Indicator In Wildlife Forensic Crime Investigation Biology Essay

geographical region indicator from detection of elements for forensic

geographical identification. The animals from Felidae family were

selected for the study and elemental analysis was done on three selected

species i.e. panthera leo, panthera pradus fusca and panthera tigris

tigris from different geographical areas of Gujarat state using Scanning

Electron Microscope-Energy Dispersive Spectra (SEM-EDS), and Energy

Dispersive X-Ray Fluorescence (EDXRF). After the entire elemental

analysis, the results have revealed that the animals of all geographical

regions have shown to have different levels of metallic contents from nil

to the highest for Sulfur, Calcium , Iron, potassium and manganese but

barium has shown a significant demarcation among animals of all three

species and observed only in the animals from Zunagarh region up to

9.879%. Which is further confirmed by the presence of these metals from

the soil and water sample analysis of the subsequent region and found to

be absent in Indroda park and Kankaria region. Our study provides a

geographical bio-marker in forensic wildlife investigation related to the

corresponding areas.

Suggested Reviewers: Rajnesh Kumar Sharma Ph. D. D. Sc.

Professor, Zoology and Wildlife, Kurukshetra University

rkszookuk@yahoo.co.in

As professor sharma is working on wild animals and have a vast experience

in this field

Gautam Talukdar Ph. D. D. Sc.

Scientist, Wildlife research, Wildlife research institute

gautam@wii.gov.in

Dr. Gautam is working in the field of wildlife forensics from last 2

decades and have a vast experience in this field

shukla malay M S. Phd.

Scientist, Wildlife research, Wildlife research institute

malayshukla29@gmail.com

Dr. shukla is working as an scientist in forensic cell of wildlife

research institute from last 10 years and has examined more than 500

cases related to wildlife crime

To

The Editor In-Chief

Egyptian Journal of Forensic Sciences

Sub: Regarding Submission of manuscript entitled "Animal hair as geographical region

indicator in wildlife forensic crime investigation".

Respected Sir/ Madam

I hereby want to say that the manuscript entitled "Animal hair as geographical region

indicator in wildlife forensic crime investigation" is original research and done by following

the wildlife laws and under permission for the same from principal chief conservator of forests

and chief wildlife warden of the state. No stress is given to the animals during the study. We,

author are providing you all the rights regarding copyright of the manuscript and confirm that

this research is not published or in consideration for publishing by any mean i.e. abstract or as

research paper. Please consider it for the publication. Authors are thankful for your kind

consideration.

Thanking you

Yours sincerely

Dr. Satendra K. Yadav

Research Scientist,

Forensic Wildlife Research Laboratory

Gujarat Forensic sciences University

Sameerforensics@gmail.com

+91 99789 11007

Cover Letter

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

Animal hair As Geographical Region Indicator In Wildlife Forensic Crime Investigation

Dahiya M.S. and Yadav S. K.*

Director, Institute of forensic science and Dy. Director, DFS Gujarat State, India

*Forensic Wildlife Biotechnology Research Laboratory, IFS GFSU, India

(Address for the correspondence Ph-+91-99789-11007, e-mail: sameerforensics@gmail.com)

Abstract:

This work presents a study of the feline hairs as a geographical region indicator from detection of

elements for forensic geographical identification. The animals from Felidae family were selected

for the study and elemental analysis was done on three selected species i.e. panthera leo,

panthera pradus fusca and panthera tigris tigris from different geographical areas of Gujarat

state using Scanning Electron Microscope-Energy Dispersive Spectra (SEM-EDS), and Energy

Dispersive X-Ray Fluorescence (EDXRF). After the entire elemental analysis, the results have

revealed that the animals of all geographical regions have shown to have different levels of

metallic contents from nil to the highest for Sulfur, Calcium , Iron, potassium and manganese but

barium has shown a significant demarcation among animals of all three species and observed

only in the animals from Zunagarh region up to 9.879%. Which is further confirmed by the

presence of these metals from the soil and water sample analysis of the subsequent region and

found to be absent in Indroda park and Kankaria region. Our study provides a geographical biomarker

in forensic wildlife investigation related to the corresponding areas.

Keywords: Wildlife Forensic, hair examination, elemental Analysis.

Title Page (with Author Credentials)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

Wild life plays an important role in eco balance, In fact there are more economical and social

importance associated with the wild animals. The use of hair for studying environmental factors

and work culture have been explored by many researchers from all over the world, some

researchers have established a purest association between hair and the environment.[1,2,3].

Microelements play vital role in the functioning of all living organisms and also in pathological

status. Cu, Zn, Mn, Fe ,Ca, Ba and many other elements are necessary for animals and in humans

[4]. One of the substrata accessible for investigation of these elements are hairs. Feeding habits,

external environment and metabolism associated processes leads to the origin of these elements

in hair. These elements have been selected for biomonitoring worldwide in global environment

monitoring system (GEMS) of the united nations environment programme in EPA 2001[5]. In

recent years, elemental study of the hair composition has become the subject of interest for

forensic investigation of chronic toxicity due to heavy metals, toxicants, ecological aspects and

clinical aspects.[6-12].

Many studies have been done on hair for elemental analysis has revealed about the level

of trace elements in hair reflects, the degree of environmental concentration like soil, water, food

and metabolism. Higher levels of these elements in the environmental concentration like soil,

water, food intake and metabolism lead the organism for strong natural selection for tolerance.

As environmental contaminants are of great importance in forensic analysis in cases where hair

is used as an evidence since last century.[13] The concentration of these elements varies between

each geographical region which leads to different levels for accumulation of these elements in

hairs. Animal hairs shows significant variation in the levels of these elements then humans

because of their extended exposure to soil contaminants through feeds. [1, 14]in the

contemporary studies on forensic science, clinical and toxicological studies involves elemental

analysis of hair has been done with the specific interest on the chronic toxicity among animals

due to heavy metals of elements present in the soil for longer durations[15,16]. Hairs of panthera

leo (Lion), panthera pradus fusca (Leopard) and panthera tigris tigris (Tiger) were used in the

present study for the production of forensically important geographical markers from elemental

profiling ,soil and water samples. In the present study an attempt has been made to correlate the

environmental conditions with that of hair with the elemental analysis on them would be

ultimately useful for the determination of the geographical area or the inhabitation of the

animals.

*Manuscript (without Author Credentials)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

Materials and Methods

Reagents: Absolute alcohol, SDS, HNO3, H2O2, Rectified Spirit used were analytical grade

reagents procured from Hi-Media and Merck specialties. Carbon tape, aluminum adhesive

and gold-palladium coating material was taken from ZEISS SEM supplies.

Sample collection : Hairs of lion, leopard and tigers were collected from Indroda nature park,

Gandhinagar (23011‟32.70"N, 720 39‟03.04"E); Kamala Nehru Zoological Garden

Kankaria, Ahmedabad (23000‟34.11"N, 72035‟58.17"E); Sakkarbaug Zoo (21032‟27.05"N,

70028‟06.53"E), Wildlife Rescue Center Junagadh (21032‟56.82"N, 70028‟23.44"E) wide

letter No. WLP/28/B/4651-54/2012-13 from the Office of the Principle Chief Conservator

of Forests and Chief Wildlife Warden, Gujarat State. 10 hair of each of 90 animals were

taken from the cage and enclosure using forceps with gloves to avoid contamination and

were packed in zip-lock bags followed by paper envelops. Soil and water samples of the

same were collected from 30 different sites from the area selected for the present study.

Geographical locations were identified by "Bhaskaracharya Institute of Space Applications

and Geo-Informatics", Gandhinagar, INDIA.

Sample preparation: Hair samples were washed with absolute alcohol followed by washing in

2% SDS, rinsed , dried in oven at 40 degree for about 24 hours followed by packing in fresh

zip-lock bags and stored at 00 C till further analysis. For EDXRF hair samples were

dissected into 2 mm pieces and placed on sample holder using forceps. For EDS hair

samples were dissected in pieces of 5 mm size and places on sample holder followed by

Gold-Palladium coating at 5 milibar vacuum and 5 mA current.

The systematic examination using various instruments resulted in detection of the elements

with their corresponding quantities. These have shown the constituent concentrations of the

elements that are present both in soil samples as well as in hair samples.

EDXRF Analysis: samples were analyzed under Energy Dispersive X-Ray Fluorescence

Spectrometer (EDXRFS) by Shimdzu EDX Series EDX-720/800 HS with 10 mm collimeter

was used for the elemental analysis. The instrument was calibrated using standard

calibration metal cubes provided with the instrument and then the hair samples were placed

for analysis. Readings of the elements were taken on two channels, first from sodium to

scandium whereas the second channel by Titanium to Uranium[17,18]. Comparative and

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

percentage analysis was observed for Sulfur, Iron, Calcium, Copper and barium. The

analytical equipment was tested with the both interlaboratory and intralaboratory controls

before use.

SEM-EDS analysis: the samples were examined under Scanning Electron Microscopy [SEM]

using Zeiss EVO 18 Special edition for the elemental analysis using gold palladium coating

for 120 seconds at 5 mA current for plasma. The SEM-EDS was taken with WD of 9.5 mm

gun beam on 20kV using Energy dispersive Spectrum detector provided by Oxford

Instruments and the elements were analyzed using Dot-Id setup for the elements S, Cu, Fe,

K, Ca and Ba.

Results

For getting the clarity of the micro and macro nutrients that are essential for the smooth

functioning of the cells and the system in general for any living organism. To achieve this, the

soil samples, those have been collected from the ambience of the specific animals that are under

study were examined for the quality and quantity of various elements which include the macro

and micro nutrients. These samples have been examined for barium, calcium, iron, silicon,

titanium and manganese. The soil samples from Indroda and Kankaria region have been

examined. Among the soil samples of Indroda region showed no trace of barium, silicon. There

were predominantly high concentrations of calcium and iron, less significant quantities of

manganese and titanium were noted.

From the above table no. 1 and figure 2, it is possible to establish that soil samples from

Junagadh region showed significantly higher concentrations of the barium and silicon. These

have been much consistent and uniform in all the sample groups. In case of calcium and iron, it

has been shown that there was least significance in the difference in the concentration of these

two elements. The same types of the results were available for titanium and manganese as they

also did not show much of the significant differences

In case of hair samples from Indroda and Kankaria Region, it was found to have contained

no trace of barium at all, so barium does not have any presence both in the ambient atmosphere

and so also in case of hair samples as shown in figure 3.

In case of iron and calcium that was less significant concentration of these metals was noted.

Sulfur and potassium were also found to have in different concentrations in hair samples. This

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

also could not be taken into consideration for the co-relation of elements present in ambient (soil,

and the hairs) as shown in detail in table no. 2

In case of Junagadh region, presence of barium was found in significant quantities. The

same kind of significant levels of concentrations of barium were observed in the hair samples of

animals of this region shown in figure 4.

The major elements such as iron and calcium were found in higher concentration levels,

both in the soil as well as in hair samples of that region. This may not be taken for consideration

as they don‟t show much of significant differences which are showed in detail in table no. 3.

Comparative analysis of different elements i.e. sulfur is found more prominent in animals

from Indroda and Kankaria region whereas iron has shown relatively slight low concentration

levels in hairs of animals from Indroda region than in Junagadh region and same observations

were observed in elemental profile of the soil samples for iron as shown in figure 5 and 6

respectively.

Discussion

The concentrations of the various elements in the soil and hair samples are in concurrence with

the given conditions. In other words, the quantity of the elements present in the hair samples are

in perfect concurrence with that of the soil samples. This will be more indicative and informative

about the place of the animal inhabitation purely based on the hair elemental analysis. In the

same way the absence of the elements in the ambience i.e. the soil samples were found to be

absent in the hair samples. This denotes the geographical location/inhabitation of the animal with

the aid of the hair samples through the elemental analysis. The results of this study is in

concurrence with the studies done on the hair samples of human beings in industrialized area by

Ramakrishna et.al. in 1996. In that study also the elemental concentrations in the hair samples

who were exposed to the industrialized areas, the levels were in perfect tender with the

concentration of such elements in the ambience. Dietz et. al.(2001),Sakuma et. al.(2010) showed

the results in which the elemental concentrations in the hair samples of the individuals were

found to be similar to the results of the present study in which the elements were detected with

the ambience and soil. Though, these studies involve the presence of elements such as sulfur,

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

calcium, iron, titanium, manganese, copper, cadmium, these elements may have the persistence

in the endogenous system[19]. But the present study revealed the element like as barium which is

remarkably significant due to its absence in vital systems and can be used to establish relation

between both the ambience and the hair samples.

The findings and the results of the work is purely an outcome of the study done in the

Gujarat region. These results are open for the future researchers to throw more light in this area

and enhance the results by extending this to larger geographical areas. The presence of the

elements finds more importance so far as the geographical area is concerned. This may be well

correlated among the soil and hair samples.

Conclusion

Based on this study, it is possible to deduce that the elemental analysis of hair would be a

significant indicator of geographical origin or inhabitation of animals just based on the hair

examination without the invasion of the tissue of such animals under study.

Acknowledgements

The authors are thankful to Mr. S.K. Goyal, PCCF Gujarat state and wildlife chief

warden for providing the permission of the research and also thankful to concerning Zoo

Authorities for providing necessary facilities and assistance during the research work.