Table Of ContentHAEMOGLOBIN POLYMORPHISM IN ASIAN ELEPHANT ELEPHAS MAXIMUS WITH
SPECIAL REFERENCE TO ELEPHANT POPULATION IN SOUTH INDIA1
Hemant S. Datye 2 A.M. Bhagawat3 and V. Krishnamurthy2
(With a text-figure
)
Polyacryl Amide Gel Electrophoresis (PAGE) was used to study haemoglobin patterns in Indian elephants
Elephasmaximus. Blood samples werecollectedfrom 12 captive elephants atMudumalai Sanctuary, TamilNadu. Out
of 12 elephants, 10 (83.33%) showed two band Hb profile whereas 2 individuals (16.66%) showed a solitaiy
haemoglobin (Hb) band, suggesting homozygosity ofgene expression.
Introduction mation it was felt that the use ofPAGE may help in
establishing biochemical polymorphic pattern, if
Biochemical differentiation ofspecies specific any, in the Indian elephants. The frequency of oc-
proteins can be established by using electrophoretic currence of polymorphic locii and possible genetic
techniques (Selanderetal. 1969,Yoshidaetal. 1971 inter-relations may also be ascertained by the
and De Smet and William 1978). Ferguson (1980) analysis of the data on PAGE.
hadrecognizedthatalthough this techniquewas use- We are also working on 3 other proteins, viz.
ful in establishing differences between forms, it was LDH, Esterase and SDH to assess polymorphism at
not successful in bringing out similarities between specific gene locii. The results of these studies will
We
them. The limitations ofthe technique are overcome be published in due course. also plan to collect
in the comparative blood sample study of various blood samples from the elephant populations of
forms collected in an identical setofconditions with central and northern India to find out genetic af-
appropriatecontrols andtheobjectiveanalysis ofthe finities or differences, which will help in suggesting
data. Hb, being a stable molecule, is very useful in sub-specific status of the populations proposed ear-
biochemical studies. There is no record of Hb pat- lier by different authors.
terns in elephants in available literature. P.E.P.
MaterialandMethods
Deraniyagala (1955) has proposed 7 living sub-
species of the Asian elephant Elephas maximus, of Blood samples of 12 elephants were collected
which two are from India. This classification was by Dr. V. Krishnamurthy, Project Officer, Indian
based on morphological differences, amount of Elephant Project, BNHS. Blood was withdrawn
depigmentation andproportion oftuskers totuskless from the veins on theback ofthe earusing sterilized
males (Maknas) and tushed males (Aliyas). syringes and 16 gauge needles. Samples (5-10 ml
PVC
Further, he has suggested that the Asian each) were transferred to sterilized bottles and
elephant is polymorphic and its wide range of in- allowed to thaw at room temperature for a few
dividual variation has obscured the existence of a minutes to separate the serum. Then all the bottles
number of subspecies which have been revealed by were transferred to a liquid nitrogen cylinder(LN2
)
studying the intensity with which,certain variations to freeze them at-192°C. The samples were then
are localized to different areas. In Sri Lanka and transported with the cylinder to R.J. College, Bom-
India, 10 main varieties and numerous sub-varieties bay forlaboratory analysis. Hb was solubilised from
have been recognized from the earliest limes, sub- isolated RBC’s after a treatment with hypotonic
divisions being based on shape, size, colour, voice, saline (0.02%).
behaviour, strength, body odour, diet and suscep- Haemoglobins were separated by PAGE under
tibility to certain diseases. In the light of this infor- precisely controlled factors like gel concentration
pH
(7.5%), buffer system (Tris-glycine, 8.4), volt-
mA
1Accepted July 1989. age 250 mV, current 4 per tube, ambient
2Bombay Natural History Society, Hombill House, Shaheed temperature (4°C ± 1° C), duration ofrun etc. Some
BhagatSingh Road, Bombay 400 023. of the samples were run successively to check the
3Department of Biological Sciences, RJ. College, Ghatkopar,
Bombay 400 086. constancy ofall factors which helped in the analysis
18 JOURNAL, BOMBAY NATURAL HIST. SOCIETY, Vol. 88
Table
1
HAEMOGLOBINVARIANTS INTERMS OFRM VALUES,THEIR OCCURRENCEANDFREQUENCIES
INA SMALLDOMESTICATEDELEPHANTPOPULATIONIN SOUTHINDIA
Rm % %
values with No. of bands population frequency
S.D. (occurrence) ofbands
0.345 ±0.015 1 8.33 4.54
0.355 ± 0.005 2 16.66 9.09
0.355 ±0.015 1 8.33 4.54
0.37 ± 0.01 1 8.33 4.54
0.37 ± 0.02 1 8.33 4.54
0.375 ± 0.005 1 8.33 4.54
0.38 ± 0.02 1 8.33 4.54
0.385 ± 0.025 1 8.33 4.54
0.395 ± 0.005 1 8.33 4.54
0.56 ± 0.01 1 8.33 4.54
0.57 ± 0.02 1 8.33 4.54
0.575 ±0.015 2 16.66 9.09
0.59 ± 0.02 1 8.33 4.54
0.595 ±0.015 2 16.66 9.09
0.605 ±0.015 3 25.00 13.63
0.61 ±0.01 1 8.33 4.54
0.615 ±0.015 1 8.33 4.54
Total no. of specimens - 12 Total no. of bands— 22
Individual Animal
C Consolidated Hb pattern and S.D.
:
F Frequency (•/.)
;
Fig. 1. Diagrammatic representation ofhaemoglobin profile ofa small domesticated elephant population in south India
of the results. The dye Bromophenol Blue was lated as a ratio of the distance travelled by Hb from
loaded on the gel columns set in neutral glass tubes the base to the distance travelled by the marker in
Rm
along with samples and served as a marker. For the same run. The mean with a standard devia-
staining the gels after electrophoresis to identify Hb tion from the mean for each identified Hb band was
fractions, we followed the procedure of Ornstein recorded. Hb profiles for each individual were
(1967). Relative mobility (Rm) of Hb was calcu- prepared by plotting Rm values and a consolidated
HAEMOGLOBIN POLYMORPHISM IN ASIAN ELEPHANT 19
Hb profile for the population was constructed. Per- However, it is likely that the Hb2 is also an expres-
centage occurrence of a fraction in the population sion ofan independentgene locii with a slightly less
and percentage frequency of each band was calcu- frequency in the population.
lated. The band of the highest mobility was num- Polymorphic expression of gene locii for Hb
bered Hbl and band of lower mobility was num- has been reported for rodents (Pradhan et al. 1984,
bered Hb2 as suggested by Ferguson (1980). Selander et al. 1969), birds (Sane et al. 1986). The
Hb profile with an individual variation of single
Resultsand Discussion
band to as many as a total of 15 bands on consolida-
Table 1 showsrecordsoftherelativemobilities tion are reported for rodents, whereas for birds the
of haemoglobin fractions for 12 individual. The number varies from one to six on consolidation.
number of occurrences of a band in the population Thus in comparison, a two band pattern on con-
and its percentage frequency is given against each solidation in elephants appear to be more conserva-
band. The same data are represented in Fig. 1 as a tive.
consolidated diagrammatic profile for the samples
Acknowledgements
studied.
It is clear from Fig. 1 that out of 12 individual, We thank the Tamil Nadu forest department
10 showed 2 pattern and 2 individual showed a and Mr. Neelkantan, Wildlife Warden,Mudumalai
solitary band suggesting homozygosity of gene ex- Sanctuary forpermitting us to collectblood samples
pression for the Hbj locii. Thus it is evident that from someofthecaptive elephants; Mr. Ajay Desai,
16.66% ofthepopulation have homozygousexpres- Scientist, BNHS, who offered active participation
sion, whereas 83.33% population show two banded and gave practical suggestions on collection of
expression. The Hbi pattern with the highest blood samples; the BNHS for providing the LN
2
mobility is strongly expressed as compared to Hb2 cylinder for transportation of samples to Bombay;
A
pattern. single band profile for any protein is con- the Principal, R.J. College, Ghatkopar, Bombay, for
sidered as an expression of homozygous gene locii providing the laboratory and other facilities and C.
specific for such protein (Ferguson 1980). However G. Rajendran andRuben Solomon,R.J. College, for
in the present study a homozygous pattern was their great help in conducting the experiments.
found only for the locii of Hbi and not Hb2.
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&
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