2022•Zenodo (CERN European Organization for Nuclear Research)Open access

Responses of soil enzymes to different heavy metals

Sonia Sethi, Saksham Gupta

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Abstract

ABSTRACT Presence of heavy metals (HM) in agricultural soil is a major hazard to the soil-plant system. In the present study, soil samples (0-5 cm depth) were taken in order to determine the effects of heavy metal pollution on soil enzymes like dehydrogenase, alkaline phosphatase (APA), urease (UA) protease, cellulose, invertase, beta glucosidase and amylase. Results showed that Cadmium (Cd) significantly inhibited the four enzyme activities and Zinc (Zn) inhibited urease and calatase activities. Lead (Pb) was not significantly inhibitory than the other heavy metals for the four enzyme activities and was shown to have a protective role on calatase activity in the combined presence of Cd, Zn and Pb. Overall, all the heavy metals were found to have an inhibitory effect on the soil enzymatic activities. Hence our results suggested that enzymatic activities may be used as a sensitive indicator for assessing changes in soil environment quality. Keywords: Heavy metals, Dehydrogenase activity, Urease activity, Invertase activity, Cellulase activity REFERENCES Filip, Z. (2002). International approach to assessing soil quality by ecologically-related biological parameters. Agric. Ecosyst. Environ, 88: 169–174. Sandaa, R.A., Torsvik, V., Enger´, E. (2001). Influence of long-term heavy-metal contamination on microbial communities in soil. Soil Biology & Biochemistry, 33: 287–295. Akmal, M., Wang, H.Z., Wu, J. J. (2005). Changes in enzymes activity, substrate utilization pattern and diversity of soil microbial communities under cadmium pollution. Journal of Environmental Sciences, 17(5): 802–807. Moreno, J.L., Sanchez-Marín, A., Hernández, T., García, C. (2006). Effect of cadmium on microbial activity and a ryegrass crop in two semiarid soils. Environ. Manage, 37 (5):626–633. Hinojosa, M.B., Carreira, J.A., Garcia-Ruiz, R. (2004). Soil moisture pre-treatment effects on enzyme activities as indicators of heavy metal-contaminated and reclaimed soils. Soil Biol Biochem, 36:1559–1568. Moreno, J.L., Garcia, C., Landi, L., Falchini, L., Pietramellara, G., Nannipieri, P.(2001). The ecological dose value (ED50) for assessing Cd toxicity on ATP content and dehydrogenase and urease activities of soil. Soil Biol. Biochem, 33(4-5): 483. Wyszkowska, J., Kucharski, J. (2003). Biochemical and physicochemicalproperties of soil contaminated with the heavy metals. Zesz. Prob. Nauk Rol. 492: 435(in Polish). Tyler, G., (1981). Heavy metals in soil biology and biochemistry. In: PAUL, E.A.; LADD, I.N. (eds) Soil Biochemistry. New York, Marcel Dekker Inc. 371–401. Casida, L.E., Klein, D.A., Santoro, T. (1964). Soil dehydrogenase activity. Soil Science, 98:371-376. Chanda Mallaiah. (2013). Studies on the persistence and degradation of endosulfan in the soil ecosystem of tropical climate. Biolife, 1(3); 116-122. Ladd, J.N., Butler, J.H.A. (1972). Short-term assays of soil proteolytic enzyme activities using proteins and dipeptide derivatives as substrates. Soil Biol. Biochem, 4:19-30. Deng, S.P., Tabatabai, M.A. (1994). Cellulase activity in soils. Soil Biol. Biochem, 26: 1347–1354. Ross, D. J. (1983). N.Z. J. Sci, 26: 339–346. Tabatabai, M.A., Brenner, J.M. (1969). Use of p-nitrophenyl phosphatefor assay of soil phosphatase activity. Soil Biol. Biochem, 4: 479-487. Eivazi, F., Tabatabai, M.A. (1988). Glucosidases and galactosidases in soils. Soil Biol. Biochem,20: 601-606. Dick, R.P., Breakwell, D.P., Turco, R.F. (1996). Soil enzyme activities and biodiversity measurements as integrative microbiological indicators. In: Dick R.P., Lal R., Lowery B., Rice Ch.W., Stott D.E. (eds.): Methods of assessing soil quality. SSSA Spec. Publ. No. 49, Madison: 247–271. Malley, C., Nair, J., Ho, G. (2005). Impact of heavy metals on enzymatic Activity of substrate and on composting worms Eisenia fetida. Biores Technol, 97: 1498–1502. Nweke, C.O., Ntinugwa, V., Obah, I.F., Ike, S.C., Eme, G.E., Opara, E.C., Okolo, J.C., Nwanyanwu, V. (2007). In vitro effects of metals and pesticides on dehydrogenase Activity in microbial community of cowpea (Vigna unguiculata) rhizoplane. Afr J Biotechnol, 6: 290-295. Lorenz, N., Hintemann, T., Kramarewa, T., Katayama, A., Yasuta, T., Marschner, P., Kandeler, E. (2006). Response of microbial activity and microbial community composition, in soils to long-term arsenic and cadmium exposure. Soil Biol Biochem, 38:1430–1437 Effron, D., de la Horra, A.M., Defrieri, R.L., Fontanive, V., Palma, P.M. (2004). Effect of cadmium, copper and lead on different soil enzymatic activities in a native forest soil. Comm Soil Sci Plant Anal, 35:1309-1321. Renella, G., Mench, M., Landi, L., Nannipieri, P. (2005). Microbial activity and hydrolase synthesis in long-term Cd-contaminated soils. Soil Biol Biochem, 37:133–139. Khan, S., Cao, Q., Heshman, A.E.L., Xia, Y., He, J. (2007). Soil enzymatic activities and microbial community structure with different application rates of Cd and Pb. J Environ Sci, 19:834-840. Vig, K., Megharaj, M., Senthunathan, N., Naidu, R. (2003). Bioavailability and toxicity of cadmium to microorganisms and their activities in soil: A Review. Adv Environ Res, 8:121–135. Geiger, G., Brandi, H., Furner, G., Schulin, R. (1998). The effect of copper on the activity of cellulase and b -glucosidase in the presence of montmorillonite or Al-montmorillonite. Soil Biol Biochem, 30:1537–1544. Esterbauer, H., Hayn, M., Abuja, P.M., Claeyssens, M. (1991). Structure of cellulolytic enzymes. In: Leatham GF, Himmel ME (eds) Enzymes in biomass conversion. American Chemical Society, Washington, pp 301–312. Wood, T..M, Garcia-Campayo, V. (1990). Enzymology of cellulose degradation. Biodegradation, 1:147–161. Teeri, T.T., Koivula, A., Linder, M., Reinikainen, T., Ruohonrn, L., Srisodsuk, M., Claeyssens, M., Jones, T.A. (1995). Modes of action of two Trichoderma reesei cellobiohydrolases. In Petersen SB, Sevensson B, Pedersen S (Eds.) Carbohydrate bioengineering. pp. 211-225. Rogers, J.E., Li, S.W. (1985). Effects of metals and other inorganic ions on soil microbial activity, soil dehydrogenase assay as a simple toxicity test. Bull Environ Contam Toxicol, 34:858–865. Balyaeva, O.N., Haynes, R.J., Birukova, O.A. (2005). Barley yield and soil microbial and enzyme activities as affected by contamination of two soils with lead, zinc or copper. Biol Fertil Soils,. 41:85–94. Nagwasiri Pride Ndasi, Nidhi Sahu, Sanjay Thul, B. Chandrashekhar, Abhinav Sharma, Ngassoum Martine Benoit and Ram Avatar Pandey. (2014). Biodegradation of phenanthrene by rhizobacteria isolated from local grass growing at hydrocarbon contaminated soil in Cameroon. Biolife, 2(2); 420-441. Nowak, J., Niedzwiecki, E., Dziel, M. (1999). Wpływ metali ciężkich na zmiany aktywności enzymatycznej gleby. Rocz. Gleboz. 50(1/2), 61 (in Polish, with English abstract). Landi, L., Renella, G., Moreno, J.L., Falchini, L., Nannipieri, P. (2000). Influence of cadmium on the metabolic quotient, L: D Glutamic acid respiration ratio and enzyme activity:microbial biomass ratio under laboratory conditions. Biol. Fert. Soils 32(1):8. Shen, G., Lu, Y., Zhou, Q., Hang, J. (2005). Interaction of polycyclic aromatic hydrocarbons and heavy metals on soil enzyme. Chemosphere, 61:1175–1182. Zheng, C.R., Tu, C., Chen, H.M. (1999). Effect of combined heavy metal pollution on nitrogen mineralization potential, urease and phosphatase activities in a Typic Udic Ferrisol. Pedosphere, 9:251-258. Yang, Z.X. Liu, S.G. (2000). Effect of single element and compound pollution of Cd, Zn and Pb on soil enzyme activities. Soil Environ Sci, 9:15–18. Šíša, R. (1993). Enzymová aktivita půdy jako ukazatel její biologické aktivity. Rostl. Výr, 39: 817–825.

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ABSTRACT Presence of heavy metals (HM) in agricultural soil is a major hazard to the soil-plant system. In the present study, soil samples (0-5 cm depth) were taken in order to determine the effects of heavy metal pollution on soil enzymes like dehydrogenase, alkaline phosphatase (APA), urease (UA) protease, cellulose, invertase, beta glucosidase and amylase. Results showed that Cadmium (Cd) significantly inhibited the four enzyme activities and Zinc (Zn) inhibited urease and calatase activities. Lead (Pb) was not significantly inhibitory than the other heavy metals for the four enzyme activities and was shown to have a protective role on calatase activity in the combined presence of Cd, Zn and Pb. Overall, all the heavy metals were found to have an inhibitory effect on the soil enzymatic activities. Hence our results suggested that enzymatic activities may be used as a sensitive indicator for assessing changes in soil environment quality. Keywords: Heavy metals, Dehydrogenase activity, Urease activity, Invertase activity, Cellulase activity REFERENCES Filip, Z. (2002). International approach to assessing soil quality by ecologically-related biological parameters. Agric. Ecosyst. Environ, 88: 169–174. Sandaa, R.A., Torsvik, V., Enger´, E. (2001). Influence of long-term heavy-metal contamination on microbial communities in soil. Soil Biology & Biochemistry, 33: 287–295. Akmal, M., Wang, H.Z., Wu, J. J. (2005). Changes in enzymes activity, substrate utilization pattern and diversity of soil microbial communities under cadmium pollution. Journal of Environmental Sciences, 17(5): 802–807. Moreno, J.L., Sanchez-Marín, A., Hernández, T., García, C. (2006). Effect of cadmium on microbial activity and a ryegrass crop in two semiarid soils. Environ. Manage, 37 (5):626–633. Hinojosa, M.B., Carreira, J.A., Garcia-Ruiz, R. (2004). Soil moisture pre-treatment effects on enzyme activities as indicators of heavy metal-contaminated and reclaimed soils. Soil Biol Biochem, 36:1559–1568. Moreno, J.L., Garcia, C., Landi, L., Falchini, L., Pietramellara, G., Nannipieri, P.(2001). The ecological dose value (ED50) for assessing Cd toxicity on ATP content and dehydrogenase and urease activities of soil. Soil Biol. Biochem, 33(4-5): 483. Wyszkowska, J., Kucharski, J. (2003). Biochemical and physicochemicalproperties of soil contaminated with the heavy metals. Zesz. Prob. Nauk Rol. 492: 435(in Polish). Tyler, G., (1981). Heavy metals in soil biology and biochemistry. In: PAUL, E.A.; LADD, I.N. (eds) Soil Biochemistry. New York, Marcel Dekker Inc. 371–401. Casida, L.E., Klein, D.A., Santoro, T. (1964). Soil dehydrogenase activity. Soil Science, 98:371-376. Chanda Mallaiah. (2013). Studies on the persistence and degradation of endosulfan in the soil ecosystem of tropical climate. Biolife, 1(3); 116-122. Ladd, J.N., Butler, J.H.A. (1972). Short-term assays of soil proteolytic enzyme activities using proteins and dipeptide derivatives as substrates. Soil Biol. Biochem, 4:19-30. Deng, S.P., Tabatabai, M.A. (1994). Cellulase activity in soils. Soil Biol. Biochem, 26: 1347–1354. Ross, D. J. (1983). N.Z. J. Sci, 26: 339–346. Tabatabai, M.A., Brenner, J.M. (1969). Use of p-nitrophenyl phosphatefor assay of soil phosphatase activity. Soil Biol. Biochem, 4: 479-487. Eivazi, F., Tabatabai, M.A. (1988). Glucosidases and galactosidases in soils. Soil Biol. Biochem,20: 601-606. Dick, R.P., Breakwell, D.P., Turco, R.F. (1996). Soil enzyme activities and biodiversity measurements as integrative microbiological indicators. In: Dick R.P., Lal R., Lowery B., Rice Ch.W., Stott D.E. (eds.): Methods of assessing soil quality. SSSA Spec. Publ. No. 49, Madison: 247–271. Malley, C., Nair, J., Ho, G. (2005). Impact of heavy metals on enzymatic Activity of substrate and on composting worms Eisenia fetida. Biores Technol, 97: 1498–1502. Nweke, C.O., Ntinugwa, V., Obah, I.F., Ike, S.C., Eme, G.E., Opara, E.C., Okolo, J.C., Nwanyanwu, V. (2007). In vitro effects of metals and pesticides on dehydrogenase Activity in microbial community of cowpea (Vigna unguiculata) rhizoplane. Afr J Biotechnol, 6: 290-295. Lorenz, N., Hintemann, T., Kramarewa, T., Katayama, A., Yasuta, T., Marschner, P., Kandeler, E. (2006). Response of microbial activity and microbial community composition, in soils to long-term arsenic and cadmium exposure. Soil Biol Biochem, 38:1430–1437 Effron, D., de la Horra, A.M., Defrieri, R.L., Fontanive, V., Palma, P.M. (2004). Effect of cadmium, copper and lead on different soil enzymatic activities in a native forest soil. Comm Soil Sci Plant Anal, 35:1309-1321. Renella, G., Mench, M., Landi, L., Nannipieri, P. (2005). Microbial activity and hydrolase synthesis in long-term Cd-contaminated soils. Soil Biol Biochem, 37:133–139. Khan, S., Cao, Q., Heshman, A.E.L., Xia, Y., He, J. (2007). Soil enzymatic activities and microbial community structure with different application rates of Cd and Pb. J Environ Sci, 19:834-840. Vig, K., Megharaj, M., Senthunathan, N., Naidu, R. (2003). Bioavailability and toxicity of cadmium to microorganisms and their activities in soil: A Review. Adv Environ Res, 8:121–135. Geiger, G., Brandi, H., Furner, G., Schulin, R. (1998). The effect of copper on the activity of cellulase and b -glucosidase in the presence of montmorillonite or Al-montmorillonite. Soil Biol Biochem, 30:1537–1544. Esterbauer, H., Hayn, M., Abuja, P.M., Claeyssens, M. (1991). Structure of cellulolytic enzymes. In: Leatham GF, Himmel ME (eds) Enzymes in biomass conversion. American Chemical Society, Washington, pp 301–312. Wood, T..M, Garcia-Campayo, V. (1990). Enzymology of cellulose degradation. Biodegradation, 1:147–161. Teeri, T.T., Koivula, A., Linder, M., Reinikainen, T., Ruohonrn, L., Srisodsuk, M., Claeyssens, M., Jones, T.A. (1995). Modes of action of two Trichoderma reesei cellobiohydrolases. In Petersen SB, Sevensson B, Pedersen S (Eds.) Carbohydrate bioengineering. pp. 211-225. Rogers, J.E., Li, S.W. (1985). Effects of metals and other inorganic ions on soil microbial activity, soil dehydrogenase assay as a simple toxicity test. Bull Environ Contam Toxicol, 34:858–865. Balyaeva, O.N., Haynes, R.J., Birukova, O.A. (2005). Barley yield and soil microbial and enzyme activities as affected by contamination of two soils with lead, zinc or copper. Biol Fertil Soils,. 41:85–94. Nagwasiri Pride Ndasi, Nidhi Sahu, Sanjay Thul, B. Chandrashekhar, Abhinav Sharma, Ngassoum Martine Benoit and Ram Avatar Pandey. (2014). Biodegradation of phenanthrene by rhizobacteria isolated from local grass growing at hydrocarbon contaminated soil in Cameroon. Biolife, 2(2); 420-441. Nowak, J., Niedzwiecki, E., Dziel, M. (1999). Wpływ metali ciężkich na zmiany aktywności enzymatycznej gleby. Rocz. Gleboz. 50(1/2), 61 (in Polish, with English abstract). Landi, L., Renella, G., Moreno, J.L., Falchini, L., Nannipieri, P. (2000). Influence of cadmium on the metabolic quotient, L: D Glutamic acid respiration ratio and enzyme activity:microbial biomass ratio under laboratory conditions. Biol. Fert. Soils 32(1):8. Shen, G., Lu, Y., Zhou, Q., Hang, J. (2005). Interaction of polycyclic aromatic hydrocarbons and heavy metals on soil enzyme. Chemosphere, 61:1175–1182. Zheng, C.R., Tu, C., Chen, H.M. (1999). Effect of combined heavy metal pollution on nitrogen mineralization potential, urease and phosphatase activities in a Typic Udic Ferrisol. Pedosphere, 9:251-258. Yang, Z.X. Liu, S.G. (2000). Effect of single element and compound pollution of Cd, Zn and Pb on soil enzyme activities. Soil Environ Sci, 9:15–18. Šíša, R. (1993). Enzymová aktivita půdy jako ukazatel její biologické aktivity. Rostl. Výr, 39: 817–825.

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ABSTRACT Presence of heavy metals (HM) in agricultural soil is a major hazard to the soil-plant system. In the present study, soil samples (0-5 cm depth) were taken in order to determine the effects of heavy metal pollution on soil enzymes like dehydrogenase, alkaline phosphatase (APA), urease (UA) protease, cellulose, invertase, beta glucosidase and amylase. Results showed that Cadmium (Cd) significantly inhibited the four enzyme activities and Zinc (Zn) inhibited urease and calatase activities. Lead (Pb) was not significantly inhibitory than the other heavy metals for the four enzyme activities and was shown to have a protective role on calatase activity in the combined presence of Cd, Zn and Pb. Overall, all the heavy metals were found to have an inhibitory effect on the soil enzymatic activities. Hence our results suggested that enzymatic activities may be used as a sensitive indicator for assessing changes in soil environment quality. Keywords: Heavy metals, Dehydrogenase activity, Urease activity, Invertase activity, Cellulase activity REFERENCES Filip, Z. (2002). International approach to assessing soil quality by ecologically-related biological parameters. Agric. Ecosyst. Environ, 88: 169–174. Sandaa, R.A., Torsvik, V., Enger´, E. (2001). Influence of long-term heavy-metal contamination on microbial communities in soil. Soil Biology & Biochemistry, 33: 287–295. Akmal, M., Wang, H.Z., Wu, J. J. (2005). Changes in enzymes activity, substrate utilization pattern and diversity of soil microbial communities under cadmium pollution. Journal of Environmental Sciences, 17(5): 802–807. Moreno, J.L., Sanchez-Marín, A., Hernández, T., García, C. (2006). Effect of cadmium on microbial activity and a ryegrass crop in two semiarid soils. Environ. Manage, 37 (5):626–633. Hinojosa, M.B., Carreira, J.A., Garcia-Ruiz, R. (2004). Soil moisture pre-treatment effects on enzyme activities as indicators of heavy metal-contaminated and reclaimed soils. Soil Biol Biochem, 36:1559–1568. Moreno, J.L., Garcia, C., Landi, L., Falchini, L., Pietramellara, G., Nannipieri, P.(2001). The ecological dose value (ED50) for assessing Cd toxicity on ATP content and dehydrogenase and urease activities of soil. Soil Biol. Biochem, 33(4-5): 483. Wyszkowska, J., Kucharski, J. (2003). Biochemical and physicochemicalproperties of soil contaminated with the heavy metals. Zesz. Prob. Nauk Rol. 492: 435(in Polish). Tyler, G., (1981). Heavy metals in soil biology and biochemistry. In: PAUL, E.A.; LADD, I.N. (eds) Soil Biochemistry. New York, Marcel Dekker Inc. 371–401. Casida, L.E., Klein, D.A., Santoro, T. (1964). Soil dehydrogenase activity. Soil Science, 98:371-376. Chanda Mallaiah. (2013). Studies on the persistence and degradation of endosulfan in the soil ecosystem of tropical climate. Biolife, 1(3); 116-122. Ladd, J.N., Butler, J.H.A. (1972). Short-term assays of soil proteolytic enzyme activities using proteins and dipeptide derivatives as substrates. Soil Biol. Biochem, 4:19-30. Deng, S.P., Tabatabai, M.A. (1994). Cellulase activity in soils. Soil Biol. Biochem, 26: 1347–1354. Ross, D. J. (1983). N.Z. J. Sci, 26: 339–346. Tabatabai, M.A., Brenner, J.M. (1969). Use of p-nitrophenyl phosphatefor assay of soil phosphatase activity. Soil Biol. Biochem, 4: 479-487. Eivazi, F., Tabatabai, M.A. (1988). Glucosidases and galactosidases in soils. Soil Biol. Biochem,20: 601-606. Dick, R.P., Breakwell, D.P., Turco, R.F. (1996). Soil enzyme activities and biodiversity measurements as integrative microbiological indicators. In: Dick R.P., Lal R., Lowery B., Rice Ch.W., Stott D.E. (eds.): Methods of assessing soil quality. SSSA Spec. Publ. No. 49, Madison: 247–271. Malley, C., Nair, J., Ho, G. (2005). Impact of heavy metals on enzymatic Activity of substrate and on composting worms Eisenia fetida. Biores Technol, 97: 1498–1502. Nweke, C.O., Ntinugwa, V., Obah, I.F., Ike, S.C., Eme, G.E., Opara, E.C., Okolo, J.C., Nwanyanwu, V. (2007). In vitro effects of metals and pesticides on dehydrogenase Activity in microbial community of cowpea (Vigna unguiculata) rhizoplane. Afr J Biotechnol, 6: 290-295. Lorenz, N., Hintemann, T., Kramarewa, T., Katayama, A., Yasuta, T., Marschner, P., Kandeler, E. (2006). Response of microbial activity and microbial community composition, in soils to long-term arsenic and cadmium exposure. Soil Biol Biochem, 38:1430–1437 Effron, D., de la Horra, A.M., Defrieri, R.L., Fontanive, V., Palma, P.M. (2004). Effect of cadmium, copper and lead on different soil enzymatic activities in a native forest soil. Comm Soil Sci Plant Anal, 35:1309-1321. Renella, G., Mench, M., Landi, L., Nannipieri, P. (2005). Microbial activity and hydrolase synthesis in long-term Cd-contaminated soils. Soil Biol Biochem, 37:133–139. Khan, S., Cao, Q., Heshman, A.E.L., Xia, Y., He, J. (2007). Soil enzymatic activities and microbial community structure with different application rates of Cd and Pb. J Environ Sci, 19:834-840. Vig, K., Megharaj, M., Senthunathan, N., Naidu, R. (2003). Bioavailability and toxicity of cadmium to microorganisms and their activities in soil: A Review. Adv Environ Res, 8:121–135. Geiger, G., Brandi, H., Furner, G., Schulin, R. (1998). The effect of copper on the activity of cellulase and b -glucosidase in the presence of montmorillonite or Al-montmorillonite. Soil Biol Biochem, 30:1537–1544. Esterbauer, H., Hayn, M., Abuja, P.M., Claeyssens, M. (1991). Structure of cellulolytic enzymes. In: Leatham GF, Himmel ME (eds) Enzymes in biomass conversion. American Chemical Society, Washington, pp 301–312. Wood, T..M, Garcia-Campayo, V. (1990). Enzymology of cellulose degradation. Biodegradation, 1:147–161. Teeri, T.T., Koivula, A., Linder, M., Reinikainen, T., Ruohonrn, L., Srisodsuk, M., Claeyssens, M., Jones, T.A. (1995). Modes of action of two Trichoderma reesei cellobiohydrolases. In Petersen SB, Sevensson B, Pedersen S (Eds.) Carbohydrate bioengineering. pp. 211-225. Rogers, J.E., Li, S.W. (1985). Effects of metals and other inorganic ions on soil microbial activity, soil dehydrogenase assay as a simple toxicity test. Bull Environ Contam Toxicol, 34:858–865. Balyaeva, O.N., Haynes, R.J., Birukova, O.A. (2005). Barley yield and soil microbial and enzyme activities as affected by contamination of two soils with lead, zinc or copper. Biol Fertil Soils,. 41:85–94. Nagwasiri Pride Ndasi, Nidhi Sahu, Sanjay Thul, B. Chandrashekhar, Abhinav Sharma, Ngassoum Martine Benoit and Ram Avatar Pandey. (2014). Biodegradation of phenanthrene by rhizobacteria isolated from local grass growing at hydrocarbon contaminated soil in Cameroon. Biolife, 2(2); 420-441. Nowak, J., Niedzwiecki, E., Dziel, M. (1999). Wpływ metali ciężkich na zmiany aktywności enzymatycznej gleby. Rocz. Gleboz. 50(1/2), 61 (in Polish, with English abstract). Landi, L., Renella, G., Moreno, J.L., Falchini, L., Nannipieri, P. (2000). Influence of cadmium on the metabolic quotient, L: D Glutamic acid respiration ratio and enzyme activity:microbial biomass ratio under laboratory conditions. Biol. Fert. Soils 32(1):8. Shen, G., Lu, Y., Zhou, Q., Hang, J. (2005). Interaction of polycyclic aromatic hydrocarbons and heavy metals on soil enzyme. Chemosphere, 61:1175–1182. Zheng, C.R., Tu, C., Chen, H.M. (1999). Effect of combined heavy metal pollution on nitrogen mineralization potential, urease and phosphatase activities in a Typic Udic Ferrisol. Pedosphere, 9:251-258. Yang, Z.X. Liu, S.G. (2000). Effect of single element and compound pollution of Cd, Zn and Pb on soil enzyme activities. Soil Environ Sci, 9:15–18. Šíša, R. (1993). Enzymová aktivita půdy jako ukazatel její biologické aktivity. Rostl. Výr, 39: 817–825.

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