1. COLLECTION SITE MYEIK, MYANMAR
Species Identification of Burmese Flying Foxes Using Non-Invasive Molecular Techniques David R. Castro, Rebecca M. Ritter, Craig S. Tepper & Tammy L. Mildenstein Department of Biology, Cornell College, Mount Vernon, IA, USA
INTRODUCTION
RESULTS
Flying foxes are important for forest maintenance and regeneration, because they are considered ‘‘keystone’’ pollinators and seed dispersers (Figure 1). However, flying foxes are threatened by habitat loss, overhunting, and other anthropogenic stressors. While flying foxes can be identified by morphology, many species are morphologically similar. We are using non-invasive DNA isolation techniques to identify species present at specific locations. Understanding population dynamics allows conservation managers to develop strategies to protect these important members of the environment.
Full length sequences (486 bp), were entered into the National Center for Biotechnology Information (NCBI) database for identification (Figure 4). A total of five samples have been cloned and sequenced. Four samples had a 98% sequence identity match to Pteropus hypomelanus and one sample had a 98% sequence identity match to Cynopterus sphinx (Figure 5).
Stool Sample
gDNA
Pteropus
giganteus
Pteropus hypomelanus
Cynopterus
sphinx
Amplification
of Cyt b
Figure 4. NCBI database
comparison
Figure 1. Examples of bat species present
in Myanmar.
COLLECTION SITE MYEIK, MYANMAR
Taken by Shantanu Kuveskar
Cyt b
Cynopterus sphinx
N=1
Taken by Nick Baker
Copyright © Ecology Asia 2018
Pteropus hypomelanus
N=4
Clone
Analysis
Copyright © 2018 Apa Digital AG λ
Eco RI
Eco 52I
Uncut
Apa I
Figure 2. Map of Myanmar, the red dot
represents our collection site.
Figure 5. Species present in Myeik, Myanmar
21.2 kb
DISCUSSION
7.4 kb
METHODS
3.48 kb
The presence of P. hypomelanus roosting in Myeik (4/5 samples) is consistent with local observations. However, the identification of C. sphinx at the same roosting site is unusual because this species has not been reported in Myeik.
Non-invasive sampling methods can be used to isolate DNA for species identification in order to map population distributions. This information can help conservation managers determine appropriate strategies to protect these bats.
3.0 kb
3.5 kb
DNA was isolated from fecal samples collected under roosting sites in Myeik, Myanmar (Figure 2). A fragment of the mitochondria cytochrome-b gene, known to be conserved within species (Waits and Paetkau 2005), was amplified using PCR. Amplified cytochrome-b fragments were run on agarose gels, purified, and cloned into pGEM-T vectors prior to sequencing (Figure 3).
1500 bp
1000 bp
486 bp
Cyt b
500 bp
Figure 3. DNA isolation, PCR amplification,
and cloning
Cornell Summer Research Institute/Faculty Research Development Grant