the start of the next call.
To determine whether echolocation calls could be separated into three independent groups corresponding to
each taxon, a Discriminant Function Analysis was performed using “MASS” and “Lattice” packages implemented
in the R platform (Vanables & Ripley 2002; Sarkar 2008). λ-Wilks’ values were obtained with a MANOVA to test
for statistical significance of DFA models. This analysis was performed using ‘Dplyr’ package implemented in the R
platform, and the differences were considered statistically significant when p-values were less than 0.05 (Wickham
et al. 2017). The standardized discriminant function coefficients were used to determine the contribution each
variable made to the ability of DFA to classify calls.
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Results
Phylogenetic relationships and genetic distances
The cyt-b dataset showed considerable variation, containing 419 variable sites, 378 of which were parsimoniously
informative. Chilean samples were recovered in two distinct clades, with high nodal support. The first clade is
composed of specimens of M. c. chiloensis and M. c. arescens, which were recovered as sister taxa (Figure 2). The
second clade is composed of M. atacamensis specimens, which are sister taxa of M. oxyotus (Figure 2). All Chilean
Myotis species are allocated into a major monophyletic cluster called the albescens-group, which in this phylogeny
is represented by M. albescens, M. atacamensis, M. attenboroughi, M. bakeri, M. c. chiloensis, M. c. arescens, M.
clydejonesi, M. dinellii, M. dominicensis, M. larensis, M. lavali, M. levis, M. martiniquensis, M. nesopolus, M.
nigricans, M. nyctor, and M. oxyotus.
FIGURE 2. Phylogenetic tree resulting from the Bayesian Inference of cytochrome-b sequences (1,140 bp) of Neotropical
Myotis species. Chilean samples are colored, being M. arescens (red), Myotis atacamensis (blue), and Myotis chiloensis (green).
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The genetic distance based on the cyt-b gene sequences between M. c. chiloensis and M. c. arescens ranged
from 1.6 to 2.2% based on the K2p model, while the intraspecific variation is less than 0.02% for both taxa. Myotis
atacamensis shows a genetic distance of 3.6% ± 0.5 in relation to its sister taxa, M. oxyotus. The genetic distance
between M. atacamensis and other Chilean Myotis is greater than 10%. The average genetic distance among species
of Neotropical Myotis, except M. arescens and M. chiloensis, ranged from 3.1 to 18%.
Morphological variation
FIGURE 3. Dorsal (upper) and ventral (below) pelage of Myotis arescens (A, B; USNM 319784), Myotis atacamensis (C, D;
MVZ 116638), and Myotis chiloensis (E, F; FMNH 24029 [neotype]).
Based on morphological comparisons between the samples and the type specimens of South American Myotis, we
inferred that there three differentiated OTUs, corresponding to M. atacamensis (OTU1), M. c. arescens (OTU2),
and M. c. chiloensis (sensu stricto, OTU3). Qualitative morphological analyses distinguish these three taxa, both
from external and cranial characters. Myotis atacamensis has long, silky, and tricolored dorsal fur, with dark-brown
bases (2/5 of total hair length), pale yellowish middle portions (2/5 of total hair length), and yellowish-brown tips
(1/5 of total hair length), with a strong contrast between bases and tips; ventral fur strongly bicolored, with dark-
brown bases (2/3 of total hair length) and whitish-gray tips (1/3 of total hair length; Figure 3). Myotis c. chiloensis
presents a remarkably different fur color pattern, with the dorsal fur woolly, medium to long, and cinnamon-brown,
unicolored or with bases (1/3 of total hair length) slightly darker than tips; ventral fur is bicolored, with medium-
brown bases (1/2 of total hair length) and slightly paler tips (Figure 3). Myotis c. arescens has a very different fur
color from both species, with dorsal fur woolly, medium to long, and sharply bicolored, with dark-brown bases (ca.
1/2 of total hair length) and cinnamon-brown tips; ventral fur is strongly bicolored, with dark-brown bases (2/3 of
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total hair length) and pale yellowish tips (1/3 of total hair length; Figure 3). In general, M. c. arescens has dorsal fur
more similar to that found in M. c. chiloensis, whereas the ventral fur resembles that of M. atacamensis, but with tips
more yellowish, which perhaps leads to misidentification when the dorsal and ventral color pattern are not analyzed
together. In M. c. chiloensis the contrast between dorsal and ventral is missing or barely evident, while in the other
two species the contrast is marked. In addition to fur coloration, there are other external morphological characters
that distinguish these three taxa (see Taxonomy section below).
The skull shape of M. c. arescens resembles M. c. chiloensis (Figure 4), however, it is narrower and has a
more elongated rostrum, the braincase is lower in profile, and the occiput is higher in relation to the braincase roof
formed by the parietals. In M. c. arescens, the second upper premolar (P3) is subtly smaller than the first premolar
(P2), different from that found in M. c. chiloensis, where P2 and P3 are the same size. Myotis atacamensis has a
remarkably different skull shape from that found in M. c. arescens and M. c. chiloensis (Figure 4). The skull is tiny
and delicate, the braincase more globular in dorsal view and higher in profile view; the frontal bone has a sharp
decline forward, towards the nasal; and the rostrum is narrower.
FIGURE 4. Dorsal (upper) and ventral (middle) and lateral skull views of Myotis arescens (USNM 319784), Myotis atacamensis
(MVZ 116638), and Myotis chiloensis (FMNH 24029 [neotype]). Scale bar = 10 mm.
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The morphometric analyses revealed complete separation between M. atacamensis, M. c. chiloensis, and M.
c. arescens in morphospace, indicating these three species have distinct skull sizes and shapes (Figure 5). The first
principal component (PC1) accoutered for 96% of the total craniometric variation, and represents overall skull size
(Figure 5A, B). Along this axis, scores of these three species overlap slightly. However, these taxa overlap broadly
along the second principal component (PC2 = 2%), which represents overall skull shape. A similar result was
obtained from the discriminant functions analysis (Figure 5C, D), which recovered complete separation of taxa along
the first axis (DF1 = 95%) and overlap along the second axis (DF2 = 5%). Measurements referring to the length of
the skull and rostrum (GLS, CBL, CCL, CIL, BAL) were the ones that contributed most to the discrimination of the
species in the multivariate statistical models (Table 2). There are significant differences in skull size among the three
species (λ-Wilks = 0.012; F = 10.95; p < 0.001), between M. atacamensis and M. c. chiloensis (p = 0.02); between
M. atacamensis and M. c. arescens (p < 0.01), and between M. c. chiloensis and M. c. arescens (p < 0.01).
FIGURE 5. Plots showing dispersion points and vector correlation of skull measurements of Principal Component Analysis (A,
B) and Discriminant Function Analysis (C, D) for Myotis species from Chile.
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Myotis atacamensis is the smallest species (FA 30.6–34.1 mm; GLS 12.6–13.5 mm) and does not overlap with
the other two species in almost all measurements (e.g., FA, GLS, IOB, BAC, MAL; Table 3). On the other hand,
M. c. chiloensis and M. c. arescens may overlap in some cranial measurements (e.g., FA, GLS, BCB; Table 3).
Still, M. c. arescens (FA 34.5–37.8 mm; GLS 13.6–14.8 mm) is smaller overall than M. c. chiloensis (FA 36.3–40.;
GLS 13.8–15.3), especially when considering the averages of cranial size, and some measurements can be used in
discrimination (e.g., IOB and BAC). This size difference, added to the qualitative differences in morphology, is
sufficient to separate these two taxa.
TABLE 2. Vector correlation loadings with original variables of principal components (PC1 and PC2) and discriminant
functions (DF1 and DF2) for selected samples of M. atacamensis, M. chiloensis, and M. arescens. See Table 1 for variable
abbreviations.
Measurements PC 1 PC 2 DF1 DF2
GLS 0.332 -0.270 0.115 -0.136
CCL 0.350 -0.219 0.124 -0.100
CBL 0.354 -0.240 0.124 -0.122
CIL 0.374 -0.213 0.132 -0.113
BAL 0.331 -0.204 0.116 -0.142
MAB 0.210 0.059 0.075 -0.076
BCB 0.156 0.136 0.056 -0.048
IOB 0.189 0.496 0.073 -0.026
POB 0.132 0.080 0.048 -0.043
BAC 0.165 0.395 0.064 0.000
BAM 0.210 0.514 0.081 -0.023
MTL 0.189 0.054 0.068 -0.059
M1M3 0.103 0.072 0.037 -0.038
MAL 0.335 0.156 0.122 -0.089
MAN 0.200 0.087 0.072 -0.072
TABLE 3. Selected measurements (mm) of M. atacamensis, M. chiloensis, and M. arescens. Descriptive statistics include
the mean, range (in parentheses), and sample size. See Table 1 for variable abbreviations.
Measurements Myotis arescens Myotis atacamensis Myotis chiloensis
Mean (min–max) N Mean (min–max) N Mean (min–max) N
FA 36.2 (34.5–37.8) 8 32.2 (30.6–34.1) 6 38.5 (36.3–40.0) 30
EL 15.7 (14.7–16.5) 4 14.0 (13.0–14.5) 5 15.0 (14.0–15.5) 27
LDF 7.2 (6.5–7.5) 6 7.7 (7.0–8.8) 3 7.7 (6.5–8.5) 10
LVF 6.5 (5.4–7.0) 6 6.3 (5.9–6.7) 3 6.5 (5.6–7.2) 10
GLS 14.0 (13.6–14.8) 19 12.9 (12.6–13.5) 8 14.8 (13.8–15.3) 45
CCL 12.3 (11.9–12.9) 19 11.2 (10.9–11.8) 8 13.0 (12.7–13.2) 45
CBL 13.0 (12.7–13.6) 19 11.9 (11.5–12.5) 8 13.7 (13.2–14.0) 45
CIL 13.1 (12.8–13.8) 19 12.0 (11.7–12.6) 8 13.9 (13.1–14.4) 45
BAL 11.9 (11.5–12.5) 19 10.8 (10.5–11.4) 8 12.6 (12.1–12.9) 44
ZYG (8.5–8.6) 2 (6.7–6.8) 2 (9.1–9.2) 2
MAB 7.1 (6.7–7.5) 18 6.4 (6.3–6.7) 6 7.5 (6.8–7.8) 46
BCB 6.7 (6.4–7.0) 19 6.2 (6.1–6.5) 7 7.0 (6.3–7.4) 47
IOB 4.2 (3.9–4.5) 17 3.7 (3.6–3.8) 8 4.7 (4.5–5.0) 46
POB 3.5 (3.4–3.8) 18 3.1 (3.0–3.2) 8 3.8 (3.5–4.0) 47
BAC 3.3 (3.2–3.6) 19 2.9 (2.8–3.0) 7 3.8 (3.6–4.0) 46
BAM 5.3 (5.0–5.6) 19 4.7 (4.6–4.9) 8 5.8 (5.4–6.1) 47
MTL 5.2 (5.1–5.5) 19 4.6 (4.4–5.0) 8 5.6 (5.2–5.9) 47
M1–3 3.0 (2.6–3.1) 19 2.6 (2.6–2.8) 8 3.2 (3.0–3.3) 47
MAL 9.8 (9.6–10.3) 19 8.8 (8.5–9.3) 5 10.6 (10.1–11.0) 45
MAN 5.6 (5.3–5.8) 19 4.9 (4.8–5.3) 7 6.0 (5.6–6.3) 46
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Bioacoustic variation
Quantitative acoustic analysis distinguished the three taxa based on their echolocation calls. Multivariate DFA
gave an overall classification of 93% of the calls. MANOVA showed that the model was significant (λ-Wilks’ =
0.104, F = 129.700, p < 0.001) and that 90% of the total acoustic variation was explained by the first discriminant
function. Echolocation calls emitted by M. c. chiloensis and M. c. arescens were correctly identified 97% and 96%
of the time and grouped separately from M. atacamensis, although there is subtle overlap (Figure 6). The most
important acoustic parameters for discrimination among these three taxa were the end frequency (EF), minimum
(MinF), and peak frequencies (PF). The DFA indicated a segregation of M. atacamensis from both M. c. chiloensis
and M. c. arescens along the first axis, which is responsible for 90% of the variation (Figure 6). On the other hand,
M. c. chiloensis and M. c. arescens overlap widely along the first axis, but are separated along the second axis,
which represents 10% of the variation. This result indicates a greater similarity of echolocation calls between M. c.
chiloensis and M. c. arescens than of those species with M. atacamensis.
FIGURE 6. Plots showing dispersion points and vector correlation of bioacoustic parameters of Discriminant Function Analysis
for Myotis species from Chile.
All three species produce echolocation calls characterized by short (< 6 ms) and single downward frequency-
modulated pulses (Figure 7). However, M. atacamensis emits pulses at a higher frequency range (EF 47.74 ± 2.89;
MinF 50.10 ± 4.27; PF 58.32 ± 3.64), compared to M. c. arescens (EF 39.75 ± 1.90; MinF 40.64 ± 2.11; PF 47.52 ±
4.40), while M. c. chiloensis has the lower frequency range (EF 38.53 ± 1.61; MinF 39.54 ± 1.66; PF 45.30 ± 3.56)
(Table 4).
TABLE 4. Descriptive statistics (mean ± SD) of the echolocation call of M. atacamensis, M. arescens and M. chiloensis.
Acoustic parameters Myotis arescens
(N = 162 pulses)
Myotis atacamensis
(N = 176 pulses)
Myotis chiloensis
(N = 105 pulses)
Duration (ms) 5.02 ± 0.71 3.61 ± 0.66 5.21 ± 0.87
Start frequency (kHz) 95.45 ± 9.92 85.74 ± 10.57 92.17 ± 11.73
End frequency (kHz) 39.75 ± 1.90 47.75 ± 2.89 38.54 ± 1.62
Bandwidth (kHz) 55.70 ± 9.78 37.99 ± 11.88 53.63 ± 11.75
Slope (kHz/ms) 11.41 ± 3.00 11.18 ± 5.00 10.68 ± 3.36
Peak frequency (kHz) 47.53 ±4.41 58.32 ± 3.64 45.31 ± 3.56
Minimum frequency (kHz) 40.65 ± 2.11 50.10 ± 4.27 39.54 ± 1.66
Maximum frequency (kHz) 69.53 ± 12.26 73.53 ± 5.76 68.57 ± 12.41
Interpulse interval (ms) 72.85 ± 12.11 77.77 ± 17.93 110.47 ± 17.18
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FIGURE 7. Oscillograms (upper) and spectrograms (below) of echolocation calls from Chilean Myotis.
Taxonomy
Molecular analyses indicated M. c. chiloensis and M. c. arescens as sister taxa, both reciprocally monophyletic, but
with a low genetic distance (< 3%). M. atacamensis was recovered in a distinct clade, phylogenetically related to
M. oxyotus. Morphological analyses, both in shape and size, indicate the existence of three phenotypically distinct
taxa. Similarly, the pattern of variation in echolocation calls also recovers bioacoustic divergences among the
three geographically delimited OTUs. Although there are possible overlaps in the distribution of these three taxa,
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which may be biasing the results obtained in the multivariate analyses based on the echolocation parameters, the
congruence among molecular, morphological, and bioacoustic evidence supports the recognition of three distinct
Myotis species in Chile. Therefore, we consider Myotis arescens as a valid species and distinct from other Chilean
congeners, M. atacamensis and M. chiloensis.
Myotis chiloensis (Waterhouse, 1840)
Type. Neotype FMNH 24029, adult female collected by J. Vera in 1923; skull partially damaged, mandible, and
skin (see LaVal 1973).
Type locality. Cucao, Chiloé Island, Los Lagos, Chile.
Distribution. Occurs in Southern Chile, eastward into western Argentina and southward to Tierra del Fuego,
occupying the Hyper-oceanic Temperate and Patagonian bioclimatic zones, in Deciduous Forest, Evergreen Forest,
Valdivian Rainforest, and Magellanic Moorland (Ossa & Rodríguez-San Pedro 2015; Novaes 2019). Records are
from sea level to ca. 1,400 m a.s.l.
Diagnosis. Medium to large size (FA 36.3–40.0 mm; GLS 13.8–15.3 mm); dorsal fur long (6.5–8.5 mm), woolly,
unicolored or subtly bicolored, with medium-brown bases (near Mummy Brown) and tips generally Brussels Brown
or Cinnamon Brown; ventral fur bicolored, with Mummy Brown bases and Dresden Brown tips; dorsal surface of
the uropatagium naked; fringe of hairs on the distal border of the uropatagium absent; plagiopatagium connected
to the feet by a broad band of membrane. Sagittal crest usually absent; broader skull; braincase high in profile and
elongated in dorsal view; braincase roof formed by the parietal bone is straight; forehead subtly sloping in lateral
view; broad and short rostrum; posterior region of the braincase rounded and quite projected beyond the limit of the
occipital condyles; mastoid processes narrow.
Description and comparisons. Dental formula is I 2/3, C 1/1, PM 3/3, M 3/3 (2x) = 38, and the teeth are robust
and well developed. The second upper premolar (P3) is approximately the same size as the first upper premolar (P2),
aligned in the toothrow, and visible labially. Skull medium to large with braincase elongated in dorsal view; parietals
slope subtly forward to frontal bone; braincase roof is straight; mastoid processes narrow and poorly developed;
rostrum long and broad; the sagittal crest is usually absent and lambdoidal crests are present and low; and the
occipital region is rounded and projected beyond the posterior surfaces of the occipital condyles.
Ears comparatively short in size, not reaching the nostrils when extended forward. Membranes and ears are
Mummy Brown. Plagiopatagium is connected to the feet at the level of the toes by a broad band of membrane;
a fringe of scattered hairs on the distal border of the uropatagium is absent. Dorsal surface of the uropatagium
virtually naked. Woolly and medium to long fur; dorsal hairs generally Brussels Brown or Cinnamon Brown, with
the basal half subtly darker. Ventral fur bicolored, with Mummy Brown bases (1/2 of total hair length) and Dresden
Brown tips (1/2 of total hair length).
Considering either the assemblage of Myotis that occurs along mountainous habitats or in the lowland portion
of the western side of the Andes (M. albescens, M. atacamensis, M. arescens, M. bakeri, M. keaysi, and M. oxyotus),
M. chiloensis can be distinguished from all by the set of diagnostic traits. It is morphologically closer to M. arescens,
from which it can be distinguished by its darker ventral fur (near Dresden Brown in chiloensis and near Pale Olive
Buff on the tips in arescens), comparatively shorter ears (in arescens the ears reach the nostrils when extended
forward), fringe of hairs on the distal border of the uropatagium absent (present in arescens), and broader rostrum
(BAC ≥ 3.6 in chiloensis, BAC ≤ 3.6 in arescens). Furthermore, M. chiloensis is larger than M. arescens in general,
with only the smallest individuals reaching sizes similar to those in the largest arescens individuals (Table 3).
Myotis chiloensis can be distinguished from M. atacamensis and M. bakeri by its general smaller size (e.g.,
FA > 36 in chiloensis, FA < 35 in atacamensis and bakeri); woolly fur with dorsal hair cinnamon-brown weakly
contrasting between bases and tips, and venter brownish; in M. atacamensis and M. bakeri the fur is silky, dorsal
hairs with strong contrast between bases (blackish) and tips (yellowish), and venter whitish.
From M. albescens, it can be distinguished by the darker ventral fur color being entirely brownish with little
contrast between base and tip in M. chiloensis; while M. albescens have the throat yellowish, grading to whitish
towards the abdomen and sides of the body, with hairs strongly bicolored (blackish base and whitish tips). In
addition, M. albescens has a more globular braincase (see Moratelli & Oliveira 2011); whereas in M. chiloensis the
braincase is elongated.
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Myotis chiloensis can be distinguished from M. oxyotus by its shorter and woolly fur, with dorsal hairs cinnamon-
brown with bases slightly darker, and ventral fur weakly bicolored, Dresden Brown in color; whereas in M. oxyotus
the fur is silky and very long (LDF > 8 mm), bicolored, with blackish bases and generally Mummy Brown or
Brownish Olive tips, and ventral fur strongly bicolored with blackish bases and tips ranging from Pale Pinkish-Buff
to Deep Olive-Buff. In M. chiloensis the parietals slope subtly to the frontal bones and the braincase is elongated
in dorsal view; whereas in M. oxyotus the parietals slope steeply to the frontal bones and the braincase is inflated
in dorsal view. Myotis chiloensis can be distinguished from M. keaysi by its fur color, with dorsal hairs cinnamon-
brown unicolored or with bases slightly darker, and ventral fur weakly bicolored, Dresden Brown in color; whereas
in M. keaysi the dorsal fur is reddish-brown (from Cinnamon Brown to Ochraceous Tawny) with the base being
remarkably darker, and ventral fur with Clove Brown bases and tips ranging from Ivory Yellow to Light Drab. In
relation to skull morphology, it differs from M. keaysi by sagittal crest absent, mastoid process narrower and poorly
developed, and parietals sloping subtly to frontal bones; in M. keaysi a sagittal crest is always present, mastoid
process larger and well-developed, and parietals slope steeply to frontal bones.
Myotis atacamensis (Lataste, 1892)
Type. Neotype USNM 391786, adult female collected by W. Mann and S. Mann in January 1944; skull partially
damaged, mandible, and skin (see Novaes et al. 2022b).
Type locality. Near Minimini, Tarapacá, Chile.
Distribution. Occurs from Arequipa, in western Peru, to northern Chile, being associated with Desertic and
Xeric Tropical formations in elevations from 990 to 3,475 m, where it inhabits vegetational formations such as
Absolute Desert, Desertic Shrubland, and Highland low-Shrublands.
Diagnosis. Small size (FA 30.6–34.1 mm; GLS 12.6–13.6); dorsal fur long (7–9 mm), silky, and tricolored,
with dark-brown bases (near Bone Brown), pale yellowish middle portions (near Pale Olive-Buff), and yellowish-
brown tips (near Light Ochraceous Buff); ventral fur strongly bicolored, with Bone Brown bases and whitish-gray
tips (near Pallid Brownish Drab); dorsal surface of the uropatagium covered by thin fur not extending beyond the
knees; presence of a fringe of sparse hairs on the distal border of the uropatagium; plagiopatagium connected to the
feet by a broad band of membrane. Sagittal crest usually absent; elongated and narrow skull; braincase remarkably
inflated and high in profile; braincase roof formed by the parietal bone is straight; forehead steeply sloping in lateral
view; narrow and short rostrum; posterior region of the braincase rounded and quite projected beyond the limit of
the occipital condyles; mastoid processes narrow
Description and comparisons. Dental formula is I 2/3, C 1/1, PM 3/3, M 3/3 (2x) = 38, and the teeth are
small. Skull small; forehead steeply sloping with inflated braincase; braincase roof is straight; mastoid processes
narrow and poorly developed; rostrum narrow and comparatively short; the sagittal crest and lambdoidal crests are
usually absent; and the occipital region is rounded and conspicuously projected beyond the posterior surfaces of the
occipital condyles. The second upper premolar (P3) is a little smaller than the first upper premolar (P2), aligned in
the toothrow and visible labially in all individuals.
Ears are moderate in size, almost reaching the nostrils when extended forward. Membranes and ears are Mummy
Brown or lighter. Plagiopatagium is connected to the feet at the level of the toes by a broad band of membrane; a
fringe of scattered hairs on the distal border of the uropatagium is present and visible only under magnification.
Dorsal surface of the uropatagium barely furred, with hairs not extending beyond to the knees. Silky and long fur;
dorsal hairs tricolored, with Bone Brown bases (2/5 of total hair length), Pale Olive-Buff middle portions (2/5 of
total hair length), and Light Ochraceous Buff (1/5 of total hair length), however, the contrast between tip and middle
portion of the hair is quite subtle. Ventral fur strongly bicolored, with Bone Brown bases (2/3 of total hair length)
and Pallid Brownish Drab tips (1/3 of total hair length).
Myotis atacamensis differs from all South American congeners by the tricolored dorsal fur. From Chilean
Myotis, M. atacamensis can be distinguished from M. arescens and M. chiloensis by its general smaller size (Table
3); silky, brighter, and longer fur, with dorsal hair tricolored and strongly contrasting between bases (dark brown)
and tips (yellowish), and whitish-gray venter; in M. chiloensis, the fur is woolly and shorter, dorsal hairs with
weak contrast between bases (medium-brown) and tips (cinnamon-brown) and medium-brown venter; whereas
in M. arescens, the fur is woolly and shorter, dorsal hairs with clear contrast between bases (dark-brown) and tips
(cinnamon-brown), and pale-yellowish venter.
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Myotis atacamensis is morphologically closer to M. bakeri, from which it can be distinguished by slightly lighter
dorsal fur (near Light Ochraceous Buff on the tips in atacamensis and near Buckthorn Brown on the tips in bakeri);
presence of a fringe of hairs along the trailing edge of uropatagium (absent in M. bakeri); and narrower skull (e.g.,
POB < 3.5 in atacamensis, POB ≥ 3.5 in bakeri); in addition, the base of dorsal fur is dark brown in M. atacamensis
and blackish in M. bakeri. From M. albescens, M. atacamensis can be distinguished by the conspicuously paler
dorsal fur color (dorsal fur dark to medium-brown on the bases [4/5 of the total fur length] and yellowish on the tips
[1/5] in albescens, conveying a brownish general appearance to the dorsum; in contrast with bases and tips strongly
contrasting and general yellowish appearance in atacamensis). Additionally, M. albescens have the throat yellowish,
grading to whitish towards the abdomen and perianal area, and a more globular braincase (see Moratelli & Oliveira
2011); whereas in M. atacamensis the entire venter, from the throat to the abdomen, is whitish-gray and the skull is
narrower, and the braincase is less globular. In addition, M. atacamensis is conspicuously smaller than M. albescens
in external and cranial measurements (see Moratelli & Oliveira 2011).
Myotis atacamensis can be distinguished from M. oxyotus by the dorsal fur paler and with strong contrast between
bases and tips, and the ventral fur whitishgray; whereas the dorsal fur is darker, and the ventral fur is yellowish in M.
oxyotus. Myotis atacamensis is smaller than M. oxyotus in virtually all external and cranial measurements (e.g., GLS
12.6–13.6 mm in atacamensis, 14.1–15.0 mm in oxyotus; POB 3.0–3.2 mm in atacamensis, 3.3–3.9 mm in oxyotus;
MAL 8.5–9.3 mm in atacamensis, 9.9–11.2 mm in oxyotus). Myotis atacamensis can be distinguished from M.
keaysi by its general smaller size (FA 30.6–34.1 mm in atacamensis, 38.5–43.4 mm in keaysi; GLS 12.6–13.5 mm in
atacamensis, 13.9–14.7 mm in keaysi), paler, longer, and silky fur (shorter, woolly, and either brownish or reddish-
brown in keaysi). In relation to skull morphology, it differs from M. keaysi by sagittal crest usually absent, mastoid
process narrower and poorly developed, and shorter rostrum. In addition to these characters, M. atacamensis differs
from its aforementioned South American congeners, except M. albescens and M. arescens, by the presence of a
fringe of scattered hair on the distal edge of the uropatagium.
Myotis arescens Osgood, 1943
Type. Holotype FMNH 24396, adult male collected by J. A. Wolffsohn on January 1, 1925; skin only.
Type locality. Hacienda Limache, Valparaíso, Chile.
Distribution. There are records from Central Chile, in the regions of Valparaíso, Coquimbo, Maule, and
Araucanía, associated with Mediterranean vegetational formations, such as Desertic low-Shrubland, Sclerophyllous
Shrublands, Sclerophyllous Forest, and Broadleaf Rainforest, from sea level to 1,020 m a.s.l.
Diagnosis. Medium-sized species (FA 34.5–37.8 mm; GLS 13.6–15.0 mm); dorsal fur medium to long (6.5–7.5
mm), woolly, and remarkably bicolored, with dark-brown bases (near Bone Brown) and tips generally Brussels
Brown or Cinnamon Brown; ventral fur strongly bicolored, with Bone Brown bases and Pale Olive Buff tips;
dorsal surface of the uropatagium barely furred; fringe of hairs on the distal border of the uropatagium present;
plagiopatagium connected to the feet by a broad band of membrane. Sagittal crest usually absent; broader skull;
braincase high in profile and elongated in dorsal view; braincase roof formed by the parietal bone is straight;
forehead subtly sloping in lateral view; broader and short rostrum; posterior region of the braincase rounded and
quite projected beyond the limit of the occipital condyles; mastoid processes narrow.
Description and comparisons. Dental formula is I 2/3, C 1/1, PM 3/3, M 3/3 (2x) = 38, and the teeth are robust
and well developed. Skull medium to large with braincase elongated in dorsal view; parietals slope subtly forward to
frontal bones; braincase roof is straight; mastoid processes narrow and poorly developed; rostrum long and narrow;
the sagittal crest absent and lambdoidal crests present and low; and the occipital region is rounded and projected
beyond the posterior surfaces of the occipital condyles. The second upper premolar (P3) smaller than first upper
premolar (P2), aligned in the toothrow and visible labially.
Ears comparatively large and reach the nostrils when extended forward. Membranes and ears are Mummy
Brown. Plagiopatagium is connected to the feet at the level of the toes by a broad band of membrane; a fringe of
scattered hairs on the distal border of the uropatagium is present. Dorsal surface of the uropatagium is barely furred,
with hairs reaching knee level or just beyond. Woolly and medium to long fur; dorsal pelage sharply bicolored with
Bone Brown bases and tips ranging from Brussels Brown to Cinnamon Brown. Ventral fur strongly bicolored, with
Bone Brown bases (1/2 of total hair length) and pale-yellowish tips (near Pale Olive Buff).
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Myotis arescens is morphologically closer to M. chiloensis, from which it can be distinguished by lighter ventral
fur (near Pale Olive Buff on the tips in arescens and near Dresden Brown in chiloensis); comparatively longer
ears (in chiloensis the ears not reaching the nostrils when extended forward); and narrower rostrum (BAC ≤ 3.6 in
arescens, BAC ≥ 3.6 in chiloensis,). Furthermore, M. arescens is smaller than M. chiloensis in general (Table 3).
Myotis arescens can be distinguished of M. atacamensis and M. bakeri by its general larger size (e.g., FA > 34.5
in arescens, FA < 34.5 in atacamensis and bakeri); woolly fur with cinnamon-brown dorsal hairs, and venter pale-
yellowish; whereas in M. atacamensis and M. bakeri the fur is silky, dorsal hairs with strong contrast between bases
(dark brown or blackish) and tips (yellowish), and venter whitish.
In relation to M. albescens, M. arescens can be distinguished by the conspicuously bicolored dorsal fur, with
Bone Brown bases and tips generally Brussels Brown or Cinnamon Brown; whereas in M. albescens the dorsal fur
is almost entire Bone Brown with yellowish on the tips (1/5 of hair length). In addition, M. albescens has a more
globular braincase (see Moratelli & Oliveira 2011); whereas in M. arescens the braincase is elongated.
Myotis arescens can be distinguished from M. oxyotus by its shorter and woolly fur, with dorsal hairs cinnamon-
brown with darker bases, and ventral fur bicolored, with pale-yellowish appearance; whereas in M. oxyotus the fur
is silky and very long (LDF > 8 mm), with blackish bases and generally Mummy Brown or Brownish Olive tips,
and ventral fur with blackish bases and tips ranging from Pale Pinkish-Buff to Deep Olive-Buff. In M. arescens
the parietals slope subtly to frontal bones and braincase is elongated in dorsal view, whereas in M. oxyotus the
parietals slope steeply to frontal bones and braincase is inflated in dorsal view. Myotis arescens can be easily
distinguished from M. keaysi by its ventral fur color, pale-yellowish in arescens and Ivory Yellow or Light Drab
in keaysi; it differs from M. keaysi by sagittal crest absent, mastoid process narrower and poorly developed, and
parietals sloping subtly to frontal bones; in M. keaysi sagittal crest is always present, mastoid process larger and
well-developed, and parietals slope steeply to frontal bones. In addition, M. arescens differs from its aforementioned
South American congeners, except M. atacamensis, by the long ear, reaching or almost reaching the nostrils when
extended forward.
Discussion
Myotis has a complex evolutionary history in the Neotropics, arising from a recent and rapid diversification caused
by dispersal and isolation events (Stadelmann et al. 2007; Larsen et al. 2012b; Novaes et al. 2021a). A recent study
using a double digestion RAD-seq method found a strong population structure with pronounced isolation-by-distance
in M. chiloensis (sensu lato; including specimens of M. arescens), revealing high degrees of heterozygosis between
southern and northern populations (Lilley et al. 2020). This study also presents a coalescent analysis indicating that
M. chiloensis populations may still not have reached secondary contact after the Last Glacial Maximum (Lilley et
al. 2020).
The results of Lilley et al. (2020) corroborate our findings indicating low genetic distance between M. chiloensis
and M. arescens (ca. 2%), probably caused by a recent separation. On the other hand, the conspicuous phenotypic
difference between these taxa indicates that they are undergoing different evolutionary pressures, following their
adaptive trajectories independently. This hypothesis can also be supported by bioacoustic data, which indicates
distinct echolocation calls patterns between M. arescens and M. chiloensis. This scenario reveals M. arescens as a
monophyletic taxon, morphologically diagnosable, and bioacoustically distinct, therefore, fulfilling the requirements
to be considered a full species from the Phylogenetic Species Concept (see Zachos 2016).
The recognition of M. arescens as a full species corroborates previous evidence that indicated the existence of
cryptic morphological complexes in Chile, including forms that did not fit into the two Myotis species currently
recognized for the region, M. atacamensis and M. chiloensis (i.e., Novaes et al. 2018). LaVal (1973) recognized
two morphologically distinct forms in M. chiloensis, one pale population from central Chile, and the other brownish
population from southern Chile. However, LaVal (op. cit.) decided to keep M. arescens as a junior synonym of M.
chiloensis, indicating that there is no phenotypic discontinuity, but a clinal variation. On the other hand, Novaes et
al. (2018) indicated that individuals originally identified as M. chiloensis from Coquimbo and Limache (mentioned
by LaVal as the pale form) do not fit morphometrically within populations of M. chiloensis from Argentina and
southern Chile and suggested that there is still hidden diversity of Myotis in central Chile. Therefore, we confirm
that the atypical forms of M. chiloensis mentioned by LaVal (1973) and Novaes et al. (2018) are, in fact, M.
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arescens. Similarly, individuals identified as M. atacamensis from Coquimbo, Valparaiso, and Metropolitan regions
(Rodriguez-San Pedro et al. 2014, 2015, 2020) would correspond to M. arescens.
It is possible that the difficulty in identifying M. arescens, both in field and museums, was due to the combination
of fur color and ear length, which resembles M. atacamensis, and the cranial shape similar to M. chiloensis, although
smaller. However, multivariate analyses based on the variation in echolocation calls segregates M. chiloensis and M.
arescens, corroborating the results obtained by the morphological analyses. The echolocation pulses of bats are vital
for spatial orientation and prey search, so there tends to be considerable variation even between phylogenetically
close species (Schuller & Moss 2004; Chornelia et al. 2022). Such variation may be due to changes in the shape of
skull and other head structures, resulting in changes in frequency, as well as selective pressures due to specialization
in the ecological niche (Heller & von Helversen 1989; Jones & Holderied 2007). This variation in bat calls has
been useful in species delimitation and recognition of new lineages in cryptic species complexes (Pavan et al. 2018;
Chornelia et al. 2022). This is the first study using bioacoustics as additional evidence for the species delimitation
of Neotropical Myotis.
We hypothesize that morphological difference of M. arescens in relation to M. chiloensis is due to a strong force
of natural selection, which has led the Myotis species that occupy the arid and semi-arid zones of the western Andes
(M. arescens, M. atacamensis, and M. bakeri) to develop a similar phenotypic pattern in fur color, ear shape, and
general size. In fact, our results indicate that the high phenotypic similarity between M. atacamensis and M. bakeri
is the result of a convergent evolution, since these two species are not phylogenetically related.
The allopatry among the three Myotis species from Chile suggests a spatial segregation resulting from adaptive
specializations related to the occupied habitats. However, it is possible that these species have wider distributions,
with the possibility of overlap in areas of transitional habitats. Indeed, Lilley et al. (2020) present evidence of
possible hybridization between M. arescens and M. chiloensis, especially in the contact zones of what would be
the southern and northern limits of the distribution of these two species, respectively. Therefore, we suggest that
further studies with Chilean Myotis should focus on defining the distributional limits of these species, in addition to
investigating the probable biogeographic events and processes involved in the evolution of the species.
Acknowledgments
We are grateful to the curators of the collections referred to in the text for providing access to the specimens: M. Ruedi
(MHNG, Geneva, Switzerland), N. Simmons and E. Westwig (AMNH, New York, USA), J. Wible, S. McLaren (CM,
Pittsburgh, USA), B. Patterson, W. Stanley and R. Banasiak (FMNH, Chicago, USA), R.M. Timm and M. Eifler
(Museum of Natural History at KU, Lawrence, USA), J.A. Esselstyn (LSUMZ, Baton Rouge, USA), C. Conroy
(MVZ, Berkeley, USA), J.K. Braun (OMNH, Norman, USA), and K. Helgen, D. Lunde, and L. Gordon (USNM,
Washington D.C., USA). We thank J.L. Allendes, D.E. Lobos and C.A. Beltrán for their valuable collaboration in
field data collection. We also thank Servicio Agrícola y Ganadero, División Protección de los Recursos Naturales
Renovables (Diproren) for the capture permits in Chile. This work was partially supported by the Smithsonian
Institution (USA) through grants to DEW. RM has received financial support from CNPq (313963/2018-5) and
FAPERJ (E-26/200.967/2021). RLMN has received support from FAPERJ (E-26/204.243/2021; E26/200.631/2022
and E26/200.395/2022).
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APPENDIX 1. Cytochrome-b sequences used in the phylogenetic analyses. The information presented for the taxonomic
terminals is the result of re-identification of the specimens and does not necessarily coincide with the original identifications
provided by the authors and GenBank. Abbreviations for specimen deposit institutions are: Carnegie Museum of Natural
History, Pittsburg, USA (CM); Field Museum of Natural History, Chicago, USA (FMNH), Museum of Vertebrate Zoology,
University of California, Berkeley, USA (MVZ); University of Nebraska State Museum (UNSM, Lincoln, USA);
Smithsonian National Museum of Natural History, Washington, DC, USA (USNM); Texas Tech University, Lubbock,
USA (TTU). Pontificia Universidad Católica del Ecuador, Quito, Ecuador (QCAZ); Biology Department of Tunghai
University, Taichung, Taiwan (THUMB).
Species #GenBank Voucher Locality Source
Myotis albescens JX130463 TTU 85088 Pastaza, Ecuador Larsen et al. (2012a)
Myotis albescens JX130522 TTU 85091 Pastaza, Ecuador Larsen et al. (2012a)
Myotis albescens AF376839 FMNH 162543 Tarija, Bolivia Ruedi & Mayer (2001)
Myotis albescens JX130503 TTU 99124 Boquerón, Paraguay Larsen et al. (2012a)
Myotis albescens JX130504 TTU 99818 Ñeembucú, Paraguay Larsen et al. (2012a)
Myotis arescens OP270161 - Araucanía, Chile Present study
Myotis arescens OP270162 - Araucanía, Chile Present study
Myotis arescens OP270166 - Coquimbo, Chile Present study
Myotis arescens AM261888 - Santiago, Chile Stadelmann et al. (2007)
Myotis attenboroughi JN020573 UNSM-ZM
29470
St. George Parish, Tobago Larsen et al. (2012b)
Myotis attenboroughi JN020574 UNSM-ZM
29483
St. George Parish, Tobago Larsen et al. (2012b)
Myotis armiensis JX130435 TTU 39146 Chiriquí, Panamá Larsen et al. (2012a)
Myotis armiensis JX130514 TTU 85060 Tungurahua, Ecuador Larsen et al. (2012a)
Myotis armiensis MW025269 QCAZ 17245 Napo, Ecuador Carrión & Cook (2020)
Myotis atacamensis OP270158 - Arica, Chile Present study
Myotis atacamensis OP270159 - Arica, Chile Present study
Myotis atacamensis OP270160 - Arica, Chile Present study
Myotis bakeri AM261882 MVZ 168933 Olmos, Peru Stadelmann et al. (2007)
Myotis chiloensis OP270163 - Los Lagos, Chile Present study
Myotis chiloensis OP270164 - Los Lagos, Chile Present study
Myotis chiloensis OP270165 - Los Lagos, Chile Present study
Myotis clydejonesi JX130520 TTU 109227 Sipaliwini, Suriname Larsen et al. (2012a)
Myotis dinellii JX130475 TTU 66489 Córdoba, Argentina Larsen et al. (2012a)
Myotis dominicensis JN020555 TTU 31507 St. Joseph’s Parish, Dominica Larsen et al. (2012b)
Myotis dominicensis JN020556 TTU 31508 St. Joseph’s Parish, Dominica Larsen et al. (2012b)
Myotis dominicensis AF376848 - St. Joseph’s Parish, Dominica Ruedi & Mayer (2001)
Myotis elegans JX130479 TTU 84380 Atlantida, Honduras Larsen et al. (2012a)
Myotis elegans JX130480 TTU 84138 Atlantida, Honduras Larsen et al. (2012a)
Myotis keaysi JX130516 QCAZ 11380 Chimborazo, Ecuador Larsen et al. (2012a)
Myotis keaysi JX130517 QCAZ 11383 Chimborazo, Ecuador Larsen et al. (2012a)
Myotis larensis JN020569 TTU 48161 Guárico, Venezuela Larsen et al. (2012a)
Myotis larensis JX130529 TTU 48162 Guárico, Venezuela Larsen et al. (2012a)
Myotis larensis JX130535 CM 78645 Guárico, Venezuela Larsen et al. (2012a)
Myotis lavali AF376864 MVZ 185681 Paraíba, Brazil Ruedi & Mayer (2001)
Myotis levis AF376853 FMNH 141600 São Paulo, Brazil Ruedi & Mayer (2001)
Myotis martiniquensis AM262332 - Martinique Island Stadelmann et al. (2007)
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APPENDIX 1. (Continued)
Species #GenBank Voucher Locality Source
Myotis martiniquensis JN020557 MNHN 2005-895 GranďRivière, Martinique Larsen et al. (2012b)
Myotis martiniquensis JN020558 MNHN 2005-896 Le Morne Rouge, Martinique Larsen et al. (2012b)
Myotis midastactus MW323450 USNM 584502 Santa Cruz, Bolivia Novaes et al. (2021b)
Myotis moratellii JX130572 QCAZ 9179 El Oro, Ecuador Larsen et al. (2012a)
Myotis moratellii MZ345120 USNM 513482 Los Ríos, Ecuador Novaes et al. (2021b)
Myotis nesopolus JN020575 - Bonaire, Netherlands Antilles Larsen et al. (2012b)
Myotis nesopolus JN020576 - Bonaire, Netherlands Antilles Larsen et al. (2012b)
Myotis nesopolus JN020577 - Bonaire, Netherlands Antilles Larsen et al. (2012b)
Myotis nigricans JX130455 TTU 95992 San Pedro, Paraguay Larsen et al. (2012a)
Myotis nigricans JX130496 TTU 99743 Presidente Hayes, Paraguay Larsen et al. (2012a)
Myotis nigricans JX130498 TTU 99046 Alto Paraguai, Paraguay Larsen et al. (2012a)
Myotis nyctor JN020563 TTU 109225 St. Thomas Parish, Barbados Larsen et al. (2012a)
Myotis nyctor JN020564 TTU 109226 St. Thomas Parish, Barbados Larsen et al. (2012b)
Myotis nyctor JN020565 TTU 109229 St. Thomas Parish, Barbados Larsen et al. (2012b)
Myotis oxyotus AF376865 FMNH 129208 Lima, Peru Ruedi & Mayer (2001)
Myotis pilosatibialis JX130526 TTU 35360 San Luis Potosí, Mexico Larsen et al. (2012a)
Myotis pilosatibialis JX130518 TTU 35631 San Luis Potosí, Mexico Larsen et al. (2012a)
Myotis pilosatibialis JX130519 TTU 60981 Santa Ana, El Salvador Larsen et al. (2012a)
Myotis riparius JX130492 TTU 102883 Esmeraldas, Ecuador Larsen et al. (2012a)
Myotis riparius JX130473 CM 68443 Para, Suriname Larsen et al. (2012a)
Myotis riparius JX130474 CM 78659 Bolívar, Venezuela Larsen et al. (2012a)
Myotis riparius JX130485 TTU 99645 Paraguarí, Paraguay Larsen et al. (2012a)
Myotis ruber AF376867 MVZ 185999 São Paulo, Brazil Ruedi & Mayer (2001)
Myotis simus JX130481 TTU 46348 Huánuco, Peru Larsen et al. (2012a)
Outgroups
Myotis emarginatus MK799667 FMNH 178892 Ajlun, Jordan Patterson et al. (2019)
Submyotodon latirostris KP187906 THUMB 30036 Heping, Taiwan Ruedi et al. (2015)
Kerivoula papillosa MG194454 FMNH 205343 Luzon I, Philippine Island Sedlock et al. (2020)
APPENDIX 2. Specimens examined and localities of occurrence for Myotis species deposited in the following biological
collections: Muséum d’Histoire Naturelle (MHNG, Geneva, Switzerland), American Museum of Natural History
(AMNH, New York, USA), Carnegie Museum of Natural History (CM, Pittsburgh, USA), Field Museum of Natural
History (FMNH, Chicago, USA), Museum of Natural History of the Kansas University (KU, Lawrence, USA), Museum
of Natural Science, Louisiana State University (LSUMZ, Baton Rouge, USA), Museum of Vertebrate Zoology (MVZ,
Berkeley, USA), Sam Noble Oklahoma Museum of Natural History (OMNH, Norman, USA), and Smithsonian National
Museum of Natural History (USNM, Washington D.C., USA).
Myotis albescens (N = 109): Argentina: Tucumán, La Cocha, Dique San Ignacio (OMNH 18877); Tucumán, Leales, Dique San
Ignacio (OMNH 18878); Santiago del Estero, Pellegrini, Santo Domingo (OMNH 23772, 23773, 23774). Peru: Ayacucho,
Río Apurimac, Hacienda Luisiana (LSUMZ 16621, 16622); Ayacucho, San José, Río Santa Rosa (LSUMZ 16623–16625);
Cusco, Quispicanchi (FMNH 68471, 68473–68478); Huánuco, Leonicio Prado, 1 km S of Tingo Maria (CM 98854);
Huánuco, Río Huallaga, ca. 4 km NE of Tingo Maria (LSUMZ 14265); Loreto, Río Curaray (AMNH 71643); Loreto,
Maynas, Puerto Indiana, Amazon River (AMNH 73235, 73237, 73239, 73242); Loreto, Maynas, Orosa, Amazon River
(AMNH 74018, 74019, 74021); Loreto, San Jacinto (KU 158160); Madre de Dios, Manú, Maskoitania, 13.4 km NW
Atalaya, left bank of Rio Alto Madre de Dios (FMNH 174919, 174921); Madre de Dios, Manú, Quebrada Aguas Calientes,
left bank, Rio Alto Madre de Dios, 2.75 km E of Shintuya (FMNH 170275); Madre de Dios, Mouth of Rio La Torre, south
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bank of Rio Tambopata (LSUMZ 24562); Madre de Dios, Pakitza (USNM 564391, 564392, 566560–566563); Pasco,
Oxapampa (AMNH 230746–230748, 230750–230757); Pasco, San Juan (USNM 364442–364480); Pasco, unknown
localities (AMNH 213428, 213430); Ucayali, Balta, Río Curanja (LSUMZ 12272, 12274–12279); Ucayali, Coronel
Portillo, Yarinacocha (LSUMZ 12253, FMNH 62178–62188).
Myotis arescens (N = 31): Chile: Valparaíso Region, Quillota, Hacienda Limache (FMNH 24075, 24076, 24077, 24078, 24079,
24396 [holotype]); Valparaíso Region, Petorca, Papudo, Aconcagua (FMNH 23635, 23636); Valparaíso Region, Zapallar,
Coastal Town (USNM 391784); Coquimbo Region, Elqui, Paiguano (FMNH 23610, 23611, 23612, 23583, 23584, 23585,
23587, 23591, 23593, 23594, 23596, 23597, 23599, 23600, 23602, 23604); Maule Region, Curicó (MHNG 1883-64, 1883-
65, 1883-66, 1883-67, 1883-68, 1883-69).
Myotis atacamensis (N = 12): Chile: Tarapacá Region, Minimini (USMN 391786); Tarapacá Region, Iquique, Los Canchones
(FMNH 23618, MHNG 1748-43, 1748-44, 1748-45, 1748-46). Peru: Arequipa, Chucarapi, Tambo Valley (MVZ 116638;
FMNH 50783, 51063); Arequipa, Arequipa, Patasagua, 3 km W of Tiabaya (FMNH 49790, 49791, 49792).
Myotis bakeri (N = 4): Peru: Lambayeque, 12 km N of Olmos (LSUMZ 21306, 21307); Lima, 7 km SE of Chilca (MVZ
137906, 137907 [holotype]).
Myotis chiloensis (N = 47): Argentina: Chubut (MVZ 150842, 150847–150851, 150853–150858); Neuquen (MVZ 150862–
150869, 150883, 150884, 150892, 150894–150898, 162181–162183); Rio Negro (MVZ 150902–150907, 150917, 150918,
150927, 150929, 152150, 152153, 154461). Chile: Valdivia, Rinihue (FMNH 23613, 23614); Chiloe, Chiloe Island (FMNH
24029 [neotype]).
Myotis oxyotus (N = 19): Argentina: Tucumán, Tafí Viejo, 5 km SW Siambón (OMNH 36218). Peru: Ancash, 31 km E
of Pariacoto by road (LSUMZ 22131); Cajamarca, Celendin, Hacienda Limón (FMNH 19969); Cusco, Iquente (USNM
195146); Cusco, Marcapata (FMNH 66375, 66376); Cusco, Santa Ana (USNM 194452, 194453, 195141, 195147, 195149);
Cusco, 6 miles N of Paucartambo (MVZ 116008); Huancavelica, Rumicruz (AMNH 60598); Huánuco, Ambo (FMNH
24864–24866); Junín, Rio Palca, 15 Km W of San Ramon (USNM 507204); Lima, Bujama Baja, 95 km south of Lima by
road (AMNH 216118); Lima, Huaros, Bosque de Zarate, San Bartolomé (FMNH 129208).
... rupestris) are also distributed in Chile. Finally, Novaes et al. (2022) recently reviewed the species of Myotis present in Chile, concluding that M. arescens is distinct from M. chiloensis. These recent studies bring to 167 the species of living mammals recorded in Chile. ...
... Along with Oligoryzomys longicaudatus, O. flavescens s. l. is the second species of the genus Oligoryzomys known for Chile (refer to D'Elía et al. 2020, in which the nominal forms O. magellanicus and O. yatesi are included as synonyms of O. longicaudatus), and the mammal species number 168 with records in the country (D'Elía et al. 2020;Ojeda et al. 2021;Novaes et al. 2022;Rodríguez-San Pedro et al. 2022). This new record, together with those presented in recent years and the new species that have recently been proposed based on specimens collected in Chile (see a synthesis in D'Elía et al. 2020; see the proposal of candidate species of Octodon in Cadenillas and D'Elía 2021b), indicate that the mammalian fauna of Chile is still not completely characterized. ...
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The Chilean mammal fauna is one of the best known of South America. In spite of this, in the last decade several new species have been described based on specimens collected in the country, while other species previously known elsewhere have been recorded for the rst time in Chile. Here we keep on this trend by recording for the rst time for Chile a species of long-tailed mouse of the genus Oligoryzomys. This mention is based on genetic (cytochrome b gene sequences) and morphological data gathered from several specimens collected at four loca- lities of Quebrada de Camarones, Región de Arica y Parinacota in northern Chile. At one of these localities a specimen was live-trapped; while in the other three localities several osteological remains were recovered from owl pellets. The morphologic and genetic information robustly indicate that the revised specimens belong to the genus Oligoryzomys. The phylogenetic analyses show that the trapped specimens belong to O. flavescens s. l. However, it remains unsolved to which of two main lineages of O. flavescens s. l., O. flavescens s. s. or O. occidentalis, belongs the specimens from Camarones. Here we increase the known species richness of Chilean living mammals by showing that northernmost Chile is inhabited by O. flavescens s. l. The possibility that the specimens from Camarones represent an undescribed species cannot be ruled out. These new records indicate, once again, that much remains to be learn about basic aspects of the Chilean mammals, including which species form the local assemblages.
... Hasta 2022, se han reportado 150 especies de mamíferos en Chile, de las cuales 16 son murciélagos correspondientes a nueve géneros y cuatro familias (Furipteridae, Phyllostomidae, Vespertilionidae y Molossidae) , Ossa et al. 2018, Novaes et al. 2022. ...
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El área contiene una muestra representativa de la comunidad de murciélagos del suroeste de Perú y norte de Chile. Incluye 18 de las 20 especies registradas para esta región. IUCN: En Peligro (Tomopeas ravus y Myotis atacamensis), Vulnerable (Amorphochilus schnablii), Casi Amenazada (Platalina genovensium) y Datos Deficientes (Promops davisoni), las cuatro primeras Amenazadas o Casi Amenazadas en Perú. Asimismo, Eumops chiribaya y Lasiurus arequipae cumplirían con IUCN para ser consideradas como amenazadas. Tres de las especies reportadas son endémicas de Perú (T. ravus, E. chiribaya y L. arequipae) y ocho son endémicas del Desierto Pacífico de Perú y norte Chile (las enlistadas anteriormente y Mormopterus kalinowskii). Existen colonias reproductivas de especies amenazadas, refugios permanentes. El área sufre de vandalismo y disminución de la calidad del hábitat.
... Hasta 2022, se han reportado 150 especies de mamíferos en Chile, de las cuales 16 son murciélagos correspondientes a nueve géneros y cuatro familias (Furipteridae, Phyllostomidae, Vespertilionidae y Molossidae) , Ossa et al. 2018, Novaes et al. 2022. ...
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El área posee una alta diversidad de murciélagos (cinco especies) en relación a la riqueza de la provincia de San Juan, donde solo se registraron seis especies. Se reportan dos especies migratorias: Tadarida brasiliensis (Molossidae), Lasiurus blossevillii (Vespertilionidae), además de colonias maternales de Myotis dinellii y Tadarida brasiliensis. En el interior del área existen comunidades humanas con intensas actividades productivas como la ganadería, agricultura y turismo. Los murciélagos registrados en el área son insectívoros por lo que ejercen una importante actividad reguladora de las poblaciones de insectos perjudiciales para los bosques nativos, la agricultura y vectores de enfermedades.
... Hasta 2022, se han reportado 150 especies de mamíferos en Chile, de las cuales 16 son murciélagos correspondientes a nueve géneros y cuatro familias (Furipteridae, Phyllostomidae, Vespertilionidae y Molossidae) , Ossa et al. 2018, Novaes et al. 2022. ...
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Region description
... Hasta 2022, se han reportado 150 especies de mamíferos en Chile, de las cuales 16 son murciélagos correspondientes a nueve géneros y cuatro familias (Furipteridae, Phyllostomidae, Vespertilionidae y Molossidae) , Ossa et al. 2018, Novaes et al. 2022. ...
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Este libro es el corolario de muchos años de trabajo y dedicación de los miembros de la RELCOM (Red Latinoamericana y del Caribe para la Conservación de los Murciélagos), cuyo objetivo es poner a disposición de la comunidad los resultados de una actividad de carácter regional, que se viene realizando de manera ininterrumpida desde 2011, cuando el primer AICOM fue reconocido. En 2009, la RELCOM elaboró una “Estrategia para la conservación de los murciélagos en Latinoamérica y el Caribe”, donde se identificaron las amenazas que sufren los murciélagos de la región. Esto despertó la necesidad de crear una figura como grupo para proteger a los murciélagos a través de una propuesta regional. Y es así que surgen las Áreas y Sitios de Importancia para la Conservación de los Murciélagos (AICOMs-SICOMs), inspiradas en las AICAs (Áreas de Importancia para la Conservación de las Aves). El reconocimiento de AICOMs y SICOMs surge como una herramienta para que, de algún modo, pueda ser utilizada por los diferentes países que conforman la red, para orientar los planes de conservación en localidades donde especies y poblaciones de murciélagos se encuentren amenazadas. Si bien no es un instrumento legal, sienta bases para el desarrollo de políticas nacionales y regionales que avancen en ese sentido.
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