Bacterial and Enzymatic Bioassays for Toxicity Testing in the Environment

  • Gabriel Bitton
  • Ben Koopman
Part of the Reviews of Environmental Contamination and Toxicology book series (RECT, volume 125)


More than 50,000 chemicals, most of which are xenobiotics, are in common use and new ones are continually and regularly added to the inventory. Serious concern has been raised over the release of these xenobiotics or their metabolites (Liu et al. 1990) into the environment. Their deleterious effect on the environment can be assessed via acute and chronic toxicity tests, using mostly fish and invertebrate bioassays (Peltier and Weber 1985). However, due to the large number of chemicals to be tested, ecotoxicologists and environmental scientists and engineers are now using short-term toxicity assays which are mostly based on inhibition of the activity of enzymes, bacteria, fungi, algae, and protozoa (Bitton 1983; Bitton and Dutka 1986; Dutka and Bitton 1986; Bitton et al. 1989; Liu and Dutka 1984). Microbial bioassays have been used for screening the toxicity of wastewater effluents and for monitoring the quality of reclaimed water (Grabow et al. 1985).


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alleman JE (1988) Respiration-based evaluation of nitrification inhibition using enriched Nitrosomonascultures. In: Scholze RJ, Smith ED, Bandy JT, Yu YC, Basilico JV (eds) Biotechnology for degradation of toxic chemicals in hazardous wastes sites. Noyes Dat Corp., Park Ridge, NJ. pp 642–650.Google Scholar
  2. Alsop GM, Waggy GT, Conway RA (1980) Bacterial growth inhibition test. J Water Pollut Control Fed 52: 2452–2456.Google Scholar
  3. Anderson AC, Abdelghani AA (1980) Toxicity of selected arsenical compounds in short term bacterial bioassays. Bull Environ Contam Toxicol 24: 289–294.CrossRefGoogle Scholar
  4. Atkinson DS, Switzenbaum MS (1988) Microtox assessment of anaerobic bacterial toxicity. In: Scholze RJ, Smith ED, Bandy JT, Yu YC, Basilico JV (eds) Biotechnology for degradation of toxic chemicals in hazardous wastes sites, Noyes Dat Corp., Park Ridge, NJ. pp 622–641.Google Scholar
  5. Benjamin MM, Woods SL, Ferguson JF (1984) Anaerobic toxicity and biodegradability of pulp mill waste constituents. Water Res 18: 601–607.CrossRefGoogle Scholar
  6. Bewley RJF, Stotzky G (1983) Effects of cadmium and zinc on microbial activity in soil: Influence of clay minerals. Part II: Metals added simultaneously. Sci Total Environ 31: 57–69.CrossRefGoogle Scholar
  7. Bitton G (1983) Bacterial and biochemical tests for assessing chemical toxicity in the aquatic environment: A review. CRC Crit Rev Environ Control 13: 51–67.CrossRefGoogle Scholar
  8. Bitton G, Dutka BJ, eds (1986) Toxicity testing using microorganisms, Vol. 1. CRC Press, Boca Raton, FL.Google Scholar
  9. Bitton G, Dutka BJ, Hendricks CW (1989) Microbial toxicity tests. In: Warren-Hicks W, Parkhurst BR, Baker SS (eds) Ecological assessment of hazardous waste sites, U.S. E.P.A., Corvallis, OR, Report EPA 600/3–89/013. pp 644–666.Google Scholar
  10. Bitton G, Dutton RJ, Koopman B (1988) Cell permeability to toxicants: An important parameter in toxicity tests using bacteriia. CRC Crit Rev Environ Control 18: 177–188.CrossRefGoogle Scholar
  11. Bitton G, Koopman B (1986) Biochemical tests for toxicity screening. In: Bitton G, Dutka BJ (eds) Toxicity testing using microorganisms, CRC Press, Boca Raton, FL. pp 27–55.Google Scholar
  12. Blaise C (1991) Microbiotests in aquatic ecotoxicology: Characteristics, utility, and prospects. Environ Toxicol Water Qual 6: 145–155.CrossRefGoogle Scholar
  13. Liaise C, van Coillie R, Bermingham N, Coulombe G (1987) Comparaison des reponses toxiques de trois indicateurs biologiques (bacteries, algues, poissons) exposes a des effluents de fabriques de pates et papiers. Rev Intern Sciences Eau 3: 9–17.Google Scholar
  14. Blaise C, Legault R, Bermingham N, van Coillie R, Vasseur P (1986) A simple microplate algal assay technique for aquatic toxicity assessment. Toxicity Assess 1: 261–281.CrossRefGoogle Scholar
  15. Blaise C, Sergy G, Wells P, Bermingham N, van Coillie R (1988) Biological testing—Development, application, and trends in Canadian Environmental Protection Laboratories. Tox Assess 3: 385–406.CrossRefGoogle Scholar
  16. Brezonik PL, Patterson JW (1972) Activated sludge ATP: Effects of environmental stresses. J San Eng Div, Proc Am Soc Civil Eng 97: 813–821.Google Scholar
  17. Brouwer HT, Murphy T, McArdle L (1990) A sediment-contact bioassay with Photobacterium phosphoreum. Environ Toxicol Chem 9: 1353–1358.CrossRefGoogle Scholar
  18. Bulich AA (1979) Use of luminescent bacteria for determining toxicity in aquatic environments. In: Markings LL, Kimerle RA (eds) Aquatic toxicology. Am Soc Test Mat, Philadelphia, PA.Google Scholar
  19. Bulich AA (1986) Bioluminescent assays. In: Bitton G, Dutka BJ (eds) Toxicity testing using microorganisms, Vol 1. CRC Press, Boca Raton, FL. pp 57–74.Google Scholar
  20. Burns RG, ed (1978) Soil enzymes. Academic Press, London.Google Scholar
  21. Burton GA (1989) Evaluation of seven toxicity tests and their relationships to stream parameters. Tox Assess 4: 149–159.CrossRefGoogle Scholar
  22. Calleja AJ, Baldasana M, Mulet A (1986) Toxicity analysis of leachates from hazardous wastes via Microtox and Daphnia magna. Tox Assess 1: 73–83.CrossRefGoogle Scholar
  23. Calleja AJ, Baldasana M, Mulet A (1986) Toxicity analysis of leachates from hazardous wastes via Microtox and Daphnia magna. Tox Assess 1: 73–83.CrossRefGoogle Scholar
  24. Carlisle SM, Trevors JT (1986) Effects of the herbicide glyphosate on nitrification, denitrification, and acetyl reduction in soil. Water Air Soil Pollut 29: 189–203.CrossRefGoogle Scholar
  25. Cenci G, Morozzi G., Caldini G (1985) Injury by heavy metals in Escherichia coli. Bull Environ Contam Toxicol 34: 188–195.PubMedCrossRefGoogle Scholar
  26. Chang FH, Broadbent FE (1982) Influence of trace metals on some soil nitrogen transformations. J Environ Qual 11: 1–4.CrossRefGoogle Scholar
  27. Clarke SM, Barrick CW, Samoiloff MR (1990) A bioassessment battery for use in an industrial setting: A new management approach. Tox Assess 5: 153–166.CrossRefGoogle Scholar
  28. Christensen GM, Olson D, Reidel B (1982) Chemical effects on the activity of eight enzymes: A review and a discussion relevant to environmental monitoring. Environ Res 29: 247–255.PubMedCrossRefGoogle Scholar
  29. Costerton JW, Cheng KJ (1975) The role of the bacterial cell envelope in antibiotic resistance. J Antimicrob Chemo 1: 363–377.CrossRefGoogle Scholar
  30. Curtis C, Lima A, Lorano SJ, Veith GD (1982) Evaluation of a bacterial bioluminescence bioassay as a method for predicting acute toxicity of organic chemicals to fish. In: Pearson JG, Foster RB, Bishop WE (eds) Aquatic toxicity and hazard assessment, STP #766. Am. Soc. Test. Mat., Philadelphia, PA. pp 170–178.CrossRefGoogle Scholar
  31. Dermer OC, Curtis VS, Leach FR (1980) Biochemical Indicators of Subsurface Pollution. Ann Arbor Sci. Pub., Ann Arbor, MI.Google Scholar
  32. Devillers J, Steiman R, Seigle-Murandi F, Prevot P, Andre C, Benoit-Guyot JL (1990) Combination of single-species laboratory tests for the assessment of the ecotoxicity of p-benzoquinone. Tox Assess 5: 405–416.CrossRefGoogle Scholar
  33. Douglas LA, Bremner JM (1971) A rapid method of evaluating different compounds as inhibitors of urease activity in soils. Soil Biol Biochem 3: 309–315.CrossRefGoogle Scholar
  34. Dutka BJ, Bitton G, eds (1986) Toxicity Testing using Microorganisms, Vol 2. CRC Press, Boca Raton, FL.Google Scholar
  35. Dutka BJ, Gorrie JF (1989) Assessment of toxicant activity in sediments by the ECHA Biocide Monitor. Environ Pollut 57: 1–7.PubMedCrossRefGoogle Scholar
  36. Dutka BJ, Jones K, Kwan KK, Bailey H, McInnis R (1988) Use of microbial and toxicant screening tests for priority site selection of degraded areas in water bodies. Water Res 22: 503–510.CrossRefGoogle Scholar
  37. Dutka BJ, Kwan KK (1988) Battery of screening tests approach applied to sediment extracts. Tox Assess 3: 303–314.CrossRefGoogle Scholar
  38. Dutton RI, Bitton G, Koopman B (1988) Enzyme biosynthesis versus enzyme activity as a basis for microbial toxicity testing. Tox Assess 3: 245–253.CrossRefGoogle Scholar
  39. Dutton RJ, Bitton G, Koopman B, Agami O (1990) Effect of environmental toxicants on enzyme biosynthesis: A comparison of ß-galactosidase, a-glucosidase and tryptophanase. Arch Environ Contam Toxicol 19: 395–398.PubMedCrossRefGoogle Scholar
  40. Dutton RJ, Bitton G, Koopman B, Agami O (1990) Inhibition of ß-galactosidase biosynthesis in Escherichia coli: Effect of alterations of the outer membrane permeability to environmental toxicants. Tox Assess. 5: 253–264.CrossRefGoogle Scholar
  41. Elnabarawy MT, Robideau RR, Beach SA (1988) Comparison of three rapid toxicity test procedures: Microtox, Polytox and activated sludge respiration inhibition. Tox Assess 3: 361–370.CrossRefGoogle Scholar
  42. EPA (1986a) Permit guidance manual on hazardous waste land treatment demonstrations. Office of Solid Waste, U.S. E.P.A.; Washington, D. C. Report EPA530/SW-86–032.Google Scholar
  43. EPA (1986b) Waste/soil treatability studies for four complex wastes. Robert S. Kerr Environ. Res. Lab., U.S. E.P.A., Ada, OK. Report EPA/600/6–86/003a.Google Scholar
  44. Giesy JP, Craney RL, Newsted JL, Rosiu CJ, Benda A, Kreis RG, Horvath FJ (1988a) Comparison of three sediments bioassay methods using Detroit River sediments. Environ Toxicol Chem 7: 483–498.CrossRefGoogle Scholar
  45. Giesy JP, Rosiu CJ, Craney RL, Newsted JL, Benda A, Kreis RG, Horvath FJ (1988b) Toxicity of Detroit River sediments interstitial water to the bacterium Photobacterium phosphoreum. J Great Lakes Res 14: 502–513.CrossRefGoogle Scholar
  46. Guibault GG, Kramer DN (1964) Fluorimetric determination of lipase, acylase, a-and ’y-chymotrypsin and inhibitors of these enzymes. Anal Chem 36: 409412.Google Scholar
  47. Grabow WOK, Morgan WSG, Slabbert JL (1985) Bioassays used for evaluating the quality of reclaimed water in Southern Africa. Water Qual Bull 10: 29–35.Google Scholar
  48. Hancock REW (1984) Alterations in outer membrane permeability. Ann Rev Microbiol 13: 1–34.Google Scholar
  49. Hinwood AL, McCormick MJ (1987) The effect of ionic solutes on EC50 values measured using the Microtox Test. Toxicity Assess 2: 449–461.CrossRefGoogle Scholar
  50. Glolland HT, Coppage DL, Butler PA (1967) Use of fish brain acetylcholinesterase to monitor pollution by organophosphorus pesticides. Bull Environ Contam Toxicol 2: 156–162.CrossRefGoogle Scholar
  51. Acob F, Monod J (1961) Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol 3: 318–356.CrossRefGoogle Scholar
  52. Ohnson LD, Young JC (1983) Inhibition of anaerobic digestion by organic priority pollutants. J Water Pollut Control Fed 55: 1441–1449.Google Scholar
  53. Caiser KLE, Ribo JM (1988) Photobacterium phosphoreum toxicity bioassay. II. Toxicity data compilation. Tox Assess 3:195–237.CrossRefGoogle Scholar
  54. Catayama K (1984) Inhibition by copper ion of the activity of ß-galactosidase and dehydrogenase of activated sludge. Japan J Water Poll Res 7: 100–107.CrossRefGoogle Scholar
  55. Katayama-Hirayama K (1986) Inhibition of the activities of ß-galactosidase and dehydrogenase of activated sludge by heavy metals. Water Res 20: 491–494.CrossRefGoogle Scholar
  56. Kennicutt MC (1980) ATP as an indicator of toxicity. Water Res 14: 225–228.CrossRefGoogle Scholar
  57. King EF, Dutka BJ (1986) Respirometric techniques. In: Bitton G, Dutka BJ (eds) Toxicity testing using microorganisms, Vol 1. CRC Press, Boca Raton, FL. pp 75–113.Google Scholar
  58. Koopman B, Bitton G, Dutton R J, Logue C L (1988) Toxicity testing in wastewater systems: Application of a short-term assay based on induction of the lacoperon in E. coli. Water Sci Technol 20 (11/12): 137–143.Google Scholar
  59. Koopman B, Bitton G, Delfino JJ, Mazidji C, Voiland G, Neita D (1989) Toxicity screening in wastewater systems. Final report (contract no. WM-222) to the Florida Dept. of Environ. Regul., Tallahassee, FL.Google Scholar
  60. Kwan KK, Dutka BJ (1990) Simple two-step sediment extraction procedure for use in genotoxicity and toxicity bioassays. Tox Assess 5: 395–404.CrossRefGoogle Scholar
  61. Lampinen J, Korpela M, Saviranta P, Kroneld R, Karp M (1990) Use of Escherichia colicloned with genes encoding bacterial luciferase for evaluation of chemical toxicity. Tox Assess 5: 337–350.CrossRefGoogle Scholar
  62. Levi Y, Henriet C, Coutant JP, Lucas M, Leger G (1989) Monitoring acute toxicity in rivers with the help of the Microtox test. Water Supply 7: 25–31.Google Scholar
  63. Liang CN, Tabatabai MA (1978) Effects of trace elements on nitrification in soils. J Environ Qual 7: 291–293.CrossRefGoogle Scholar
  64. Liu D (1987) Agar plate method for rapid screening of chemical toxicity. Tox Assess 2: 463–468.CrossRefGoogle Scholar
  65. Liu D, Dutka BJ, eds (1984) Toxicity Screening Procedures using Bacterial Systems. Marcel Dekker, NY.Google Scholar
  66. Liu D, Kwasnieska K (1981) An improved agar plate method for rapid assessment of chemical inhibition to microbial populations. Bull Environ Contam Toxicol 27: 289–294.PubMedCrossRefGoogle Scholar
  67. Liu D, Maguire RJ, Dutka BJ, Pacepavicius GJ (1990) Rationale for including metabolites in chemical toxicity bioassay. Tox Assess 5: 179–188.CrossRefGoogle Scholar
  68. Logue CL, Koopman B, Brown GK, Bitton G (1989) Toxicity screening in a large, municipal wastewater system. J Water Pollut Cont Fed 61: 632–640.Google Scholar
  69. Martin MH, Duncan EM, Coughtrey RI (1982) The distribution of heavy metals in a contaminated woodland ecosystem. Environ Pollut Sci B 3: 147–157.CrossRefGoogle Scholar
  70. Mathes K, Schulz-Berendt VM (1988) Ecological risk assessment of chemicals by measurements of nitrification combined with a computer simulation model of the N-cycle. Tox Assess 3: 271–286.CrossRefGoogle Scholar
  71. Mazidji CN, Koopman B, Bitton G, Voiland G (1990) Use of Microtox and Ceriodaphniabioassays in wastewater fractionation. Tox Assess 5: 265–277.CrossRefGoogle Scholar
  72. Morel JL, Bitton G, Koopman B (1988) Use of Microtox for assessing copper com- plexation with organic ligands. Arch Environ Contam Toxicol 17: 493–496.PubMedCrossRefGoogle Scholar
  73. Mount DI, Anderson-Carnahan L (1987) Methods for aquatic toxicity identification evaluations: Phase I toxicity characterization procedures. U.S. E.P.A., Duluth, MN. Report EPA 600 (draft document).Google Scholar
  74. Munkittrick KR, Power EA, Sergy GA (1991) The relative sensitivity of Microtox, daphnid, rainbow trout and fathead minnow acute lethality tests. Environ Toxicol Water Qual 6: 35–62.CrossRefGoogle Scholar
  75. Nakae T (1986) Outer membrane permeability of bacteria. Crit Rev Microbiol 13: 162.CrossRefGoogle Scholar
  76. Nikaido H, Vaara M (1985) Molecular basis of bacterial outer membrane permeability. Microbiol Rev 49: 1–32.PubMedGoogle Scholar
  77. Nikaido H (1976) Outer membrane of Salmonella typhimurium: Transmembrane diffusion of some hydrophobic substances. Biochim Biophys Acta 433: 118–132.PubMedCrossRefGoogle Scholar
  78. Obst U, Holzapfel-Pschorn A, Wiegand-Rosinus M (1988) Application of enzyme assays for toxicological water testing. Tox Assess 3: 81–91.CrossRefGoogle Scholar
  79. Owen WF, Stuckey DC, Healy JB, Young LY, McCarty PL (1979) Bioassay for monitoring biochemical methane potential and anaerobic toxicity. Water Res 13: 485–492.CrossRefGoogle Scholar
  80. Paran JH, Sharma S, Qureshi AA (1990) A rapid and simple toxicity assay based on growth rate inhibition of Pseudomonas fluorescens. Tox Assess 5: 351–365.CrossRefGoogle Scholar
  81. Parker CE, Pribyl EJ (1984) Assessment of bacterial ATP response as a measurement of aquatic toxicity, In: Liu D, Dutka BJ (eds) Toxicity screening procedures using bacterial systems, Marcel Dekker, NY. pp 283–293.Google Scholar
  82. Parkin GF, Speece RE, Yang CHJ, Kocher WM (1983) Response of methane fermentation systems to industrial toxicants. J Water Pollut Cont Fed 55: 44–53.Google Scholar
  83. Peltier WH, Weber CI (1985) Methods for measuring the acute toxicity of effluents to freshwater and marine organisms, 3rd ed. U.S. E.P.A., Cincinnati, OH. Report EPA-600/4–85/013.Google Scholar
  84. Persaud D, Lomas TD, Hayton A (1987) The in-place pollutant program, Vol III: Phase 1 studies. Water Res. Branch, Ontario Ministry of the Environment, Canada.Google Scholar
  85. Plotkin S, Ram NM (1984) Multiple bioassays to assess the toxicity of a sanitary landfill leachate. Arch Environ Contam Toxicol 13: 197–206.PubMedCrossRefGoogle Scholar
  86. Polybac Corp (1986) Polytox rapid toxicity test procedure. Polybac application procedure, Allentown, PA.Google Scholar
  87. Qureshi A, Flood KW, Thompson SR, Janhurst SM, Inniss CS, Rokosh DA (1982) Comparison of a luminescent bacterial test with other bioassays for determining toxicity of pure compounds and effluents. In: Pearson JG, Foster RB, Bishop WE (eds) Aquatic toxicology and hazard assessment, 5th Conf., STP No. 766. Am. Soc. Test. Mat., Philadelphia. pp 179–195.CrossRefGoogle Scholar
  88. Reidel B, Christensen G (1979) Effect of selected water toxicants and other chemicals upon adenosine triphosphatase activity in vitro. Bull Environ Contam Toxicol 23: 365–368.CrossRefGoogle Scholar
  89. Reinhartz A, Lampert I, Herzberg M, Fish F (1987) A new short-term, sensitive bacterial assay kit for the detection of toxicants. Tox Assess 2: 193–206.CrossRefGoogle Scholar
  90. Ribo JM, Kaiser KLE (1983) Effect of chemicals to photoluminescent bacteria and their correlations with acute and sublethal effects on other organisms. Chemosphere 12: 1421–1442.CrossRefGoogle Scholar
  91. Ribo JM, Rogers F (1990) Toxicity of mixtures of aquatic contaminants using the luminesecent bacterial bioassay. Tox Assess 5: 135–152.CrossRefGoogle Scholar
  92. Rhodes AN, Hendricks CW (1990) A continuous flow method for measuring effects of chemicals on soil nitrification. Tox Assess 5: 77–89.CrossRefGoogle Scholar
  93. Ross PE, Henebry MS (1989) Use of four microbial tests to assess the ecological hazard of contaminated sediments. Tox Assess 4: 1–21.CrossRefGoogle Scholar
  94. Sanchez PS, Sato MIZ, Paschoal CMRB, Alves MN, Furlan EV, Martins MT (1988) Toxicity assessment of industrial effluents from Sao Paulo state, Brazil, using short-term microbial assays. Tox Assess 3: 55–80.CrossRefGoogle Scholar
  95. Schiewe MH, Hawk EG, Actor DI, Krahn MM (1985) Use of bacterial bioluminescence assay to assess toxicity of contaminated marine sediments. Can J Fish Aquat Sci 42: 1244–1248.CrossRefGoogle Scholar
  96. Sergy G (1987) Recommendations on aquatic biological tests and procedures for environmental protection, conservation, and protection. Tech. Dev. and Tech. Serv. Branch, Env. Prot., Conserv. Prot., Environ. Canada, Edmonton, Alberta, 102 pp.Google Scholar
  97. Seyfried PL, Morgan CB (1983) Effect of cadmium on lake water bacteria as determined by the luciferase assay of adenosine triphosphate. In: Bishop NE, Caldwell RD, Heidelph BB (eds) Aquatic toxicology and hazard assessment, 6th Symposium, STP No. 802, Amer. Soc. Test. Mat., Philadelphia, PA. pp 425–441.CrossRefGoogle Scholar
  98. Shapiro HM (1990) Flow cytometry in laboratory microbiology: New directions. Am Soc Microb News 56: 584–588.Google Scholar
  99. Slabbert JL (1986) Improved bacterial growth test for rapid water toxicity screening. Bull Environ Contam Toxicol 37: 565–569.PubMedCrossRefGoogle Scholar
  100. Symons BD, Sims RC (1988) Assessing detoxification of a complex hazardous waste, using the Microtox bioassay. Arch Environ Contam Toxicol 17: 497–505.PubMedCrossRefGoogle Scholar
  101. Trevors JT, Mayfield CI, Innis WE (1981) A rapid toxicity test using Pseudomonas fluorescens. Bull Environ Contam Toxicol 28: 433–439.Google Scholar
  102. Trevors JT (1986) Bacterial growth and activity as indicators of toxicity. In: Bitton G, Dutka BJ (eds) Toxicity testing using microorganisms, Vol 1. CRC Press, Boca Raton, FL. pp 9–25.Google Scholar
  103. True CJ, Hayward AA (1990) Relationships between Microtox test results, extraction methods and physical and chemical compositions of marine sediment samples. Tox Assess 5: 29–45.CrossRefGoogle Scholar
  104. Tyler G (1976) Heavy metal pollution, phosphatase activity, and mineralization of organic phosphorus in forest soils. Soil Biol Biochem 8: 327–332.CrossRefGoogle Scholar
  105. Wainwright M (1978) A review of the effects of pesticides on microbial activity in soils. J Soil Sci 29: 287–298.CrossRefGoogle Scholar
  106. Wang X, Yu X, Bartha R (1990) Effect of bioremediation on polycyclic hydrocarbon residues in soil. Environ Sci Technol 24:1086–1089.CrossRefGoogle Scholar
  107. Williamson KJ, Johnson DG (1981) A bacterial bioassay for assessment of wastewater toxicity. Water Res 15: 383–390.CrossRefGoogle Scholar
  108. Xu H, Dutka BJ (1987) ATP-TOX system: A new rapid sensitive bacterial toxicity screening system based on the determination of ATP. Tox Assess 2: 149–166.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag New York Inc. 1992

Authors and Affiliations

  • Gabriel Bitton
    • 1
  • Ben Koopman
    • 1
  1. 1.Department of Environmental Engineering SciencesUniversity of FloridaGainesvilleUSA

Personalised recommendations