Case report Establishing the identity of the massacred tigress in a case of wildlife crime Sandeep Kumar Gupta *, Jyotsna Bhagavatula, Kumarasamy Thangaraj, Lalji Singh ** Centre for Cellular and Molecular Biology, Hyderabad, India 1. Introduction Despite a very strict wildlife protection act, the poaching rate of tigers (Panthera tigris) and leopards (Panthera pardus) is still increasing in several countries for body parts for Chinese medicinal products and ornaments. Developments in molecular biology have strengthened wildlife forensic science by virtue of the availability of molecular markers such as mitochondrial DNA (mtDNA) [1–3] and microsatellites [4]. The application of mitochondrial cyto- chrome-b gene-based species identification has helped tremen- dously in identification of forensic samples in wildlife offences [5]. Microsatellites are the best available marker for parentage testing and linkage analysis [6]. Control region sequences of mtDNA provide improved phylogenetic resolution grouping in the big cats i.e. Panthera spp. [7]. There are 10 different haplotypes of tigers in India based on control region and coding regions of mtDNA, which can be used to find maternal lineage [8]. In the present study, we were successful in providing evidence that the source of the claw is of the missing progeny of the same tiger family of the zoological park. 2. Case report 2.1. Case history In 2000, a carcass of a female tiger was found outside of its cage in the ‘‘Tiger Safari’’ of a Zoo Park in India. The death of the tigress was a highlight in media for several days and a wildlife crime was reported, which was pending for the last four years. Unfortunately, no biological sample was made available for us from the deceased tigress for the DNA profiling. After four years, a team of Police Officials of the same city had seized a claw, encased in silver frame, and pieces of decomposed hide from a local person. These samples were sent to our laboratory to establish whether they belong to the illegally taken tigress. 2.2. DNA analysis to establish the identity 2.2.1. DNA isolation from claw and skin Samples taken from: a thin stratum of the ventral portion of the claw and pieces of the decomposed skin were separately washed twice with normal saline. The washed material was subjected to DNA extraction [5]. The DNA extracted from the putrefied hide was degraded, hence not found suitable for genotyping. The DNA extracted from the claw was used for genotyping and further analysis. The DNA sample of the biological parents and a male sibling of the victim tigress were obtained from our DNA bank. Forensic Science International: Genetics xxx (2010) xxx–xxx ARTICLE INFO Article history: Received 24 December 2009 Received in revised form 21 May 2010 Accepted 24 May 2010 Available online xxx Keywords: Biological remnant Wildlife crime Sibling Biological parent STRs Mitochondrial DNA markers ABSTRACT We report a case study, where we have established the identity from a challenging biological sample of a deceased tigress by parentage analysis. A wildlife crime was committed in one of the zoological parks in India in the year 2000, where one young tigress was killed for its claws. This was of media interest for several days and remained an unsolved case for four years. A framed claw and decomposed tiger hide were seized from the accused in 2005. Biological samples of the victim tigress was not available for further forensics examination, therefore; DNA samples of the biological parents and a male sibling were used to establish the identity of the claw using STRs and mitochondrial DNA markers. Our analysis indicates that the seized claw belongs to the victim tigress. ß 2010 Elsevier Ireland Ltd. All rights reserved. * Corresponding author. Present address: Wildlife Institute of India, Dehra Dun 248 001, India. Tel.: +91 135 2640111/15x353; fax: +91 135 2640117. ** Corresponding author. Tel.: +91 40 27192634; fax: +91 40 27160252/27160591. E-mail addresses: skg@wii.gov.in (S.K. Gupta), lalji@ccmb.res.in (L. Singh). G Model FSIGEN-606; No. of Pages 2 Please cite this article in press as: S.K. Gupta, et al., Establishing the identity of the massacred tigress in a case of wildlife crime, Forensic Sci. Int. Genet. (2010), doi:10.1016/j.fsigen.2010.05.004 Contents lists available at ScienceDirect Forensic Science International: Genetics journal homepage: www.elsevier.com/locate/fsig 1872-4973/$ – see front matter ß 2010 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.fsigen.2010.05.004