MSc.

The study was carried out at Parambikulam Tiger Reserve during 2012-
2013 with an objective of DNA barcoding the insectivorous bats to unravel the
taxonomic ambiguity. The methods employed were the phenol-chloroform
extraction (Sambrook et al., 1989) or the GeniPurelM Mammalian Genomic DNA
Purification Kit (GeNei™). The sequences were compared with those registered
in NeB! databank (blast.ncbi.nlm.nih.gov). The phylogeny reconstruction and the
calculation of genetic distances were done using the MEGA 5.0 (Tamura et al.,
2011).
In this study, three bat species such as Hipposideros speoris, Megederma
spasma and Rhinolophus rouxii, were identified using the partial cytochonne b
sequence from nine samples of individuals. But the species, Megaderma spasma
and Hipposideros speoris showed a genetic distance between two percent and
11 % while compared with the Genbank sequences of those species. Bradly and
Baker (2001) considered that a genetic distance more than 10% or between two
percent and 11% at cytochrome b is an indication of species level divergence or
valid species. So the high sequence divergence in Megaderma spasma and
Hipposideros speoris of present study with the same species from South Asia and
South India gives an idea on the existence of conspecific population or valid
species in these localities. The high sequence divergence may possible as they
were collected from two different geographic locations and they result from
geographic barriers separating genetic flow. To identify this complexity, the
present study recommends additional study concerning specific status and for that
more specimens should be collected from each geographical location. Because of
this high genetic distance or sequence divergence between the sequences of
Megaderma spasma and Hipposideros speoris of present study and that of
retrieved from Genbank were clustered as sister clades in their respective
phylogenetic neighbour-joining tree. The two sequences of Megaderma spasma,

KAUNHM2012318 and KAUNHM2012320, and one sequence of Hipposideros
speoris, KAUNHM2012314, were identified as a haplotypes.
In the case of Rhinolophus rouxii sequences, the phylogenetic analysis put
KAUNHM20114 and KAUNHM201185 together in one branch and they again
clustered to the GenBank sequences of 80 kHz phonic type of Rhinolophus rouxii.
The genetic distance also confirms this. But the third sequence of Rhinolophus
rouxii from the present study, KAUNHM2012310, clustered with the sequences
of 90 kHz phonic type of Rhinolophus rouxii and that showed a genetic distance
of 9.5% when compared with two other Rhinolophus rouxii sequences of present
study. So, the study clearly indicates the existence of sibling species of
Rhinolophus rouxii and Rhinolophus indorouxii in the study area and this support
the study ofChattopadhyay et al. (2012).