Using genetics and morphology to examine species diversity of old world bats: report of a recent collection from Malaysia

2008·16.1 MB·English

by  Khan Anwarali

Using Genetics and Morphology to Examine Species Diversity of
Old World Bats: Report of a Recent Collection from Malaysia
Faisal AliAnwarali Khan, Vicki J. Swier, Sergio Solari, Peter A. Larsen, Besar Ketol, Wahap Marni,
Sivanathan Ellagupillay, Maklarin Lakim, Mohammad Tajuddin Abdullah, and Robert J. Baker
Abstract
A three-week field survey was conducted to sample bat species diversity in Malaysia.
The survey sampled five localities during August 2006, including one in Peninsular Malaysia
and four in Borneo. A total of 259 specimens representing at least 50 species were recorded.
Three sampling nights were spent at each locality; four harp traps and 15 mist nets were set
most nights. The highest species diversity was recorded in Mount Penrisen (Borneo) with 27
species, whereas the lowest diversity was at Monggis Sub Station and Bako National Park
(Borneo) with 13 species each. We added four new geographic records for Sarawak, Malaysian
Borneo: Arielulus cuprosus, Hesperoptenus tomesi, Kerivoula lenis, and Myotis adversus; and
the second record of Hipposideros doriae in Sarawak. Other findings included a colony of
earwigs collected from an Eonycteris major and the documentation of highly developed facial
glands on male Hipposideros ater and H. galeritus. Species were identified using traditional
skin and skull characteristics as well as mitochondrial cytochrome-# gene sequences that were
compared to existing GenBank records. This multifaceted approach, using genetic and mor¬
phological identifications, has provided greater resolution to species identification as well as a
better understanding of the diversification of Old World Bats from Malaysia. Although in most
cases genetic data validated the present taxonomy, there were still a number of cases of cryptic
species suggesting an underestimation of the actual biodiversity. This study has demonstrated
the value of genetic data in providing an independent test for species boundaries among the 50
species of bats collected (about 40% of the known Malaysian bat fauna).
Key words: Borneo, Chiroptera, cytochrome-#, diversification, Peninsular Malaysia
Introduction
The first faunal list of bats from Malaysia (includ¬ ment of capture techniques (e.g., harp traps; Francis
ing Peninsular Malaysia, Sabah, and Sarawak) was 1989), and the resolving power (to delimit species) of
published by Chasen (1940) and contained 84 species. methodologies and species concepts for species recog¬
Several subsequent compilations listing the Malaysian nition (e.g., DNA sequencing: Avise and Walker 1999;
chiropteran fauna were published, each with alternative Bradley and Baker 2001; Baker and Bradley 2006).
numbers of bat species (80 spp. in Malaysia and Sin¬ Given this, it is clear that an integrated approach using
gapore, Medway 1969; 92 spp. in Borneo, Payne et al. morphology and genetics has the potential to provide
1985; 307 spp. within the Indomalayan range, Corbet better resolution of species and species boundaries (e.g.,
and Hill 1992). The most recent taxonomic synthesis Francis et al. 2007; Mayer et al. 2007). It is effective
identified 125 species with Malaysian distributions to incorporate these methods into field surveys aimed
(Simmons 2005). at describing species diversity.
Comparing the number of species compiled with¬ We assigned species level names to our specimens
in these faunal lists provides insight into the advance¬ through comparison of morphology to published spe-
2 Occasional Papers, Museum of Texas Tech University
cies descriptions, as well as by comparing the sequences history collections of the Texas Tech University Mu¬
of a portion of the mitochondrial cytochrome-# gene seum, with a focus on mammals and genomic resources.
to those deposited in GenBank. A goal of these expeditions was to provide students
an educational experience through hands-on scientific
Herein, we report our results for the 2006 Ma¬ studies as well as exposure to unique ecosystems and
laysian Sowell Expedition. Since 2001, Mr. James E. cultures (see inside front cover).
Sowell has funded expeditions to enhance the natural
Materials and Methods
Sampling sites.—The expedition by Texas Tech is the oldest national park in Sarawak. It is located
University (TTU) and the Universiti Malaysia Sarawak 37 km northeast of Kuching city and includes 27 km2
(UNIMAS) to Peninsular Malaysia and Borneo was of the Muara Tebas Peninsula. This park is bordered
conducted from 3 to 19 August 2006. Sampling locali¬ to the north by the South China Sea with the coastline
ties (Fig. 1) are as follows: providing secluded beaches and rock formations. Bako
National Park is home to at least 34 species of bats
Krau Wildlife Reserve, Pahang, Peninsular (Anwarali et al. 2007). Seven different vegetation
Malaysia (03°35'45.6"N, 102°10'54.2"E; elevation 72 types are present: mixed dipterocarp forest, riverine
m; 3-6 August).—This is likely the most extensively or alluvial forest, heath forest, mangrove forest, beach
studied site for bat diversity in Peninsular Malaysia vegetation, cliff vegetation, and shrubland (Hazebroek
(e.g., Zubaid 1993; Hodgkison 2001; Campbell 2004; and Abang Kashim 2000).
Kingston et al. 2006) as it is part of a large research site
maintained by the Department of Wildlife and National Kubah National Park (Kubah NP), Sarawak,
Parks (DWNP). This locality is reported to have the Malaysian Borneo (01°36'42.3"N, 110°26,39.3ME;
highest diversity of insectivorous bats in the tropics, elevation 8 m, 14-16 August).—This park is located
with more than 52 species from a single 3 km2 area approximately 22 km southeast of Kuching city and
surrounding the Kuala Lompat field station (Kingston covers about 22 km2. Mount Serapi (911m) is Kubah’s
et al. 2006). In total, there are at least 69 species of highest point. Kubah is widely covered by mixed
bats in the reserve. Our study site consisted mainly of dipterocarp forest and is well known for its frog pond,
undisturbed dipterocarp forest and a small stream. which includes a high diversity of frog species (Haze¬
broek and Abang Kashim 2000). This small stagnant
Monggis Sub Station of Kinabalu National Park, pond was one of our major mist netting sites.
Sabah, Malaysian Borneo (06°12T.5"N, 116°45,7.5,,E;
elevation 330 m; 7-10 August).—This research station Mount Penrisen, Sarawak, Malaysian Borneo
is an extension of Kinabalu National Park that was not (01°07'41.4"N, 110°13'06.7"E; elevation 857 m, 17-19
open to visitors at the time of our survey. Our sampling August).—Mount Penrisen, locally known as Borneo
was part of a collaborative work between the Kinabalu Heights and located within the Padawan district, is ap¬
Park management and UNIMAS to provide a check¬ proximately 70 km south of Kuching. This site was
list of small mammals that occur in the vicinity of the the highest elevation surveyed during our expedition.
station. The majority of this site consisted of mixed Most of the vegetation at Penrisen was covered by
dipterocarp forest with intermittent thickets of bamboo primary mixed dipterocarp forest with patches of sec¬
and small streams. ondary forest (near cleared areas for logging and road
construction purposes). Our two major collecting sites
Bako National Park, Sarawak, Malaysian Borneo at this location were near the edge of a newly developed
(01°42'59.5"N, 110°26'39.3"E; elevation 8 m; 11-13 golf course and within the primary forest located on a
August).—Established in 1957, Bako National Park plateau near the Sarawak-Kalimantan border.
Anwarali Khan et al.—Species Diversity of Malaysian Bats 3
Figure 1. Map in upper right corner showing South East Asia with Peninsular Malaysia (A) and
Malaysian Borneo (B) in gray The five collecting localities described herein are Peninsular Malaysia:
1 = Krau Wildlife Reserve; and Malaysian Borneo: 2 = Monggis Sub Station; 3 = Bako National
Park; 4 = Kubah National Park; 5 = Mount Penrisen.
4
Occasional Papers, Museum of Texas Tech University
Field methods.—Fifteen mist nets (2,6,9,12, and other families). PCR products were purified using the
30.5 m long) and four sets of four-bank harp traps were QIAquick PCR Purification Kit (Qiagen Inc., Chats-
set most nights. Nets and traps were set by 1730 h, and worth, California). DNA sequencing was performed
closed between 0200 h and 0700 h the following day. using ABI Big Dye chemistry chain terminators version
Nets were checked every 20 minutes during the first six 3.1 and fragments were electrophoresed on an ABI
hours, and then checked every 2-3 hours before closing 3100-Avant Genetic Analyzer (PE Applied Biosystems,
them the next day. Harp traps were checked three to Foster City, California). The first 404 or 450bp of the
four times each night. Scoop, hand nets, and short nets cyt-6 gene were sequenced in this study using L14724
(2 m) were used to collect specimens at several roosting (Irwin et al. 1991) and the internal primer MVZ04
locations, especially within culverts. (Smith and Patton 1993). Additionally, we used three
internal primers: HF041 (5’-CCT-CAR-AAR-GAT-
Specimens were identified in the field following ATT-TGG-CCT-CAT-GG-3 ’) for the genera Eonycteris,
identification keys by Payne et al. (1985), Corbet and Penthetor, Rhinolophus, and Emballonura; HF042
Hill (1992), and Kingston et al. (2006). After assign¬ (5,-RAA-RGA-YAT-YTG-TCC-TCA-TGG-3’) for
ing species identification, three individuals per species Hipposideros and Megaderma', and EPH520 (5’-RAA-
were taken as voucher specimens. An additional three KGG-GAG-TAG-AAA-GTG-GAA-GGC-3 ’) for spe¬
specimens were collected for species that exhibited cies of Vespertilionidae and Miniopterus as alternative
substantial morphological variation suggesting more primers for MVZ04 (Smith and Patton 1993).
than a single species existed within the sampled popu¬
lation. Standard measurements were taken from each Validating field identifications.—Field identifica¬
individual, which was then prepared as a museum tions of specimens were re-evaluated using skin and
voucher specimen, either as a skin and skeleton or as skull characteristics which were compared to published
a fluid preserved specimen. Tissue samples from liver descriptions as well as cyt-6 gene sequences. Se¬
and muscle were preserved in lysis buffer (Longmire quences were viewed in a Neighbor Joining (NJ) tree
et al. 1997) and 95% ethanol, and blood samples were to confirm the genetic distinction of each species as
collected using Nobuto blood filter strips (Advantec well as to assess variation among conspecifics through
Inc.). Phoretic insects and ectoparasites found on the genetic distances. Genetic distances were calculated
specimens were preserved in 70% ethanol. Karyo¬ following the Kimura 2-parameter model using MEGA
types were prepared in the field (Baker et al. 2003) for software version 4.0 (Kumar et al. 2004). Specimens
individuals from each species. Animals were handled identified as conspecifics that exhibited high genetic
following the standards of the American Society of divergences (> 5%), were further verified by examin¬
Mammalogists (Gannon et al. 2007) and the Texas Tech ing external and cranio-dental characters (e.g., Payne
Animal Care and Use Committee (Permit # 02217-02). et al. 1985; Corbet and Hill 1992; Francis 2008). This
Voucher specimens and duplicates of tissue samples combination of data allowed the documentation of the
(liver and muscle) were deposited in the Natural Sci¬ genetic variation within putative species identified us¬
ence Research Laboratory, Museum of Texas Tech ing morphology, an operational method suggested for
University (NSRL-TTU), Natural History Museum of application of the Genetic Species Concept (see Bradley
Universiti Malaysia Sarawak (UNIMAS), Institute of and Baker 2001; Baker and Bradley 2006). Under this
Biological Diversity, Bukit Rengit, Pahang (IBD), and operational method, a genetic distance of > 5% in the
Kinabalu National Park (Kinabalu NP). cyt-6 gene is used as an indicator for speciation events
because many sister species of mammals described
Molecular methods.—Methodology for DNA based on morphological variation have >5% genetic
extraction and Polymerase Chain Reaction (PCR) fol¬ distance value. We interpret this as indicating that a >
lowed Larsen et al. (2007). The entire cytochrome-6 5% genetic distance value for the cytochrome-b gene
(cyt-6) gene was amplified using primer pair L14724- sequence data documents that two clades may have
H15915r (Irwin etal. 1991). The optimized annealing been isolated from each other for sufficient time to
temperature for each bat family was determined using have speciated by the Bateson-Dobzhansky-Muller
gradient PCR at the following temperatures: 40.2°C model (Baker and Bradley 2006). We provide GenBank
(Rhinolophidae, Vespertilionidae), and 50.0°C (all accession numbers for representatives of each species
Anwarali Khan et al.—Species Diversity of Malaysian Bats 5
reported herein. Genetic divergences associated with phylogenetic and phylogeographic analyses will be
geographic variation between Peninsular Malaysia and provided in subsequent publications.
Bornean specimens are discussed briefly, as detailed
Species Accounts
Species accounts were compiled using vari¬ (1985), Corbet and Hill (1992), Abdullah et al. (1997;
ous published data as well as UNIMAS, DWNP, and 2003), Yasuma and Andau (2000), Csorba et al. (2003),
Southeast Asian Mammal Databank databases (Biotani Hall (2004), Kingston et al. (2006), Francis (2008),
et al. 2006). Natural history and species designations Southeast Asian Mammal Databank (SAMD), unpub¬
presented herein were reviewed for each species based lished UNIMAS Museum records, and unpublished
primarily on the data derived from our identifications. DWNP specimen records.
Unless otherwise noted, the taxonomic arrangement
herein follows Simmons (2005). Main departures Remarks.—Species group or tribe was noted
from that work include Csorba et al. (2003) for Rhi- when available. Key diagnostic features that distin¬
nolophidae and Hoofer and Van Den Bussche (2003) guish the species from similar congenerics, the type of
for Vespertilionidae, including the recognition of the traps used, and collecting habitats (also derived from
family Miniopteridae. Common names in English were Payne et al. 1985; Corbet and Hill 1992; Csorba et al.
from Simmons (2005) whereas Malay names were from 2003; Kingston et al. 2006; Francis 2008) are included
Medway (1969) and Khan (1992), and are included in in this section. Other information involving genetics
parentheses. Information was organized into the fol¬ and/or morphology was included where appropriate.
lowing categories: Status of reproduction for females was: lactating,
post lactating, with embryo (pregnant), or carrying a
Specimens examined.—Total number of speci¬ juvenile. The following abbreviations were used: FA=
mens examined given in parentheses, followed by forearm; Mt. = Mount; and NP = National Park.
locality and TTU catalogue numbers (for voucher
specimens, tissues, and karyotypes that are deposited Family Pteropodidae
at NSRL) or TK numbers (for tissues and karyotypes at Aethalops aequalis G. M. Allen 1938
NSRL but voucher specimens deposited at IBD, Kina¬ Borneo Fruit Bat (Cecadu Bukit)
balu NP, or UNIMAS). For each species, a GenBank
accession number was assigned to one specimen (in Specimens examined (5).— Mt. Penrisen: TTU
parentheses following a TTU or TK number). 108348 (GenBank EU521595), TTU 108352, TTU
108357, TTU 108409, TTU 108381.
Type locality.—The known type locality of spe¬
cies according to country, region, state (if available), Type locality.—Malaysia (Borneo), Sabah,
and specific locality (if available) is described. Mount Kinabalu.
Malaysian distribution.—Species distribution in Malaysian distribution.—Recorded from high¬
Malaysia is described in the following order: country, lands in Sabah and Sarawak. Specimens were collected
region, state (if available) and specific locality (if avail¬ from Mt. Penrisen in this study.
able). State and specific localities are not provided for
species that are widely distributed either in Peninsular Remarks.—Aethalops aequalis is commonly
Malaysia or Borneo. We provided specific localities found at higher elevations in hill and mountain forests
for those species that do not have many collection (Payne et al. 1985). This species was caught using
records, and specific localities for those species with mist nets across small streams. Compared to other
new records from this study to assist in future faunal fruit bats, A. aequalis is fragile and requires careful
surveys. Distribution in Malaysia was retrieved from handling and quick release if it is not to be prepared
the following sources: Lim et al. (1974), Payne et al. as a voucher specimen. This species was previously
6 Occasional Papers, Museum of Texas Tech University
recognized as Aethalops alecto (Payne et al. 1985). Balionycteris seimundi (Kloss 1921)
Studies (Kitchener et al. 1993) suggest that only Pen¬ Malayan Spotted-winged Fruit Bat
insular Malaysia populations, as well as Sumatra, Java, (Cecadu Sayap Bertitik Malaya)
Bali, and Lombok populations, should be recognized as
alecto (Pygmy Fruit Bat) whereas Borneo populations Specimens examined (2).—Krau: TK 152011
are referable to aequalis. Although morphological (GenBank EU521597), TTU 108175.
characters differentiate aequalis from alecto, allozy-
mic variation between them is low (see Kitchener et Type locality.—Malaysia (Peninsular Malaysia),
al. 1993). Cytochrome-^ gene sequences within our Pahang, Gunung Tahan.
sample from Borneo averaged 0.2%. Pending genetic
data from Peninsular Malaysia populations as well Malaysian distribution.—Recorded from Penin¬
as specimens from Sabah, we follow Kitchener et al. sular Malaysia (all states).
(1993) for geographic boundaries of A. alecto and A.
aequalis. Remarks.—Balionycteris seimundi represents the
smallest fruit bat of Peninsular Malaysia (FA: 39-43
Balionycteris maculata (Thomas 1893) mm; weight: 13-14.1 g). We propose that the common
Spotted-winged Fruit Bat (Cecadu Sayap Bertitik) name for this species be the Malayan Spotted-winged
Fruit Bat in English and Cecadu Sayap Bertitik Ma¬
Specimens examined (7).—Bako NP: TTU laya in Malay. See B. maculata species account for
108265 (GenBank EU521596), TTU 108266, TTU details.
108299; KubahNP: TTU 108312, TTU 108317, TTU
108321; Mt. Penrisen: TTU 108364. Cynopterus brachyotis Muller 1838
Lesser Short-nosed Fruit Bat (Cecadu Pisang)
Type locality.—Malaysia (Borneo), Sarawak.
Specimens examined (18).—Monggis: TTU
Malaysian distribution.—Recorded from Bor¬ 108245 (GenBank EU521598), TTU 108246, TK
neo. 152094, TTU 108247, TTU 108248, TTU 108257,
TTU 108258, TTU 108259; Bako NP: TTU 108280;
Remarks.—Balionycteris maculata is the smallest KubahNP: TTU 108320, TTU 108323, TTU 108324,
fruit bat in Borneo (FA: 40-45 mm; weight: 10-15 g). TTU 108330, TTU 108338; Mt. Penrisen: TTU 108411,
This species was caught using mist nets near rock cliffs TTU 108401, TTU 108402, TTU 108382.
in mangrove forests, over ponds, and in open areas near
hill edges. A diagnostic feature of this bat is the obvious Type locality.—Dewei River in Borneo, also
white to pinkish spots on its wings. These frugivorous known as Tewei River in Naga Cave.
bats are commonly found in lowland dipterocarp forest
and occasionally at higher elevations. Two subspecies, Malaysian distribution.—Recorded from Penin¬
B. maculata seimundi and B. maculata maculata, have sular Malaysia and Borneo from sea level to 1600 m.
been recognized in Peninsular Malaysia and Borneo,
respectively (Kloss 1921), a taxonomic arrangement Remarks.—Cynopterus brachyotis was netted
followed by many authors (e.g., Payne et al. 1985; mainly in primary forest, secondary forest, and agricul¬
Hodgkison and Kunz 2006; Francis 2008). We found tural areas. We identified two forms (large FA: 60-68
a genetic distance of 12% in the cyt-b gene sequences mm; small FA: 54-59 mm) of C. brachyotis (reported
between populations in Borneo and Peninsular Ma¬ in Abdullah 2003; Campbell et al. 2004), but these
laysia. Following application of the 5% species level size groups were not congruent with mitochondrial
separation proposed by Baker and Bradley (2006), DNA data. The level of cyt-b genetic distances within
and the morphological variation between Kalimantan Peninsular Malaysia (10.3%) and Borneo (9.5%) sug¬
(Indonesian Borneo) and Sumatra (Indonesia) popula¬ gested two independent lineages; however, both genetic
tions (Maryanto 2003), we recognize these populations lineages include individuals that overlap in forearm
as separate species. sizes.
Anwarali Khan et al.—Species Diversity of Malaysian Bats 7
Cynopterus horsfieldii Gray 1843 ond digit of the wing. We caught Eonycteris major at
Horsfield’s Fruit Bat (Cecadu Pisang Besar) Kubah NP in a net placed in front of a banana tree, and
in Mt. Penrisen this species was caught at the edge of a
Specimens examined (9).—Krau: TK 152002 steep cliff where bats were flying near stands of banana
(GenBank EU521599), TK 152003,TTU 108170,TTU trees. This species is the largest nectar-feeding bat in
108212, TTU 108214, TTU 108215, TK 152058, TTU Malaysia, with a long muzzle that is slender and curved.
108223, TTU 108224. Eonycteris major also has an enlarged gland near the
anus. Forearm length distinguishes E. major (71-80
Type locality.—Indonesia, Java. mm) from the smaller E. spelaea (62-70 mm; Payne
et al. 1985). Both of these species roost in limestone
Malaysian distribution.—Recorded from Penin¬ caves (SAMD database). Low cyt-b genetic distance
sular Malaysia (all states) and Borneo from sea level (0.1%) was observed among Bornean individuals. A
to 1400 m. specimen, TTU 108410 (18 August 2006; Mt. Pen-
risen), was carrying earwigs (Scizochelisoches sp.; Fig.
Remarks.— This species was caught using mist 2) on its dorsum near the base of the tail. This is the
nets at Krau across forest trails. In Borneo, C. horsfiel¬ first record for earwigs on this bat species. Specimens
dii was captured at high elevations (900 m) in primary of these earwigs are housed in the NSRL.
forest (Jayaraj et al. 2005; 2006). Forearm length (68-
76 mm) is a good character to separate this species from Eonycteris spelaea (Dobson 1871)
C. brachyotis in the field (Payne et al. 1985). Unlike Lesser Dawn Bat (Cecadu Gua Kecil)
C. brachyotis, there is low cyt-b genetic distance (2%)
observed within and between Peninsular Malaysian Specimens examined (7).—Krau: TK 152009
(samples from this study) and Bornean (samples from (GenBank EU521601), TTU 108211; Monggis: TTU
UNIMAS museum collections) individuals. Habitats 108240, TK 152091; Mt. Penrisen: TTU 108373, TTU
where C. horsfieldii occurs in Borneo should be given 108378, TTU 108387.
high conservation priority, as this species is rare or at
least not commonly reported when compared to Pen¬ Type locality.—Burma, Tenasserim, Moulmein,
insular Malaysia samples. Two of four adult female Farm Caves.
specimens were pregnant: TK 152002 (3 August 2006)
and TK 152058 (6 August 2006); and two were carrying Malaysian distribution.—Recorded from the low¬
young: TK 152003 (6 August 2006) and TTU 108223 lands of Peninsular Malaysia (all states) and Borneo.
(7 August 2006).
Remarks.—We netted this species near banana
Eonycteris major K. Andersen 1910 trees. Eonycteris spelaea can be distinguished from
Greater Dawn Bat (Cecadu Gua Besar) their larger counterpart E. major by forearm length
and shorter muzzle size (see E. major species account).
Specimens examined (4).—Kubah NP: Like E. major, this species also has an enlarged ex¬
TTU108327 (GenBank EU521600); Mt. Penrisen: ternal gland lateral to the anus. Eonycteris spelaea
TTU 108371, TTU 108372, TTU 108410. is distributed throughout most of Southeast Asia and
often found in colonies of thousands in caves within
Type locality.—Malaysia (Borneo), Sarawak, agricultural areas (McClure et al. 1967; Heaney et al.
Mount Dulit. 1989). Genetic distance value of < 0.8% was observed
within Peninsular Malaysia and Borneo individuals,
Malaysian distribution.—Eonycteris major is whereas genetic distance value of 1% was observed
endemic to Borneo. between Peninsular Malaysia and Borneo individuals.
A pregnant female, TK 152085 (8 August 2006), was
Remarks.—The genus Eonycteris is distinguished recorded with an embryo length of 31 mm.
from all other fruit bats by the lack of a claw on the sec¬
8 Occasional Papers, Museum of Texas Tech University
Figure 2. The dorsal rump of a female Eonycteris major (TTU
108410) with a colony of earwigs (Order: Dermaptera: Chelisochidae;
Scizochelisoches sp.). The tail of the bat is visible in the upper right
quadrant of the photo.
Macroglossus minimus (E. Geoffroy 1810) were caught alongside C. brachyotis. Macroglossus
Dagger-toothed Long-nosed Fruit Bat minimus can be morphologically distinguished from
(Cecadu Madu Bakau) M. sobrinus by the absence of a median groove in the
upper lip, and its smaller FA length (FA < 42 mm vs.
Specimens examined (11).—Monggis: TTU FA> 42 mm; Payne et al. 1985; Francis 2008). Recent
108225 (GenBank EU521602), TTU 108226, TK literature and SAMD database indicate M. minimus oc¬
152070, TK 152071, TTU 108227, TTU 108231, TTU curs throughout Southeast Asia, whereas M. sobrinus
108232; Mt. Penrisen: TTU 108350, TTU 108366, TTU is restricted to Peninsular Malaysia, Sumatera, and
108374, TTU 108376. Java. Low cyt-b genetic distance (1%) was observed
within Bornean populations of M. minimus. A speci¬
Type locality.—Indonesia, Java. men, TTU 108366 (male), from Mt. Penrisen, Borneo,
was identified as M. minimus based on the absence of a
Malaysian distribution.—Recorded from both median groove in the upper lip and FA= 42 mm. This
Peninsular Malaysia (all states) and Borneo. individual is genetically closer to M. sobrinus rather
than M. minimus. Genetic distance between M. mini¬
Remarks.—Macroglossus minimus and M. sobri- mus (Borneo) and M. sobrinus (Peninsular Malaysia)
nus were netted near banana trees and in most cases they averaged 2% and was the lowest for any sister species
Anwarali Khan et al.—Species Diversity of Malaysian Bats 9
relationship within our data for the family Pteropodi- ears. Megaerops ecaudatus has low cyt-b genetic dis¬
dae from Malaysia. Although they may represent two tance within our Borneo samples (< 2%). Comparison
species, a genetic distance value of 2% is typical for of DNA sequences with others from GenBank showed
conspecific bat populations (Bradley and Baker 2001; high genetic distances (7% and 11%) between Penin¬
Baker and Bradley 2006) and we question the recogni¬ sular Malaysian (EF105533) and Bornean (EF105536;
tion of these geographically isolated populations as dis¬ Mount Murud) cyt-b sequences, suggesting the pres¬
tinct species. Three of our five female specimens were ence of two species.
pregnant; TK 152070 (7 August 2006), TTU 108231 (7
August 2006), and TTU 108350 (17 August 2006). Penthetor lucasi (Dobson 1880)
Lucas’s Short-nosed Fruit Bat
Macroglossus sobrinus K. Andersen 1911 (Cecadu Hitam Pudar)
Greater Long-nosed Fruit Bat (Cecadu Madu Bukit)
Specimens examined (10).—Bako NP: TTU
Specimens examined (4).—Krau: TTU 108174 108284 (GenBank EU521605), TTU 108287; Kubah
(GenBank EU521603), TK 152047, TTU 108206, NP: TTU 108313, TTU 108318, TTU 108319, TTU
TTU 108207. 108322, TTU 108329, TTU 108346; Mt. Penrisen:
TTU 108363, TTU 108377.
Type locality.—Malaysia (Peninsular Malaysia),
Perak, Gunung Igari (Mount Igari; 610 m). Type locality.—Malaysia (North Borneo), Sara¬
wak.
Malaysian distribution.—Recorded from Penin¬
sular Malaysia (all states). Malaysian distribution.—Recorded from Penin¬
sular Malaysia and Borneo.
Remarks.—Macroglosus sobrinus can be dis¬
tinguished from their smaller counterpart M. minimus Remarks.—Penthetor lucasi was netted near rock
by larger forearm length, body size, and presence of crevices (Bako NP and Mt. Penrisen) and ponds (Kubah
median groove in the upper lip (see M. minimus species NP). We verified the occurrence of this species at an
account for details). elevation (857 m) higher than the previously reported
upper limit of 600 m (Payne et al. 1985; Jayaraj et
Megaerops ecaudatus (Temminck 1837) al. 2006). Within our sample there appears to be two
Temminck’s Tailless Fruit Bat genetically distinct clusters with about 5% genetic
(Cecadu Tiada Berekor) divergence, indicating separate lineages. Sequences
in GenBank (EF 105541-EF105544) referred as Pen¬
Specimens examined (4).—Mt. Penrisen: TK thetor lucasi from Peninsular Malaysia and Borneo
152245 (GenBank EU521604), TTU 108349, TTU do not cluster with our sequences (25-27% genetic
108412, TTU 108413. divergence), suggesting misidentification as they are
closer to our Macroglossus sequences (2-4% genetic
Type locality.—Indonesia, West Sumatra, Pa- divergence). One of 10 adult female specimens, TTU
dang. 108313 (14 August 2006), was post lactating.
Malaysian distribution.—Recorded from Penin¬ Family Rhinolophidae
sular Malaysia (all states) and Borneo. Rhinolophus affinis Horsfield 1823
Intermediate Horseshoe Bat (Kelawar Ladam Hutan)
Remarks.—Megaerops ecaudatus can be netted
from lowland as well as mountain forests (Hodgkison Specimens examined (4).—Mt. Penrisen: TTU
2004). The external morphology of this species is simi¬ 108353 (GenBank EU521607), TTU 108360, TTU
lar to Cynopterus and Penthetor but with an absence 108361, TTU 108362.
of a tail in Megaerops. Other diagnostic characters
include tubular nostrils and absence of white edged Type locality.—Indonesia, Java.
10 Occasional Papers, Museum of Texas Tech University
Malaysian distribution.—Recorded from Penin¬ malayanus. Despite a wide distribution, R. borneensis
sular Malaysia (all states) and Borneo. is locally rare in Peninsular Malaysia when compared
to Borneo (SAMD). Low cyt-b genetic distance (0.3%)
Remarks.—Megaphyllus species group (Csorba was observed within Bornean populations.
et al. 2003). Rhinolophus affinis is distinguished from
other similar species (R. ferrumequinum, R. rouxi, Rhinolophus luctus Temminck 1834
and R. sinicus) by the broadly rounded connecting Woolly Horseshoe Bat (Kelawar Ladam Terbesar)
process, as well as the tall and triangular noseleaf with
a straight-sided lancet (Csorba et al. 2003; Francis Specimens examined (6).—Kubah NP: TTU
2008). All of our specimens were caught using harp 108342 (GenBank EU521609), TTU 108343, TTU
traps along a forest trail. Coloration for this cave 108345; Mt. Penrisen: TTU 108407, TTU 108408,
dwelling species ranges from bright orange to brown. TTU 108395.
Bright orange facial color on the rostrum was thought
to be a diagnostic character for this species but after Type locality.—Indonesia, Java, Tapos.
examination of several other specimens with similar
noseleaf characters and measurements, we concluded Malaysian distribution.—Recorded from Penin¬
that the orange pelage represented intraspecific varia¬ sular Malaysia (all states) and Borneo. Our record is the
tion. Approximately 4% in cyt-Z> genetic distance was second record for Mt. Penrisen after Medway (1977).
observed between Bornean individuals from this study
and a single individual from Thailand (TK 2116). Remarks.—Trifoliatus species group (Csorba et
al. 2003). In Kubah NP, this species was found roosting
Rhinolophus borneensis Peters 1861 inside a culvert, alongside individuals of R. sedulus\
Bornean Horseshoe Bat (Local name not available) whereas in Mt. Penrisen they were caught within a for¬
est along a cliff edge. Rhinolophus luctus is the largest
Specimens examined (5).—Monggis: TTU species of horseshoe bat in Malaysia with forearm size
108228 (GenBank EU521608); Kubah NP: TTU of 63-67 mm (Francis 2008).
108308; Mt. Penrisen: TTU 108385, TTU 108388,
TTU 108393. Rhinolophus philippinensis Waterhouse 1843
Large-eared Horseshoe Bat
Type locality.—Malaysia (Borneo), Sabah, (Local name not available)
Labuan Island.
Specimens examined (2).—Mt. Penrisen: TTU
Malaysian distribution.—Recorded from Pen¬ 108359 (GenBank EU521610), TTU 108380.
insular Malaysia (Perlis, Kampung Juara, and Tioman
Island) and Borneo. Type locality.—Philippines, Luzon.
Remarks.—Megaphyllus species group (Csorba Malaysian distribution.—Recorded from Borneo
et al. 2003). Rhinolophus borneensis is distinguished (Sabah: Gomantong Cave, Madai Caves, Sapulut, and
from other similar species (R. stheno, R. celebensis, Tapadong Cave; Sarawak: Bintulu, Lobang, Long
and R. malayanus) by its larger forearm size, nar¬ Lama, Niah Caves, and Puteh Caves). Our record is
rower anterior swellings of upper skull, and the me¬ new for Mt. Penrisen.
dian septa of its lancet being wider than the middle
pocket (Csorba et al. 2003; Francis 2008). In Kubah Remarks.—Philippinensis species group (Csorba
NP, we netted these bats near a small frog pond and et al. 2003). Rhinolophus philippinensis was netted
also found them roosting inside a culvert. Orange and near a pond in Mt. Penrisen. Scarcity of records in
brown color forms were present within our specimens. Borneo suggests this species may be rare. Rhinolo¬
Rhinolophus borneensis can be distinguished easily in phus philippinensis is distinguished from other similar
Borneo as they do not occur in sympatry with species species by a FA length of 48-53 mm and by a peculiar
of similar external morphology such as R. stheno or R. noseleaf structure (i.e., wide internarial region and a