Skip to main content
Log in

Effects of lead nitrate on the activity of metabolic enzymes during early developmental stages of the African catfish, Clarias gariepinus (Burchell, 1822)

  • Original Paper
  • Published:
Fish Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

Glucose-6-phosphate dehydrogenase (G6PDH), lactate dehydrogenase (LDH) and pyruvate kinase (PK) are key metabolic enzymes. G6PDH has been used as a biomarker of pollution-induced carcinogenesis in fish. LDH has been used as marker of lesions in toxicology and clinical chemistry, and PK catalyses the conversion of phosphoenol pyruvate to pyruvate, with regeneration of ATP. The effect of different concentrations of lead nitrate on the activity of these enzymes in two different early ontogenetic stages (embryonic and free embryonic stage) of the African catfish Clarias gariepinus was investigated. Embryo homogenates were used for measurements of G6PDH, LDH and PK activity spectrophotometrically at 340 nm and 25°C. The ontogenetic variations of the three enzymes during early ontogeny, from the 30 h to the 168 h post-fertilisation stage (PFS) (beginning of exogenous feeding), were studied. There was a significant decrease in activities of all three enzymes from 30 h-PFS to 96 h-PFS, followed by a significant increase in G6PDH and LDH. PK showed insignificant fluctuations in activity. Different patterns of enzyme activities were recorded due to exposure to different lead nitrate concentrations (100 μg/l, 300 μg/l and 500 μg/l). In the pre-hatching stage (30 h-PFS) the activity of the three enzymes increased at exposure to 100 μg/l lead nitrate and then decreased with increasing dose. In the post-hatching stages (48 h-PFS–168 h-PFS) G6PDH activity increased and LDH activity decreased with increasing lead concentrations. Unlike G6PDH and LDH, the PK enzyme fluctuated during the post-hatching stages and did not reveal a specific trend of response (increase or decrease) with increasing lead concentrations. Therefore, the measurement of G6PDH and LDH activities, but not PK activity, could be useful biomarkers of intoxication to reveal the embryotoxic potential of lead nitrate in fish embryos. The post-hatching stages of the African catfish are more sensitive than the pre-hatching stage (30 h-PFS) is, probably due to the protective capacity provided by the hardened chorion. The interaction and the main effects of age and lead doses were found to be highly significant, referring to the great impact of lead on these enzyme systems with increasing early development.

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

Access this article

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

  • Adham KG (2002) Sublethal effects of aquatic pollution in Lake Maryut on the African sharptooth catfish, Clarias gariepinus (Burchell, 1822). J Appl Ichthyol 18:87–94

    Article  CAS  Google Scholar 

  • Adham KG, Hassan IF, Taha N, Amin T (1999) Impact of hazardous exposure to metals in the Nile and Delta lakes on the catfish, Clarias lazera. Environ Monit Assess 54:107–124

    Article  CAS  Google Scholar 

  • Ahmed MA (1996) Environmental studies on the effect of some heavy metals pollution on tilapia fish in Egypt. Master’s dissertation, Cairo University

  • Almeida JA, Diniz YS, Marques SFG, Faine LA, Ribas BO, Burneiko RC, Novelli ELB (2002) The use of the oxidative stress responses as biomarkers in Nile tilapia (Oreochromis niloticus) exposed to in vivo cadmium contamination. Environ Int 27:673–679

    Article  PubMed  CAS  Google Scholar 

  • Almeida JA, Novelli ELB, Silva MD, Alves R (2001) Environmental cadmium exposure and metabolic responses of the Nile tilapia, Oreochromis niloticus. Environ Pollut 114:169–175

    Article  PubMed  CAS  Google Scholar 

  • Bainy ACD, Saito E, Carvalho PSM, Junqueira VBC (1996) Oxidative stress in gill, erythrocytes, liver and kidney of Nile tilapia (Oreochromis niloticus) from a polluted site. Aquat Toxicol 34:151–162

    Article  CAS  Google Scholar 

  • Barnhoorn IEJ (1996) Effects of manganese on the haematology of the Oreochromis mossambicus and the bioaccumulation of metals in Labeo umbratus. Master’s dissertation, Rand Afrikaans University, South Africa

  • Barnhoorn IEJ, van Vuren JHJ (2004) The use of different enzymes in feral freshwater fish as a tool for the assessment of water pollution in South Africa. Ecotox Environ Safe 59:180–185

    Article  CAS  Google Scholar 

  • Berges JA, Ballantyne JS (1991) Size scaling of whole body maximal enzyme activities in aquatic crustaceans. Can J Fish Aquat Sci 48:2385–2394

    CAS  Google Scholar 

  • Bergmeyer H, Bergmeyer J, Grabl M (1983) Methods of enzymatic analysis. Verlag Chemie, Weinheim

    Google Scholar 

  • Casillas E, Myers M, Ames WE (1983) Relationship of serum chemistry values to liver and kidney histopathology in English sole (Parophrys vetulus) after acute exposure to carbon tetrachloride. Aquat Toxicol 3:61–78

    Article  CAS  Google Scholar 

  • Christensen GM (1975) Biochemical effects of methylmercuric chloride, cadmium chloride, and lead nitrate on embryos and alevins of brook trout, Salvelinus fontinalis. Toxicol Appl Pharmacol 32:191–197

    Article  PubMed  CAS  Google Scholar 

  • Christensen GM, Olson D, Riedel B (1982) Chemical effects on the activity of 8 enzymes—a review and a discussion relevant to environmental monitoring. Environ Res 29:247–255

    Article  PubMed  CAS  Google Scholar 

  • Clarke ME, Calvi C, Domeier M, Edmonds M, Walsh PJ (1992) Effects of nutrition and temperature on metabolic enzyme activities in larval and juvenile red drum, Sciaenops ocellatus and lane snapper, Lutjanus synagris. Mar Biol 112:31–36

    Article  CAS  Google Scholar 

  • Coppes Z (1992) Lactate-dehydrogenase in teleosts—the role of ldh-C4 isozyme. Comp Biochem Physiol B 102:673–677

    Article  PubMed  CAS  Google Scholar 

  • Das PC, Ayyappan S, Das BK, Jena JK (2004a) Nitrite toxicity in Indian major carps: sublethal effect on selected enzymes in fingerlings of Catla catla, Labeo rohita and Cirrhinus mrigala. Comp Biochem Physiol C 138:3–10

    CAS  Google Scholar 

  • Das PC, Ayyappan S, Jena JK, Das BK (2004b) Acute toxicity of ammonia and its sub-lethal effects on selected haematological and enzymatic parameters of mrigal, Cirrhinus mrigala (Hamilton). Aquacult Res 35:134–143

    Article  CAS  Google Scholar 

  • De Graaf GJ, Janssen H (1996) Artificial reproduction and pond rearing of the African catfish Clarias gariepinus in sub-Saharan Africa. FAO Fish Tech Pap 362:1–73

    Google Scholar 

  • Degroot SJ (1987) Culture of Clarias species. Aquaculture 63:1–366

    Article  Google Scholar 

  • Diwan AP, Dhakad NK (1995) Embryology of fishes. Anomal Publications PVT, New Delhi

    Google Scholar 

  • Elumalai M, Antunes C, Guilhermino L (2002) Effects of single metals and their mixtures on selected enzymes of Carcinus maenas. Water Air Soil Pollut 141:273–280

    Article  CAS  Google Scholar 

  • Fennouh S, Casimiri V, Geloso-Meyer A, Burstein C (1998) Kinetic study of heavy metal salt effects on the activity of L-lactate dehydrogenase in solution immobilized on an oxygen electrode. Biosens Bioelectron 13:903–909

    Article  PubMed  CAS  Google Scholar 

  • Forstner H, Hinterleitner S, Mahr K, Wieser W (1983) Towards a better definition of metamorphosis in coregonus Sp—biochemical, histological, and physiological data. Can J Fish Aquat Sci 40:1224–1232

    Google Scholar 

  • Gayet JC, Haouz A, Gelosomeyer A, Burstein C (1993) Detection of heavy-metal salts with biosensors built with an oxygen electrode coupled to various immobilized oxidases and dehydrogenases. Biosens Bioelectron 8:177–183

    Article  CAS  Google Scholar 

  • Gill TS, Tewari H, Pande J (1990) Use of the fish enzyme system in monitoring water quality—effects of mercury on tissue enzymes. Comp Biochem Physiol C 97:287–292

    Article  PubMed  CAS  Google Scholar 

  • Gilli MRMS, Ocampos D, Pugine SMP, Lavandeira L, Cerqueira CM, Rosa R, Bacila M (2000) The effect of metal ions, oxalate and amino acids on the activity of pyruvate kinase from Curimbata (Prochilodus lineatus) epaxial muscle. Arch Vet Sci 5:73–79

    Google Scholar 

  • Gupta PK, Sastry KV (1981) Alterations in the activities of 3 dehydrogenases in the digestive system of 2 teleost fishes exposed to mercuric chloride. Environ Res 24:15–23

    Article  PubMed  CAS  Google Scholar 

  • Hallare AV, Schirling M, Luckenbach T, Kohler HR, Triebskorn R (2005) Combined effects of temperature and cadmium on developmental parameters and biomarker responses in zebrafish (Danio rerio) embryos. J Thermal Biol 30:7–17

    Article  CAS  Google Scholar 

  • Hansen JI, Mustafa T, Depledge M (1992) Mechanisms of copper toxicity in the shore crab, Carcinus maenas. 2. Effects on key metabolic enzymes, metabolites and energy charge potential. Mar Biol 114:259–264

    Article  CAS  Google Scholar 

  • Hardewig I, Portner HO, van Dijk P (2004) How does the cold stenothermal gadoid Lota lota survive high water temperatures during summer? J Comp Physiol B 174:149–156

    Article  PubMed  CAS  Google Scholar 

  • Heath AG (1996) Water pollution and fish physiology. Lewis, Boca Raton

    Google Scholar 

  • Hinterleitner S, Platzer U, Wieser W (1987) Development of the activities of oxidative, glycolytic and muscle enzymes during early larval life in 3 families of fresh water fish. J Fish Biol 30:315–326

    Article  CAS  Google Scholar 

  • Honkanen JO (2004) Ecotoxicological testing of organic chemicals on early life stages of salmonid fish at environmentally realistic temperatures. PhD thesis, University of Joensuu

  • Hunaiti AA, Soud M (2000) Effect of lead concentration on the level of glutathione, glutathione-S-transferase, reductase and peroxidase in human blood. Sci Total Environ 248:45–50

    Article  PubMed  CAS  Google Scholar 

  • Isani G, Cattani O, Carpene E, Cortesi P (1994) Kinetic properties of liver and muscle pyruvate kinase of a marine teleost, sea bass (Dicentrarchus labrax L). Comp Biochem Physiol B 107:617–624

    Article  Google Scholar 

  • Jackim E, Hamlin JM, Sonis S (1970) Effects of metal poisoning on 5 liver enzymes in killifish (Fundulus heteroclitus). J Fish Res Bd Can 27:383–390

    CAS  Google Scholar 

  • Jiminez BD, Stegeman JJ (1990) Detoxification enzymes as indicator of environmental stress on fishes. Am Fish Soc Symp 8:69–79

    Google Scholar 

  • Kiessling A, Kiessling KH, Storebakken T, Asgard T (1991) Changes in the structure and function of the epaxial muscle of rainbow trout (Oncorhynchus mykiss) in relation to ration and age. 2. Activity of key enzymes in energy metabolism. Aquaculture 93:357–372

    Article  CAS  Google Scholar 

  • Kletzien RF, Harris PKW, Foellmi LA (1994) Glucose-6-phosphate-dehydrogenase—a housekeeping enzyme subject to tissue-specific regulation by hormones, nutrients, and oxidant stress. FASEB J 8:174–181

    PubMed  CAS  Google Scholar 

  • Kurutas EB, Tuncer I (2000) A mouse model for evaluating the induction of liver glucose-6-phosphate dehydrogenase by halothane. Turk J Vet Anim Sci 24:511–515

    Google Scholar 

  • Lashein FE (1996) The effect of heavy metals cadmium and lead on embryogenesis and larval development of the teleost fish Ctenopharyngodon idellus. PhD thesis, South Vally University

  • Lemos D, Salomon M, Gomes V, Phan V, Buchholz E (2003) Citrate synthase and pyruvate kinase activities during early life stages of the shrimp Farfantepenaeus paulensis (Crustacea, Decapoda, Penaeidae): effects of development and temperature. Comp Biochem Physiol B 135:707–719

    Article  PubMed  CAS  Google Scholar 

  • Leopold JA, Loscalzo J (2000) Cyclic strain modulates resistance to oxidant stress by increasing G6PDH expression in smooth muscle cells. Am J Physiol 279:H2477–H2485

    CAS  Google Scholar 

  • Leopold JA, Zhang YY, Scribner AW, Stanton RC, Loscalzo J (2003) Glucose-6-phosphate dehydrogenase overexpression decreases endothelial cell oxidant stress and increases bioavailable nitric oxide. Arterioscler Thromb Vasc Biol 23:411–417

    Article  CAS  Google Scholar 

  • Long SM, Ryder KJ, Holdway DA (2003) The use of respiratory enzymes as biomarkers of petroleum hydrocarbon exposure in Mytilus edulis planulatus. Ecotox Environ Safe 55:261–270

    Article  CAS  Google Scholar 

  • Luckenbach T, Triebskorn R, Muller E, Oberemm A (2001) Toxicity of waters from two streams to early life stages of brown trout (Salmo trutta f. fario L.), tested under semi-field conditions. Chemosphere 45:571–579

    Article  PubMed  CAS  Google Scholar 

  • Mekkawy IAA, Lashein FE (2003) The effect of lead and cadmium on LDH and G-6-PDH isoenzyme patterns exhibited during the early embryonic development of the teleost fish, Ctenopharyngodon idellus with emphasis on the corresponding morphological variations. Presented at the 26th annual fish larva conference, Bergen, Norway, 22–26 July 2003. Institute of Marine Research, 1870 Nordnes.

  • Mekkawy IAA, Osman AGM (2006) Ultrastructural studies of the morphological variations of the egg surface and envelopes of the African catfish Clarias gariepinus (Burchell, 1822) before and after fertilization with a discussion of fertilization mechanism. Sci Mar 70:23–40

    Article  Google Scholar 

  • Melancon MJ (1995) Bioindicators used in aquatic and terrestrial monitoring. Lewis, Boca Raton, FL

    Google Scholar 

  • Mishra R, Shukla SP (1997) Impact of endosulfan on lactate dehydrogenase from the freshwater catfish Clarias batrachus. Pest Biochem Physiol 57:220–234

    Article  CAS  Google Scholar 

  • Nathanailides C (1996) Metabolic specialization of muscle during development in cold-water and warmwater fish species exposed to different thermal conditions. Can J Fish Aquat Sci 53:2147–2155

    Article  Google Scholar 

  • Nguyen LTH, Janssen CR (2002) Embryo-larval toxicity tests with the African catfish (Clarias gariepinus): comparative sensitivity of endpoints. Arch Environ Contam Toxicol 42:256–262

    Article  PubMed  CAS  Google Scholar 

  • Nguyen LTH, Janssen CR, Volckaert FAM (1999) Susceptibility of embryonic and larval African catfish (Clarias gariepinus) to toxicants. Bull Environ Contam Toxicol 62:230–237

    Article  PubMed  CAS  Google Scholar 

  • Nogae I, Johnston M (1990) Isolation and characterization of the Zwf1 gene of Saccharomyces cerevisiae, encoding glucose-6-phosphate-dehydrogenase. Gene 96:161–169

    Article  PubMed  CAS  Google Scholar 

  • Ocampos D, Rosa C, Rodrigues E, Rosa R (1987) Purification of muscle pyruvate kinase and lactate dehydrogenase and stability of pyruvate kinase from fish (Mugil lisa and Chaetoditherus faber). Arq Biol Technol 29:699–705

    Google Scholar 

  • Pandey S, Parvez S, Sayeed I, Haque R, Bin-Hafeez B, Raisuddin S (2003) Biomarkers of oxidative stress: a comparative study of river Yamuna fish Wallago attu (Bl. & Schn.). Sci Total Environ 309:105–115

    Article  PubMed  CAS  Google Scholar 

  • Pandolfi PP, Sonati F, Rivi R, Mason P, Grosveld F, Luzzatto L (1995) Targeted disruption of the housekeeping gene encoding glucose-6-phosphate-dehydrogenase (G6pd)—G6pd is dispensable for pentose synthesis but essential for defense against oxidative stress. EMBO J 14:5209–5215

    PubMed  CAS  Google Scholar 

  • Pelletier D, Blier PU, Dutil JD, Guderley H (1995) How should enzyme activities be used in fish growth studies. J Exp Biol 198:1493–1497

    PubMed  CAS  Google Scholar 

  • Pelletier D, Dutil JD, Blier P, Guderley H (1994) Relation between growth rate and metabolic organization of white muscle, liver and digestive-tract in Cod, Gadus morhua. J Comp Physiol B 164:508–508

    Article  Google Scholar 

  • Pickering A, Pottinger T (1995) Biochemical effects of stress. Elsevier Science, New York

    Google Scholar 

  • Powers DA (1989) Fish as model systems. Science 246:352–358

    Article  PubMed  CAS  Google Scholar 

  • Rajanna B, Chetty CS, McBride V, Rajanna S (1990) Effects of lead on K+-para-nitrophenyl phosphatase activity and protection by thiol reagents. Biochem Int 20:1011–1018

    PubMed  CAS  Google Scholar 

  • Randall RF, Anderson PJ (1975) Purification and properties of pyruvate kinase of sturgeon muscle. Biochem J 145:569–573

    PubMed  CAS  Google Scholar 

  • Rashed MN (2001) Monitoring of environmental heavy metals in fish from Nasser Lake. Environ Int 27:27–33

    Article  PubMed  CAS  Google Scholar 

  • Richter C, Gogvadze V, Laffranchi R, Schlapbach R, Schweizer M, Suter M, Walter P, Yaffee M (1995) Oxidants in mitochondria—from physiology to diseases. Biochem Biophys Acta 1271:67–74

    PubMed  Google Scholar 

  • Rosety-Rodriguez M, Ordonez F, Rosety I, Rosety J, Rosery M (2005) Erythrocyte antioxidant enzymes of gilthead as early-warning bio-indicators of oxidative stress induced by malathion. Haema 8:237–240

    CAS  Google Scholar 

  • Salvemini F, Franze A, Iervolino A, Filosa S, Salzano S, Ursini MV (1999) Enhanced glutathione levels and oxidoresistance mediated by increased glucose-6-phosphate dehydrogenase expression. J Biol Chem 274:2750–2757

    Article  PubMed  CAS  Google Scholar 

  • Sastry KV, Gupta PK (1980) Alterations in the activities of a few dehydrogenases in the digestive system of 2 teleost fishes exposed to lead nitrate. Ecotox Environ Safe 4:232–239

    Article  CAS  Google Scholar 

  • Sastry KV, Rao DR (1984) Effect of mercuric chloride on some biochemical and physiological parameters of the freshwater murrel, Channa punctatus. Environ Res 34:343–350

    Article  PubMed  CAS  Google Scholar 

  • Segner H, Verreth J (1995) Metabolic enzyme activities in larvae of the African catfish, Clarias gariepinus—changes in relation to age and nutrition. Fish Physiol Biochem 14:385–398

    Article  CAS  Google Scholar 

  • Shaklee JB, Champion MJ, Whitt GS (1974) Developmental genetics of teleosts—biochemical analysis of lake Chubsucker ontogeny. Dev Biol 38:356–382

    Article  PubMed  CAS  Google Scholar 

  • Singh J, Husain R, Tandon SK, Seth PK, Chandra SV (1974) Biochemical and histopathological alterations in early manganese toxicity in rats. Environ Physiol Biochem 4:16–23

    PubMed  CAS  Google Scholar 

  • Singh RK, Sharma B (1998) Carbofuran-induced biochemical changes in Clarias batrachus. Pest Sci 53:285–290

    Article  CAS  Google Scholar 

  • Singhal RK, Anderson ME, Meister A (1987) Glutathione, a 1st line of defense against cadmium toxicity. FASEB J 1:220–223

    PubMed  CAS  Google Scholar 

  • Somero S, Childress J (1985) Scaling of oxidative and glycolytic enzyme activities in fish muscle. Springer, Berlin

    Google Scholar 

  • SPSS (1998) SPSS for Windows. SPSS, Chicago

    Google Scholar 

  • Stephensen E, Svavarsson J, Sturve J, Ericson G, Adolfsson-Erici M, Forlin L (2000) Biochemical indicators of pollution exposure in shorthorn sculpin (Myoxocephalus scorpius), caught in four harbours on the southwest coast of Iceland. Aquat Toxicol 48:431–442

    Article  PubMed  CAS  Google Scholar 

  • Stouthart A, Spanings FAT, Lock RAC, Bonga SEW (1994) Effects of low water pH on lead toxicity to early life stages of the common carp (Cyprinus carpio). Aquat Toxicol 30:137–151

    Article  CAS  Google Scholar 

  • Strik J, De Iond H, Van Rijn J, Wuite T (1975) Toxicity of chromium(VI) in fish, with special defences to organ weight, liver and plasma enzyme activities, blood parameters and histological alteration. Elsevier Scientific, Amsterdam

    Google Scholar 

  • Strmac M, Braunbeck T (2002) Cytological and biochemical effects of a mixture of 20 pollutants on isolated rainbow trout (Oncorhynchus mykiss) hepatocytes. Ecotox Environ Safe 53:293–304

    Article  CAS  Google Scholar 

  • Tian WN, Braunstein LD, Apse K, Pang JD, Rose M, Tian XN, Stanton RC (1999) Importance of glucose-6-phosphate dehydrogenase activity in cell death. Am J Physiol 276:C1121–C1131

    PubMed  CAS  Google Scholar 

  • Verreth J, Eding E, Rao G, Huskens F, Segner H (1993) A review of feeding practices, growth and nutritional physiology in larvae of the Catfish Clarias gariepinus and Clarias batrachus. J World Aquacult Soc 24:135–144

    Google Scholar 

  • Volckaert FAM, Hellemans BA, Galbusera P, Ollevier F, Sekkali B, Belayew A (1994) Replication, expression and fate of foreign DNA during embryonic and larval development of the African catfish Clarias gariepinus. Mol Mar Biol Biotechnol 3:57–69

    PubMed  CAS  Google Scholar 

  • Winzer K, Van Noorden CJF, Kohler A (2001) Quantitative cytochemical analysis of glucose-6-phosphate dehydrogenase activity in living isolated hepatocytes of European flounder for rapid analysis of xenobiotic effects. J Histochem Cytochem 49:1025–1032

    PubMed  CAS  Google Scholar 

  • Wu RSS, Lam PKS (1997) Glucose-6-phosphate dehydrogenase and lactate dehydrogenase in the green-lipped mussel (Perna viridis): possible biomarkers for hypoxia in the marine environment. Water Res 31:2797–2801

    Article  CAS  Google Scholar 

  • Zollner H (1993) Handbook of enzyme inhibitors. VCH, Weinheim

    Google Scholar 

Download references

Acknowledgements

We would like to thank K. Kuntze for her continuous help during the measurement of the enzyme activities; colleagues at the chemical laboratory of the IGB for their continuous advice; Mr. Gaertner for help during the trip to the Netherlands, and colleagues at the Department of Fish Culture and Fisheries in Wageningen for their help concerning the reproduction of Clarias gariepinus.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Alaa G. M. Osman.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Osman, A.G.M., Mekkawy, I.A., Verreth, J. et al. Effects of lead nitrate on the activity of metabolic enzymes during early developmental stages of the African catfish, Clarias gariepinus (Burchell, 1822). Fish Physiol Biochem 33, 1–13 (2007). https://doi.org/10.1007/s10695-006-9111-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10695-006-9111-8

Keywords

Navigation