Journal of Threatened
Taxa | www.threatenedtaxa.org | 26 December 2022 | 14(12): 22277–22292
ISSN 0974-7907 (Online) | ISSN 0974-7893
(Print)
https://doi.org/10.11609/jott.8040.14.12.22277-22292
#8040 | Received 07 June 2022 | Final
received 01 September 2022 | Finally accepted 02 December 2022
Systematics of the enigmatic and
narrowly endemic toad genus Bufoides Pillai
& Yazdani, 1973: rediscovery of Bufoides
kempi (Boulenger, 1919)
and expanded description of Bufoides meghalayanus (Yazdani & Chanda, 1971) (Amphibia: Anura: Bufonidae) with notes on
natural history and distribution
R.S. Naveen 1, S.R. Chandramouli 2, Gautam Kadam 3, S. Babu 4, P.V. Karunakaran
5, H.N. Kumara 6 & N. Parthasarathy 7
1,3,4,5,6 Sálim Ali Centre for Ornithology and
Natural History, Anaikatty, Coimbatore, Tamil Nadu
641108, India.
1,2,7 Department of Ecology and
Environmental Sciences, School of Life Sciences, Pondicherry University,
Puducherry 605014, India.
1 naveen89240@gmail.com, 2 findthesnakeman@gmail.com
(corresponding author), 3 gautamkadam7wild@gmail.com,
4 sanbabs@gmail.com (corresponding
author), 5 karunakaran.pv@gmail.com, 6 honnavallik@gmail.com,
7 nparthasarathypu@gmail.com
Editor: Raju Vyas,
Vadodara, Gujarat, India. Date of
publication: 26 December 2022 (online & print)
Citation: Naveen, R.S., S.R. Chandramouli, G. Kadam, S. Babu,
P.V. Karunakaran, H.N. Kumara
& N. Parthasarathy (2022). Systematics
of the enigmatic and narrowly endemic toad genus Bufoides
Pillai & Yazdani, 1973: rediscovery of Bufoides
kempi (Boulenger, 1919)
and expanded description of Bufoides meghalayanus (Yazdani & Chanda, 1971) (Amphibia: Anura: Bufonidae) with notes on
natural history and distribution. Journal of Threatened Taxa 14(12): 22277–22292. https://doi.org/10.11609/jott.8040.14.12.22277-22292
Copyright: © Naveen et al. 2022. Creative Commons Attribution
4.0 International License. JoTT allows unrestricted use, reproduction, and
distribution of this article in any medium by providing adequate credit to the
author(s) and the source of publication.
Funding: National Mission on Himalayan Studies (GBPNI/NMHS-2017-18/MG 32,
dated:28/3/2018).
Competing interests: The authors
declare no competing interests.
Author details: R.S. Naveen works on herpetofauna &
ichthyofauna and obtained his MSc in ecology from Pondicherry University. S.R. Chandramouli obtained
his PhD in ecology and environmental sciences from Pondicherry University works
on systematics, ecology, biogeography and conservation of herpetofauna of
India, with special focus on the Andaman and Nicobar Islands. Gautam Kadam is interested in the study
of spiders and herpetofauna and holds a MSc in biodiversity and wildlife
conservation from Mumbai University. S. Babu is a senior scientist at SACON, Tamil Nadu and
his research focuses on the conservation of birds, landscape ecology and remote
sensing & GIS. Honnavalli N. Kumara is
a principal scientist in SACON. His research focuses on various aspects of
conservation. Dr. P.V. Karunakaran
is a senior principal scientist at SACON, conducting biodiversity and natural
resource based studies and strategic planning in different landscapes of India.
Prof. N. Parthasarathy is a faculty at the
department of Ecology and Environmental Sciences, engaging in teaching of
ecology of terrestrial biome, forest botany, medicinal plant resources,
functional ecology, and dynamics of tropical forests across the forests of
Western and Eastern Ghats, Andamans and Coromandel coast of India.
Author contributions: SRC and SB conceived and designed
the study, RSN & GK conducted field studies, SRC, RSN, & SB generated
and analysed the data, SRC, RSN, & SB wrote the manuscript, SB, PVK, &
HNK procured funds, SB, PVK, HNK, & NP supervised the study, reviewed,
& edited the manuscript draft, and all authors approved the final draft.
Acknowledgements: This publication is an outcome of
the project entitled ‘Characterization of Community Reserves and Assessment of
their Conservation Values in Meghalaya’ funded by the National Mission on
Himalayan Studies (GBPNI/NMHS-2017-18/MG 32, dated: 28.03.2018). Our sincere
thanks are due to the additional principal chief conservator of forests
(Wildlife) and chief wildlife warden (CWLW), Meghalaya, and officers of the
Department of Forest and Environment, Government of Meghalaya for facilitating
permission from the Community Reserve Management Committees to carry out the
field study (No. FWC/Research/15/603-04 dated 31st May, 2019). We are indebted
to the management committees of each community reserves who have given us
permission for this work. We thank the technology platform services, C-CAMP,
National Centre for Biological Sciences for their expertise in sequencing a
sample used in this study. We thank Soni R Marak for his assistance with fieldwork. We thank director, SACON for smooth execution
of the project.
Abstract: Bufoides kempi (Boulenger,
1919) known only from the two historical syntype specimens until now was
rediscovered after more than a century from near its type locality in the Garo
Hills, Meghalaya, northeastern India. Analysis of mitochondrial 16S rRNA gene
reveals congenericity between B. kempi and B. meghalayanus
with an inter-specific genetic divergence of 4.67%. Description of B. kempi is expanded based on the six male and two female
specimens collected during this study. We provide the first description of
calls for this genus, notes on their breeding biology and larval morphology.
Additional specimens of B. meghalayanus
collected during this study are described to supplement its
characterization.
Keywords: Amphibians, breeding
biology, calls, Garo Hills, Khasi Hills, larval morphology, new records,
syntype specimens.
Introduction
The cosmopolitan anuran family Bufonidae Gray, 1825 is represented in India by nine genera
comprising 33 species spread across several biogeographic regions (Frost 2022).
One of the most poorly-known among them is the genus Bufoides
Pillai & Yazdani (1973) which comprises two species namely B. meghalayanus (Yazdani & Chanda 1971) and B. kempi (Boulenger 1919) (after
Chandramouli & Amarasinghe
2016). Among them, the type species, B. meghalayanus
is fairly better-known in terms of its distribution, biology, ecology, and
natural history (Yazdani & Chanda 1971; Pillai & Yazdani 1973; Das et
al. 2009; Deuti et al. 2012). On the other hand,
there has been no information on any of the above aspects for the
previously-described species B. kempi. B. kempi was originally described as Nectophryne
kempi from ‘above Tura, 2,500 ft’ based on two
specimens, an adult and a subadult (ZSI 18481a,b), from which the species is known
till date (Boulenger 1919). It was later transferred
to the genus Pedostibes Günther, 1875 by
Barbour (1938) and subsequently transferred to Bufoides
by Chandramouli & Amarasinghe
(2016) based on morphological characters. As a part of an on-going project
documenting faunal diversity in community reserves of Meghalaya, we
rediscovered B. kempi from near its
type locality and located additional specimens of B. meghalayanus
whose descriptions are expanded based on new data.
Materials
and Methods
The study was conducted in the
northeastern Indian state of Meghalaya. Surveys were conducted in ten different
locations spread across the Garo, Khasi, and Jaintia Hills of Meghalaya. Specifically, the
type localities of the two known species viz. “above Tura, Garo Hills” (Boulenger 1919) for B. kempi
and “Mawblang plateau” (Yazdani & Chanda 1971)
for B. meghalayanus and the vicinity of these
localities were surveyed intensively, in addition to the other sites to locate
the target species. Additional sites within Meghalaya (Deuti
et al. 2012), apart from these two localities from where Bufoides
is known until now were also surveyed. Field sampling was carried out from
March–May & October 2021. A total of seven specimens of B. kempi, comprising six adult males and one adult female
along with a subadult female were collected from Eman
Asakgre (25.37° N, 90.55° E, 100–250 m). Likewise,
three adult specimens of Bufoides meghalayanaus were collected from a hill stream in the
Khasi Hills (25.23° N, 91.73° E, 1,100–1,250 m). Other locations of B. kempi and B. meghalayanus
were marked with a GPS. Encounter rates of these species, expressed as the
number of individuals encountered over an hour’s duration of field sampling, is
presented as an index of their abundance.
The following measurements were
recorded to the nearest 0.02 mm from the specimens with a dial caliper:
snout–vent length (SVL, from the tip of the snout to the anterior margin of the
cloaca), axilla–groin distance (AG, from the posterior margin of the forelimb
at its insertion point on the body to the anterior margin of the hind limb at
its insertion point on the body), head length (HL, from the posterior edge of
the mandible to the tip of the snout), head width (HW, the maximum width of the
head at the angle of the jaws), head depth (HD, the maximum depth of the head),
body width (BW, the maximum width of the body at the trunk), eye diameter (ED,
the greatest horizontal diameter of the orbit), eye–nostril distance (EN, from
the anterior border of the orbit to the middle of the nostril), eye–snout
distance (ES, from the anterior border of the orbit to the tip of the snout),
upper eyelid width (UEW, the maximum width of the upper eyelid), interorbital
distance (IO, distance between the upper eyelids), internarial distance (IN,
distance between the nostrils), upper arm length (UAL, from the axilla to
elbow), lower arm length (LAL, from the posterior margin of the elbow to the
base of the outer metacarpal tubercle), palm length (PAL, from the posterior border
of the outer metacarpal tubercle to tip of the 3rd finger), femur
length (FEL, from the cloaca to the knee), tibia length (TBL, from knee to
heel), foot length (FOL, from inner metatarsal tubercle to the tip of the 4th
toe). Webbing formulae follows Savage & Heyer
(1997). A principal component analysis was conducted based on 18 morphometric
measurements (standardised to their SVL; Table 1) of
the two Bufoides species to examine their
morphometric distinction from each other. Calls were recorded with the camera
as videos and the audio (as .mp4 at an audio sampling rate of 48 kHz) was
extracted and analysed with Adobe Audition 6 and
Adobe Soundbooth CS3. Two specimens, SACON VA 157
(female) and VA 159 (male) were radiographed to study osteological characters
of B. kempi. A brief description of its
osteology is provided following the terminologies of Noble (1931).
Eggs of B. kempi
observed in tree holes were collected and reared for 11 days, and the growth of
the larvae was monitored with preservation of samples across various
developmental stages. The following measurements of the tadpoles: HBL―
head-body length; HBW― head-body width; HBD― head-body depth; TOT― total
length; TAIL― tail length; IO― inter-orbital distance; and TH― tail-fin height
were recorded with a stereo microscope following Chandramouli
& Kalaimani (2014). Staging of tadpoles follow Gosner (1960) and terminologies follow McDiarmid
& Altig (1999). Labial tooth-row formula for the
larvae follow Rödel (2000).
Total genomic DNA was extracted
from one specimen of B. kempi (SACON VA 180)
with a DNA extraction and purification kit, following the manufacturer’s
protocols. 16S rRNA gene was amplified using the primers 16sAR-L
(5’-CGCCTGTTTATCAAAAACAT-3’) and 16sB R-H (5’-CCGGTCTGAACTCAGATCACGT 3’),
respectively (Kocher et al. 1989). Amplifications were performed in a Applied
Bio Systems Veriti 96 well thermal cycler: 20 µl
reactions with 4 µl of 5X Phusion HF buffer, 0.4 µl of 10 mM
dNTP, 0.2 µl of Phusion DNA Polymerase,
0.1 µl each of forward and reverse primers, 2.0 µl of DNA template and 13.2 µl
of nuclease free water with the following procedure: initial denaturation of
DNA at 95 °C for 5 min, 35 cycles of: denaturation at 95 °C for 1 m, annealing
at 55 °C for 1 min, extension at 72 °C for 1 m and at last, final extension at
72 °C for 10 min. The amplicon was checked by running it through an agarose gel
electrophoresis for a clear band of the desired region in the amplified PCR
product. The amplified PCR product was purified and sequenced commercially
(National Centre for Biological Sciences, Bengaluru). The sequence thus
obtained (NCBI voucher no: OP920605) was aligned along with ten other taxa from
Bufonidae, comprising the genera Adenomus,
Beduka, Blythophryne,
Bufoides, Bufotes,
Duttaphrynus, and Pedostibes
with Hyla arborea
as the outgroup taxon. The sequences were aligned with MUSCLE (Edgar 2004) in MEGA 6.0 (Tamura et al. 2013). This
alignment of 491 bp was exported in FASTA and MEGA
formats, and was then used to determine uncorrected pairwise genetic distances
between the samples with MEGA 6. The FASTA alignment was converted to PHYLIP
format in the Alignment Transformation Environment (ALTER) website
(www.sing.ei.uvigo.es/ ALTER) and was subjected to a maximum likelihood (ML)
analysis in RAxML GUI v. 1.3 (Stamatakis
2006) using the general time reversible model, GTR GAMMA, (as RAxML uses only the general time reversible (GTR) model of
sequence evolution) with 500 bootstrap replicates. Likewise, for the Bayesian
analysis, the FASTA alignment was converted to NEXUS format and analysed in MrBayes 3.1.2 (Ronquist & Huelsenbeck 2003)
by running it for three million MCMC iterations initially until the standard
deviation of the split frequencies reached a value of ≤ 0.001. Else, the
analysis was continued for another 10000–100000 generations until the standard
deviation of ≤ 0.001 was obtained for the split frequencies. Initial 20% of the
trees were discarded as ‘burn-in’. The tree files generated were then
visualized using Fig Tree v. 1.4.0.
Results
Our analyses of molecular data
(both maximum likelihood - ML and Bayesian - BI) recovered the two species
allocated to the genus Bufoides to form a
monophyletic group; with the two species B. meghalayanus
and B. kempi showing a congeneric, sister
relationship to each other with high support (87 & 1.0 in ML and BI,
respectively). The ML and BI analyses recovered the genera Blythophryne
Chandramouli et al., 2016 & Beduka
Dubois et al., 2021 to be close to the genus Bufoides,
as assessed earlier (Chandramouli et al. 2016)
although with low support (36 & 0.63 in ML & BI, respectively).
Pairwise genetic divergence between B. meghalayanus
and B. kempi was found to be moderate (4.67 %
at 16s rRNA) supporting their specific distinction from each other (Figures
1a,b). The PCA conducted based on 18 morphometric variables clearly separates
the two species into two discrete clusters (Figure 2, Table 1).
Systematics
Bufoides kempi (Boulenger,
1919)
Nectophrryne kempi Boulenger, 1919
Pedostibes kempi – Barbour, 1938
Bufoides kempi – Chandramouli &
Amarasinghe, 2016
Syntypes:
Two specimens; an adult (29.8 mm SVL) and a subadult (17.4 mm) (ZSI 18481 a&b, respectively)
Other
material studied: SACON VA 157 (an adult female) and, VA 181(a subadult
female), and SACON VA156; VA 158 –160; VA 164 & VA 180 six adult males
collected from Eman Asakgre
(25.37°N, 90.54°E, 200 m asl.), Garo Hills, Meghalaya
(Image 1).
Diagnosis:
(after Chandramouli & Amarasinghe
2016)
A
semi-arboreal to rupicolous Bufoides
from the Garo Hills diagnosed by: small to medium body size (SVL 24.1–32.36
mm); presence of irregular, non-keratinized cranial ridges (pre and post
orbital); short, ovoid parotoid glands; absence of an
externally visible tympanum; moderate degree of webbing between toes (two
phalanges of toe IV free); partial webbing between fingers, and the presence of
small, slightly dilated, rounded terminal digital discs at the tips of both
fingers and toes. Dorsum black with mossy green shade along the flanks in
males, females predominantly green with black reticulations; a pale white
venter; eggs partially pigmented and laid in strings within water-filled tree
holes (phytotelmata).
Description
and variation: (based on the newly collected material) Table 2
Female (SVL
32.36 mm) slightly larger than males (mean SVL 26.38 mm ± 0.88, n = 6). Head
flat, fairly large, and distinct, (HL:SVL 0.31), broader than long, slightly
more wider in the female (HL:HW 0.82) than in males (HL:HW 0.91), with an
obtusely pointed to rounded snout tip. Trunk short (AG:SVL 0.4) and slightly
gracile in males (AG:BW1.45) than in females (AG:BW 1.81). Eyes fairly large
(ED:HL 0.33) their diameter shorter than the snout length (ED:ES 0.71).
Nostrils situated closer to the snout tip than to the eyes (EN:ES 0.74). Upper
eyelids wide, (mean UEW 2.92± 0.12) rugose with keratinized pustules, narrower
than the interorbital space (IO:UEW 1.79). Inter-orbital space broader than
inter-narial space (IO:IN 1.59). Upper arms short (UAL:SVL 0.23), nearly as
long as the lower arms (UAL:LAL 1.02); palm slightly shorter than the upper
arms (UAL:PAL 0.88). Fingers partially webbed, webbing formula I0-1II2-3III3-2IV;
relative length of fingers III>IV>II>I. Outer metacarpal tubercle
large and evident. Finger tips with slightly expanded rounded discs. Femur
relatively short (FEL:SVL 0.38), tibia slightly longer than femur (FEL:TBL
0.91); foot about as long as the femur (FEL:FOL 0.99). Toes moderately webbed,
webbing formula: I0-0II0-0.5III0.5-2IV2-1V,
a relatively large inner and a slightly smaller ovoid metatarsal tubercle at
the base of the foot. Toe tips with discs as broad as the toes; tarsal ridge
not discernible. Vocal sac not discernibly distinct in males. Skin rugose in
texture with keratinized granules.
Colouration in life
Males were
generally dark grey in colour with traces of mossy
green along the flanks and yellow patches near the axilla, belly and groin on
the ventro-lateral region. Females are predominantly
mossy green with an irregular black hour-glass pattern on the dorsum. Limbs
visibly barred with black. Venter pale and much lighter than the dorsum (Image
2).
Osteology
Skull large
and triangular, with an obtusely pointed snout tip. Pre and post-orbital ridges
discernible. Fronto-parietals fairly broad and
hexagonal in shape. Nasal bones of the skull short, nearly as long as broad.
Vertebral column with eight procoelous presacral
vertebrae; the first four relatively larger than the following. Sacral
diapophyses broad, flattened, and expanded laterally. Urostyle
cylindrical, about half the length of the presacral vertebral column, lacking
lateral expansions. Ilia curved laterally, as long as the urostyle.
Ischium protruding posteriorly. Pectoral girdle arciferal. Humerus
longer than radio-ulna. Phalangeal formula of the fingers: 2-2-3-3. Femur long,
nearly as long as the tibiofibula; tarsus about 3/4th
the length of tibiofibula. Phalangeal formula of the
toes 2-2-3-4-3 (Image 3).
Breeding
biology and natural history
A total of
17 individuals were seen in the following precise locations surrounding Eman Asakgre community reserve,
South Garo Hills, the details of which are mentioned below (Image 4).
Chibanda cave (25.36° N, 90.53° E, 122 m)
The cave was about 6 m below the
ground level, surrounded by moist evergreen forest. A small creek was flowing
into the cave and the surface of the rocks and boulders in the area was covered
with moss and were wet. The canopy cover provided about 70% shade to the
ground. The first individual, a subadult male was found under a boulder near
the mouth of the cave. Second individual was seen inside the cave in a deep
narrow Horizontal crevice of a limestone rock. Odorrana
chloronota, Amolops
assamensis, Limnonectes
khasianus, and Ingerana
borealis were some of the anuran species that were observed in sympatry
with B. kempi at this location. During the
night surveys in the subsequent months, seven more individuals were spotted in
total. Some individuals were observed on leaves of shrubs without exhibiting
any specific behaviour between 1900–2200 h. Later in
October, an adult male was sighted in a tree hole filled with rain water at a
height of about 2 m above the ground and a subadult female was recorded on low
lying shrubs at about one foot above the ground.
Dhangit cave (25.36° N, 90.52° E, 220 m)
This cave, surveyed in May was at
a depth of about 12 m, surrounded by moist evergreen forest; the terrain was
rocky and filled with boulders covered with moss. No individuals were found
inside the cave, however two males were observed near the cave about 150 m away
which were calling actively from a cavity filled with rain water (5 cm deep,
water temperature 21° C) measuring 10 cm in diameter at about 1.2 m off the
ground at around 2200 h possibly trying to attract the attention of a receptive
female nearby. Two males were observed to show aggression by kicking each other while calling and one of them was
seen kicking the other with its hind feet repeatedly. Upon further search in
the region, a tree cavity with about 30–40 eggs laid in strings was found. Eggs
from this cavity were collected and maintained in a plastic jar with water from
the same cavity for the next 11 days.
Cehise Stream (25.34° N, 90.51° E, 250
m)
The area surrounding a small
stream flowing near the village of Eman Dura Banda
was surveyed in May. This area had a rocky terrain and was covered with moist
evergreen forest, with a tall canopy. An adult male was found resting under a
boulder.
Eman Asakgre
Community Reserve (25.37° N, 90.54° E, 108 m)
Congregation of four males was
seen along with a female in a buttress root cavity at a height of 0.6 m above
the ground, measuring about 15 cm in diameter, filled with 10 cm of rain water.
The water temperature here was 23.5° C, the humidity of the location was 80%.
The group was first spotted at about 1730 h in amplexus with four males and a
female. One of the four males was seen mounted ventrally while the other three
males were mounted dorsally and laterally. Amplexus was axillary. The episode
lasted till about 2100 h by the time the female laid egg strings with 30–40
partially pigmented eggs that measured about 2 mm diameter. Once the eggs were
laid both males and the female started leaving the cavity and no further
attendance was observed.
Description of calls
The call of B. kempi described here was composed of syllables of ‘treek.. treek...’
that lasted for a duration of 2.1 s and was composed of three distinct notes,
each of which comprise seven─ten pulses. The mean
duration of each note was 6.33 ms, with a mean
interval of 1.05 s in between. Maximum amplitude of the call was -1 dB with a
dominant frequency of 2.5 kHz (Image 5).
Larval description: (Table 3)
On day three, 39 larvae emerged
which were reared subsequently for the next eight days during which their
development was documented. Stage-wise descriptions of the larvae are presented
in detail below.
Stage 20: (n = 3)
The larvae reached stage 20 on
day two after emerging from the eggs. At this stage, they measured 9.75 mm ±
0.31 in total length, with a head-body length of 5.77 mm ± 0.37; streamlined
and narrow in form (HBW 1.91 mm ± 0.3); slightly higher than broad (HBH 2.44 mm
± 0.48); tail a little shorter than the
head-body (3.98 mm ± 0.66); and with an average tail height of 1.91 mm ± 0.46.
Eyes and mouthparts not discernible at this stage.
Stage 22: (n = 1)
The larvae reached stage 22 on
day four after emergence from the eggs. At this stage, the body & tail
elongated a little more, with the larva measuring 10.6 mm in total length, with
a head-body length of 5.3 mm and a relatively longer tail (5.3 mm) which equalled the HBL. Head-body oval & narrow (HBW 2.76 mm;
HBH 2.0 mm). Tail-fin relatively well developed than in the earlier stage,
longer and broader, with a height of 2.34 mm. Eyes dorsal in position and
traces of gills discernible at this stage.
Stage 30: (n = 2)
At this stage, the larvae grew a
little longer, measuring 13.28 mm in length, with a 7.9 mm long tail and 5.39
mm long head-body. Head-body ovoid, broader (HBW 2.76 mm) than deep (HBH
2.33 mm). Oral disc discernible at this stage with keratodont
and keratinized jaw sheaths; oral tooth-row formula 1/1+1//1/1. Tail fins
transparent and high 2.32 mm. Rudimentary hindlimb buds visible at the
posterior end of the head-body. Eyes well developed than in the earlier stage
and in lateral position, with an inter-orbital space of 2.62 mm (Image 6).
Distribution
During the present study, B. kempi was recorded from the above four locations, of
which, two are quite close-by and the type locality, Tura, lies at about 64 km
northwest of the present study sites. However, our surveys at locations north
of the Garo ridge at a higher elevation such as Mandalgre
(25.50 °N, 90.37 °E, 1,019 m) and Daribokgre (25.48
°N, 90.31 °E, 1,123 m) could not locate this species. Further surveys at the
vicinity of the current study sites and locations to the north of the Garo
ridge in the lower reaches are necessary to determine whether the species
occurs in those areas as well (Figure 3).
Abundance
The rate of encounter of B. kempi was 0.53/hour, or two hours of effort to locate
one individual in this region. The encounter rate across the survey duration of
57 hours ranged from 0.17–2.0 (Table 4).
Bufoides meghalayanus (Yazdani & Chanda, 1971)
Ansonia meghalayana Yazdani & Chanda, 1971
Holotype: ZSIC A 6969, an
adult from Mawblang, Cherrapunji,
Khasi Hills, Meghalaya.
Material studied: SACON VA 215,
SACON VA 251 and SACON VA 252 three adult males from a hill stream in the Khasi
Hills, Meghalaya (Image 7).
Diagnosis and comparison: A
semi-arboreal to rupicolous Bufoides
from the Khasi Hills diagnosed by: small–medium body size (SVL 31. –33.5 mm);
presence of irregular, non-keratinized cranial ridges (pre and post orbital);
short, elongated parotoid glands (vs. ovoid in B. kempi); absence of an externally visible tympanum;
well-developed webbing between toes, with only half a phalange free of webbing
on toe IV (vs. relatively poor, with two phalanges free of webbing on toe IV in
B. kempi); partial, but better developed
webbing between fingers than B. kempi and the
presence of small, slightly dilated, rounded terminal digital discs at the tips
of both fingers and toes. Dorsum black with mossy green along the flanks in
males, females predominantly green with black reticulations; a dark grey venter
with small white spots (vs. pale white in B. kempi).
Description and Variation: (based
on the newly collected material) Table 2
Body small (31.99 mm ± 0.76),
trunk relatively short (AG:SVL 0.38) and stout (AG:BW 1.0). Head large (HL:SVL
0.37); slightly broader than long (HL:HW 0.85); and half as deep as long (HL:HD
2.08); snout tip obtusely pointed in dorsal view. Eyes large (ED:HL 0.35);
snout slightly longer than eye (ED:ES 0.84); interorbital space about one and
half times the width of the upper eyelid (IO:UEW 1.46) and nearly twice the
internarial distance (IO:IN 2.09). Tympanum absent. Upper arm short (UAL:SVL
0.21); lower arm slightly longer than upper arm (UAL:LAL 0.93); palm a little
longer (UAL:PAL 0.77). Fingers partially webbed, webbing formula I0-1II0-2III1-1IV;
relative length of fingers III>IV>II>I. Outer metacarpal tubercle
large & evident. Thigh relatively short (FEL:SVL 0.39); tibia a little
longer than the thigh (FEL:TBL 0.94); foot nearly as long as the thigh (FEL:FOL
0.98). Toes partially webbed, webbing formula: I0-0II0-0III0-0.5V0.5-0V,
a relatively large inner and a slightly smaller ovoid metatarsal tubercles at
the base of the foot. Toe tips with discs as broad as the toes; tarsal ridge
not discernible. Finger and toe-tips bearing slightly expanded terminal discs
lacking circum-marginal grooves. Dorsal colouration
uniform black, with irregular feeble yellow markings; venter grey with fine
white spots.
Description of calls
The call of B. meghalayanus recorded during this study comprised of a
series of high pitched syllables of ‘ti-tuk’
that lasted for a duration of 20 s and was composed of eight separated notes,
each of which comprise two pulses. The mean duration of each note was 0.28 ms, with a mean interval of 2.45 s in between. Maximum
amplitude of the call was -7 dB with a dominant frequency of 1.0 kHz (Image 8).
Distribution: (Table 4)
During this study, B. meghalayanus was recorded from a few locations in the
Khasi hills within an altitudinal range of 1060–1240 m that are mapped in
Figure 3. Additional localities were provided by Deuti
et al. (2012).
Abundance
The rate of encounter of B. meghalayanus was 0.875 / hour, or about an hour of
effort to locate one individual in this region. The encounter rate across the
survey duration of 38 h ranged from 0.67–5.33 (Table 4).
Discussion
B. kempi was described by Boulenger in 1919 based on the two specimens collected by
S.W. Kemp, and presented to him by Nelson Annandale from ‘above Tura, 2,500 ft’
in the Garo Hills. Since its description, no further records or observations of
this species have been made until now. Studies conducted in this region have uncovered
several new and noteworthy species but B. kempi
remained elusive to scientists until now (Datta-Roy
et al. 2013; Deuti et al. 2012; Biju et al. 2016; Giri et al. 2019). Although Das et al. (2009) mentioned a
specimen (MFA 10134) of Bufoides collected
from Tura, Garo Hills, no taxonomic assessment of this specimen has been made
until now, which still remains unidentified. Therefore, with the results of the
present study, we announce the authentic rediscovery of B. kempi after a period of more than a century (1919–2022)
from near the type locality, Garo Hills. First ever field observations on its
ecology, behaviour, breeding biology, and natural
history have been presented here. Observation on their breeding in phytotelmata and multiple males participating in amplexus
with a single female have been made for the first time. Das & Dutta (2007)
noted the absence of any larval descriptions for B. kempi,
which has now been provided for the first time, across three developmental
stages. Based on our field observations, the distribution of B. kempi mapped here shows that it is restricted to the
lower reaches of a small hillock in the western part of Meghalaya, south of the
Garo Hills, across an elevation range of 100─250 m. above which B. kempi was not detected despite intensive surveys. In
comparison, B. meghalayanus was found to be a
strictly montane species occurring only on the hilltops between elevations of
1,000–1,240 m. Further surveys north of the Garo ridge could possibly uncover
additional locations characterized by lowland evergreen forests with perennial
streams and rock boulders, that could potentially be occupied by B. kempi and we recommend additional surveys in such
localities in the future.
Unlike B. kempi,
for B. meghalayanus, the original descriptions
of the species as well as the genus were comprehensive in terms of both
morphology & natural history (Yazdani & Chanda 1971; Pillai &
Yazdani 1973). Subsequent studies have supplemented information on its
morphology (Das et al. 2009), osteology (Chandramouli
& Amarasinghe 2016); ecology and distribution (Deuti et al. 2012). Das et al. (2009) after examining the
specimen reported by Pawar & Birand
(2001) from Mizoram, opined that it is not conspecific with B. meghalyanus. Hence, we do not include that record
within the range of B. meghalyanus. Therefore,
the Mizoram population of Bufoides reported by
Das et al. (2009) still needs a proper taxonomic assessment.
Availability of Data
Specimens collected and studied
are deposited in the collections of SACON. DNA sequence generated in this study
has been deposited in the genbank under the NCBI
voucher number OP920605.
Table 1. Eigenvalues and the
proportion of variance explained by each of the principal component.
|
PC 1 |
PC 2 |
PC 3 |
PC 4 |
PC 5 |
PC 6 |
PC 7 |
PC 8 |
AG |
0.13 |
-0.05 |
0.66 |
-0.25 |
-0.26 |
-0.16 |
0.19 |
0.13 |
BW |
0.73 |
-0.38 |
-0.01 |
0.25 |
-0.25 |
0.11 |
-0.02 |
0.09 |
HL |
0.24 |
0.32 |
-0.13 |
-0.46 |
-0.29 |
-0.10 |
0.05 |
-0.07 |
HW |
0.51 |
0.20 |
-0.02 |
-0.02 |
0.31 |
0.01 |
0.04 |
0.10 |
HD |
0.11 |
0.09 |
-0.19 |
0.08 |
0.02 |
-0.24 |
-0.15 |
-0.25 |
ED |
0.16 |
0.08 |
0.06 |
0.10 |
-0.02 |
0.01 |
-0.36 |
0.23 |
EN |
0.08 |
0.26 |
0.09 |
0.13 |
-0.16 |
0.22 |
-0.38 |
-0.49 |
ES |
0.04 |
0.20 |
-0.22 |
0.16 |
-0.20 |
0.32 |
0.39 |
-0.18 |
UEW |
0.04 |
0.08 |
-0.12 |
-0.09 |
0.16 |
0.13 |
-0.22 |
0.14 |
IO |
-0.06 |
0.22 |
-0.12 |
0.16 |
-0.05 |
0.19 |
-0.03 |
0.33 |
IN |
-0.07 |
0.27 |
0.03 |
0.08 |
-0.17 |
0.34 |
-0.15 |
0.54 |
UAL |
-0.06 |
0.18 |
0.53 |
0.54 |
0.05 |
-0.08 |
-0.14 |
-0.14 |
LAL |
-0.01 |
0.14 |
0.01 |
0.22 |
-0.13 |
-0.29 |
0.48 |
0.12 |
PAL |
0.11 |
0.06 |
-0.17 |
0.38 |
0.35 |
-0.13 |
0.26 |
0.01 |
FEL |
0.14 |
0.54 |
0.01 |
-0.03 |
-0.15 |
-0.10 |
0.09 |
-0.17 |
TBL |
-0.03 |
0.29 |
-0.01 |
0.04 |
0.09 |
-0.40 |
-0.10 |
0.31 |
TAR |
0.06 |
0.14 |
0.33 |
-0.18 |
0.45 |
0.51 |
0.28 |
-0.07 |
FOL |
0.19 |
0.07 |
0.05 |
-0.22 |
0.44 |
-0.21 |
-0.14 |
-0.04 |
Eigenvalue |
0.01 |
0.01 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
% variance |
46.40 |
24.50 |
11.91 |
6.90 |
5.37 |
2.64 |
1.47 |
0.81 |
Table 2. Morphometric
measurements of Bufoides kempi and B. meghalayanus
at SACON.
Species |
Bufoides kempi |
||||||||
Voucher no: |
VA 164 |
VA 159 |
VA 158 |
VA 160 |
VA 180 |
VA 156 |
MEAN |
±SD |
VA 157 |
Sex |
M |
M |
M |
M |
M |
M |
|
|
F |
SVL |
25.3 |
29.3 |
26.8 |
24.1 |
32.7 |
26.3 |
27.4 |
3.1 |
32.4 |
AG |
8.5 |
11.3 |
10.9 |
7.8 |
13.9 |
10.6 |
10.5 |
2.2 |
13.8 |
BW |
6.2 |
9.5 |
7.3 |
5.3 |
12.5 |
5.5 |
7.7 |
2.8 |
7.6 |
HL |
7.7 |
9.5 |
8.06 |
8.7 |
10.4 |
8.4 |
8.8 |
1.0 |
9.9 |
HW |
8.8 |
10.6 |
8.8 |
9.2 |
13.9 |
9.2 |
10.1 |
2.0 |
12.1 |
HD |
4.1 |
4.0 |
4 |
4.2 |
5.1 |
3.6 |
4.2 |
0.5 |
5.5 |
ED |
2.8 |
3.1 |
2.9 |
2.3 |
3.8 |
2.5 |
2.9 |
0.5 |
3.3 |
EN |
2.6 |
2.9 |
3.4 |
3.1 |
3.3 |
2.2 |
2.9 |
0.4 |
3.5 |
ES |
4.1 |
4.7 |
3.7 |
4.6 |
4.9 |
2.9 |
4.1 |
0.8 |
4.5 |
UEW |
2.6 |
2.9 |
2.2 |
2.7 |
2.8 |
2.7 |
2.7 |
0.3 |
3.2 |
IO |
4.9 |
4.9 |
4.3 |
4.7 |
4.9 |
4.0 |
4.6 |
0.4 |
5.9 |
IN |
2.9 |
2.9 |
2.4 |
2.6 |
2.2 |
2.2 |
2.5 |
0.3 |
4.1 |
UAL |
6.5 |
6.2 |
7.7 |
5.4 |
8.9 |
6.2 |
6.8 |
1.3 |
7.2 |
LAL |
6.3 |
6.2 |
6.1 |
5.8 |
8.1 |
5.6 |
6.4 |
0.9 |
7.4 |
PAL |
7.2 |
6.9 |
6.3 |
6.7 |
9.4 |
6.2 |
7.1 |
1.2 |
8.9 |
FEL |
9.6 |
10.1 |
9.7 |
10.2 |
12.2 |
9.4 |
10.2 |
1.0 |
12.3 |
TBL |
11.2 |
11.1 |
10.9 |
10.6 |
13.3 |
11.2 |
11.4 |
1.0 |
13.1 |
TAR |
5.4 |
7.6 |
6.2 |
6.2 |
8.9 |
7.3 |
6.9 |
1.3 |
7.5 |
FOL |
8.8 |
10.2 |
9.3 |
9 |
12.7 |
10.4 |
10.1 |
1.4 |
13.0 |
F1 |
1.7 |
2.3 |
1.2 |
1.4 |
2.3 |
1.6 |
1.8 |
0.5 |
2.5 |
F2 |
2.8 |
2.6 |
2.4 |
1.7 |
3.1 |
2.4 |
2.5 |
0.5 |
4.9 |
F3 |
5.4 |
4.4 |
4.2 |
3.6 |
5.9 |
3.5 |
4.5 |
1.0 |
7.1 |
F4 |
3.7 |
4.0 |
2.9 |
2.6 |
4.9 |
2.8 |
3.5 |
0.9 |
5.5 |
T1 |
1.3 |
1.4 |
1.2 |
0.9 |
1.5 |
1.3 |
1.3 |
0.2 |
2.1 |
T2 |
2.5 |
4.1 |
1.8 |
2.7 |
2.7 |
2.1 |
2.7 |
0.8 |
2.9 |
T3 |
4.9 |
3.8 |
2.6 |
4.1 |
4.0 |
3.3 |
3.8 |
0.8 |
3.9 |
T4 |
7.4 |
6.0 |
4.7 |
5.9 |
6.6 |
4.8 |
5.9 |
1.0 |
7.7 |
T5 |
4.0 |
2.4 |
2.7 |
4.2 |
3.9 |
2.9 |
3.3 |
0.8 |
4.9 |
Species |
Bufoides meghalayanus |
||||
Voucher no: |
VA 215 |
VA 251 |
VA 252 |
MEAN |
±SD |
Sex |
M |
M |
M |
|
|
SVL |
31.3 |
33.5 |
31.2 |
31.9 |
1.3 |
AG |
10.7 |
13.2 |
12.1 |
12.3 |
1.4 |
BW |
12.0 |
14.1 |
10.9 |
12.3 |
1.6 |
HL |
9.7 |
12.9 |
12.7 |
11.8 |
1.8 |
HW |
12.0 |
16.8 |
12.9 |
13.9 |
2.5 |
HD |
5.4 |
6.2 |
5.4 |
5.7 |
0.5 |
ED |
3.2 |
5.3 |
3.8 |
4.1 |
1.1 |
EN |
2.1 |
4.9 |
3.6 |
3.5 |
1.4 |
ES |
4.2 |
5.4 |
5.1 |
4.9 |
0.6 |
UEW |
3.2 |
4.1 |
3.1 |
3.5 |
0.6 |
IO |
4.3 |
5.7 |
5.1 |
5.0 |
0.7 |
IN |
1.1 |
3.2 |
2.9 |
2.4 |
1.2 |
UAL |
5.3 |
8.2 |
6.4 |
6.6 |
1.5 |
LAL |
6.5 |
7.4 |
7.6 |
7.1 |
0.6 |
PAL |
8.6 |
9.7 |
7.5 |
8.6 |
1.1 |
FEL |
9.6 |
14.4 |
13.2 |
12.4 |
2.5 |
TBL |
11.9 |
14.4 |
13.4 |
13.2 |
1.3 |
TAR |
6.4 |
9.3 |
6.9 |
7.6 |
1.5 |
FOL |
11.9 |
14.3 |
11.8 |
12.7 |
1.4 |
F1 |
2.1 |
2.0 |
1.8 |
1.9 |
0.2 |
F2 |
3.2 |
3.2 |
3.1 |
3.2 |
0.1 |
F3 |
5.4 |
4.7 |
4.4 |
4.8 |
0.5 |
F4 |
4.3 |
4.4 |
3.9 |
4.2 |
0.3 |
T1 |
2.1 |
2.4 |
1.6 |
2.0 |
0.4 |
T2 |
3.2 |
2.9 |
3.2 |
3.1 |
0.2 |
T3 |
5.4 |
4.5 |
3.7 |
4.5 |
0.8 |
T4 |
9.7 |
6.6 |
5.9 |
7.4 |
2.0 |
T5 |
6.5 |
5.0 |
3.8 |
5.1 |
1.3 |
Table 3. Larval measurements of Bufoides kempi.
Stage |
20 |
20 |
20 |
Mean |
±SD |
21 |
30 |
31 |
31 |
Mean |
HBL |
5.4 |
6.14 |
5.8 |
5.77 |
0.37 |
5.3 |
5.34 |
5.28 |
5.5 |
5.39 |
TOT |
10.1 |
9.56 |
9.58 |
9.75 |
0.31 |
10.6 |
11.64 |
12.2 |
14.36 |
13.28 |
TAIL |
4.7 |
3.42 |
3.78 |
3.98 |
0.66 |
5.3 |
6.3 |
6.92 |
8.86 |
7.89 |
HBW |
1.7 |
2.3 |
1.74 |
1.91 |
0.34 |
2.76 |
2.36 |
2.72 |
2.8 |
2.76 |
TH |
1.6 |
2.44 |
1.7 |
1.91 |
0.46 |
2.34 |
1.88 |
2.44 |
2.2 |
2.32 |
IO |
0.3 |
0.3 |
0.3 |
0.00 |
0.00 |
0.4 |
1.46 |
2.64 |
2.6 |
2.62 |
HBH |
2.82 |
2.6 |
1.9 |
2.44 |
0.48 |
2.0 |
2.24 |
2.24 |
2.42 |
2.33 |
Table 4. Abundance estimates of B.
kempi and B. meghalayanus.
Bufoides kempi |
||||||||||||
Site |
Lat. (0N) |
Long. (0E) |
Elevation (m asl.) |
time |
Duration (h) |
No. of ind. |
ER |
Microhabitat |
Forest type |
Habitat |
Canopy cover |
Ambient tempe-rature
(◦C) |
Eman Asakgre |
25.40 |
90.54 |
225 |
day |
3 |
1 |
0.33 |
Under boulder |
Evergreen |
Dry stream |
90 |
26 |
Eman Asakgre |
25.36 |
90.53 |
122 |
day |
6 |
1 |
0.17 |
Under boulder |
Evergreen |
Stream |
90 |
27 |
Eman Asakgre |
25.37 |
90.54 |
202 |
day |
6 |
5 |
0.83 |
Tree hole |
Evergreen |
Forest |
90 |
26 |
Eman Asakgre |
25.36 |
90.53 |
122 |
night |
6 |
2 |
0.33 |
Tree hole |
Evergreen |
Forest |
90 |
23 |
Eman Asakgre |
25.36 |
90.53 |
122 |
night |
5 |
1 |
0.20 |
on leaf |
Evergreen |
Forest |
90 |
23 |
Eman Asakgre |
25.36 |
90.53 |
220 |
night |
6 |
2 |
0.33 |
Tree hole |
Evergreen |
Forest |
90 |
23 |
Eman Asakgre |
25.36 |
90.53 |
220 |
night |
6 |
2 |
0.33 |
on leaf |
Evergreen |
Forest |
90 |
23 |
Eman Asakgre |
25.36 |
90.53 |
220 |
night |
6 |
4 |
0.67 |
on leaf/under boulder |
Evergreen |
Forest |
90 |
23 |
Eman Asakgre |
25.34 |
90.51 |
250 |
night |
6 |
1 |
0.17 |
Under boulder |
Evergreen |
Forest |
90 |
23 |
Eman Asakgre |
25.37 |
90.54 |
202 |
night |
6 |
3 |
0.50 |
on leaf |
Evergreen |
Forest |
90 |
23 |
Eman Asakgre |
25.36 |
90.53 |
122 |
night |
1 |
2 |
2.00 |
Tree hole/on leaf |
Evergreen |
Forest |
90 |
23 |
|
|
|
|
|
57 |
24 |
0.53 |
|
|
|
|
|
Bufoides meghalayanus |
||||||||||||
Stream behind Mawsmi cave |
25.25 |
91.72 |
1200 |
Day/Night |
1 |
0 |
0 |
|
Montane |
Stream |
70 |
21 |
Stream behind Mawsmi cave |
25.25 |
91.72 |
1200 |
Day/Night |
1 |
0 |
0 |
|
Montane |
Stream |
70 |
21 |
Stream 1 behind Mablang village |
25.24 |
91.74 |
1200 |
Day/Night |
1 |
0 |
0 |
|
Montane |
Stream |
80 |
21 |
Stream 1 behind Mablang village |
25.23 |
91.74 |
1200 |
Day/Night |
2 |
1 |
0.67 |
Rock Crevice |
Montane |
Stream |
80 |
21 |
Stream behind Mawsmi cave |
25.25 |
91.72 |
1200 |
Night |
1 |
0 |
0 |
|
Montane |
Stream |
70 |
20 |
Stream 1 behind Mablang village |
25.23 |
91.74 |
1200 |
Day/Night |
4 |
0 |
0 |
|
Montane |
Stream |
80 |
20 |
Stream behind Mawsmi cave |
25.25 |
91.72 |
1200 |
Day/Night |
5 |
0 |
0 |
|
Montane |
Stream |
70 |
20 |
Stream 2 behind Mablang village |
25.23 |
91.74 |
1200 |
Day/Night |
4 |
0 |
0 |
|
Montane |
Stream |
80 |
20 |
Stream 2 behind Mablang village |
25.23 |
91.74 |
1200 |
Day |
4 |
0 |
0 |
|
Montane |
Stream |
80 |
20 |
Stream 1 on-route to Thangkarank park |
25.239 |
91.73 |
1200 |
Day |
6 |
4 |
1.33 |
Rock Crevice |
Montane |
Dry stream |
90 |
18 |
Stream 2 on-route to Thangkarank park |
25.23 |
91.74 |
1200 |
Day/Night |
6 |
32 |
10.67 |
Rock Crevice / Pandanus tree |
Montane |
Dry stream |
95 |
21 |
Stream 1 on-route to Thangkarank park |
25.23 |
91.73 |
1200 |
Day |
6 |
19 |
3.17 |
Rock Crevice |
Montane |
Dry stream |
90 |
22 |
|
|
|
|
|
38 |
56 |
0.875 |
|
|
|
|
|
For figures &
images - - click here for full PDF
References
Barbour, T.
(1938). Notes on Nectophryne. Proceedings of the Biological
Society of Washington 51: 191–196.
Biju, S.D.,
G. Senevirathne, S. Garg, S. Mahony, R.G. Kamei, A.
Thomas, Y.S. Shouche, C.J. Raxworthy,
M. Meegaskumbura & I.V. Bocxlaer
(2016). Frankixalus, a New Rhacophorid Genus of Tree hole
Breeding Frogs with Oophagous Tadpoles. PLoS
ONE 11(1): e0145727. https://doi.org/10.1371/journal.pone.0145727
Boulenger, G.A. (1919). Descriptions of three new
batrachians from the Garo Hills, Assam. Records of the Indian Museum 16:
207–208. https://doi.org/10.5962/bhl.part.25921
Chandramouli, S. R., K. Vasudevan, S. Harikrishnan, S. K. Dutta, S. J. Janani, R. Sharma, I. Das
& R. K. Aggarwal (2016). A new genus and species of arboreal toad with phytotelmonous
larvae, from the Andaman Islands, India (Lissamphibia,
Anura, Bufonidae). ZooKeys 555: 57–90.
Chandramouli, S.R. & A.A.T. Amarasinghe (2016). Taxonomic reassessment of the
arboreal toad genus Pedostibes Günther 1876 (Anura: Bufonidae) and some allied oriental bufonid genera. Herpetologica 7: 137–147. https://doi.org/10.1655/HERPETOLOGICA-D-15-00053
Chandramouli, S.R. & A. Kalaimani (2014). Description of the larvae of
Günther’s toad Duttaphrynus hololius (Günther, 1876) (Anura:
Bufonidae) with notes on development and oral
ultra-structure. Alytes, 31: 3–12.
Das, I. &
S.K. Dutta (2007). Sources of larval identities for amphibians of India. Hamadryad
31: 152–181.
Das, I., D. Rangad, R.K.L. Tron, K. Deuti
& R.N.K. Hooroo (2009). Rediscovery of the endangered
Khasi Hills rock toad, Bufoides meghalayana in Meghalaya, Northeastern India. Froglog 92: 1–4.
Datta-Roy, A., I. Das, A.M. Bauer,
R.K.L. Tron & P. Karanth (2013). Lizard Wears Shades. A Spectacled Sphenomorphus (Squamata: Scincidae),
from the Sacred Forests of Mawphlang, Meghalaya,
North-east India. Zootaxa, 3701 (2): 257–276. https://doi.org/10.11646/zootaxa.3701.2.7
Deuti, K., S. Rey & S.K. Dey (2012). Status survey of the Khasi Hills rock toad (Bufoides meghalayana)
at Cherrapunjee, Meghalaya. Records of Zoological
Survey of India III: 21–25. https://doi.org/10.26515/rzsi/v111/i3/2011/158851
Edgar, R. C.
(2004). MUSCLE:
multiple sequence alignment with high accuracy and high throughput. Nucleic
acids research 32(5): 1792–1797. https://doi.org/10.1093/nar/gkh340
Frost, D.R.
(2022). Amphibian
Species of the World: an Online Reference. Version 6.1 (Date of access 10 Mar
2022). Electronic Database accessible at
https://amphibiansoftheworld.amnh.org/index.php. American Museum of Natural
History, New York, USA. https:// doi.org/10.5531/db.vz.0001
Giri, V.B., R. Chaitanya, S. Mahony,
S. Lalrounga, C. Lalrinchhana,
A. Das, V. Sarkar, P. Karanth & V. Deepak (2019). On the systematic status of the
genus Oriocalotes Günther, 1864 (Squamata: Agamidae: Draconinae) with the
description of a new species from Mizoram state, Northeast India. Zootaxa 4638 (4): 451–484. https://doi.org/10.11646/zootaxa.4638.4.1
Gosner, K.L. (1960). A simplified table for staging
anuran embryos and larvae with notes on identification. Herpetologica,
16: 183–190.
Kocher, T.D.,
W.K. Thomas, A. Meyer, S.V. Edwards, S. Pääbo, F.X. Villablanca & A.C. Wilson (1989). Dynamics of mitochondrial DNA
evolution in animals: amplification and sequencing with conserved primers. Proceedings
of the National Academy of Sciences of the United States of America, 86:
6192–6200. https://doi.org/10.1073/pnas.86.16.6196
McDiarmid, R.W. & R. Altig (1999). Tadpoles: the biology of anuran larvae.
University of Chicago Press, Chicago, 476pp.
Noble, G.K.
(1931). The
Biology of the Amphibia. McGraw Hill Book Company, London: 577pp.
Pillai, R.S.
& G.M. Yazdani (1973). Bufoides, a new genus for the
rock-toad, Ansonia meghalayana Yazdani &
Chanda, with notes on its ecology and breeding habits. Journal of the
Zoological Society of India 25: 65–70.
Ronquist, F. & J.P. Huelsenbeck (2003). MrBayes
3: Bayesian phylogenetic inference under mixed models. Bioinformatics
19: 1572–1574. https://doi.org/10.1093/bioinformatics/btg180
Rödel, M.-O. (2000). Herpetofauna of West Africa.
Volume I: Amphibians of the West African Savanna. Frankfurt am Main
(Edition Chimaira): 332 pp.
Savage, J.M.
& W.R. Heyer (1997). Digital webbing formulae for
anurans: a refinement. Herpetological Review 28: 131.
Stamatakis, A. (2006). RAxML-VI-HPC:
Maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed
models. Bioinformatics 22: 2688–2690. https://doi.org/10.1093/bioinformatics/btl446
Tamura, K.,
G. Stecher, D. Peterson, A. Filipski,
& S. Kumar (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular
Biology and Evolution 30 (12): 2725–2729. https://doi.org/10.1093/molbev/mst197
Yazdani, G.M. & S.K. Chanda
(1971). A new toad, Ansonia
meghalayana (family Bufonidae)
from Meghalaya (Assam), India, with observations on its breeding on Pandanus
furcatus Roxb. (Pandanales: Pandanaceae). Journal
of the Assam Science Society 14: 76–80.