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Lead toxicity: Biochemical and physiological imbalance in nine freshwater teleosts
Toxicology Letters, 4( 1979) 155-161 o Elsevier/North-Holland Biomedical Press
155
LEAD TOXICITY: BIOCHEMICAL AND PHYSIOLOGICAL IMBALANCE IN NINE FRESHWATER TELEOSTS
S.A. SHAFFI Department
of Science, R.C.E., N.C.E.R.T.,
Bhopal-
462013
(India)
(Received April 30th, 1979) (Accepted May 12th, 1979)
SUMMARY
The effect of various concentrations (5, 10, 15 and 20 ppm) of lead nitrate on liver, muscle, brain and kidney glycogen and on serum glucose and lactate was studied in 9 species of fish. Glycogenolysis was higher in major carp species than in catfish or murrels. The lowest level of glycogen was in muscle, followed by liver, kidney and brain. There was an inverse relationship between the concentration of lead nitrate and the fall in liver, muscle and brain glycogen. There was a direct relationship between the concentration of lead nitrate and the levels of serum glucose and lactate, which were again higher in the major carps.
INTRODUCTION
Pollution may lead to a concentration in rivers and estuaries of heavy metals to which fish have not previously been exposed [ 7,9] . Fish are of great nutritional significance in India and the implication of lead poisoning from their consumption would be considerable [ 2,3] . The effect of lead nitrate (5, 10, 15 and 20 ppm) on liver, muscle, brain and kidney glycogen, serum glucose and lactate levels in 9 nutritionally and economically important species of fish, viz. Lubeo rohitu (Ham), Cirrhinu mrigula (Ham), Cutlu c&u (Ham), Clarius butruchus (Linn), Heteropneustes fossilis Bloch, Mystus seenghalu (Sykes), Chunnu s triutus Bloch, Chunnu punctutus Bloch and Channu marulius (Ham). MATERIAL AND METHODS
Healthy specimens of L. rohitu, C. mrigula, C. catla, C. batrachus, H. fossilis, M. seenghala, C. striutus, C. punctatus, and C. marulius (size range 18-20 cm) were obtained locally and acclimatized in the laboratory. Six fish of each species were sacrificed for initial estimation of liver, muscle, brain and kidney
156
glycogen, serum glucose and serum lactate were maintained in concentrations of lead for 3 h, then were removed and decapitated kidneys removed. The collection of serum samples and estimation of glycogen, serum were as previously described.
levels. Equal numbers of fish nitrate of 5, 10, 15 and 20 ppm and the liver, muscle, brain and [ 5,12,16], preparation of tissue glucose and serum lactate [ 10-151
TABLE I VARIATIONS IN TISSUE GLYCOGEN CONTENT DUE TO LEAD INTOXICATION - MAJOR CARPS Tissue
Control
Lead in ppm
% of fall
5
10
15
20
4986.94 + 364.28
4458.10 t210.19
3649.78 k324.38
1986.10 t160.38
1064.17 lt95.10
70.60
Muscle
3236.19 t290.18
2854.39 i195.14
2150.44 +240.58
1380.50 t110.56
640.16 k74.10
80.00
Brain
1250.14 +135.36
964.38 *62.16
710.20 +120.90
489.12 t 38.94
380.11 t29.10
69.60
840.82 t64.38
1420.13 t150.66
1924.10 ~124.38
940.44 t 68.94
315.29 f 28.36
62.50
4492.56 lt412.10
3650.36 c290.38
2150.19 5210.64
1444.38 $195.10
750.56 f 102.44
80.00
Muscle
3085.38 k210.56
2586.10 5205.19
1680.12 *145.90
950.10 t131.98
425.00 t39.10
74.80
Brain
1176.12 * 96.88
924.38 k125.19
454.34 t72.10
315.24 k45.50
73.00
778.66 t69.38
1360.90 k114.12
1760.86 k135.11
1080.54 +98.19
270.22 ?:19.46
65.00
4437.24 1t170.38
3856.44 k23.0.44
2938.50 i270.10
1420.50 t84.88
624.10 ~42.17
85.30
Muscle
2932.18 t21O.J.Q
2438.10 i180.38
1890.38 t195.10
820.55 f 104.36
324.99 k20.10
85.00
Brain
1025.29 * 140.58
840.58 f 84,56
690.56 * 75.01
390.58 k70.24
245.48 * 18.96
76.00
730.54 *95.10
1096.48 268.96
1428.50 zk139.21
520.48 k46.48
194.12 k34.10
73.00
C. mrigala Liver
Kidney
L. rohita Liver
Kidney
C. catla Liver
Kidney
628.19 kQ5.48
Values @g/g wet wt. of tissue) are mean i: SE of 6 replicates.
157 RESULTS
No visible signs of toxic reaction were detected, except for the formation of a thin mucus layer at the opercular region, more marked in the major carp species (C. catla, L. rohita, C. mrigala) than in catfish (M. seenghala, H. fossilis, C. batrachus) OPI murrels (C. ~unc~a~~s, C. ~ar~Z~~s, C. striatus). The scarring and shrinking of liver and kidney was common to all 9 species. TABLE II VARIATIONS - CATFISH Tissue
IN TISSUE GLYCOGEN CONTENT DUE TO LEAD INTOXICATION
ColltXOl
Lead in ppm
% of fall
5
10
15
20
2530.16 f 195.78
1950.48 k215.10
1520.18 i84.50
1250.11 t50.12
56.10
1810.38 +99.10
1624.98 k95.16
1196.66 *85.10
950.24 258.11
650.46 t38.16
64.44
Brain
720.56 82.10
510.44 48.56
390.32 48.16
295.52 24.12
248.19 34.18
65.41
Kidney
515.94 *30.19
850.39 k56.56
1025.40 e48.56
690.18 259.17
160.64 +-l&12
68.90
2530.88 +120,x2
2150.48 ~150.48
1636.38 294.16
1210.19 *76.10
1090.30 k84.22
56.90
1690.44 i80.36
1410.56 k99.10
1090.44 t115.11
744.16 *38.45
440.72 129.11
73.00
Brain
490.68 546.10
386.49 k29.36
298.40 *.25.x0
240.92 i20.38
195.77 t19.50
60.20
Kidney
470.29 k52.11
696.38 184.44
892.56 k36.46
424.84 k40.99
130.12 k12.10
72.20
M. seenghata Liver 2372.99 -r165.24
1874.82 *134.10
1340.88 278.18
955.10 ~64.17
600.48 k-38.28
72.21
1430.58 ~110.10
1180.38 t124.16
948.26 i94.10
624.19 2 29.38
345.22 219.10
75.00
Brain
605.24 t84.12
474.14 t66.34
324.90 * 36.10
240.15 k20.19
185.11 k24.11
68.00
Kidney
390.19 t28.10
540.18 k48.96
764.98 i 58.10
328.70 * 29.16
130.45 *19.84
66.00
C. batrachus Liver 2850.24 t172.18 Muscle
H. fossilk Liver Muscle
Muscle
Values @g/g wet wt. of tissue) are mean f SE of 6 replicates.
158
TABLE
III
VARIATIONS IN TISSUE GLYCOGEN - SNAKE-HEADED FISH Tissue
Control
CONTENT
DUE TO LEAD
INTOXICATION
%-of fall
Lead in ppm 5
10
15
20
C. striatus Liver
3410.77 k210.98
3184.54 5150.12
2786.37 *113.40
2040.12 2129.32
1609.10 ~56.24
53.62
Muscle
2396.46 ~184.38
1970.50 +240.30
1540.64 k95.18
1120.36 k104.58
950.04 f 74.45
42.22
Brain
987.35 + 102.40
834.56 *79.11
620.18 t64.64
540.56 k50.19
480.34 *41.30
50.35
640.12 +64.18
790.58 i58.11
980.56 579.10
425.19 +38.10
250.20 +18.16
60.31
C. punctatus Liver 3296.21 + 380.86
2988.15 5175.10
2380.50 *135.58
1720.56 k115.34
1290.18 k64.10
2285.66 k220.18
1896.24 k129.38
1356.48 k84.10
1020.44 +85.36
780.24 t38.12
Brain
905.51 k95.16
768.40 +94.80
620.30 e65.10
515.36 -r95.18
450.64 k28.12
Kidney
595.05 563.78
720.18 k52.12
890.58 k59.19
380.18 k40.11
215.22 t21.18
63.80
C. marulius Liver 3017.18 + 294.44
2715.80 k134.10
1940.16 i105.74
1540.35 ?r75.10
1140.26 k74.16
62.21
1920.10 k118.38
1380.34 i90.38
990.10 k56.74
690.32 k46.12
67.73
Kidney
Muscle
Muscle
2139.42 2180.58
60.89 65.80
Brain
840.82 f 94.54
610.48 +60.38
549.18 * 64.58
440.24 i44.38
325.29 *19.19
61.00
Kidney
565.99 t38.90
684.38 +42.42
920.62 A90.10
295.11 +12.36
190.17 +21.19
48.60
Values
(pg/g
wet wt. of tisSue) are mean + SE of 6 replicates.
Lead intoxication led to significant variations in glycogen content of tissues and in the levels of serum glucose and lactate (Tables I-V). There was an inverse relationship between the concentration of lead nitrate and the fall in glycogen content of liver, muscle and brain by comparison with initial values. The lower level of glycogen was in muscle, followed by liver, kidney and brain. In kidney there was an elevation in glycogen content at 5 and 10 ppm but a
159 TABLE IV VARIATIONS IN SERUM GLUCOSE LEVEL DUE TO LEAD INTOXICATION NINE FRESHWATER TELEOSTS Name of the fish
Control
Lead in ppm
Rise in folds
5
10
15
20
1085.32 k54.16
1250.38 k84.10
1536.49
1950.18 A98.10
2387.00 k112.15
3.4
c210.11
L. rohita
1324.84 Ago.38
1896.36 k124.10
2638.10 k194.38
3510.56 f 240.58
4601.60 t380.16
2.2
C. catla
906.12 t47.10
1530.24 k79.14
2270.15 k145.12
3128.14 t198.17
3714.68 k240.12
4.1
590.40 k34.10
650.12 k50.18
820.44 f 74.10
990.94 * 64.36
1239.75 *80.12
2.1
H. fossilis
730.82 *45.54
850.38 k72.10
960.56 261.50
1025.56 k95.38
1168.40 k90.16
1.6
M. seenghala
470.90 i29.19
610.70 k48.10
850.12 k84.10
1120.48 k79.86
1363.48 f 59.46
2.9
415.59 k34.16
490.10 k44.10
560.38 k52.17
625.84 k90.44
705.54 k58.17
1.7
C. punctatus
520.40 t42.10
680.18 t56.28
740.54 t64.10
864.38 574.16
988.16 k94.22
1.9
C. marulius
380.12 t21.38
420.84 535.10
590.44 k29.36
790.12 k80.12
950.41 + 69.50
2.5
Major carps C. mrigala
Catfish C. batrachus
Murrels C. striatus
IN
Values @g/ml serum) are mean + SE of 6 replicates.
decrease at 15 and 20 ppm. There was a direct relationship between the concentration of lead and the level of serum glucose and lactate. The greatest fall in tissue glycogen was recorded in the major carps (Table I), followed by catfish (Table II) and snake-headed fish or murrels (Table III). Serum glucose was highest in the major carps (Table IV) followed by catfish and murrels (Table IV). Levels of serum lactate were higher in the major carps (Table V) than in murrels (Table V) or catfish (Table V). DISCUSSION
Heavy metal intoxication in fish has been associated with disruption of metabolic activities, suppression of breeding potential, impaired survival of young, depressed growth at all stages, mucus formation on the body and at
160 TABLE
V
VARIATIONS IN SERUM LACTATE NINE FRESHWATER TELEOSTS Name of the fish
Control
LEVEL
DUE TO LEAD
INTOXICATION
Lead in ppm
Rise in folds
5
10
15
20
134.36 k12.10
190.11 k24.12
524.39 *40.38
756.12 r72.16
1046.40 k24.14
6.2
L. rohita
109.12 k18.24
236.22 +28.19
490.80 k39.18
648.48 k49.12
830.80 k64.10
9.6
C. catla
96.01 +lO.ll
350.19 ~34.18
680.64 k54.10
950.90 e74.18
1190.40 299.10
12.4
44.09 t10.42
92.19 ill.10
150.11 k24.28
195.96 t17.18
259.64 i19.10
3.5
H. fossilis
40.24 k9.44
68.40 +13.10
125.84 +15.10
180.72 k12.48
204.30 59.48
M. seenghala
36.15 k8.44
84.96 +24.10
195.22 t16.98
230.18 k24.10
363.60 k31.40
7.9
74.40 ?9.80
120.36 k11.10
164.10 k18.10
195.44 +16.84
259.24 k22.10
5.9
68.36 k7.10
105.19 t8.78
190.22 +20.12
250.33 + 25.10
367.28 k42.10
5.1
52.19 f 10.40
130.30 k14.10
210.56 k11.50
334.09 ~38.10
410.84 k45.80
Major carps C. mrigala
Catfish C. batrachus
Murrels C. striatus
C. punctatus C. marulius
Values
(pg/mi
serum)
IN
-
are mean + SE of 6 replicates.
the opercular region, breakdown of gas exchange, tissue hypoxia, and accumulation or imbalance in various physiological and biochemical processes [1,4,6,8,9,11,12,14-161. Deposition of mucus at the opercular region may interfere with gas exchange and reduce the oxygen transport to various internal organs, resulting in acidosis and the accumulation of glucose and lactate. Disturbance of the buffering system would enhance glycogenolysis [12,15-171. The fall in tissue glycogen content and the rise in serum glucose and lactate may be related to the cardiotoxicity of lead [ 12,161. The rise in renal glycogen observed at 5 and 10 ppm may represent an attempt by the kidney to conserve glycogen. At higher levels, lead intoxication leads to visible changes in kidney tubules with impaired function [ 21. The fall in the glycogen content of kidney agrees well with impaired function.
161
The variations in glycogen content and in serum glucose and lactate may be related to the fact that the major carp species obtain oxygen only through gills, whereas the other two groups possess accessory respiratory organs. The biochemical compartmentation of fish tissues may also be involved [ 13,181. ACKNOWLEDGEMENTS
The author is grateful to Prof. J.S. Rajput, Dr. G.K. Lehri, Dr. C.K. Ashok Kumar and Dr. Sandhya for encouragement and for many useful suggestions. The financial assistance rendered by U.G.C., New Delhi is gratefully acknowledged. REFERENCES 1 D.T. Burton, AH. Jones and J. Cairns, Acute zinc toxicity to Rainbow trout: Confirmation of the hypothesis that death is related to tissue hypoxia, J. Fish. Res. Bd. Can., 29 (1972) 1463-1466. J.J. Chisolm, Lead poisoning, Sci. Amer., 10 (1976) 3715-3724. S.K. Hall, Lead pollution and poisoning. Environ. Sci. Tech., 6 (1972) 30-35. G.M. Hughes, Polluted respiratory physiology in A.P.M. Lockwood (Ed.) Effects of Pollutants on Aquatic Organisms, Vol. 2, Cambridge Univ. Press, London, 1976, p. 163. AK. Jafri and S.A. Shaffi, Effect of asphyxiation on the serum and kidney alkaline phosphatase activity in Clarias batrachus (Linn) Broteria XLIV (1975) 59-62. V.G. Jhingran, Ecotoxicological hazards of the inland fisheries of India, Natl. Coll. Biochem. Fish., (1977) 15-16. AD. Kumar, Modern concepts of ecology, Vikas Publicity House, New Delhi, 1977, 101-140. S.D. Lewis and W.M. Lewis, The effect of zinc and copper on the osmolality of blood serum of the channel cat fish, Zctalurus punctatus Rafinesque and Golden Shiner Notemigonous crysoleucas Mitchill, Trans. Am. Fish. Sot., 100 (1971) 639-643. Effects of Pollutants on Aquatic Organisms, Cambridge Univ. Press, 9 A.P.M. Lockwood, London, 1976, pp. 7-34. distribution of glycogen, lactate and 10 S.A. Shaffi and M. Habibulla, Differential pyruvate in different regions of the adult rat brain, Ind. J. Exp. Biol., 15 (1977) 309-310. in three 11 S.k Shaffi, Changes in tissue glycogen content due to cadmium intoxication fresh water teleosts, Cm-r. Sci., 47 (1978) 868-870. 12 S.A. Shaffi, Variations in tissue glycogen content, serum lactate and glucose levels due to copper intoxication in three fresh water teleosts, Cm-r. Sci., 47 (1978) 954-956. compartmentation of fish tissues, I. Brain energy reserve and 13 S.A. Shaffi, Biochemical its metabolic products. Acta Physiol., 99 (1978) 35-40. on tissue glycogen 14 S.A. Shaffi and M.A. Qayyum, Effect of cadmium intoxication content in a fresh water cat fish H. fossilis Bloch, Zool. Jb. Anat., 99 (1978) 129-132. on some fresh water fishes, I. Variations in 15 S.A. Shaffi, Effect of zinc intoxication tissue glycogen content, Arch. Gen. Exp. Zool., (1979) (In Press). on fresh water fishes, II. Accumulation of 16 S.A. Shaffi, Effect of zinc intoxication metabolic products, Toxicol. Lett., 3 (1979) 319-323. toxicity to fresh water fishes. Toxicol. Lett., 4 (1979) 17 S.A. Shaffi, The acute heptachlor 31-37. compartmentation of fish tissues, II. Non-specific phos18 S.A. Shaffi, Biochemical phomono esterases in brain, Experientia, (1979) (In Press).
155
LEAD TOXICITY: BIOCHEMICAL AND PHYSIOLOGICAL IMBALANCE IN NINE FRESHWATER TELEOSTS
S.A. SHAFFI Department
of Science, R.C.E., N.C.E.R.T.,
Bhopal-
462013
(India)
(Received April 30th, 1979) (Accepted May 12th, 1979)
SUMMARY
The effect of various concentrations (5, 10, 15 and 20 ppm) of lead nitrate on liver, muscle, brain and kidney glycogen and on serum glucose and lactate was studied in 9 species of fish. Glycogenolysis was higher in major carp species than in catfish or murrels. The lowest level of glycogen was in muscle, followed by liver, kidney and brain. There was an inverse relationship between the concentration of lead nitrate and the fall in liver, muscle and brain glycogen. There was a direct relationship between the concentration of lead nitrate and the levels of serum glucose and lactate, which were again higher in the major carps.
INTRODUCTION
Pollution may lead to a concentration in rivers and estuaries of heavy metals to which fish have not previously been exposed [ 7,9] . Fish are of great nutritional significance in India and the implication of lead poisoning from their consumption would be considerable [ 2,3] . The effect of lead nitrate (5, 10, 15 and 20 ppm) on liver, muscle, brain and kidney glycogen, serum glucose and lactate levels in 9 nutritionally and economically important species of fish, viz. Lubeo rohitu (Ham), Cirrhinu mrigula (Ham), Cutlu c&u (Ham), Clarius butruchus (Linn), Heteropneustes fossilis Bloch, Mystus seenghalu (Sykes), Chunnu s triutus Bloch, Chunnu punctutus Bloch and Channu marulius (Ham). MATERIAL AND METHODS
Healthy specimens of L. rohitu, C. mrigula, C. catla, C. batrachus, H. fossilis, M. seenghala, C. striutus, C. punctatus, and C. marulius (size range 18-20 cm) were obtained locally and acclimatized in the laboratory. Six fish of each species were sacrificed for initial estimation of liver, muscle, brain and kidney
156
glycogen, serum glucose and serum lactate were maintained in concentrations of lead for 3 h, then were removed and decapitated kidneys removed. The collection of serum samples and estimation of glycogen, serum were as previously described.
levels. Equal numbers of fish nitrate of 5, 10, 15 and 20 ppm and the liver, muscle, brain and [ 5,12,16], preparation of tissue glucose and serum lactate [ 10-151
TABLE I VARIATIONS IN TISSUE GLYCOGEN CONTENT DUE TO LEAD INTOXICATION - MAJOR CARPS Tissue
Control
Lead in ppm
% of fall
5
10
15
20
4986.94 + 364.28
4458.10 t210.19
3649.78 k324.38
1986.10 t160.38
1064.17 lt95.10
70.60
Muscle
3236.19 t290.18
2854.39 i195.14
2150.44 +240.58
1380.50 t110.56
640.16 k74.10
80.00
Brain
1250.14 +135.36
964.38 *62.16
710.20 +120.90
489.12 t 38.94
380.11 t29.10
69.60
840.82 t64.38
1420.13 t150.66
1924.10 ~124.38
940.44 t 68.94
315.29 f 28.36
62.50
4492.56 lt412.10
3650.36 c290.38
2150.19 5210.64
1444.38 $195.10
750.56 f 102.44
80.00
Muscle
3085.38 k210.56
2586.10 5205.19
1680.12 *145.90
950.10 t131.98
425.00 t39.10
74.80
Brain
1176.12 * 96.88
924.38 k125.19
454.34 t72.10
315.24 k45.50
73.00
778.66 t69.38
1360.90 k114.12
1760.86 k135.11
1080.54 +98.19
270.22 ?:19.46
65.00
4437.24 1t170.38
3856.44 k23.0.44
2938.50 i270.10
1420.50 t84.88
624.10 ~42.17
85.30
Muscle
2932.18 t21O.J.Q
2438.10 i180.38
1890.38 t195.10
820.55 f 104.36
324.99 k20.10
85.00
Brain
1025.29 * 140.58
840.58 f 84,56
690.56 * 75.01
390.58 k70.24
245.48 * 18.96
76.00
730.54 *95.10
1096.48 268.96
1428.50 zk139.21
520.48 k46.48
194.12 k34.10
73.00
C. mrigala Liver
Kidney
L. rohita Liver
Kidney
C. catla Liver
Kidney
628.19 kQ5.48
Values @g/g wet wt. of tissue) are mean i: SE of 6 replicates.
157 RESULTS
No visible signs of toxic reaction were detected, except for the formation of a thin mucus layer at the opercular region, more marked in the major carp species (C. catla, L. rohita, C. mrigala) than in catfish (M. seenghala, H. fossilis, C. batrachus) OPI murrels (C. ~unc~a~~s, C. ~ar~Z~~s, C. striatus). The scarring and shrinking of liver and kidney was common to all 9 species. TABLE II VARIATIONS - CATFISH Tissue
IN TISSUE GLYCOGEN CONTENT DUE TO LEAD INTOXICATION
ColltXOl
Lead in ppm
% of fall
5
10
15
20
2530.16 f 195.78
1950.48 k215.10
1520.18 i84.50
1250.11 t50.12
56.10
1810.38 +99.10
1624.98 k95.16
1196.66 *85.10
950.24 258.11
650.46 t38.16
64.44
Brain
720.56 82.10
510.44 48.56
390.32 48.16
295.52 24.12
248.19 34.18
65.41
Kidney
515.94 *30.19
850.39 k56.56
1025.40 e48.56
690.18 259.17
160.64 +-l&12
68.90
2530.88 +120,x2
2150.48 ~150.48
1636.38 294.16
1210.19 *76.10
1090.30 k84.22
56.90
1690.44 i80.36
1410.56 k99.10
1090.44 t115.11
744.16 *38.45
440.72 129.11
73.00
Brain
490.68 546.10
386.49 k29.36
298.40 *.25.x0
240.92 i20.38
195.77 t19.50
60.20
Kidney
470.29 k52.11
696.38 184.44
892.56 k36.46
424.84 k40.99
130.12 k12.10
72.20
M. seenghata Liver 2372.99 -r165.24
1874.82 *134.10
1340.88 278.18
955.10 ~64.17
600.48 k-38.28
72.21
1430.58 ~110.10
1180.38 t124.16
948.26 i94.10
624.19 2 29.38
345.22 219.10
75.00
Brain
605.24 t84.12
474.14 t66.34
324.90 * 36.10
240.15 k20.19
185.11 k24.11
68.00
Kidney
390.19 t28.10
540.18 k48.96
764.98 i 58.10
328.70 * 29.16
130.45 *19.84
66.00
C. batrachus Liver 2850.24 t172.18 Muscle
H. fossilk Liver Muscle
Muscle
Values @g/g wet wt. of tissue) are mean f SE of 6 replicates.
158
TABLE
III
VARIATIONS IN TISSUE GLYCOGEN - SNAKE-HEADED FISH Tissue
Control
CONTENT
DUE TO LEAD
INTOXICATION
%-of fall
Lead in ppm 5
10
15
20
C. striatus Liver
3410.77 k210.98
3184.54 5150.12
2786.37 *113.40
2040.12 2129.32
1609.10 ~56.24
53.62
Muscle
2396.46 ~184.38
1970.50 +240.30
1540.64 k95.18
1120.36 k104.58
950.04 f 74.45
42.22
Brain
987.35 + 102.40
834.56 *79.11
620.18 t64.64
540.56 k50.19
480.34 *41.30
50.35
640.12 +64.18
790.58 i58.11
980.56 579.10
425.19 +38.10
250.20 +18.16
60.31
C. punctatus Liver 3296.21 + 380.86
2988.15 5175.10
2380.50 *135.58
1720.56 k115.34
1290.18 k64.10
2285.66 k220.18
1896.24 k129.38
1356.48 k84.10
1020.44 +85.36
780.24 t38.12
Brain
905.51 k95.16
768.40 +94.80
620.30 e65.10
515.36 -r95.18
450.64 k28.12
Kidney
595.05 563.78
720.18 k52.12
890.58 k59.19
380.18 k40.11
215.22 t21.18
63.80
C. marulius Liver 3017.18 + 294.44
2715.80 k134.10
1940.16 i105.74
1540.35 ?r75.10
1140.26 k74.16
62.21
1920.10 k118.38
1380.34 i90.38
990.10 k56.74
690.32 k46.12
67.73
Kidney
Muscle
Muscle
2139.42 2180.58
60.89 65.80
Brain
840.82 f 94.54
610.48 +60.38
549.18 * 64.58
440.24 i44.38
325.29 *19.19
61.00
Kidney
565.99 t38.90
684.38 +42.42
920.62 A90.10
295.11 +12.36
190.17 +21.19
48.60
Values
(pg/g
wet wt. of tisSue) are mean + SE of 6 replicates.
Lead intoxication led to significant variations in glycogen content of tissues and in the levels of serum glucose and lactate (Tables I-V). There was an inverse relationship between the concentration of lead nitrate and the fall in glycogen content of liver, muscle and brain by comparison with initial values. The lower level of glycogen was in muscle, followed by liver, kidney and brain. In kidney there was an elevation in glycogen content at 5 and 10 ppm but a
159 TABLE IV VARIATIONS IN SERUM GLUCOSE LEVEL DUE TO LEAD INTOXICATION NINE FRESHWATER TELEOSTS Name of the fish
Control
Lead in ppm
Rise in folds
5
10
15
20
1085.32 k54.16
1250.38 k84.10
1536.49
1950.18 A98.10
2387.00 k112.15
3.4
c210.11
L. rohita
1324.84 Ago.38
1896.36 k124.10
2638.10 k194.38
3510.56 f 240.58
4601.60 t380.16
2.2
C. catla
906.12 t47.10
1530.24 k79.14
2270.15 k145.12
3128.14 t198.17
3714.68 k240.12
4.1
590.40 k34.10
650.12 k50.18
820.44 f 74.10
990.94 * 64.36
1239.75 *80.12
2.1
H. fossilis
730.82 *45.54
850.38 k72.10
960.56 261.50
1025.56 k95.38
1168.40 k90.16
1.6
M. seenghala
470.90 i29.19
610.70 k48.10
850.12 k84.10
1120.48 k79.86
1363.48 f 59.46
2.9
415.59 k34.16
490.10 k44.10
560.38 k52.17
625.84 k90.44
705.54 k58.17
1.7
C. punctatus
520.40 t42.10
680.18 t56.28
740.54 t64.10
864.38 574.16
988.16 k94.22
1.9
C. marulius
380.12 t21.38
420.84 535.10
590.44 k29.36
790.12 k80.12
950.41 + 69.50
2.5
Major carps C. mrigala
Catfish C. batrachus
Murrels C. striatus
IN
Values @g/ml serum) are mean + SE of 6 replicates.
decrease at 15 and 20 ppm. There was a direct relationship between the concentration of lead and the level of serum glucose and lactate. The greatest fall in tissue glycogen was recorded in the major carps (Table I), followed by catfish (Table II) and snake-headed fish or murrels (Table III). Serum glucose was highest in the major carps (Table IV) followed by catfish and murrels (Table IV). Levels of serum lactate were higher in the major carps (Table V) than in murrels (Table V) or catfish (Table V). DISCUSSION
Heavy metal intoxication in fish has been associated with disruption of metabolic activities, suppression of breeding potential, impaired survival of young, depressed growth at all stages, mucus formation on the body and at
160 TABLE
V
VARIATIONS IN SERUM LACTATE NINE FRESHWATER TELEOSTS Name of the fish
Control
LEVEL
DUE TO LEAD
INTOXICATION
Lead in ppm
Rise in folds
5
10
15
20
134.36 k12.10
190.11 k24.12
524.39 *40.38
756.12 r72.16
1046.40 k24.14
6.2
L. rohita
109.12 k18.24
236.22 +28.19
490.80 k39.18
648.48 k49.12
830.80 k64.10
9.6
C. catla
96.01 +lO.ll
350.19 ~34.18
680.64 k54.10
950.90 e74.18
1190.40 299.10
12.4
44.09 t10.42
92.19 ill.10
150.11 k24.28
195.96 t17.18
259.64 i19.10
3.5
H. fossilis
40.24 k9.44
68.40 +13.10
125.84 +15.10
180.72 k12.48
204.30 59.48
M. seenghala
36.15 k8.44
84.96 +24.10
195.22 t16.98
230.18 k24.10
363.60 k31.40
7.9
74.40 ?9.80
120.36 k11.10
164.10 k18.10
195.44 +16.84
259.24 k22.10
5.9
68.36 k7.10
105.19 t8.78
190.22 +20.12
250.33 + 25.10
367.28 k42.10
5.1
52.19 f 10.40
130.30 k14.10
210.56 k11.50
334.09 ~38.10
410.84 k45.80
Major carps C. mrigala
Catfish C. batrachus
Murrels C. striatus
C. punctatus C. marulius
Values
(pg/mi
serum)
IN
-
are mean + SE of 6 replicates.
the opercular region, breakdown of gas exchange, tissue hypoxia, and accumulation or imbalance in various physiological and biochemical processes [1,4,6,8,9,11,12,14-161. Deposition of mucus at the opercular region may interfere with gas exchange and reduce the oxygen transport to various internal organs, resulting in acidosis and the accumulation of glucose and lactate. Disturbance of the buffering system would enhance glycogenolysis [12,15-171. The fall in tissue glycogen content and the rise in serum glucose and lactate may be related to the cardiotoxicity of lead [ 12,161. The rise in renal glycogen observed at 5 and 10 ppm may represent an attempt by the kidney to conserve glycogen. At higher levels, lead intoxication leads to visible changes in kidney tubules with impaired function [ 21. The fall in the glycogen content of kidney agrees well with impaired function.
161
The variations in glycogen content and in serum glucose and lactate may be related to the fact that the major carp species obtain oxygen only through gills, whereas the other two groups possess accessory respiratory organs. The biochemical compartmentation of fish tissues may also be involved [ 13,181. ACKNOWLEDGEMENTS
The author is grateful to Prof. J.S. Rajput, Dr. G.K. Lehri, Dr. C.K. Ashok Kumar and Dr. Sandhya for encouragement and for many useful suggestions. The financial assistance rendered by U.G.C., New Delhi is gratefully acknowledged. REFERENCES 1 D.T. Burton, AH. Jones and J. Cairns, Acute zinc toxicity to Rainbow trout: Confirmation of the hypothesis that death is related to tissue hypoxia, J. Fish. Res. Bd. Can., 29 (1972) 1463-1466. J.J. Chisolm, Lead poisoning, Sci. Amer., 10 (1976) 3715-3724. S.K. Hall, Lead pollution and poisoning. Environ. Sci. Tech., 6 (1972) 30-35. G.M. Hughes, Polluted respiratory physiology in A.P.M. Lockwood (Ed.) Effects of Pollutants on Aquatic Organisms, Vol. 2, Cambridge Univ. Press, London, 1976, p. 163. AK. Jafri and S.A. Shaffi, Effect of asphyxiation on the serum and kidney alkaline phosphatase activity in Clarias batrachus (Linn) Broteria XLIV (1975) 59-62. V.G. Jhingran, Ecotoxicological hazards of the inland fisheries of India, Natl. Coll. Biochem. Fish., (1977) 15-16. AD. Kumar, Modern concepts of ecology, Vikas Publicity House, New Delhi, 1977, 101-140. S.D. Lewis and W.M. Lewis, The effect of zinc and copper on the osmolality of blood serum of the channel cat fish, Zctalurus punctatus Rafinesque and Golden Shiner Notemigonous crysoleucas Mitchill, Trans. Am. Fish. Sot., 100 (1971) 639-643. Effects of Pollutants on Aquatic Organisms, Cambridge Univ. Press, 9 A.P.M. Lockwood, London, 1976, pp. 7-34. distribution of glycogen, lactate and 10 S.A. Shaffi and M. Habibulla, Differential pyruvate in different regions of the adult rat brain, Ind. J. Exp. Biol., 15 (1977) 309-310. in three 11 S.k Shaffi, Changes in tissue glycogen content due to cadmium intoxication fresh water teleosts, Cm-r. Sci., 47 (1978) 868-870. 12 S.A. Shaffi, Variations in tissue glycogen content, serum lactate and glucose levels due to copper intoxication in three fresh water teleosts, Cm-r. Sci., 47 (1978) 954-956. compartmentation of fish tissues, I. Brain energy reserve and 13 S.A. Shaffi, Biochemical its metabolic products. Acta Physiol., 99 (1978) 35-40. on tissue glycogen 14 S.A. Shaffi and M.A. Qayyum, Effect of cadmium intoxication content in a fresh water cat fish H. fossilis Bloch, Zool. Jb. Anat., 99 (1978) 129-132. on some fresh water fishes, I. Variations in 15 S.A. Shaffi, Effect of zinc intoxication tissue glycogen content, Arch. Gen. Exp. Zool., (1979) (In Press). on fresh water fishes, II. Accumulation of 16 S.A. Shaffi, Effect of zinc intoxication metabolic products, Toxicol. Lett., 3 (1979) 319-323. toxicity to fresh water fishes. Toxicol. Lett., 4 (1979) 17 S.A. Shaffi, The acute heptachlor 31-37. compartmentation of fish tissues, II. Non-specific phos18 S.A. Shaffi, Biochemical phomono esterases in brain, Experientia, (1979) (In Press).