ffournal of Genetics V o l u m e 63, N u m b e r 1 April 1977

HAPTOGLOBIN AND ACID PHOSPHATASE GENE DISTRIBUTIONS IN THE

DHANGARS OF MAHARASHTRA, INDIA

B . N . Mukherjee, S.K. Das, I<.C. Malhotra and S.R. D a s

A a t h r o p o m e t r y and H u m a n Genetics Unit, Indian Statistical Institute
Calcutta, India

The D h a n g a r s are a s e m i - n o m a d i c people, with a total population of about

3.5 millions. T h e y are found scattered over all the 26 districts of Riaharashtra.
T h e y speak M a r a t h i and are largely concentrated in the so-called "famine tracts"
of Maharashtra, comprising the districts of Nasik, Foona, Satara, I'(olhapur,
Sangll and Ahrnednagar. The average annual rainfall in this area is below 25 c m .

The D h a n g a r s are not a single m a t i n g unit but consist of 22 e n d o g a m o u s

groups, forming a caste cluster. T h e s e e n d o g a m o u s groups differ f r o m one
another in m a n y respects. Their population figures v a r y f r o m several lakhs to
a few thousands. Again, s o m e are restricted to a very small area, while others
are scattered in several districts, thereby cutting across different ecological
regions. S o m e of t h e m have b e c o m e settled hut dwellers and food growers, while
other-s still live a m o r e or less n o m a d i c life. Of the latter s o m e m o v e only within
a small territory, while a few travel far and wide covering h u n d r e d s of miles
every year. Variations are also noticed in the occupations of the various groups.
T h e D a n g e D h a n g a r s found in the coastal strip are a pastoral people and practise
shifting cultivatior~4 the Hatkars and a few others are shepherds ; the Sangars
are w e a v e r s ot woollen blankets ; the I'(hatik D h a n g a r s sell meat, but do not
rear sheep, and the Khutekars rear sheep and weave. S o m e of the groups prefer
consanguineous marriages, but others strictly prohibit it.

T h e presence of as m a n y as 22 e n d o g a m o u s groups in contiguous areas, with

different occupations, but a c o m m o n name, stimulated the late P r o f e s s o r Iravat[
K a r v e of the D e c c a n College (Poona) and the authors to take up a detailed genetic
survey of these people. Results on s e r u m haptog[obin and red cell acid phosphatase
are reported here.

MATERIALS AND METHODS

A total of 1241 blood s a m p l e s w e r e collected by finger pricks f r o m 15 D h a n g a r

groups, namely, llande, I(hutekars, Hatkars, Shegar, K u r m a r , Unikankan, Dange,
Hande, Ladshe, Hattikankan, Zede, M e n d h e , Halmat, Sanger and Telang[ living
in the several districts of M a h a r a r a s h t r a already mentioned. T h e blood s a m p l e s
w e r e preserved in an iced thermoilask in the field until they r e a c h e d the laboratory
at Calcutta by air w h e r e the sera and haemolysates w e r e p r e p a r e d and deep frozen

39
40 H a p t o g t o b i n and A c i d P h o s p h a t a s e genes

at - Z 0 ~ for storage. W h e r e a s h a p t o g l o b i n w a s s t u d i e d in a l l the 15 g r o u p s of

the D h a n g a r s , a c i d p h o s p h a t a s e c o u l d be s t u d i e d in o n l y t en of t h e m , l e a v i n g o u t
Kande, Kumar, Dange, L a d s h e and T a l a n g i . The e n t i r e field w o r k was
o r g a n i z e d by o n e of us (KCM).

Starch gel electrophorests

F o r the a n a l y s i s ot h a p t o g l o b t n p h e n o t y p e s h o r i z o n t a l s t a r c h g e l e l e c t r o -
p h o r e s t s w a s d o n e u s i n g a d i s c o n t i n u o u s b u f f e r s y s t e m ( P o u l i k 1957). T h e t e s t
f o r the r e d c e l l a c i d p h o s p h a t a s e s y s t e m u s e d the b u f f e r s y s t e m s and r e a c t i v e
m i x t u r e d e s c r i b e d in K a r p and S u t t o n (1967). T h e a c t i v i t y of a c i d P h o s p h a t a s e
e n z y m e w a s r e s t o r e d by a d d i n g I m l . of 2 - m e r c a p t o e t h a n o l to e a c h g e l a f t e r
degasstng. E a c h w a s p l a c e d b e t w e e n m e t a l c o o l i n g p l a t e s m a i n t a i n e d at 1 0 - 1 Z ~
d u r i n g the e [ e c t r o p h o r e t i c r u b s .

Results and discussion

T h e d i s t r i b u t i o n of the p h e n o t y p e s a n d the g e n e - f r e q u e n c l e s *or h a p t o g l o b i n

a m o n g the g r o u p s a r e p r e s e n t e d in T a b l e 1, a n d t h o s e f o r a c i d p h o s p h a t a s e in
T a b l e 2. T h e C h i - s q u a r e v a l u e s f o r c o n s i s t e n c y a r e a l s o i n c l u d e d in t h e s e t a b l e s ,
t h e s e v a l u e s i n d i c a t e d s a t i s f a c t o r y a g r e e m e n t w i t h the e x p e c t e d v a l u e s in e a c h
case.

Haptoglobin :

T h e f r e q u e n c i e s of the Hp 1 g e n e in the t w e l v e of the 15 g r o u p s s t u d i e d l i e

w i t h i n the r a n g e , 0.0715-0.1913. T h e t h r e e r e m a i n i n g g r o u p s , n a m e l y , the
H a l m a t , the S a n g a r an d the T e l a n g i ( T a b l e 1) show h i g h e r Hp I f r e q u e n c i e s , in
the r a n g e of 0 . 2 7 2 7 - 0 . 3 4 6 2 . It is not c l e a r if the a p p a r e n t l y h i g h e r f r e q u e n c i e s
in t h e s e t h r e e g r o u p s c a n be a t t r i b u t e d to s a m p l i n g e r r o r s r e s u l t i n g f r o m s m a l l
sample sizes. Hp 1 gene g e n . e r a t l y s h o w s a low f r e q u e n c y in I n d i a n p o p u l a t i o n s
(55/o to a b o u t 2 0 % ) . T h e l o w e s t f r e q u e n c y of 5~ w a s f o u n d in the N a i c k e r s of
T a m i l n a d u ( A n a n t h a k r t s h n a n et a[ 1969), I~[rk et a l (196I) o b s e r v e d 34% Hp 1
g e n e f r e q u e n c y a m o n g s t the T o d a s of N i l g i r i s , w h i c h is v e r y c l o s e to the
* r e q u e n c i e s o b s e r v e d in the H a l m a t , the S a n g a r and t h e T e l e n g [ . T h e s e t h r e e
g r o u p s a r e h o m o g e n e o u s a m o n g t h e m s e l v e s (xZ = 3. 375, Z d . f . ) w i t h r e f e r e n c e
to h a p t o g l o b i n g e n e c o u n t s . T w o c a s e s (0. 16%) of a h a p t o g l o b t n a e m i a (Hp0) w e r e
n o t i c e d in t h i s s t u d y . No r a r e Hp p h e n o t y p e w a s r o u n d in the D h a n g a r s . Among
I n d i a n p o p u l a t i o n s a s i n g l e c a s e of Hp ' J o h n s o n ' w a s d e t e c t e d in a B e n g a l i
H i n d u m a l e by M u k h e r j e e and D a s (1970) in an e a r l i e r i n v e s t i g a t i o n .

T h e h a p t o g l o b i n and the a c i d p h o s p h a t a s e g e n e d i s t r i b u t i o n s h a v e b e e n t e s t e d
f o r h o m o g e n e i t y in c o n t i n g e n c y t a b l e s . T h e c o n t i n g e n c y t a b l e s w e r e p r e p a r e d not
 B . N , M u k h e r j e e et at. 41

T A B L E- i

D[strs of Phenotypes and Genes for Haptoglob[n in D h a n g a r caste Cluster of M a h a r a s h t r a {India)

No Phenotypes .Z
Sub-caste groups Tested Hp 1 Gene + S.E. X
1-1 Z-1 2-2 0 lZr equency ( l . D. F . )
........................................................................................ .........

1 KANDE an 7 0 1 6 0
% 14.29 85.71 0.0714 + 0.0688 1,0801

2 KHUTIEKAR no 117 1 25 90 1
~, 0.85 21.38 76.92 0.85 0.1164 + 0.0210 0.2659

3 HATKAR no Z46 5 52 189 0

2.03 21.14 76.83 0.1260 + 0.0149 0.4007

4 SHEGAR no 84 0 22 62 0
r 26. 19 73.81 0. 1310 + 0, 0260 i. 1732

5 KURMAR no 105 0 28 76 i
~oo 26.67 72.38 0.95 0.1534 + 0.0234 1.1893

6 UNIKANKAN no 63 1 15 47 0
I. 59 23.81 14.60 0. 1350 + 0. 0304 0, 1530

7 DANGE no 47 2 i0 35 0
go
o 4.25 21.28 74.47 0.1489 + 0,0369 I. III0

8 HAND//: no 80 0 23 57 0
o 31.63 78.37 0. 158E + 0.028"8 1.0850

9 LADSHE no 85 3 21 61 0
0 3.53 24.71 71.76 0. 1589 + 0. 0280 0. 4854

i0 H A T T [I<:ANK A N no 34 0 ii 23 0
0.00 32.35 67,65 0.1618 + 0.0446 1.2029

11 ZADE no 145 4 43 98 0
0 2.76 29.66 67.58 0.1759 + 0.0223 0.1366

12 MENDHE no 149 7 43 99 0
4.70 28.86 66.44 0. {913 + 0. 0241 0. 3680

13 H A L/v[AT r~o ii I 4 6 0
9,09 36.36 54.55 0,2727 + 0.0949 0.3329

14 SANGAR no 55 6 21 28 0
% 10.91 38,18 50.91 0.3000 + 0.0436 0.341'9

15 TELANGI no 13 0 9 4 0
~o 69,23 30,17 0.3462 + 0.0933 I. 6410

Total ~o 1241 30 328 881 2

o Z. 42 26..43 70.99 0. 16 0. 1564 + 0. 0072 0, 0158

T h e X 2 Values in the last column~-however, s h o w that the phenntype Frequencies agree wLth the
hypothetical expecI;at~,oa~
H o m o g e n e i t y Ch[-square test b.y 13 x ;~ cons table for the H p gone count in the 15-group =
38. 855, IZ. D.F. I ~ 0, 001 (IZ single groups and 1 c o m b i n e d group of Halmas Telang[
and Sangaz).
42 Haptog[obin and Acid Phosphatase genes

T A B LE- Z

Distribution of Phenotypes and genes for Acid Phosphatase [n Dhangar caste cluster of Maharashtra {India)

PHS A
Sub- caste groups No
Tested Phenotypes gene + S.E. XZ
A AB B frequency (i. D. F. )
................... , ................................................... .............................

I SHEGAR no 99 i 19 79
% 1.01 19.19 79.80 0.1061 + 0.0218 0.1120

2 KHUTEKAR no 58 3 i0 45
% 5.17 17.24 77.59 0.1379 + 0.0310 2.0463

3 H A T I~AR no 267 8 60 199

3.00 22.47 74.53 0.1424 + 0. 0151 0. 5718

4 HATTIKANIAAN no 35 1 9 25
~o 2.86 25.71 71.43 0.1572 + 0.0435 0.1932

5 UNIKANKAN no 63 Z 16 45
% 3.18 25.39 71.43 0.1588 + 0.0325 0.3165

6 MENDHE no 151 7 35 109

% 4.63 23.18 72.19 0.1622 + 0.0212 0.9655

7 HANDE no 65 2 20 43
% 3. 08 30.77 66. 15 0. 1847 + 0. 0330 0. 1250

8 ZADE no 130 5 42 83
%o 3.85 32.31 65.84 0. 2001 + 0. 0247 0.0496

9 SANGAR no 40 2 13 25
% 5.00 32.50 62.50 0.2125 + 0.0457 0.1478

10 HALMAT no 14 1 4 9
% 7.14 28.57 64.29 0.2143 + 0.0775 0.7536

......................... - ..........................................................................
no 922 32 228 662
Total 4o 3.47 24.73 71. 80 0,1584 + 0.0085 0.4695
....................................................................................................

H o m o g e n e i t y C h [ - s q u a r e t e s t by 10 x Z c o n t i n g e n c y t a b l e f o r the P H S
g e n e c o u n t in the 10 g r o u p s = 1 1 . 8 8 , 9. D.l~., 0 . 3 0 ~ P > 0.20
 B . N . M n k h e r j e e et al. 43

with the phenotype n u m b e r s but with the gene counts. T h e Halmat, the Sangar
and the Telan~[ sections, on a preliminary examination w e r e found to be h o m o -
geneous an%ong themselves. T h e y possessed significantly higher l-lpI gene
frequencies than the others. H a l m a t and the Telang~ are, however, only ii and
13 respectLvely (too small). T h e sample sizes for these three groups have been
m e r g e d together into a single c o m b i n e d group in the contingency table for the
haptogloDin gene counts. The resulting table contained 13 r o w s corresponding to
12 single groups and 1 c o m b i n e d group, and Z c o l u m n s corresponding to the gene
counts for the alleles, H p I and H p Z. The Chi-square value obtained w a s 38. 855
with 12 d.f. { P < 0 . 0001). This is highly significant and the hypothesis of h o m o -
geneity is therefore rejected. O n scrutiny of the analysis [t nnay be seen that the
c o m b i n e d group (Halmat, Sangar and Telangi} alone contributed 25. 928 to the
Chi-square value; on leaving out this c o m b i n e d group, the Ch[-square value
c o m e s d o w n to 12. 934 with ii d.f. which is not significant (P very near to
0.30). If Chi-square ~s recalculated omitting these three groups f r o m the
contingency table, a still smaller Chl-square value will be obtained. L a r g e r
samples of the Halmat, the Sangar and the Telangi groups should be studied
belore an explanation is offered for these unusual data.

H o w e v e r , the haptoglobin data suggest that twelve of the 15 D h a n g a r groups

studied here are quite homogeneous, but the other three groups which are h o m o -
geneous a m o n g themselves differ significantly f r o m the rest.

Acid Phosphatase :

Three c o m m o n phenotypes of acid phosphatase A, A B and B, have been

observed in all the D h a n g a r groups. P H S A gene frequency in the ten Dha,ngar
groups for this e n z y m e ranges f r o m 0. 1016 (Shegar) and 0. Z143 (Halmat).
T h e allele P H S C is absent or rare in Indian populations. T w o cases of C A
phenotype w e r e detected in Bengalis of Calcutta by D a s et al (1970). T h e
highest frequency (0.4153) of the P H S A gene so far o h s e r v e d in India has been
a m o n g the Arora, a caste in Punjab in northern India {Blake es al 1971).

Data f r o m ten D h a n g a r groups which included the H a l m a t (14) and the

Sangar (40) in w h o m acid phosphatase have also been studied for h o m o g e n e i t y
by g Chi-square test on a contingency table. T h e r e w,~re 1O rows for the 10
groups and 2 c o l u m n s for the genes P H Z A and P H S B. The Chi-square value
of 11.88 with 9. d.f. obtained in this w a y w a s insignificant (0.50 P > 0. Z0)
T h e ten groups are, therefore, indistinguishable in respect of acid phosphatase.

Phosphatase gene frequencies :

The H a l m a t and the Sangar, which w e r e differentiated f r o m the rest (along

with Telangi) in the H p gene distribution, however, s h o w e d homogeneity with
the other 8 groups in their acid phosphatase gene frequencies.
44 Haptoglobin and Acid Phosphatase genes

AC K N O W L E D GEMENTS

Financial support f r o m the University Grants C o m m i s s i o n is gratefully

acknowledged. Additional support c a m e f r o m the Ind'lan Statistical Institute
(Calcutta) and D e c c a n College ~oona). W e recallwith gratitude the help of
the late Professor Irawati K a r v e w h o iniated this "work. W e are also grateful
to m a n y friends and officials of the G o v e r n m e n t of M a h a r a s h t r a for timely help.
Sri S a m a t Banerjee and Sr[ Badal D e y provided able technical assistance.

SUMMAR Y

A total of 1241 blood s a m p l e s f r o m 15 D h a n g a r groups - a s e m i - n o m a d i c

population, w e r e studied by starch get electrophoresis for haptoglobin and acid
phosphatas e types.

T h e gene frequencies for H p I in 12 of the 15 groups studied are between

0.0715 - o. 1913 and they are h o m o g e n e b u ~ a m o n g themselves, but significantly
differ f r o m the remaining three. The latter three groups s h o w higher H p I gene
frequencies, in the range of 0.272? - 0.3462, and suggest that they are also quite
homogeneous.

A, A B and B phenotypes of acid phosphatase w e r e seen a m o n g all the groups.

Frequencies of the P A S A gene ranges f r o m 0.1016 to 0.2143 in the ten groups
tested for this enzyme.

REFERENCES

Ananthakrlshnan, R. and Kirk, R . L . 1969. T h e distribution of s o m e

s e r u m Protein and e n z y m e group s y s t e m s in two e n d o g a m o u s groups
in S. India. Ind. Jour. M e d . Res., 57 : 1011-1017.

Blake, N . M . , Kirk, R.L., M c D e r m l d , E.M., Kefichi, O. and Ahaja,

Y.R. 1971. The distribution of s e r u m proteins an~ e n z y m e group
s y s t e m s a m o n g North Indians. H u m . Hered. 2 1 : 4 4 0 - 4 5 7 .

Das, S.R., Mukherjee, B.N., Das, S.I~., Blake, N . M . and Kirk, R . L .

1970. The distribution of s o m e e n z y m e group s y s t e m s a m o n g Bengalis.
Indian J. Med. Res., 58: 866-875.

l~[rk, R.L., and La[, L . Y . C . 1961. The distribution of haptoglobin and

transferrln in South and South East Asia. Acta Genet. Besel. 11:97-105

Karp, C . W . and Sutton, I-I.E. 1967. S o m e n e w phenotypes of h u m a n red

cell acid phosphatase. A m e r J. H u m . Genet. 19: 54-62.
 B.N. Mukherjee eta[. 45

Mukherjee, B.N. and Das, S.K. 1970. The l-laptoglobin and Transferrin
Types in W e s t Bengal and a caseof Haptoglobin 'Johnson I. H u m . I-lered.
Z0:Z09-ZI4

Poulik, M . D . 1957. Starch gel electrophoresks in discontinuous syste1~n of

buffer. Nature, 180: 1477-1479.