Integrative taxonomy resolves three new cryptic species of small southern African horseshoe bats (Rhinolophus)

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© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28 1
Zoological Journal of the Linnean Society, 2018, XX, 1–28. With 14 figures.
Integrative taxonomy resolves three new cryptic species
of small southern African horseshoe bats (Rhinolophus)
PETER J. TAYLOR1,2*, ANGUS MACDONALD2, STEVEN M. GOODMAN3,4,
TERESA KEARNEY5,6, FENTON P. D. COTTERILL7, SAM STOFFBERG8,
ARA MONADJEM9,10, M. CORRIE SCHOEMAN2, JENNIFER GUYTON11,
PIOTR NASKRECKI12 AND LEIGH R. RICHARDS13
1SARChI Chair on Biodiversity Value & Change and Core Team Member of the Centre for Invasion
Biology, School of Mathematical & Natural Sciences, University of Venda, Private Bag X5050,
Thohoyandou 0950, South Africa
2School of Life Sciences, University of KwaZulu-Natal, Biological Sciences Building, South Ring Road,
Westville Campus, Durban, KwaZulu-Natal 3630, South Africa
3Field Museum of Natural History, 1400 South Lake Shore Drive, Chicago, IL 60605, USA
4Association Vahatra, BP 3972, Antananarivo 101, Madagascar
5Ditsong National Museum of Natural History, PO Box 413, Pretoria 0001, South Africa
6School of Animal, Plant and Environmental Sciences, University of the Witwatersrand, Private Bag 3,
Wits 2050, South Africa
7Geoecodynamics Research Hub, c/o Department of Botany and Zoology, University of Stellenbosch,
Private Bag X1 Matieland, 7602, Stellenbosch, South Africa
8Department of Botany and Zoology, University of Stellenbosch, Private Bag X1 Matieland, 7602,
Stellenbosch, South Africa
9All Out Africa Research Unit, Department of Biological Sciences, University of Swaziland, Private Bag
4, Kwaluseni, Swaziland
10Mammal Research Institute, Department of Zoology & Entomology, University of Pretoria, Private Bag
20, Hatfield 0028, Pretoria, South Africa
11Department of Ecology and Evolutionary Biology, Princeton University, 106A Guyot Hall, Princeton,
NJ 08544, USA
12Museum of Comparative Zoology, Harvard University, 26 Oxford Street, Cambridge, MA 02138, USA
13Durban Natural Science Museum, P. O. Box 4085, Durban, 4001, South Africa
Received 13 December 2017; revised 4 March 2018; accepted for publication 7 March 2018
Examination of historical and recent collections of small Rhinolophus bats revealed cryptic taxonomic diversity
within southern African populations previously referred to as R. swinnyi Gough, 1908 and R. landeri Martin, 1832.
Specimens from Mozambique morphologically referable to R. swinnyi were phylogenetically unrelated to topotypic
R. swinnyi from the Eastern Cape Province of South Africa based on cytochrome b sequences and showed distinctive
echolocation, baculum and noseleaf characters. Due to their genetic similarity to a previously reported molecular
operational taxonomic unit (OTU) from north-eastern South Africa, Zimbabwe and Zambia, we recognize the avail-
able synonym (R. rhodesiae Roberts, 1946) to denote this distinct evolutionary species. This new taxon is genetically
identical to R. simulator K. Andersen, 1904 based on mtDNA and nuclear DNA sequences but can easily be distin-
guished on morphological and acoustic grounds. We attribute this genetic similarity to historical introgression, a
frequently documented phenomenon in bats. An additional genetically distinct and diminutive taxon in the swinnyi
s.l. group (named herein, R. gorongosae sp. nov.) is described from Gorongosa National Park, central Mozambique.
*Corresponding author. Email: peter.taylor.univen@gmail.com
[Version of Record, published online 24 April 2018;
http://zoobank.org/urn:lsid:zoobank.
org:pub:65FFC8DB-4738-49FF-99F5-FC8532CC9795]
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2 P. J. TAYLOR ET AL.
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Specimens from Mozambique referable based on morphology to R. landeri were distinct from topotypic landeri from
West Africa based on mtDNA sequences, and acoustic, noseleaf and baculum characters. This Mozambique popula-
tion is assigned to the available synonym R. lobatus Peters, 1952.
ADDITIONAL KEYWORDS: Africa – Baculum – bioacoustics – cranial – DNA – dental – horseshoe bats –
morphology – Rhinolophus.
INTRODUCTION
Recent decades have seen an increase in the number
of new species descriptions of mammals, including
bats, in the region comprising Africa and the Western
Indian Ocean Islands. In the 20 years preceding 2008,
22 new species of Afro-Malagasy bats were described
(Hoffmann et al., 2009). This pace of discovery has
intensified, with a further 35 new Afro-Malagasy spe-
cies described between 2009 and 2017, 20 of them from
the African mainland (African Chiroptera Report,
2015; references listed below). These recent discover-
ies span several families, including Hipposideridae
(Goodman et al., 2016), Miniopteridae (e.g. Goodman
et al., 2008, 2011; Monadjem et al., 2013b), Molossidae
(e.g. Goodman et al., 2010; Ralph et al., 2015),
Pteropodidae (Nesi et al., 2013; Hassanin et al., 2015),
Rhinolophidae (Benda & Vallo, 2012; Taylor et al., 2012;
Kerbis Peterhans et al., 2013) and Vespertilionidae
(e.g. Monadjem et al., 2013b; Brooks & Bickham, 2014;
Decher et al., 2015; Goodman et al., 2017; Hassanin
et al., 2017).
One reason for this burst of discoveries has been
the use of multiple lines of evidence in resolving the
species diversity of regional bat faunas, referred to
as the integrative approach (Schlick-Steiner et al.,
2010). Traditional morphological evidence alone, even
established skull and dental characters, is in many
cases unable to discriminate between cryptic species,
although baculum (os penis) characters do distinguish
many cryptic species of horseshoe bats (Cotterill, 2002;
Taylor et al., 2012; Monadjem et al., 2013b). However,
the inclusion of molecular data has radically altered
the ability to distinguish morphologically similar spe-
cies. With reference to bats, the use of morphological
(including baculum), genetic and acoustic features
together provides the integrative approach to advance
taxonomy.
Molecular evidence contributes increasingly to
resolve challenging taxonomic problems in bats, and
particularly African horseshoe bats. Recent molecular
studies have uncovered several new cryptic species
and undescribed lineages of African horseshoe bats
of the genus Rhinolophus Lacépède, 1799 (Stoffberg,
Schoeman & Matthee, 2012; Taylor et al., 2012; Jacobs
et al., 2013; Dool et al., 2016). Nevertheless, at least
four species of Rhinolophus have been described based
only on morphological differences (Kock, Csorba &
Howell, 2000; Cotterill, 2002; Fahr et al., 2002; Kerbis
Peterhans et al., 2013). In combination with sonar, cra-
nial, dental and bacula characters, Taylor et al. (2012)
employed molecular techniques to diagnose five genet-
ically distinct lineages comprising the R. hildebrandtii
complex, and so characterize four distinct new species
in addition to R. hildebrandtii sensu stricto.
Echolocation calls of Rhinolophus spp. have a con-
stant frequency (CF) component (Monadjem et al.,
2010), which has also been used to distinguish differ-
ent taxa. For example, the morphologically similar R.
simulator and R. swinnyi have distinctly dissimilar
peak frequencies (Monadjem et al., 2010), but differ-
ences in echolocation frequency are not infallible diag-
nostic characters. For example, variation in constant
frequency within species and species-complexes in
this genus has been attributed inter alia to specia-
tion (Taylor et al., 2012; Jacobs et al., 2013), genetic
drift (Odendaal & Jacobs, 2011), allometric effects
(Stoffberg, Jacobs & Matthee, 2011) and environ-
mental variables such as relative humidity (Mutumi,
Jacobs & Winker, 2016).
Using several mitochondrial and nuclear DNA
markers, Dool et al. (2016) showed that both R. swin-
nyi Gough, 1908 and R. landeri Martin, 1838 were par-
aphyletic. Topotypic R. swinnyi from the Eastern Cape
Province of South Africa (type locality: Pirie Forest),
was affiliated with the R. capensis group, while two
specimens of R. cf. swinnyi (which they called cf. simu-
lator) from Zambia and northern South Africa were
affiliated more closely with nominate R. simulator. In
the case of R. landeri, two specimens from Zimbabwe
formed a sister lineage with R. alcyone, whilst two
West African specimens of R. landeri (type locality:
Equatorial Guinea) from Mali formed a distinct clade,
which was sister to the alcyone and landeri complexes.
Informed by newly collected series of R. cf. swin-
nyi and R. cf. landeri from Mozambique and South
Africa, the aim of this study was to review the sta-
tus of above mentioned cryptic species, augmented
by broader sampling, with the combined evidence of
craniometric, morphological, acoustic, bacular, dental
and molecular data. Here we diagnose the different
taxa comprising the landeri, simulator and swinnyi
species complexes, and we resurrect two distinct spe-
cies from synonymy, and name a third new species
using the integrative taxonomic approach. Of the two
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© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 3
species-groups, R. landeri s.l. has a wide but sparse
distribution encompassing much of western, eastern
and south-central Africa. On the other hand, R. swin-
nyi s.l. is more restricted in range occurring in the
eastern parts of the subcontinent from the southern
reaches of the Democratic Republic of Congo to as far
south as the Eastern Cape Province of South Africa
(Monadjem et al., 2010; African Chiroptera Report,
2015). Given the close genetic relationship between
the new R. cf. swinnyi taxon and R. simulator, and
as the two species are easily confused in the field, we
included R. simulator s.l. K. Andersen, 1904, in this
study, as well as topotypic samples of R. swinnyi and
R. landeri.
MATERIAL AND METHODS
Molecular analysis
PCR amplification and sequencing
DNA was extracted from each sample using a
Zymogen DNA extraction kit (www.zymoresearch.
com) according to the manufacturer’s instructions.
Specimen DNA was amplified via Polymerase
Chain Reaction (PCR) using primers designed to
amplify the cytochrome oxidase B gene. Mammalian
cytochrome b (Cytb) primers were used, C3FF5
(5’ACCAATGMMATGAAAAATCATCGTT’3) and
C3FF6 (5’ TCTYCATTTYWGGTTTACAAGAC’3),
to amplify and sequence Cytb regions and was
derived from Irwin, Kocher & Wilson (1991). The
PCR reactions for both markers contained 9.5 µl
H2O, 12.5 µl 2x EconoTaq buffer, 1 µl forward and
reverse Cytb primer (10 µM). The PCR thermal
cycle was [95 °C for 5 min], 30 × [(94 °C for 30 s),
(58 °C for 45 s), (72 °C for 45 s)] and [72 °C for
10 min], [hold at 15°C]. PCR products for each
sample were checked for quality and quantity
using a Nanodrop 3000 spectrophotometer and
subsequently run on a 1% agarose gel, using 1 µl
of loading dye and 5 µl of PCR product per sample.
Samples were then Sanger sequenced using an ABI
3730 capillary sequencer at Inqaba Biotechnical
Industries (Pretoria, South Africa).
Sequence alignment and editing
Between 700 and 900 base pairs (bp) of the Cytb
locus of the mtDNA cytochrome oxidase gene were
amplified and sequenced for the Rhinolophus spp.
samples. BioEdit v.7.0.9 (Hall, 1999) was used to
align and edit the sequence electropherograms. The
sequences were aligned using ClustalW (Larkin
et al. 2007), and the resultant alignment checked
and realigned by eye. Any remaining ambiguities
were compared to their respective complement
sequences, and corrected using the toggle translate
function. The final alignments were trimmed to a
length of 500 bp for the Cytb marker. Sequences
were deposited on Genbank with the following
accession numbers: MG 980648–980682.
Phylogenetic and haplotype analysis
jModeltest (Guindon & Gascuel, 2003; Darriba et al.,
2012) was used to search for the best fit model of
evolution that fitted the genetic marker dataset.
The GTR model of Tavare (1986) was chosen to con-
struct the likelihood (ML) trees in MEGA v.7 (Kumar,
Stecher & Tamura, 2016). MrBayes 3.1.2 (Ronquist
& Huelsenbeck, 2003) was used to estimate a phylo-
genetic tree and to estimate the posterior probabil-
ity of nodes using the same model of evolution with
appropriate priors. The Cytb Bayesian trees were
rooted using outgroups from other Rhinolophus spp.
and additional sequences from relevant reference col-
lections accessioned on GenBank were used as sup-
plementary data (Supporting Information, Table S1).
After re-alignment, including GenBank accessioned
sequences, likelihood and parsimony analyses were
performed. The likelihood and parsimony trees boot-
strapped for 100 iterations, and genetic distances
(corrected and uncorrected ‘p’) calculated using pair-
wise comparisons of taxa, were completed in MEGA
v.7 (Kumar et al., 2016). The Bayesian trees estimated
with specimen sequences were created using four
Markov chains of 1 000 000 generations each, sampled
every ten generations. After diagnostics indicated
that all independent runs had converged, a consen-
sus tree was constructed. The first 10 000 trees were
discarded as burn in, and the rest of the genealogies
used to construct a 50% majority-rule consensus tree.
Minimum-spanning was used to construct a haplo-
type network using the Cytb sequence data with gaps
coded as missing and with each node supplied with
the number of specimens assigned to haplotypes per
region by Popart (http://popart.otago.ac.nz).
Morphological analysis
Specimens
We examined recent collections of skins and skulls of
three small rhinolophid species (Rhinolophus swinnyi
s.l., R. simulator s.l. and R. landeri s.l.) from across
the continental range of these species, with special
emphasis on southern African subregion (Fig. 1).
The associated voucher specimens are held in the
Durban Natural Science Museum, Durban (DM), Field
Museum of Natural History, Chicago (FMNH) and
Ditsong National Museum of Natural History, Pretoria
(formerly Transvaal Museum; TMSA) (Supporting
Information, Table S1). We also included measure-
ments from relevant type specimens and other rep-
resentative series in the collections of The Natural
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4 P. J. TAYLOR ET AL.
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History Museum, London [formerly British Museum
(Natural History); (BMNH)], the Muséum National
d’Histoire Naturelle, Paris (MNHN), the Zoologisches
Museum, Berlin (ZMB), the Natural History Museum
of Zimbabwe, Bulawayo (NMZB) and the Ditsong
National Museum of Natural History (Supporting
Information, Table S1). We only selected adults for
further measurement and analysis, which we defined
based on the degree of tooth wear and extent of ossi-
fication of finger bone epiphyses (Taylor et al., 2012).
Morphometric analyses
We used two morphometric approaches: analysis of
traditional linear cranial measurements, as well as
landmarks placed on lateral images of specimen cra-
nia. We analysed variation in a sample of 124 intact
skulls using ten craniometric variables (see definitions
below) and standard external measurements obtained
from the field notes of AM, JAG, LRR, MCS, PJT and
SMG, although a few measurements were obtained
from museum specimen labels. Because the premax-
illa was often absent in museum skulls, numerous
specimens could not be measured for two standard
skull variables: greatest length of skull measured
dorsally from occiput to anterior point of skull (GSL)
and condyloincisive length from occipital condyles to
front of incisors (CIL). For this reason, herein, we did
not include these two variables in statistical analyses,
but certain comparisons were made using these two
measurements. The following ten cranial measure-
ments were taken to the nearest 0.01 mm using either
Mitutoyo or Tesa digital callipers with an accuracy of
0.01 mm: (1) condylocanine length from occipital con-
dyles to front of canines (CCL); (2) zygomatic width,
the greatest distance across the zygoma (ZW); (3) mas-
toid width, the greatest distance across the lateral pro-
jections of the mastoid processes (MW); (4) width of
maxilla between outer edges of M3 (M3M3); (5) brain-
case width measured at dorsal surface of posterior
root of zygomatic arches (BCW); (6) least interorbital
width (IOW); (7) upper toothrow length from ante-
rior surface of C to posterior surface of M3 (C1M3);
(8) greatest width across anterior lateral nasal infla-
tions (NW); (9) length from occipital condyles to front
Figure 1. Map showing localities of R. swinnyi s.l. (circles), R. landeri s.l. (triangles) and R. simulator (crosses) sampled
by this study. The locations of type specimens included in the study are denoted with arrows. See Supporting Information,
Table S1 for list of localities, specimens and Genbank sequences included in the study.
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 5
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
of nasal inflations (NL); and (10) height of nasal
inflation directly above the anterior cingulum of M2
(NH). To include data from type specimens and others
for which only five cranial measures (CCL, ZW, MW,
M3M3, C1M3) were available, principal component
analysis (PCA) was repeated for these five variables
resulting in a final sample of 160 skulls (Supporting
Information, Table S1).
Because both multivariate and univariate analy-
ses failed to detect significant sexual dimorphism in
either cranial or external measurements of the three
taxa; adult males and females were pooled. To visual-
ize intraspecific and interspecific variation in multi-
dimensional space of linear measurements, PCA was
carried out on log-transformed cranial variables using
the programme PAST (Hammer, Harper & Ryan,
2001).
Geometric morphometric analysis
We used this approach to explore intra- and interspe-
cific differences in cranial size and shape from digital
images of the left lateral profile of 118 Rhinolophus
spp. skulls (Supporting Information, Table S1). Digital
images were captured using a Canon Powershot A650
IS digital camera mounted on a tripod (×6 optical zoom,
5 megapixel resolution, 25 mm focal length, macro func-
tion). Eighteen homologous landmarks were captured
in two dimensions (2D) from cranial images using tps-
Dig, v.2.17 (Rohlf, 2013). Landmarks were described as
follows: (1) junction of maxillae and anterior margin
of C1; (2) junction of maxillae and anterior margin of
PM2; (3) posterior margin of maxillae/end of toothrow;
(4) junction of maxillae and jugal process; (5) junction
of squamosal and auditory meatus; (6) highest point
of external bullae; (7) junction of bullae and occipital
condyle; (8) longest point of condyle; (9) anterior mar-
gin of foramen magnum; (10) posterior margin of fora-
men magnum; (11) junction of sagittal and lambdoidal
crests; (12) sharpest point of parietals; (13) junction of
parietal and temporal; (14) highest point of braincase/
temporal region; (15) most posterior point of the inter-
orbital constriction; (16) base of nasal inflation; (17)
highest point of nasal inflation; and (18) junction of
maxillae and nasal inflation. A Generalized Procrustes
Analysis (GPA) of the landmark dataset and PCA of
the total shape matrix were conducted using MorphoJ
software (Klingenberg, 2011). Taxon-specific thin plate
splines illustrating global and localized cranial shape
characteristics, exaggerated 3×, were generated using
tpsREGR v.1.38 (Rohlf, 2011).
Morphological comparisons
In additional to morphometric analysis of continuous
characters, we scored the following qualitative charac-
ters: the position (external or within toothrow) and rel-
ative size of the small anterior upper premolar, and the
presence or absence of apical tufts (distinct cluster of
stiff orange hairs located in the armpit of certain male
R. landeri individuals). We provided detailed descrip-
tions of the noseleaf morphology of taxa delineated
by genetic and morphometric analyses. The terminol-
ogy of the connecting process structure and protuber-
ance and the shape of the lancet are based on Csorba,
Ujhelyip & Thomas (2003); Happold & Cotterill (2013)
and Cotterill (2013).
Baculum morphology
Thirty-three bacula were prepared from alcohol-pre-
served adult male specimens following the methods of
Hill & Harrison (1987) and Kearney et al. (2002), with
slight modification of the procedures to clear penile
tissue (specimens listed in Supporting Information,
Table S1). The glans penis was removed and hydrated
in distilled water for 24 h. The penile tissue was then
macerated for 36–48 h in a 5% KOH solution with
Alizarin red stain. The stained baculum was cleared of
the macerated tissue by hand using fine forceps. Each
baculum was placed on a stage micrometer and was
photographed in the dorsal, lateral and ventral views
using a dissecting microscope and an ocular mounted
MU500 5 M pixel digital USB microscope camera
(AmScope, USA). Eight measurements were recorded
from digital images using the ‘Length Measurement’
feature of the ToupView software program (ToupTek
Phototonics): (1) total length; (2) greatest base width
(dorsal view); (3) greatest base width (lateral view);
(4) widest region along the shaft; (5) narrowest region
along the shaft; (6) widest region of shaft tip; (7) great-
est length of the dorsal basal notch; and (8) base height.
In specimens without a distinct and/or expanded shaft
tip, the tip was regarded as occupying the uppermost
portion of the shaft (approximately 20% of the entire
shaft). Measurements were log-transformed and
subjected to PCA using IBM SPSS Statistics v. 21.0
(IBM Corp). As sample size was in several cases low,
a Kaiser–Meyer–Olkin (KMO) test was performed to
determine sampling adequacy for each variable and
the overall model (Kaiser, 1974). Line drawings of
selected bacula were made from digital images of the
respective views that were representative of the dif-
ferent taxa.
acoustic analysis
Acoustic recordings of most bats were obtained using
one of the following three time expansion bat detec-
tors: Pettersson D1000X, Pettersson D980 ultrasound
recorders (Pettersson Electronik AB, Uppsala, Sweden)
or Avisoft Ultrasound 116Hb bat detector (Avisoft
Bioacoustics, Berlin, Germany). Sampling rate was
set to 300 kHz, with 16-bit Analogue-to-Digital con-
verter. Recorded calls were analysed with BatSound
Pro software (Pettersson Electronik AB, Uppsala,
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6 P. J. TAYLOR ET AL.
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Sweden). The dominant harmonic from each call was
taken from the fast Fourier transform power spec-
trum. A Hanning window was used to eliminate effects
of background noise. Peak echolocation frequency was
measured from the maximum amplitude of the power
spectrum. Bats were recorded hand-held. The echolo-
cation calls of the two specimens of R. landeri from
Liberia were recorded using an Anabat SD2 detector
(Titley Electronics, Ballina, Australia), whilst the bats
were held, separately, in a small cloth bag. Calls were
analysed using ANALOOK software (Chris Corben,
v.4.8, http://www.hoarybat.com). The frequency of the
constant frequency (CF) component [= maximum fre-
quency F(max)] was measured.
RESULTS
Molecular analysis
Based on cytochrome b (Cytb) sequences (Figs 2, 3) ,
Rhinolophus cf. swinnyi collected in Mozambique
(Malashane Cave, Chihalatan Cave and Gorongosa
National Park) were phylogenetically distinct from
topotypical R. swinnyi from the Eastern Cape
Province, South Africa; the latter has R. capensis
as its sister species. Specimens assigned to cf. swin-
nyi from Malashane Cave and Chihalatan Cave are
genetically very close to Genbank sequences of cf.
swinnyi obtained from Zimbabwe, Zambia and the
Pafuri region of the extreme north of South Africa (see:
Dool et al., 2016), allowing us to assign this lineage to
the taxon R. rhodesiae Roberts, 1946 described from
Bezwe River in southern Zimbabwe, and previously
classified as R. s., rhodesiae (Ellerman, Morrison-
Scott & Hayman 1953). However, two cf. swinnyi from
Gorongosa National Park form a highly distinctive
group separated by uncorrected p-distances of 13.2%
and 7.2% from topotypic R. swinnyi and R. rhodesiae,
respectively (Table 1). Despite these two individuals
being relatively genetically distinct (p = 1.8%) from
each other (Figs 2, 3), we refer to them both as the new
species (described below), Rhinolophus gorongosae sp.
nov. The species is closely affiliated with topotypic R.
landeri from Liberia, West Africa, but still genetically
distant (15.0%; Table 1).
Rhinolophus cf. landeri from Malashane Cave,
Chihalatan Cave and Gorongosa National Park form
a homogeneous group that is phylogenetically dis-
tinct from West African R. landeri (Mount Nimba,
Liberia). These three localities in central and eastern
Mozambique fall in close geographic proximity to the
type locality of R. lobatus at Sena, Mozambique, on the
south bank of the Zambezi River (Peters, 1852) (Figs 1,
2; Supporting Information, Table S1). We therefore
refer this taxon to R. lobatus.
Enigmatically, individuals of R. rhodesiae are genet-
ically inseparable (Fig. 2; Table 1: p = 0.6%) from the
morphologically distinct species, R. simulator, a result
also obtained by Dool et al. (2016) informed by mito-
chondrial and nuclear evidence.
MorphoMetric variation
External morphology
External characters (Table 2) confirm Rhinolophus
swinnyi s.l. (including R. swinnyi s.s., R. gorongosae
sp. nov. and R. rhodesiae) to be smaller (total length
always <80 mm; 61–79 mm) than R. simulator (total
length 77–79 mm) and R. landeri s.l. (total length typi-
cally >80 mm; 79–87 mm), as also reflected in the PCA
of cranial variables (Fig. 4). Within R. swinnyi s.l., as
also clearly reflected in the cranial data (see follow-
ing paragraph), the genetically-distinct Gorongosa
National Park population (R. gorongosae sp.nov.) is
distinctly smaller in external measurements (e.g.
mean total length 68 mm) than both rhodesiae from
Malashane and Chihalatan Caves (mean total length
75 mm) and swinnyi s.s. from South Africa (mean
total length 73 mm). Specimens of R. lobatus from
Gorongosa National Park appear smaller externally
(total length 72–82 mm, mean 76 mm) than R. lobatus
from Malashane and Chihalatan Caves (total length
79–87 mm, mean 82 mm; Table 2) with some overlap.
While the same trend is present in tail and ear lengths,
the opposite is true for hind foot length, which is larger
in the Gorongosa National Park population than in
those from Malashane and Chihalatan Caves (Table 2).
Intraspecific craniometric variation
A PCA of ten log-transformed craniodental variables
revealed considerable overlap in size between males
and females in both R. swinnyi s.l. and R. landeri
s.l. (Supporting Information, Fig. S1). Furthermore,
means of ten craniodental variables showed no signifi-
cant sexual dimorphism in any external or cranial var-
iables for either species (data from t-tests, not shown).
For example, mean forearm length was 44.0 mm (42.8–
45.0 mm, N = 15) and 43.7 mm (42.0–45.0 mm, N = 10)
in female and male R. rhodesiae from Mozambique
(Malashane, Chihalatan Caves and Mount Inago),
respectively (t23 = 0.65, P > 0.05). In R. lobatus from
Mozambique (Malashane, Chihalatan Caves and
Muchena), mean forearm length was 46.6 mm (44.4–
49.0 mmm, N = 9) and 45.5 mm (43.4–46.0 mm,
N = 5) in females and males, respectively (t12 = 1.30,
P > 0.05). These values fall within the range reported
by Monadjem et al. (2010) for R. swinnyi s.l. but above
the mean (44.0 mm) and range of 42.0–47.8 mm
for R. landeri s.l. (Monadjem et al., 2010). In terms
of cranial size, mean CCL of R. rhodesiae females
was 15.4 mm (15.0–15.7 mm, N = 15) and for males
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 7
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Figure 2. Bayesian tree of partial Cytb sequences, with nodal support values based on Bayesian probabilities and boot-
strap values for Maximum Likelihood and Maximum Parsimony. Samples included in all analyses (molecular, morphomet-
ric and bacular) are underlined.
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8 P. J. TAYLOR ET AL.
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
15.4 mm (15.1–15.7 mm, N = 12) (t23 = 0.77, P > 0.05),
and in R. landeri, for females 16.4 mm (15.9–16.8 mm,
N = 9) and for males 16.3 mm (16.2–16.5 mm, N = 5)
(t12 = –0.12, P > 0.05). These values for CCL are close
to the published ranges and means of condylo-incisive
length (CIL) in Monadjem et al. (2010) for both spe-
cies. Based on our data, CCL is typically 0.1–0.7 mm
larger than CIL in any one skull, suggesting that our
samples referred to R. swinnyi s.l. and R. landeri s.l.
might be slightly smaller than recorded by Monadjem
et al. (2010).
Interspecific craniometric variation
A PCA based on ten log-transformed cranial variables
clearly resolved three groups commensurate with R.
swinnyi s.l., R. landeri s.l. and R. simulator s.l. (data not
shown but available from PJT on request). To include
data from type specimens for which only five cranial
measures were available, PCA was repeated with only
these five variables, which resolved the same three dis-
tinct groups (Fig. 4). Specimens of R. swinnyi s.l. could
be distinguished from R. landeri s.l. and R. simula-
tor based on their smaller size (all variable loadings
on the first component were positive and R. swinnyi
s.l. plotted to the left; Fig. 4; Table 3). Condylocanine
skull length (CCL) was always <16.0 mm in R. swin-
nyi s.l. and >16.0 mm in R. simulator and R. landeri s.l.
(Table 2). There was noticeable variation within R. swin-
nyi s.l., which occupied a much broader morphometric
space than the other two taxa (R. simulator s.l. and R.
landeri s.l.). In particular, specimens from Gorongosa
National Park, herein designated R. gorongosae sp.
nov., were distinctly smaller-sized than other members
of this group. Animals referable to R. rhodesiae, as iden-
tified by genetic analysis, overlapped considerably with
R. swinnyi s.s. (Fig. 4). The holotype of rhodesiae (TMSA
1325) from Bezwe River in Zimbabwe plotted within
the range of variation of specimens referred geneti-
cally to rhodesiae, confirming the validity of the name.
Specimens assigned to R. landeri s.s. from west, central,
north and north-eastern Africa and R. lobatus from
southern Africa overlapped considerably in size. As it
is a sub-adult, the landeri holotype from Equatorial
Guinea was distinctly smaller-sized than other landeri
individuals, and is therefore not informative. The two
co-types of lobatus from Sena River, Mozambique, plot-
ted within the range of variation of specimens referred
genetically to lobatus, confirming the validity of this
name. Although R. simulator overlapped slightly in size
with R. landeri s.l. (Fig. 4), specimens of R. simulator
grouped separately from those of R. cf. landeri on PC2;
more positive scores are associated with a dispropor-
tionately longer and narrower palate/toothrow (high
positive loading for CM3 and high negative loading for
M3M3) (Table 3).
Geometric morphometric variation
A PCA of 2D cranial landmark data recovered six par-
tially overlapping groups corresponding to R. gorongo-
sae sp. nov., R. landeri, R. lobatus, R. simulator, R.
rhodesiae and R. swinnyi (Fig. 5). The type specimens
of rhodesiae and swinnyi fell within the range of vari-
ation of the corresponding taxa defined on genetic
grounds (Fig. 5). Deformation grids provide graphical
representation of shape changes associated with PC 1
(from left to right) and those along PC 2 (bottom to
top) (Fig. 5), as well as an illustration of overall cra-
nial morphology of the different taxa as identified by
PCA (Fig. 6). The diminutive Gorongosa National Park
animals had a reduced foramen magnum as indicated
by landmarks (LMs) 8–10, a noticeable depression
along the parietal region (LM 13), low-set bullae (LMs
5–7) and a narrow and high-set nasal inflation with
a sharp slope from the nasal inflation to the maxillae
(LMs 1–18). The braincase of R. gorongosae sp. nov.,
in relation to R. rhodesiae and R. swinnyi, was nar-
rower (LMs 6 and 14). Rhinolophus swinnyi s.s. had a
dorsally expanded braincase due to the elevated posi-
tions of LMs 12–14, a broad foramen magnum (LMs
9–10), high-set bullae (LMs 5–7) and a short and
Table 1. Mean uncorrected p-distances (below diagonal) and Kimura 2-parameter model-corrected distances (above diag-
onal) based on 667 Cytb sequences between eight recognized and newly proposed species of small African Rhinolophus
bats. Within-species uncorrected p-distances provided in parentheses on the diagonals (in parentheses), except for R. lan-
deri where only one individual was available
simulator denti rhodesiae swinnyi capensis landeri lobatus gorongosae sp. nov.
simulator (0.009) 0.043 0.006 0.093 0.087 0.256 0.156 0.077
denti 0.041 (0.021) 0.043 0.077 0.070 0.243 0.141 0.102
rhodesiae 0.006 0.041 (0.002) 0.094 0.087 0.256 0.156 0.076
swinnyi 0.086 0.071 0.086 (0.011) 0.060 0.244 0.143 0.147
capensis 0.080 0.066 0.080 0.057 (0.006) 0.241 0.147 0.135
landeri 0.215 0.206 0.215 0.207 0.205 NA 0.237 0.168
lobatus 0.138 0.126 0.137 0.128 0.131 0.202 (0.003) 0.194
gorongosae sp. nov. 0.073 0.095 0.072 0.132 0.122 0.150 0.168 (0.018)
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 9
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Table 2. Summary statistics for six external and three cranial variables (in mm), mass (in g) and echolocation peak frequency for museum-type specimens and
series of taxa sampled in this study in the swinnyi, simulator and landeri groups. Except in the case of type specimens and certain series where indicated, all
measurements were taken by the authors of this paper
Taxon group or
type
Total
length
(mm)
Tail
length
(mm)
Ear
length
(mm)
Hindfoot
length
(mm)
Forearm
length
(mm)
Noseleaf
width
(mm)
Peak
frequency
(kHz)
Mass (g) Greatest
skull length
(mm)
Condylo-
canine
length
(mm)
Zygomatic
width
(mm)
R. gorongosae sp.
nov.
(holotype: DM
14820)
66.5 26 18 7.5 41.5 7 - 5.1 - 15.2 8.25
R. gorongosae sp.
nov.
67.6 ± 3.8,
N = 15,
61–78
24.7 ± 1.6,
N = 15,
22–27
17.3 ± 2.2,
N = 15,
12.5–22
8.1 ± 0.5,
N = 15,
7.5–9.0
41.3 ± 1.6,
N = 15,
38.5–44.5
7.1 ± 0.3,
N = 15,
7–8
106.2 ± 1.5,
N = 16,
103.5–
107.8
5.6 ± 0.5,
N = 15,
4.5–6.6
- 15.1 ± 0.19,
N = 7,
14.8–15.2
8.39 ± 0.16,
N = 8,
8.13–8.56
R. swinnyi Gough
1908
(co-types: TMSA
1021, 1022)
60–61 18–19 18 8 40.8–41.8
(40–40.7 in
description)
- - - 17.5 15.5 8.73–8.97,
N = 2
R. swinnyi (South
Africa)
73.0 ± 1.7,
N = 3,
71–74
22.0 ± 3.8,
N = 6,
18.3–27
18.8 ± 1.6,
N = 6,
17.2–21.6
8.5 ± 0.3,
N = 6,
8.2–9.1
43.6 ± 1.1,
N = 9,
41.7–45.0
- 105.9 ± 0.7,
N = 6,
104.9–
106.7
- 17.7 ± 0.5,
N = 10,
16.8–18.0
15.4 ± 0.45,
N = 10,
14.3–16.0
9.1 ± 0.2,
N = 11,
8.7–9.2
R. s. rhodesiae
Roberts 1946
(holotype: TMSA
1325)
74 22 15 - 41.4 (40.5 in
description)
- - - 17.6 14.9 8.34
R rhodesiae
(Chihalatan/
Malashane)
74.8 ± 5.4,
N = 20,
68–79
24.4 ± 1.5,
N = 20,
23–29
19.9 ± 0.6,
N = 20,
19–21
6 ± 0,
N = 20, 6
44.2 ± 0.9,
N = 20,
43–45
7.2 ± 0.7,
N = 18,
6.2–8.1
100.1 ± 1.0,
N-8,
99.1–102
5.7 ± 0.33,
N = 20,
4.9–6.1
17.8 ± 0.4,
N = 13,
17.2–18.6
15.4 ± 0.22,
N = 13,
15.1–15.7
8.7 ± 0.1,
N = 20,
8.46–8.96
R. landeri Martin
1838
(holotype: BMNH
1847.5.7.49)
- - - - - - - - 16.7
(broken)
- -
R. landeri (Liberia) 79–81,
N = 2
27, N = 1 18–19,
N = 2
- 42–43.9,
N = 2
- 104.3 ± 0.4,
N = 2,
104–104.6
7.8–8.5,
N = 2
18.8–18.9,
N = 2
16.3–16.6,
N = 2
9.44–9.62,
N = 2
R. l. lobatus Peters
1852
(syntypes: ZMB
24922, 24927
81.7 ± 2.9,
N = 3,
80–85
25.0 ± 1.0,
N = 3,
24–26
16.0 ± 0,
N = 3, 16
9.8 ± 0.29,
N = 3,
9.5–10
45.2 ± 0.76,
N = 3,
44.5–45
7.1 ± 0.14,
N = 3,
7–7.25
- - - - -
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10 P. J. TAYLOR ET AL.
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
steeply inclined frontal region (LMs 14–15). The nasal
inflation was posteriorly set and small (LMs 15–17).
There was a short slope from the nasal inflation to the
maxillae (LMs 17–18) and the anterior margin of the
maxillae was recessed (LM 1). Rhinolophus rhodesiae
presented with an expanded braincase (LMs 10–14),
relatively high positioned LM 12, elongated frontal
region (LMs 14–15), wide bullae (LMs 5–7) and a nar-
rowed nasal inflation (LMs 16–18).
Rhinolophus landeri from West Africa was distin-
guished from R. lobatus from south-eastern Africa by a
broader rostrum (LMs 1, 2, 4 and 18), wider and higher
set tympanic bullae (LMs 5–7), noticeably shortened
foramen magnum (LMs 9–10) and a more rounded
posterior portion of the braincase (LMs 10–14). The
taxon had a narrow nasal inflation (LMs 16–18), with
a steeply inclined slope from the highest point of the
nasal inflation to the maxillae (LMs 17–18). In relation
to R. landeri, R. lobatus had a less rounded braincase
with higher positioned lambdoidal crests (LMs 10–14),
and less steeply inclined anterior margin of the nasal
inflation (LMs 17–18). Rhinolophus simulator s.l. was
characterized by an elongated cranium, with a dis-
tinctly widened nasal inflation (LMs 16–18) and broad
tympanic bullae (LMs 5–7).
Bacular morphology
The KMO statistic = 0.88, indicating that the factors
extracted by PCA accounted for the majority of sample
variance. PCA of the eight bacular measurements delim-
ited ten groupings varying in bacular length (PC1) and
width (PC2), and which corresponded with all the taxa
recognized in this study (Tables 4, 5; Figs 7, 8). In addi-
tion, four highly distinct groupings (bacular types, num-
bered 1–4 in Table 5 and Fig. 7) were detected within
specimens referred to R. lobatus from Mozambique, and
two distinct bacular types (labelled 5–6 in Table 5 and
Fig. 7) were detected within R. simulator from South
Africa and Zimbabwe. Bacular morphometric groups
identified in Fig. 7 correspond to clearly recognizable
morpho-types as indicated by the drawings in Fig. 8.
Within swinnyi s.l. (swinnyi, rhodesiae and gorongo-
sae sp. nov.), animals classified as R. gorongosae sp.
nov. and R. swinnyi s.s. had short bacula with reduced
bases (negative scores on PC1 and PC2), the majority
of which presented with a characteristic notch on one
side of the shaft tip (position variable). A slightly wider
baculum shaft and broader tip (higher PC2 scores) dis-
tinguished R. swinnyi from R. gorongosae sp. nov. The
long, tapered baculum of R. rhodesiae, with its broad
base (high positive PC1 and PC2 scores) and the shal-
low notch in the lower shaft visible in lateral profile,
was clearly distinct from all other specimens in the
swinnyi complex (Figs 7, 8). Within landeri s.l. (landeri
and lobatus), individuals from Chihalatan Cave and
Gorongosa National Park (labelled 1 in Table 5 and
Taxon group or
type
Total
length
(mm)
Tail
length
(mm)
Ear
length
(mm)
Hindfoot
length
(mm)
Forearm
length
(mm)
Noseleaf
width
(mm)
Peak
frequency
(kHz)
Mass (g) Greatest
skull length
(mm)
Condylo-
canine
length
(mm)
Zygomatic
width
(mm)
R. lobatus
(Chihalatan/
Malashane)
82.5 ± 2.45,
N = 9,
79–87
26.4 ± 2.30,
N = 9,
23–31
19 ± 1.12,
N = 9,
17–20
6.89 ± 0.79,
N = 9,
6–8
47 ± 1.22,
N = 9,
46–49
7.5 ± 0.56,
N = 9,
6.6–8.3
106.8 ± 0.4,
N = 4,
106.5–107
7.5, N = 9,
7–9
19.0 ± 0.41,
N = 9,
18.5–19.5
16.4 ± 0.16,
N = 9,
16.2–16.5
9.5 ± 0.23,
N = 9,
9.3–9.9
R. lobatus
(Gorongosa)
75.8 ± 2.6,
N = 26,
71.5–81.5
25.2 ± 3.3,
N = 26,
21.4–28.0
17.3 ± 1.3,
N = 26,
13.7–19.5
9.7 ± 0.9,
N = 26,
7.9–11.7
45.2 ± 1.5,
N = 26,
43.0–49.5
7.6 ± 0.4,
N = 26,
7.1–8.1
105.1 ± 1.8,
N = 20,
101.9–
107.6
8.2 ± 1,
N = 26,
5.2–9.5
18.9 ± 0.33,
N = 12,
18.4–19.5
16.5 ± 0.29,
N = 12,
16.0–17.0
9.8 ± 0.21,
N = 12,
9.4–10.1
R. simulator
(holotype: BMNH
2.2.7.10)
- 25.7 20 - 41.5 (43.5 in
description)
8.3 - - 18.9 16.2 8.8
R. simulator (South
Africa)
78 ± 0.6,
N = 6,
77–79
24.8 ± 2.0,
N = 6,
22–28
19.5 ± 1.3,
N = 6,
17.2–21.1
10.0 ± 0.5,
N = 5,
9.4–10.4
44.0 ± 1.0,
N = 6,
42.7–45.8
- 83.4 ± 0.6,
N = 6,
82.6–83.8
18.9 ± 0.48,
N = 11,
18.3–19.7
16.6 ± 0.32,
N = 13,
16.1–17.2
9.0 ± 0.2,
N = 13,
8.8–9.4
*Data obtained from Taylor (2005). See Material and methods for definitions of museum acronyms.
Table 2. Continued
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 11
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Fig. 7), herein referred to as R. lobatus, had a long, wide
baculum, with a spatulate tip and asymmetric base (the
dorsal and ventral edges being of different heights).
Two individuals of lobatus from Mozambique (Pemba
and Gorongosa National Park, numbered 2 in Table 5
and Fig 7) have a long, robust baculum with distinc-
tive flanges on the ventral surface and an asymmetric
base. On the other hand, the other lobatus bacular types
(numbered 3–4 in Fig. 7) are smaller and more gracile
(Figs 7, 8). Compared to R. lobatus, the Mount Nimba
individual (R. landeri) had a shorter baculum with a
distinctly bulbous tip (Table 5; Figs 7, 8). Within R. sim-
ulator s.l., animals from Zimbabwe and South Africa
(N = 6), were distinct from specimens originating from
Stanhope Mine in South Africa (N = 3), based on their
deeper basal notch, spatula-shaped tip and less robust
base (Figs 7, 8).
Qualitative characters
Noseleaf morphology
Populations assigned to Rhinolophus swinnyi s.l. ani-
mals fell into three distinct groups based on anatomi-
cal structure of the connecting process and lancet (top
row; Fig. 9). Rhinolophus gorongosae sp. nov.possessed
a subtriangular lancet with straight to slightly con-
cave sides, and a high, rounded connecting process;
the sella has a diminutive, pointed tip. Rhinolophus
swinnyi s.s. presented with a subtriangular lancet
with concave sides and bluntly pointed tip, and a high,
rounded connecting process, with a bluntly pointed
sella tip directed downwards. Rhinolophus rhodesiae
was characterized by a hastate lancet and less erect,
low, rounded, anteriorly projecting connecting pro-
cess and pronounced sella tip. Rhinolophus landeri
from Liberia possessed a short, narrow lancet with a
pointed tip, a less erect and subtriangular connecting
process, markedly constricted sella and a downward-
facing sella tip. Rhinolophus lobatus had a tall, sub-
triangular lancet with slightly concave sides and a
rounded tip. The connecting process was erect and
subtriangular, the sella bore a deep constriction, and
the sella tip was sharper and more anteriorly directed
than the West African form. Two additional R. lobatus
forms were documented: one, from Gorongosa National
Park, with a small hastate lancet and bluntly pointed
tip, and a relatively small subtriangular connecting
process with reduced sella tip, and another group from
Pemba and Gorongosa National Park (Mozambique)
with a tall, hastate lancet, a high, subtriangular
(partially rounded) connecting process and a sharply
pointed sella tip (Fig. 9). Animals classified under R.
simulator s.l. exhibited two distinct morphologies: one
with a subtriangular lancet with concave side and a
rounded tip, and a low, rounded, wide connecting pro-
cess, with an asymmetric sella tip, and another with
a hastate lancet, pronounced bluntly pointed tip and
a low, rounded, wide connecting process; the sella tip
was broader than the other group (Fig. 9).
Dental variation
We observed subtle differences in the size and position
of the anterior premolar (PM1) among the taxa exam-
ined above (Fig. 10). Rhinolophus swinnyi, R. gorongo-
sae sp. nov. and R. rhodesiae all possessed a minute
and laterally displaced PM1. The alveolar borders of
canine and PM2 in R. swinnyi were almost in contact,
whereas in R. gorongosae sp. nov. and R. rhodesiae a
distinct gap was present. The PM1 in R. landeri and R.
lobatus was positioned in the toothrow and due to its
relatively larger size than the afore-mentioned taxa,
resulted in a larger space between the canine and PM2.
The anterior premolar of the Pemba individual is more
robust than other lobatus-type specimens and was
positioned further in the toothrow. One of the two R.
simulator taxa, (as identified by baculum and noseleaf
morphology), had a PM1 situated more external to the
toothrow, resulting in a more narrowed space between
the alveolar borders of the canine and PM2, when com-
pared to the other taxon.
acoustic analysis
Recent large series obtained in 2007 and 2015 from
two adjacent caves (1 km apart) in the Inhambane
Table 4. Variable loading matrix for PCA of bacular mor-
phological variation in three small African Rhinolophus
spp.
Character PC 1 PC 2 PC 3
Total length (dorsal) 0.867 0.349 0.183
Greatest base width (dorsal) 0.653 0.492 0.311
Greatest base width (lateral) 0.751 0.413 0.179
Greatest shaft width (dorsal) 0.188 0.194 0.934
Narrowest shaft width (dorsal) 0.351 0.878 0.167
Greatest tip width (dorsal) 0.444 0.660 0.423
Height of basal notch (dorsal) 0.471 0.281 0.223
Height of base (dorsal) 0.695 0.319 0.483
Table 3. Variable loading matrix for PCA of interspecific
variation in three small African Rhinolophus spp. (see
Fig. 5 for details)
Variable PC 1 PC 2 PC 3
Condylo-canine length (CCL) 0.337 0.509 0.254
Zygomatic width (ZW) 0.520 -0.392 0.353
Mastoid width (M) 0.380 0.061 0.562
Width of maxilla between outer
edges of M3 (M3M3)
0.519 -0.490 -0.518
Upper toothrow length (C1M3) 0.450 0.586 -0.476
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12 P. J. TAYLOR ET AL.
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Province of Mozambique, specifically the Malashane
and Chihalatan Caves, comprised both R. rhodesiae
and R. lobatus. Specimens collected from Chihalatan
in October 2007 comprised only R. rhodesiae, those
from February and May 2015 comprised only R. loba-
tus. The specimens collected in February and May
2015 from Malashane Cave comprised predominantly
R. rhodesiae with a few individuals of R. lobatus. The
Table 5. Bacula measurements for 33 individuals of three small African Rhinolophus taxa. Mean, standard deviation,
range and sample sizes are provided. All measurements recorded in mm. Numbers in superscript refer to the position of
taxa in Fig. 7
Taxon Total length
(dorsal)
Greatest
base width
(dorsal)
Greatest
base width
(lateral)
Greatest
shaft width
Narrowest
shaft width
Greatest
tip width
Height of
basal notch
(dorsal)
Height of
base
R. gorongosae sp.
nov.
(Gorongosa,
Mozambique)
2.00 ± 0.11,
N = 3
1.91–2.12
0.53 ± 0.01,
N = 3
0.52–0.54
0.48 ± 0.04,
N = 3
0.43–0.50
0.22 ± 0.02,
N = 3
0.20–0.23
0.11 ± 0.01,
N = 3
0.10–0.12
0.22 ± 0.02,
N = 3
0.20–0.23
0.11 ± 0.02,
N = 3
0.09–0.12
0.41 ± 0.02,
N = 3
0.38–0.43
R. swinnyi
(Eastern Cape /
KwaZulu-Natal,
South Africa)
2.06 ± 0.09,
N = 4
1.97–2.17
0.37 ± 0.06,
N = 4
0.45–0.60
0.37 ± 0.02,
N = 4
0.35–0.40
0.18 ± 0.02,
N = 4
0.16–0.21
0.12 ± 0.03,
N = 4
0.10–0.17
0.23 ± 0.05,
N = 4
0.17–0.29
0.07 ± 0.01,
N = 4
0.06–0.09
0.38 ± 0.08,
N = 4
0.32–0.49
R. rhodesiae
(Chihalatan Cave,
Mozambique /
KwaZulu-Natal,
South Africa)
2.59 ± 0.08,
N = 7
2.50–2.70
0.93 ± 0.06,
N = 7
0.83–0.99
0.83 ± 0.03,
N = 7
0.80–0.88
0.33 ± 0.03,
N = 7
0.28–0.37
0.19 ± 0.02,
N = 7
0.17–0.21
0.23 ± 0.02,
N = 7
0.19–0.26
0.20 ± 0.03,
N = 7
0.17–0.23
0.65 ± 0.05,
N = 7
0.59–0.74
R. landeri
(Mount Nimba,
Liberia)
2.29, N = 1 0.88, N = 1 0.78, N = 1 0.27, N = 1 0.18, N = 1 0.30, N = 1 0.15, N = 1 0.72, N = 1
R. lobatus1
(Gorongosa /
Chihalatan Cave,
Mozambique)
2.57 ± 0.07,
N = 3
2.51–2.65
0.94 ± 0.04,
N = 3
0.91–0.99
0.97 ± 0.02,
N = 3
0.96–1.00
0.29 ± 0.02,
N = 3
0.28–0.32
0.22 ± 0.02,
N = 3
0.20–0.24
0.35 ± 0.02,
N = 3
0.32–0.37
0.17 ± 0.01,
N = 3
0.17–0.18
0.92 ± 0.01,
N = 3
0.92–0.92
R. cf. lobatus2
(Gorongosa / Pemba,
Mozambique)
2.96 ± 0.05,
N = 2
2.92–2.99
1.12 ± 0.00,
N = 2
1.22 ± 0.03,
N = 2
1.20–1.24
0.39 ± 0.01,
N = 2
0.38–0.39
0.22 ± 0.03,
N = 2
0.20–0.24
0.41 ± 0.01,
N = 2
0.41–0.42
0.27 ± 0.01,
N = 2
0.27–0.28
0.91 ± 0.01,
N = 2
0.90–0.92
R. cf. lobatus3
(Gorongosa,
Mozambique)
1.98 ± 0.00,
N = 2
1.98–1.98
0.42 ± 0.02,
N = 2
0.41–0.43
0.53 ± 0.01,
N = 2
0.52–0.53
0.28 ± 0.01,
N = 2
0.27–0.29
0.16 ± 0.00,
N = 2
0.16–0.16
0.30 ± 0.01,
N = 2
0.29–0.31
0.15 ± 0.00,
N = 2
0.15–0.15
0.48 ± 0.02,
N = 2
0.47–0.50
R. cf. lobatus4
(Gorongosa /
Malashane Cave,
Mozambique)
1.98 ± 0.26,
N = 2
1.80–2.16
0.50 ± 0.03,
N = 2
0.48–0.52
0.52 ± 0.09,
N = 2
0.46–0.58
0.22 ± 0.03,
N = 2
0.20–0.24
0.13 ± 0.01,
N = 2
0.13–0.14
0.22 ± 0.04,
N = 2
0.19–0.24
0.14 ± 0.03,
N = 2
0.12–0.16
0.43 ± 0.05,
N = 2
0.40–0.47
R. simulator5
(South Africa/
Zimbabwe)
2.48 ± 0.18,
N = 6
2.20–2.69
0.82 ± 0.11,
N = 6
0.61–0.94
0.77 ± 0.05,
N = 6
0.67–0.82
0.24 ± 0.03,
N = 6
0.21–0.30
0.18 ± 0.02,
N = 6
0.14–0.20
0.29 ± 0.03,
N = 6
0.24–0.33
0.24 ± 0.05,
N = 6
0.16–0.29
0.68 ± 0.13,
N = 6
0.50–0.90
R. simulator6
(KwaZulu-Natal,
South Africa)
2.61 ± 0.15,
N = 3
2.43–2.70
0.98 ± 0.02,
N = 3
0.96–1.00
0.83 ± 0.05,
N = 3
0.78–0.88
0.24 ± 0.01,
N = 3
0.24–0.25
0.16 ± 0.00,
N = 3
0.15–0.16
0.29 ± 0.03,
N = 3
0.25–0.32
0.29 ± 0.01,
N = 3
0.28–0.31
0.90 ± 0.09,
N = 3
0.80–1.00
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 13
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larger-sized R. lobatus from Chihalatan and Malashane
Caves had CF frequencies of ca. 107 kHz. The smaller-
sized R. rhodesiae from Chihalatan and Malashane
Caves had CF frequencies of 99–102 kHz, which
did not match any known African rhinolophid spe-
cies (Monadjem et al., 2010). Peak frequencies for
Gorongosa National Park R. gorongosae sp. nov. and
R. lobatus were 103 kHz and 107 kHz, respectively.
Maximum frequencies of the two R. landeri specimens
from Mount Nimba, Liberia, were 104–105 kHz.
DISCUSSION
phylogeny, hybridization and speciation in the
R. swinnyi and R. landeRi coMplexes
Although molecular evidence combined with morpho-
logical and acoustic characters are powerful diagnostic
tools to reveal cryptic species of bats, including African
horseshoe bats of the genus Rhinolophus (Taylor et al.,
2012; Jacobs et al., 2013), a growing body of evidence
attests to the frequency of past hybridization events
between closely and even distantly related lineages
of bats (Artyushin et al., 2009; Nesi et al., 2011; Vallo
et al., 2013; Khan, Phillips & Baker, 2014); and perti-
nently Dool et al. (2016) revealed introgression of R.
ferrumequinum into R. clivosus. Molecular genetic evi-
dence from mtDNA reveals that some morphologically
distinct ‘good’ species may be genetically very similar.
Such cases of hybridization have been attributed to the
past divergences of two species, whereafter subsequent
environmental changes have led to the expansion of one
species range into that of the other (Artyushin et al.,
2009; Vallo et al., 2013). We propose that an analogous
scenario may explain the enigmatic genetic affinities
recovered for the morphologically distinctive R. simu-
lator and R. rhodesiae. These two species currently
overlap in range (Fig. 1) and are clearly differentiated
on craniometric (Figs 4–6; Table 2), bacular (Figs 7, 8;
Table 5), noseleaf (Fig. 9), dental (Fig. 10) and acoustic
(Table 2) evidence. We conclude that they are highly
unlikely to be recently diverged sister species.
Apart from this single apparent case of historical
introgression, taxa within the former swinnyi s.l. and
landeri s.l. complexes are well differentiated from each
other based on Cytb sequences, 7.2–13.2% between the
three taxa within swinnyi s.l. and 20.2% between the
two taxa within landeri s.l. (Table 1). Clearly, cryptic
species within both species complexes are paraphyl-
etic (Fig. 2), indicating that the morphological char-
acters used to define these species-groups are subject
to convergent evolution. For example, the distinctive
erect connecting process and brown apical tufts in
males of the landeri group clearly evolved indepen-
dently in R. lobatus (which is basal to all other African
taxa in our study) and R. landeri s.s., which is nested
within R. gorongosae sp. nov. This paraphyletic rela-
tionship between R. landeri from West Africa and
R. gorongosae from central Mozambique is unlikely to
be due to historical introgression given the widely dis-
junct distributions of these two species and the high
genetic distance between them (p = 15%). The range
of R. gorongosae sp. nov. is still poorly known (await-
ing further sampling) but we propose that speciation
of this taxon was associated with the establishment
of unique habitats on the inselbergs of the Gorongosa
range, isolated high above the coastal plain (see:
Moore, Cotterill & Key, 2017), where it is congruent
with the recently described endemic gecko, Afroedura
gorongosa (Branch et al., 2017). The speciation of
another recently described cryptic horseshoe bat, R.
mabuensis, was associated with isolated montane for-
est patches on inselbergs of northern Mozambique
(Mounts Mabu and Inago) and, similarly, R. smithersi
has a distribution that is largely associated with the
Soutpansberg and Waterberg ranges in South Africa
(Taylor et al., 2012). To reinforce this point, a specimen
of R.cf. swinnyi from Mount Inago has a very small
size within the range of R. gorongosae sp. nov. and,
without molecular data, is tentatively assigned to R.
gorongosae sp. nov.
Known specimens restrict R. swinnyi s.s. to the
Eastern Cape Province and the southern and central
reaches of the KwaZulu-Natal Province, both within
South Africa (Fig. 11). Rhinolophus rhodesiae occurs
from the central and northern regions of the KwaZulu-
Natal Province and Limpopo Province, extending
northwards to at least Zambia, northern Mozambique
and Tanzania (Fig 11). Based on recent collections,
R. gorongosae sp. nov.is restricted to the Gorongosa
Mountains, and possibly Mount Inago. In the case of
R. landeri s.l., R. lobatus seems to occur throughout
Mozambique (and probably more widely in southern
Africa), while R. landeri s.s. occurs in west, central,
north and north-eastern Africa (Fig. 11). Due to lack of
sampling, the ranges of these respective species and the
location of the break between them remain uncertain.
The mechanism(s) of speciation of these cryptic spe-
cies of Rhinolophus are not yet clear. The chronomet-
ric tree with dated nodes, estimated in BEAST from
six nuclear introns (Fig. S5 in Dool et al., 2016), con-
strains the formative nodes that shaped diversifica-
tion of Afrotropical Rhinolophus to the late Neogene;
the founding of the clade including simulator s.s. and
swinnyi s.s. lineages is constrained to 4.89 (6.61–3.5)
Mya, with the alcyone and landeri complexes at 4.42
(6.41–2.85) Mya. Following Dool et al. (2016) diver-
gence of R. swinnyi s.s. from other lineages of the
capensis-group (including both simulator and rhode-
siae, classified by them as ‘cf. simulator’) is estimated
at ~2.5 Mya; with isolation of R. simulator and R. cf.
simulator (= rhodesiae) estimated as Early Pleistocene
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14 P. J. TAYLOR ET AL.
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(~2 Mya), but the latter estimate might be biased by
historical introgression, as discussed above (first para-
graph of this section).
The recurrent uplift and volcanism along the
Albertine Rift System has been invoked to explain
allopatric speciation in large and small mammals, and
centred on the ‘Mbeya triple junction’ and Rungwe
volcanic province between lakes Malawi and Rukwa
(Cotterill, 2003; Faulkes et al., 2011, 2017). Recurrent
uplift of southern Africa (intensifying erosion) acted
to accentuate the relief along the eastern lowlands
from the central Plateau and Eastern Highlands
of Zimbabwe to the Eastern Cape (Partridge 1998;
Partridge & Maud, 2000; Moore et al., 2009). These
twinned geomorphic processes are also invoked to
have isolated Mount Gorongosa and neighbouring
inselbergs in Central Mozambique (Moore, Cotterill &
Key, 2017). It is possible that these regional palaeo-
climatic and geomorphological events acted in concert
to cause the allopatric speciation of horseshoe bats
within the landeri s.l. and swinnyi s.l. complexes.
Further phylogeographic studies informed by broader
sampling are required to unravel the causes of the evo-
lution of these two species complexes.
TAXONOMIC CONCLUSIONS: DESCRIPTION
AND RE-DEFINITION OF SPECIES
FaMily rhinolophidae bell, 1836
genus Rhinolophus lacépède, 1799
Rhinolophus goRongosae sp. nov.
least horseshoe bat
Holotype: Durban Natural Science Museum (DM)
No. 14820 (field number JAG196), is an adult male,
preserved in 70% ethyl alcohol, originally part of series
of specimens collected by J. A. Guyton on 25 April
2015. The cranium and baculum has been extracted
and examined for this study. The specimen has been
included in both morphometric analyses.
Type locality: Bunga Inselberg, Gorongosa National
Park, Sofala Province, Mozambique −18.599° S,
34.343° E, 212 m.
Paratypes: Eight specimens collected 24–25 April 2015
(DM 14815–14819), 2 May 2015 (DM 14828), 5 November
2015 (DM 14843) and 22 July 2015 (DM 14865).
Referred specimens having molecular identification:
DM 14815 (JAG 188) a female specimen collected
by J. A. Guyton from Mozambique, Sofala Province,
Gorongosa National Park, Bunga Inselberg, -18.599°
S, 34.343° E, 212 m; DM 14843 (JAG 228) a female
specimen collected by J. A. Guyton from Mozambique,
Sofala Province, Gorongosa National Park, -18.694° S,
34.208° E, 308 m.
Referred specimens having only morphological
identification: TMSA 49116 (JAG 31), adult male,
collected on 21 April 2013 by J. A. Guyton from
Mozambique, Sofala Province, Gorongosa National
Park, Cheringoma Plateau, Gorge Rim, Site 1, -18.635°
S, 34.808° E, 213 m. Incertae sedis: DM 14864 adult
female collected by J. A. Guyton on 21 July 2015 from
Mozambique, Sofala Province, Gorongosa National
Park, -18.465° S, 34.052° E, 1150 m; DM 11482, adult
female collected on 1 May 2009 by J. Bayliss from
Mozambique, Nampula Province, Mount Inago Forest
Camp, -15.045° S, 37.396° E 1478 m.
Etymology: The species derives its name from the
Gorongosa district of Mozambique, in particular
Gorongosa National Park, a biologically diverse region
of southern Africa.
Diagnosis: The species can be clearly distinguished
from both R. swinnyi s.s. and R. rhodesiae on molecular
grounds (Figs 2, 3) as well as by its smaller size (Fig. 4;
Table 2), distinct cranial shape (Figs 5, 6), echolocation
call peak frequency (Table 2), baculum (Figs 7, 8) and
noseleaf (Figs 9, 12) characteristics. Although some
measurements overlap, there is minimal overlap in
condylocanine skull length and zygomatic skull width
between this species (14.8–15.2 mm; 8.13–8.56 mm)
and R. swinnyi (14.3–16.0 mm; 8.7–9.2 mm) and R.
rhodesiae (15.1–15.7; 8.46–8.96 mm). The small size of
this form makes it even smaller than denti (regarded
by Csorba et al., 2003 as the smallest species in the
Ethiopian region), therefore making this new species
Africa’s smallest horseshoe bat. Comparing means for R.
gorongosae sp. nov. (Table 2) and denti (Monadjem et al.,
2010): forearm 41.3 mm cf. 43.1 mm; mass 5.6 g cf. 7.0 g.
Description: The genetically-distinct R. gorongosae sp.
nov. is similar in pelage colour but distinctly smaller
in external (mean total length 68 mm, mean forearm
length = 41 mm) and cranial (mean condylocanine
length 15.1 mm) measurements (Table 2) than both
rhodesiae (mean total length 75 mm, forearm length
44 mm, condyolocanine length 15.4 mm) and swinnyi s.s.
from South Africa (mean total length 73 mm, forearm
length 44 mm, condyolocanine length 15.4 mm). Based
on geometric morphometric results, the diminutive
R. gorongosae sp. nov. has a reduced foramen magnum,
a noticeable depression along the parietal region, low
set bullae, a narrow braincase and a narrow and high
set nasal inflation with a sharp slope from the nasal
inflation to the maxillae (Fig. 6). Lancet is subtriangular
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 15
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Figure 3. Haplotype network using minimum-spanning inference method based on 667 base pairs of Cytb.
Figure 4. PCA variation for five log-transformed cranial variables for six proposed small southern Africa Rhinolophus spp.
defined on molecular and morphological grounds.
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16 P. J. TAYLOR ET AL.
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with straight to slightly concave sides, and a high,
rounded connecting process; the sella has a diminutive,
pointed tip (Fig. 12). Baculum short with reduced base,
with a characteristic notch on one side of the shaft tip
(position variable). The slightly narrower baculum shaft
with narrower tip distinguishes R. gorongosae sp. nov.
Figure 5. Biplot showing the first and second principal components from a PCA of 2D landmark data for small southern
Africa Rhinolophus spp., including data from type specimens of R. swinnyi and R. s. rhodesiae. Deformation grids illustrate
cranial shape changes associated with each PC.
Figure 6. Overall lateral cranial morphology of small southern African Rhinolophus taxa as illustrated by deformation
grids (exaggerated 3×).
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 17
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
from R. swinnyi (Table 5; Figs 7, 8). Maxillary toothrow
with minute and laterally displaced PM1 with a distinct
gap between the alveolar borders of canine and PM2
(Fig. 10). Mean peak echolocation CF frequency is
106 kHz (104–108 kHz; Table 2).
Distribution and biology: So far, it appears that this
tiny species is restricted in its distribution to Gorongosa
National Park in Mozambique (Fig. 11), although we
provisionally refer a very small adult individual from
Mount Inago in Mozambique to this taxon. Molecular
sequences are required from a wider range of localities
to determine the range of this taxon. Given that the two
individuals sequenced from Gorongosa National Park
were distinct from each other, the possibility exists that
more than one cryptic species may be present.
Rhinolophus Rhodesiae roberts, 1946
roberts’s horseshoe bat
Synonyms: None.
Holotype: TMSA 1325, adult female, collected by
A. Roberts on 16 August 1913.
Type locality: ‘Southern Rhodesia’ (= Zimbabwe),
Bezwe River, tributary of ‘Wanetsi’ (= Nuanetsi) River,
−21.500° S, 31.167° E.
Referred specimens having molecular identifications:
FMNH 228942 (SMG 19017), female, 228943 (SMG
19018), male, 228944 (SMG 19019), female, 228945
(SMG 19020), male, 228946 (SMG 19021), female,
228948 (SMG 19023), male, 228949 (SMG 19024),
female, 228950 (SMG 19025), female, 228951 (SMG
19026), female, 228952 (SMG 19027), male, 228953
(SMG 19028), male, 228955 (SMG 19030), male,
228957 (SMG 19032), female, 228958 (SMG 19033),
female, 228959 (SMG 19034), female, 228960 (SMG
19035), female, 228961 (SMG 19036), male, 228962
(SMG 19037), male, 228964 (SMG 19039), female,
all collected on 2 May 2015 by S. M. Goodman, M. C.
Schoeman and G. le Minter from Mozambique,
Inhambane Province, Malashane Cave, 39.1 km Efrom
Inhassoro, −21,668° S, 34,847° E, and situated <2 km
from Chihalatan Cave referred to above.
Referred specimens having only morphological
identifications: DM 7080, adult male, KwaZulu-
Natal Province, Hlabeni Forest Reserve, −29.933° S,
29.766° E collected by D. Forbes on 29 July 2000; DM
12007 (adult male); DM 14034 (adult male), collected
by M. C. Schoeman from KwaZulu-Natal Province,
Pietermaritzburg, Ferncliff Nature Reserve, Ferncliff
Cave, −29.550° S, 30.320° E; DM 11270 (female),
11271 (male), 11272 (female), 11273 (female), 11275
(male), all collected by S. Stoffberg from Chihalatan
Cave, 38.2 km E of Inhassoro, Inhambane Province,
Mozambique −21.671° S, 34.864° E;. DM 11275
collected on 8 August 2006 while the other specimens
were collected on 3 September 2007; FMNH 228956
(SMG 19031), collected 2 May 2015 by S. M. Goodman,
M. C. Schoeman and G. le Minter from Mozambique,
Inhambane Province, Malashane Cave, 39.1 km E
from Inhassoro, −21,668° S, 34,847° E; DM 13450
(female), DM13451 (female), collected on 8 May 2012
by J. Bayliss at Mozambique, Niassa Province, Mount
Mecula, −12.068° S, 37.662° E.
Etymology: The name refers to the location in Southern
Rhodesia (now Zimbabwe) where the type specimen
was collected.
Figure 7. Biplot showing the first and second principal components from a PCA of eight bacular measurements for small
southern Africa Rhinolophus taxa. Numbers indicate the position of taxa referred to in Table 5.
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18 P. J. TAYLOR ET AL.
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Figure 8. Dorsal, lateral and ventral views (from left to right) of bacula of nine of the ten taxa as identified by PCA of
bacular measurements. Scale bars = 1 mm.
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 19
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Re-diagnosis and description: Roberts (1946) described
this subspecies as being slightly smaller than the
nominate R. s. swinnyi based on slightly smaller body
size, slightly longer tail, smaller ears and its bright
ochraceous colour. Since the last-mentioned character
is known to be an environmentally induced effect
in many cave-dwelling bat species, it does not serve
as a diagnostic character. In our analysis, molecular
evidence closely matched our series from Chihalatan
and Malashane Caves with Genbank sequences from
Figure 9. Noseleaf structure of representatives of various Rhinolophus taxa (not drawn to scale).
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20 P. J. TAYLOR ET AL.
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the extreme northern South Africa (Pafuri), Zimbabwe
(Dambanzara) and Zambia (Kalenda and Shimalala
Caves). Since these localities encompass the type locality
of rhodesiae (Bezwe River in Zimbabwe), and Pafuri is
only 100 km south of Bezwe, we are confident to use this
available name for this widespread taxon. The species
Figure 10. Occlusional views of the maxillary toothrows of the various Rhinolophus taxa detailed in this study. Arrows
indicate the position of the small anterior premolar.
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 21
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
can be further diagnosed by having echolocation peak
frequencies around 100 kHz (99–102 kHz, N = 8 from
Malashane Cave; Table 2), which are quite distinct from
typical swinnyi (105–107kHz, N = 6; Table 2) as well as
the new Gorongosa National Park taxon R. gorongosae
sp. nov. (104–108 kHz, N = 16; Table 2). Noseleaf
structure is distinctive, being characterised by a hastate
lancet, not as concave as true swinnyi, less erect, low,
rounded connecting process and more pronounced
posterior lobe. Bacular structure of rhodesiae is clearly
distinct from other swinnyi-like animals (see Figs 7,
8), being characterized by a distinctly longer tapered
baculum with a distinctly broader base and shallow
notch along the lower portion of the shaft that is visible in
the lateral profile. Traditional morphometrics (Table 2;
Fig. 4) do not differentiate rhodesiae from swinnyi
proper; however, rhodesiae is clearly distinguished with
minimal overlap from the distinctly smaller gorongosae
sp. nov. Although quite small, the R. rhodesiae holotype
falls within the range of variation of specimens assigned
to R. rhodesiae from Mozambique, Zimbabwe, Zambia,
northern South Africa (Pafuri, Limpopo Province), and
Zanzibar (Table 2; Fig. 4). It falls clearly outside (larger
than) the range of variation of the smaller gorongosae
sp. nov. taxon (Fig. 4). Geometric morphometric results
result in a better separation between rhodesiae and
swinnyi s.s. with only minimal overlap (Fig. 5). Once
again, the rhodesiae holotype from Bezwe River clusters
within the range of variation of the rhodesiae taxon and
outside the gorongosae sp. nov. or swinnyi taxa, thus
validating the use of this name for this taxon.
Distribution and biology: Combined molecular and
morphometric data suggest the widespread distribution
of this taxon from central and northern South Africa
through Zimbabwe, Zambia and Mozambique extending
to Zanzibar (Fig. 11). Based on specimen assignments
on morphological grounds, the species co-occurs with
R. swinnyi in central KwaZulu-Natal at Ferncliff Cave,
as well as occurring in close proximity in northern
KwaZulu-Natal, recorded at Hlabisi Forest close to
Ngome Forest where swinnyi was recorded (Fig. 11).
The widespread extent of this taxon and its occurrence
in northern South Africa is confirmed by the widespread
occurrence of a hitherto unidentified 100 kHz acoustic
type recorded in the Soutpansberg (Taylor et al., 2013),
and Pafuri Region of northern Kruger National Park
(Taylor & Parker, unpublished data).
Figure 11. Final distribution of small Rhinolophus species classified according to the present study.
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22 P. J. TAYLOR ET AL.
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Rhinolophus lobatus peters, 1852
peters’s horseshoe bat
Synonyms: R. l. angolensis (Seabra, 1898),
Angola, Hahna.
Type locality: Mozambique, Tete (= Tete Province),
Sena village, south bank of the Zambesi River, -15.679°
S, 33.809° E.
Syntypes: ZMB 375 (female, poor skin only), ZMB 2496
(female, skin in alcohol, skull extracted, broken), ZMB
24922 (adult male, complete skeleton), ZMB 24927
(female, complete skeleton), collected between 1843
and 1847 by W. C. H. Peters from Mozambique, Tette (=
Tete Province), Sena on the south bank of the Zambesi
River, -15.679° S, 33.809° E.
Referred specimens having molecular identifications:
FMNH 228936 (SMG 18959), male, 228937 (SMG
18988), female, 228938 (SMG 18989), male, 228939
(SMG 18990), male, 228940 (SMG 18991), female,
228941 (SMG 18992), male, all collected on 2 May 2015
by S. M. Goodman, M. C. Schoeman and G. le Minter
from Mozambique, Chihalatan Cave, 38.2 km E of
Inhassoro, -21,671° S, 34,864° E. FMNH: 228947 (SMG
19022), female, 228954 (SMG 19029), female, 228965
(SMG 19040), female, all collected on 2 May 2015 by
S. M. Goodman, M. C. Schoeman and G. le Minter
from Mozambique, Mozambique, Inhambane Province,
Malashane Cave, 39.1 km E of Inhassoro, -21,668° S,
34,847° E, and situated <2 km from Chihalatan Cave
referred to above. DM13905 (female), 13916 (female),
DM 14531 (male), all collected by A. Monadjem, G. le
Minter and E. Lagadec in July 2015 from Mozambique,
Sofala Province, Gorongosa National Park.
Referred specimens having non-molecular
identifications: DM 13894 (female), 13927 (male),
13938 (unknown sex), 13942 (unknown sex), 13943
(unknown sex), 13944 (unknown sex), 13945 (male),
Figure 12. Portraits of: A, the lateral facial profile showing the connecting process (marked by arrow); B, general noseleaf
morphology; and, C, in flight behaviour of Rhinolophus gorongosae sp. nov. (Photographs by P. Naskrecki.)
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 23
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Figure 14. Colony of R. lobatus showing present of orange apical tuft in flying males (Photographs by P. Naskrecki.)
Figure 13. Portraits of: A, the lateral facial profile showing the connecting process (marked by arrow); B, general noseleaf
morphology; and, C, in flight behaviour of Rhinolophus lobatus. (Photographs by P. Naskrecki.)
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24 P. J. TAYLOR ET AL.
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
14529 (male), 14531 (male), 14532 (female), collected
by A. Monadjem, G. le Minter and E. Lagadec in July
2015 from Mozambique, Sofala Province, Gorongosa
National Park. TMSA 49114 (female), TMSA 49117
(male), collected from Mozambique, Sofala Province,
Gorongosa National Park by A. J. Guyton in July
2015 from Chitengo Camp and Cheringoma Plateau
respectively. TMSA 14653 (female), 14655 (unknown
sex), 14656 (female), 14657 (female), 14662 (male),
14663 (male), 14664 (female), collected 28 – 29 July
1964 from Mozambique, Tete Province, Muchena,
-15.679° S, 33.809° E. Incertae sedis: DM 8575 (male)
from Mozambique, Cabo Delgado Province, Pemba
Island (-13.006° S, 40.524° E) and DM 8574 (female),
from Mozambique, Sofala Province, Chinizuia Forest
(-18.977° S, 35.052° E), both collected by A. Monadjem
in June 2006. TMSA 49115 (male), TMSA 49118
(male), collected from Mozambique, Sofala Province,
Gorongosa National Park by A. J. Guyton in July
2015 from Chitengo Camp and Cheringoma Plateau,
respectively.
Etymology: The Latin word lobatus means lobed,
perhaps referring to the general shape of the noseleaf
(Fig. 13).
Re-diagnosis and comparisons: This taxon is clearly
phylogenetically distinct on Cytb gene sequences (see
also: Dool et al., 2016) from topotypic landeri from
West Africa and seems to be more closely affiliated
with the capensis group of Csorba et al. (2003), i.e.
R. capensis, R. denti, R. simulator and R. swinnyi
(Fig. 2). Specimens from Chihalatan and Malashane
(N = 4) had a mean echolocation call peak frequency of
106.8 ± 0.4 kHz (Table 2), close to the 107 kHz generally
reported for southern Africa ‘landeri’ (Monadjem et al.,
2010). This contrasts with a mean maximum frequency
of 104.3 ± 0.42 kHz (N = 2) recorded for topotypic
West African (Liberian) R. landeri, making a peak
frequency of c.107 kHz a possible diagnostic criterion
for R. lobatus. However, a wider range (102–108 kHz)
and slightly lower mean frequency of 105 kHz was
reported for 20 individuals at Gorongosa (Table 2).
This variation could be indicative of further undetected
cryptic speciation in this taxon, as also indicated
by the very wide variation in baculum and noseleaf
characters, discussed in the following paragraph.
Although not easily distinguished on body or skull
size (Table 2; Fig. 4), this taxon differs from topo-
typic R. landeri by displaying a prognathic rostrum
(LMs 1–3), a shortened braincase that extends pos-
teriorly (LMs 10–14) and a broader nasal inflation
(LMs 16–18) (Figs 5, 6). Specimens of cf. lobatus from
Pemba (Mozambique) had a shorter foramen magnum
(LMs 9–10), a narrower jugal process (LM 3–4) and
a slightly broader nasal inflation (LMs 16–18), than
other Mozambique animals. The same individual from
Pemba possessed a distinctive robust baculum (Fig. 8)
and noseleaf (Fig. 9) having a markedly hastate lancet
and pronounced posterior lobe of the connecting pro-
cess. However, pending molecular data, we here refer
them to lobatus incertae sedis. Although genetically
clearly assigned to lobatus, individuals from Gorongosa
National Park are somewhat smaller in external (but
not cranial) measurements (Table 2), and also display
unique noseleaf and bacular morphologies. The nose-
leaf of a Gorongosa individual has a small hastate lan-
cet and bluntly pointed tip, and a relatively small and
more erect connecting process with semi-symmetrical
lobe (Figs 9, 13), while its baculum is highly divergent
from all other forms considered in our study (Fig. 8),
having a remarkable short baculum with wide shaft
and very small base compared with landeri from West
Africa, as well as cf. lobatus from Malashane and
Chihalatan Caves. Given the clear evidence for close
genetic identity in our study between Malashane and
Chihalatan Caves and Gorongosa National Park for
animals referred to R. lobatus, we provisionally regard
this variation in noseleaf and baculum structure to
represent polymorphic traits in this species. It is pos-
sible that these morphological types represent good
species between which introgression has occurred.
Further analysis with additional nuclear sequences is
necessary to test this hypothesis. There is also the pos-
sibility that these could represent subadults with bac-
ula that are not fully ossified, as detailed in R. adami
described by Kock et al. (2000).
Male R. lobatus from Chihalatan and Malashane
Caves, as well as Gorongosa National Park, typically
possessed a dark brown apical tuft of stiff hairs char-
acteristic of R. landeri (Fig. 14; Csorba et al., 2003;
Monadjem et al., 2010). This convergent character
could be one of the contributing factors that have led
to the historical misidentification of the species with
R. landeri.
Distribution and biology: As anticipated by Monadjem
et al. (2010), who considered that the southern (and
possibly eastern) African savannah-occurring lobatus
might prove to be distinct from the West African
forest-occurring landeri, we here refer all southern
African specimens to lobatus (Fig. 11) [see appendix of
Monadjem et al. (2010) for a full list of localities]. We
agree with Monadjem et al. (2010) that R. angolensis
Seabra, 1989 from western Angola may merit specific
status. Nevertheless, in the absence of the type
series that was destroyed in the Lisbon fire of 1978,
resolution of its status must await a detailed revision,
including molecular evidence with designation of new
type material. Likewise, the status of east and central
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THREE NEW CRYPTIC SPECIES OF RHINOLOPHUS 25
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
African populations must await further studies,
including molecular data. Until further evidence
becomes available, we suggest that it is prudent to
use the name lobatus for all savannah populations in
southern and east Africa.
Rhinolophus swinnyi gough, 1908
swinny’s horseshoe bat
Synonyms: Rhinolophus swinnyi piriensis Hewitt,
1913, South Africa, Eastern Cape Province, Pirie, near
King Williams Town.
Types: TMSA 1021 (holotype, male), 1022 (cotype,
male), collected on 22–23 February 1908 by H. H.
Swinny.
Type locality: South Africa, Eastern Cape Province,
Pondoland, Ngqeleni District, -31.667° S, 29,033° E.
Referred specimens having molecular identifications:
None.
Referred specimens having only morphological
identifications: All specimens originate from South
Africa. DM 7084, adult male collected by P. J. Taylor
on 3 March 2001 and DM 13250, 13252 and 13254
(all of unknown sex), collected by E. J. Richardson on
2 November 2010 from the Eastern Cape Province,
Insizwe Mine, (-30.804° S, 29.281° E); DM 14036
(male), collected by M. C. Schoeman on 9 May 2012
from the KwaZulu-Natal Province, Pietermaritzburg,
Ferncliff Nature Reserve, (-29.550° S, 30.320° E);
DM 14291 (female) and 14292 (female) collected by
L. R. Richards on 1 April 2014 from the KwaZulu-
Natal Province, Eshowe, Entumeni Nature Reserve,
(-28.886° S, 31.376° E); DM 14441 (female) and 14440
(male) collected by S. Stoffberg and M. C. Schoeman
on 23 July 2004 from Eastern Cape Province, Kokstad
Mine, (-30.810° S, 29.280° E); DM 15018, adult female
collected by L. R. Richards on 7 April 2016 from
Eastern Cape Province, Sandile’s Rest Trout and
Forest Country Estate, (-32.661° S, 27.298° E); TMSA
39848, adult male collected by G. Bronner on 14 March
1988 from KwaZulu-Natal Province, Ngome Forest
Reserve, (-27.833° S, 31.413° E).
Re-diagnosis and comparisons: Gough’s (1908) original
description emphasized inter alia, the very small size
of the species, the ‘mouse = grey’ colour, the position
of the anterior premolar within the toothrow, the
medium-sized ears, the shape of the connecting process
(‘forming a marked projection, rounded terminally’),
the parallel-sided edges of the sella and the moderate
lancet with strongly concave edges. Rhinolopus swinny
s.s., as defined here, confined to the Eastern Cape,
KwaZulu-Natal and possibly Mpumalanga Provinces,
South Africa, is clearly distinct on molecular grounds
from R. rhodesiae and R. gorongosae sp. nov., and
has as its closest relative, R. capensis with which it
overlaps in range in the Eastern Cape. The echolocation
peak frequency of R. swinnyi s.s. calls recorded in
the Eastern Cape [mean 106.6 ± 0.4 kHz, N = 10;
Schoeman & Jacobs (2008; Table S1)] and KwaZulu-
Natal (mean 105.6 ± 0.76 kHz; this study) Provinces,
South Africa overlap considerably, but differ clearly
from the 100 kHz calls of R. rhodesiae from central and
northern South Africa and Mozambique (and probably
more broadly).
Although R. swinnyi s.s. overlaps in external and
cranial characters with R. rhodesiae, it can be distin-
guished from the latter taxon based on both baculum
and noseleaf characters, as well as on detailed cranial
shape analysis (see above). It can be differentiated from
R. gorongosae sp. nov. on its larger size (e.g. condyle-
canine skull length 14.8–15.2 mm in R. gorongosae sp.
nov., 14.3–16.0 mm in R. swinnyi; zygomatic width
8.13–8.56 mm in R. gorongosae sp. nov., 8.7–9.2 mm
in R. swinnyi; Table 2), as well as on noseleaf, bacular
and subtle cranial shape characters (see above).
Distribution and biology: We here restrict the
distribution of R. swinnyi s.s. to South Africa, including
the Eastern Cape and KwaZulu-Natal Provinces and
possibly Mpumalanga Province (Fig. 11). However, a
recent specimen collected from the montane western
region of Swaziland has tentatively been assigned
to R. rhodesiae based on morphology and acoustics
(maximum frequency = 102 kHz), but a molecular
analysis has yet to be conducted (A. Monadjem,
unpublished data); this suggests that R. rhodesiae (and
not R. swinnyi) occurs to the north in Mpumalanga
Province. Specimens from Limpopo Province in the
north of South Africa are referred to R. rhodesiae. As
pointed out above, swinnyi and rhodesiae co-occur
at at least one locality, Ferncliff Cave, in central
KwaZulu-Natal.
ACKNOWLEDGEMENTS
We thank Professor Christiane Denys for access to the
collections of the Muséum National d’Histoire Naturelle
in Paris, and Dr Frieder Mayer for access to the
Zoologisches Museum in Berlin. Julian Bayliss provided
specimens from Mount Mecula, Mount Mabu and Mount
Inago in Mozambique. Two field trips to the Inhassoro
District of Mozambique in 2015 were financed by a
grant from FEDER, under the name POCT MOZAR, to
Centre de Veille sur les maladies émergentes en Océan
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26 P. J. TAYLOR ET AL.
© 2018 The Linnean Society of London, Zoological Journal of the Linnean Society, 2018, XX, 1–28
Indien (CRVOI). Gildas le Minter, Erwan Lagadec and
Charles Gumbi are thanked for help with fieldwork in
Mozambique. PJT acknowledges the financial support
of the University of Venda, the South African National
Research Foundation and the Department of Science
and Technology under the South African Research
Chair Initiative (SARChI) on Biodiversity Value and
Change within the Vhembe Biosphere Reserve hosted
at University of Venda and co-hosted by the Centre for
Invasion Biology at University of Stellenbosch. For assis-
tance with obtaining research and shipping permits we
acknowledge Gorongosa National Park, Department of
Scientific Services; Universidade Eduardo Mondlane,
Museu de História Natural; South African Department
of Agriculture, Forestry and Fisheries, and eZemvelo
KZN Wildlife.
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SUPPORTING INFORMATION
Additional Supporting Information may be found in the online version of this article at the publisher’s web-site:
Table S1. Sampling localities and specimens or Genbank sequences sampled for molecular (Mol), traditional
(TM) and geometric (GM), morphometric and bacular (Bac) analysis. Abbreviations of museums provided in text.
Bold face indicates type specimens
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