Skip to main content
SpringerLink
Log in

Pangolin Indexing System: implications in forensic surveillance of large seizures

  • Original Article
  • Published:
International Journal of Legal Medicine Aims and scope Submit manuscript

Abstract

Demand for pangolin scales in East Asia has increased dramatically in the past two decades, raising concern to the pangolin survival and bringing them to the brink of local extinction. Enumerating the number of individuals from the seized pangolin scales primarily goes undocumented, mostly due to the unavailability of the appropriate methods. In this study, we developed a Pangolin Indexing System, a multi-locus STR panel of eight dinucleotide microsatellites that showed promising results in individualization and assignment of scales into Chinese and Indian pangolins. The combined power of exclusion was 0.83 and 0.99 for Chinese and Indian pangolin. The select panel of eight polymorphic STRs exhibited the cumulative probability of identity 3.7 × 10−9 for Indian pangolin and 3.6 × 10−7 for Chinese pangolin and identified 51 unique genotypes from the 74 scales selected from the four pangolin seizures. The study demonstrated the first report of cross-species validation of STRs developed from Malayan pangolin to Indian pangolin and showed the potential application of Pangolin Indexing System in screening of large seizures through DNA profiling from the scales of Indian and Chinese pangolin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1

Similar content being viewed by others

References

  1. Heinrich S, Wittmann TA, Prowse TA, Ross JV, Delean S, Shepherd CR, Cassey P (2016) Where did all the pangolins go? International CITES trade in pangolin species. Glob Ecol Conserv 8:241–253

    Article  Google Scholar 

  2. Choudhary AN, Badola S, Fernandes M, Chhabra DB (2018) TRAFFIC factsheet: the scale of pangolin trade in India. TRAFFIC India. https://www.traffic.org/site/assets/files/2647/factsheet_scale_of_pangolin_trade_in_india_2009_2017. Accessed 2nd February 2020

  3. Gaubert P, Antunes A, Meng H, Miao L, Peigné S, Justy F, Njiokou F, Dufour S, Danquah E, Alahakoon J, Verheyen E, Stanley WT, O’Brien SJ, Johnson WE, Luo SJ (2018) The complete phylogeny of pangolins: scaling up resources for the molecular tracing of the most trafficked mammals on earth. J Hered 109:347–359

    Article  CAS  PubMed  Google Scholar 

  4. Baillie J, Challender D, Kaspal P, Khatiwada A, Mohapatra R, Nash H (2014) Manis crassicaudata. The IUCN Red List of Threatened Species 2014

  5. Challender D et al (2014) Manis pentadactyla. The IUCN Red List of Threatened Species 2014

  6. Challender D, Wu S, Kaspal P, Khatiwada A, Ghose A, Ching-Min SN, Laxmi ST (2019) Manis pentadactyla. The IUCN Red List of Threatened Species 2019

  7. Mahmood T et al (2019) Manis crassicaudata . The IUCN Red List of Threatened Species2019. https://doi.org/10.2305/IUCN.UK.20193.RLTS.T12761A123583998.en. Accessed 11 April 2020

  8. Khwaja H et al (2019) Pangolins in global camera trap data: Implications for ecological monitoring,Global Ecology and Conservation, 20:e00769. https://doi.org/10.1016/j.gecco.2019.e00769

  9. Zhou ZM, Zhao H, Zhang ZX, Wang ZH, Wang H (2012) Allometry of scales in Chinese pangolins (Manis pentadactyla) and Malayan pangolins (Manis javanica) and application on judicial expertise. Zool Res 33:271–275

    Article  Google Scholar 

  10. Challender D, Waterman C (2017) Implementation of CITES decisions 17.239 b) and 17.240 on pangolins (Manis spp.). https://cites.org/sites/default/files/eng/com/sc/69/E-SC69-57-A.pdf. Accessed 1st December 2019

  11. Mondol S, Navya R, Athreya V, Sunagar K, Selvaraj VM, Ramakrishnan U (2009) A panel of microsatellites to individually identify leopards and its application to leopard monitoring in human dominated landscapes. BMC Genet 10:79

    Article  PubMed  PubMed Central  Google Scholar 

  12. Mishra S, Singh S, Munjal A, Aspi J, Goyal S (2014) Panel of polymorphic heterologous microsatellite loci to genotype critically endangered Bengal tiger: a pilot study. SpringerPlus 3:4

    Article  PubMed  PubMed Central  Google Scholar 

  13. Gugala NA, Ishida Y, Georgiadis N, Roca A (2016) Development and characterization of microsatellite markers in the African forest elephant (Loxodonta cyclotis). BMC Res Notes 9:364

    Article  PubMed  PubMed Central  Google Scholar 

  14. Potoczniak M, Chermak M, Quarino L, Tobe S, Conte J (2020) Development of a multiplex, PCR-based genotyping assay for African and Asian elephants for forensic purposes. Int J Legal Med 134(1):55–62

    Article  PubMed  Google Scholar 

  15. Luo SJ et al (2006) Isolation and characterization of microsatellite markers in pangolins (Mammalia, Pholidota, Manis spp.): primer note. Mol Ecol Notes 7:269–272

    Article  Google Scholar 

  16. Butler JM (2005) Forensic DNA typing: biology, technology, and genetics of STR markers. Elsevier Academic Press, Burlington

    Google Scholar 

  17. Verma SK, Singh L (2003) Novel universal primers establish identity of an enormous number of animal species for forensic application. Mol Ecol Notes 3(1):28–31

    Article  CAS  Google Scholar 

  18. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Sympser 41:95–98

    CAS  Google Scholar 

  19. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    Article  CAS  PubMed  Google Scholar 

  20. Scandura M, Capitani C, Iacolina L, Apollonio M (2006) An empirical approach for reliable microsatellite genotyping of wolf DNA from multiple noninvasive sources. Conserv Genet 7(6):813–823

    Article  CAS  Google Scholar 

  21. Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetics software for teaching and research-an update. Bioinformatics 28:2537–2539

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Gouy A, Zieger M (2017) STRAF-A convenient online tool for STR data evaluation in forensic genetics. Forensic Sci Int Genet 30:148–151

    Article  CAS  PubMed  Google Scholar 

  23. Queller DC, Goodnight KF (1989) Estimating relatedness using genetic markers. Evolution 43:258–275

    Article  PubMed  Google Scholar 

  24. Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Earl DA, VonHoldt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conserv Genet 4(2):359–361

    Article  Google Scholar 

  26. Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Mol Ecol 14:2611–2620

    Article  CAS  PubMed  Google Scholar 

  27. Kopelman NM, Mayzel J, Jakobsson M, Rosenberg NA, Mayrose I (2015) CLUMPAK: a program for identifying clustering modes and packaging population structure inferences across K. Mol Ecol Resour 15(5):1179–1191

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Andreassen R, Schregel J, Kopatz A, Tobiassen C, Knappskog PM, Hagen SB, Kleven O, Schneider M, Kojola I, Aspi J, Rykov A, Tirronen KF, Danilov PI, Eiken HG (2012) A forensic DNA profiling system for northern European brown bears (Ursusarctos). Forensic Sci Int Genet 6(6):798–809

    Article  CAS  PubMed  Google Scholar 

  29. Mukesh LK, Sharma VP, Kumar SA, Charoo NM, Goyal SP, Sathyakumar S (2013) Loss of genetic diversity and inbreeding in Kashmir red deer (Cervus elaphushanglu) of Dachigam National Park, Jammu & Kashmir, India. BMC Res Notes 6:326

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. National Research Council (1996) The evaluation of forensic DNA evidence, National Academy Press, Washington, DC

  31. Evett IW, Weir BS (1998) Interpreting DNA evidence: statistical genetics for forensic scientists. Sinauer Associates Inc., Sunderland, Massachusetts

    Google Scholar 

  32. Balding DJ (2005) Weight of evidence for DNA profiles. John Wiley & Sons Inc., Great Britain

    Book  Google Scholar 

  33. Linacre A, Gusmao L, Hecht W, Hellmann AP, Mayr WR, Parson W, Prinz M, Schneider PM, Morling N (2011) ISFG: recommendations regarding the use of non-human (animal) DNA in forensic genetic investigations. Forensic Sci Int Genet 5(5):501–505

    Article  CAS  PubMed  Google Scholar 

  34. Cota-Larson R (2017) Pangolin species identification guide: a rapid assessment tool for field and desk. United State Agency for International Development, Bangkok: USAID Wildlife Asia Activity

Download references

Acknowledgments

The authors thank the law enforcement agencies for providing scales for forensic examination and identification.

Funding

The study was conducted with the funding support from the National Mission for Himalayan Studies, Ministry of Environment, Forest and Climate Change (MoEF&CC) (Grant No. NMHS/2017-18/LG09/02/476).

Author information

Author notes

  1. Abhishek Singh and Prajnashree Priyambada contributed equally to this work.

Authors and Affiliations

  1. Zoological Survey of India, New Alipore, Kolkata, West Bengal, 700053, India

    Abhishek Singh, Prajnashree Priyambada, Gul Jabin, Sujeet Kumar Singh, Bheem Dutt Joshi, Chinnadurai Venkatraman, Kailash Chandra, Lalit Kumar Sharma & Mukesh Thakur

  2. Gujarat Forensic Sciences University, Sector 9, Gandhinagar, Gujarat, 382007, India

    Abhishek Singh

  3. Department of Zoology, University of Calcutta, 35, B.C. Road, Kolkata, West Bengal, 700019, India

    Prajnashree Priyambada & Gul Jabin

Authors
  1. Abhishek Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Prajnashree Priyambada
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gul Jabin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sujeet Kumar Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bheem Dutt Joshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chinnadurai Venkatraman
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kailash Chandra
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Lalit Kumar Sharma
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Mukesh Thakur
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceptualization, MT; data curation, AS, PP, GJ, SKS, and BDJ; formal analysis, AS, GJ, and MT; funding acquisition, LKS, MT and KC; investigation, MT and LKS; methodology, AS, PP, GJ, SKS, and BDJ; project administration, MT and LKS; resources, MT, CV, KC, and LKS; software, AS, PP, GJ, SKS, BDJ, and MT; supervision, validation, and visualization, MT; roles/writing (original draft), AS, PP, and MT; and writing (review and editing), SKS and MT.

Corresponding author

Correspondence to Mukesh Thakur.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(DOCX 676 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Singh, A., Priyambada, P., Jabin, G. et al. Pangolin Indexing System: implications in forensic surveillance of large seizures. Int J Legal Med 134, 1613–1618 (2020). https://doi.org/10.1007/s00414-020-02362-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00414-020-02362-5

Keywords

Search

Navigation