Solid-Phase Test for Sediment Toxicity Using the Luminescent Bacterium, Vibrio Fischeri (original) (raw)

Abstract

sparkles

AI

The solid-phase test utilizing luminescent bacteria (Vibrio fischeri) provides a method for assessing the toxicity of sediment samples. This automated procedure simplifies the toxicity testing of whole sediment and related materials, allowing for efficient screening of various environments. The test can inform sediment quality assessments and helps identify the spatial extent of contamination.

Figures (11)

Table 1: Rapid summary of the test procedure

Table 1: Rapid summary of the test procedure

Figure 1: Preparation of a series of concentrations of whole sediment sample by seria! dilutions in water using a 50 % dilution series

Figure 1: Preparation of a series of concentrations of whole sediment sample by seria! dilutions in water using a 50 % dilution series

The test array consists of 3 controls (comprised of dilution water only) and 12 test concentrations. The maximum test concentration is 197,000 mg/L (19.7%, wt:v), with each successive concentration being 50% of the previous one. A schematic overview of the various stages in the test has been provided as Figure 3. Figure 2: Microtox photometer, solid-phase tubes, and the computer to run the test, calculate the endpoints, and store the data.

The test array consists of 3 controls (comprised of dilution water only) and 12 test concentrations. The maximum test concentration is 197,000 mg/L (19.7%, wt:v), with each successive concentration being 50% of the previous one. A schematic overview of the various stages in the test has been provided as Figure 3. Figure 2: Microtox photometer, solid-phase tubes, and the computer to run the test, calculate the endpoints, and store the data.

Figure 3: Summary of the Solid-Phase Test using Luminescent Bacteria

Figure 3: Summary of the Solid-Phase Test using Luminescent Bacteria

Figure 7: Plots of Gamma versus concentration and % effect versus concentration, as plotted by the Microtox Omni software. These plots are from the raw data provided in Table 3, which was also generated using the Microtox Omni software using data from a test on a field-collected sediment. where IC50,, is the calculated IC50 (or its 95% confidence limits) of the wet sediment sample, S1, through $3, are the dry weights of the sediment subsamples from Section 7.8, and S1,, through S3,, are the corresponding wet weights. These calculations can be expedited by entering the weights and IC50 values into a spreadsheet and using the necessary formulae.

Figure 7: Plots of Gamma versus concentration and % effect versus concentration, as plotted by the Microtox Omni software. These plots are from the raw data provided in Table 3, which was also generated using the Microtox Omni software using data from a test on a field-collected sediment. where IC50,, is the calculated IC50 (or its 95% confidence limits) of the wet sediment sample, S1, through $3, are the dry weights of the sediment subsamples from Section 7.8, and S1,, through S3,, are the corresponding wet weights. These calculations can be expedited by entering the weights and IC50 values into a spreadsheet and using the necessary formulae.

Table 3: Raw data used in Figure 7 plots. “It” is the light reading of a filtrate of a particular concentration of the test material. The Microtox Omni software automatically selects the data to be used in the calculation.

Table 3: Raw data used in Figure 7 plots. “It” is the light reading of a filtrate of a particular concentration of the test material. The Microtox Omni software automatically selects the data to be used in the calculation.

a valid test must be met (CV < 12 %). Additionally, it is recommended that the dose-response curve for each sample of test sediment be examined to confirm that light loss decreases as test concentration decreases, in an approximately monotonic manner. We recommend a r° value for the regression equation (provided by the Microtox Omni software, or calculated by the user) be > 0.90. If not (e.g., if one or more data points appear to be “out of place” with respect to the others), consideration should be given to repeating the test for that sample as this type of response is normally caused by pipetting errors. Finally, the results of any reference toxicity test (Environment Canada 1990, 1995) with a toxic positive control sediment which was initiated with the same lot of Bacterial Reagent as that used in the sediment toxicity test should be considered during the interpretive phase of the investigation. These results, when compared with historic test results derived by the testing facility using the same reference toxicant and test procedure (i.e., by comparison against the laboratory’s warning chart for this reference toxicity test), will provide insight into the sensitivity of the test organisms as well as the laboratory’s testing precision and performance for a reference toxicity test with V. fischeri. If the results of the reference toxicity test are outside of three standard deviations of the historic mean value of all previous tests with this reference toxicant (the “Control Limit’), all test conditions pertaining to the test should be double-checked thoroughly and consideration should be given to repeating the test (Figure 8). Figure 8; Example of a Quality Control Chart for the Microtox Solid-Phase test

a valid test must be met (CV < 12 %). Additionally, it is recommended that the dose-response curve for each sample of test sediment be examined to confirm that light loss decreases as test concentration decreases, in an approximately monotonic manner. We recommend a r° value for the regression equation (provided by the Microtox Omni software, or calculated by the user) be > 0.90. If not (e.g., if one or more data points appear to be “out of place” with respect to the others), consideration should be given to repeating the test for that sample as this type of response is normally caused by pipetting errors. Finally, the results of any reference toxicity test (Environment Canada 1990, 1995) with a toxic positive control sediment which was initiated with the same lot of Bacterial Reagent as that used in the sediment toxicity test should be considered during the interpretive phase of the investigation. These results, when compared with historic test results derived by the testing facility using the same reference toxicant and test procedure (i.e., by comparison against the laboratory’s warning chart for this reference toxicity test), will provide insight into the sensitivity of the test organisms as well as the laboratory’s testing precision and performance for a reference toxicity test with V. fischeri. If the results of the reference toxicity test are outside of three standard deviations of the historic mean value of all previous tests with this reference toxicant (the “Control Limit’), all test conditions pertaining to the test should be double-checked thoroughly and consideration should be given to repeating the test (Figure 8). Figure 8; Example of a Quality Control Chart for the Microtox Solid-Phase test

ND = Below detection limit; PEL = Probable effects level; * T ono ef al 1998 Table 4: Summary ofselected results from Sydney Harbour, NS, Canada, pollution gradier study (Zajdlik et al. 2001).

ND = Below detection limit; PEL = Probable effects level; * T ono ef al 1998 Table 4: Summary ofselected results from Sydney Harbour, NS, Canada, pollution gradier study (Zajdlik et al. 2001).

Loading...

Loading Preview

Sorry, preview is currently unavailable. You can download the paper by clicking the button above.

References (50)

  1. Anonymous. 1979. Flash ! Instrument Replaces Fish. Environmental Science and Technology. 13: 646.
  2. ASTM (American Society for Testing and Materials). 1995. Standard Guide for Conducting Sediment Toxicity Tests with Luminescent Bacteria, Draft No. 8, ASTM, Philadelphia, PA, USA .
  3. AZUR. 1997. Basic Solid Phase Test, Draft 6-11-96, 4 p., AZUR Environmental, Carlsbad, CA, USA. AZUR. 1998a. Basic Solid-Phase Test (Basic SPT), 16 p., AZUR Environmental, Carlsbad, CA, USA. AZUR. 1998b. Solid-Phase Test (SPT), 19 p., AZUR Environmental, Carlsbad, CA, USA.
  4. Benton, M.J., M.L. Malott, S.S. Knight, C.M. Cooper, and W.H. Benson. 1995. Influence of sediment composition on apparent toxicity in solid-phase test using bioluminescent bacteria. Environ. Toxicol. Chem. 14 (3): 411-414.
  5. Bombardier, M. and N. Bermingham. 1999. The sed-tox index: toxicity-directed management tool to assess and rank sediments based on their hazard -concept and application. Environ. Toxicol. Chem. 18 (4): 685-698.
  6. Brouwer, H. and T. Murphy. 1995. Volatile Sulfides and their Toxicity in Freshwater Sediments. Environ. Toxicol. Chem. 14 (2): 203-208.
  7. Brouwer, H., T. Murphy, and L. McArdle. 1990. A Sediment-contact Bioassay with Photobacterium phosphoreum. Environ. Toxicol. Chem. 9: 1353-1358.
  8. Burton, G.A., Jr., D. l. Denton, K. Ho, and D. S. Ireland. 2003. Sediment Toxicity Testing: Issues and Methods, p. 111-150, Chapter 5 in: Handbook of Ecotoxicology, 2 nd Edition, D. J. Hoffman, B. A. Rattner, G. A. Burton, Jr., and J. Cairns, Jr. (eds.), CRC Press LLC. 1290 pp.
  9. Carter, J. A., R. E. Mroz, K.L. Tay, and K.G. Doe. 1998. An evaluation of the use of soil and sediment bioassays in the assessment of three contaminated sites in Atlantic Canada. Water Quality Research Journal of Canada. 33 (2): 295-317.
  10. CEPA (Canadian Environmental Protection Act). 1999. Disposal at Sea, Part 7, Division 3, Sections 122-127 and Schedules 5 and 6, Statutes of Canada, Chapter 33.
  11. Cook, N.H. and P.G. Wells. 1996. Toxicity of Halifax Harbour Sediments: an Evaluation of the Microtox Solid-phase Test. Water Qual. Res. J. Canad. 31 (4): 673-708.
  12. Day, K.E., B.J. Dutka, K.K. Kwan, N. Batista, T.B. Reynoldson, and J.L. Metcalfe-Smith. 1995. Correlations Between Solid-phase Microbial Screening Assays, Whole-sediment Toxicity Tests with Macroinvertebrates and In Situ Benthic Community Structure. J. Great Lakes Res. 2 (2): 192-206.
  13. Denning, A., K. Doe, B. Ernst, and G. Julien. 2003. Screening Level Ecological Risk Assessment. Stephenville Upper Air Station. Stephenville, Newfoundland. Surveillance Report EPS-5- AR-03-02. Environment Canada, Dartmouth, NS. B2Y 2N6. 47 pp. plus appendices.
  14. Environment Canada. 1990. Guidance Document on the Control of Toxicity Test Precision Using Reference Toxicants, Conservation and Protection, Ottawa, ON, Report EPS 1/RM/12, 85 pp. Environment Canada. 1992. Biological Test Method: Toxicity Test Using Luminescent Bacteria (Photobacterium phosphoreum), Conservation and Protection, Ottawa, ON, Report EPS 1/RM/24, 61 pp.
  15. Environment Canada. 1995. Guidance Document on Measurement of Toxicity Test Precision Using Control Sediments Spiked with a Reference Toxicant, Environmental Protection Service, Ottawa, ON, Report EPS 1/RM/30, 56 pp.
  16. Environment Canada. 1996a. Procedure for Conducting a Microtox Solid Phase Test, Standard Operating Procedure #43, 10 p., prepared January 1996 by G. Wohlgeschaffen. Atlantic Environmental Science Centre, Moncton, NB.
  17. Environment Canada. 1996b. 1996 National Compendium Monitoring at Ocean Disposal Sites, Unpublished Report, Disposal at Sea Program, Marine Environment Division, Ottawa, ON.
  18. Environment Canada. 1997a. 1996-97 Discussion Paper on Ocean Disposal and Cost Recovery, Unpublished Report, Disposal at Sea Program, Marine Environment Division, Ottawa, ON.
  19. Environment Canada. 1997b. Biological Test Method: Test for Survival and Growth in Sediment Using the Larvae of Freshwater Midges (Chironomus tentans or Chironomus riparius), Environmental Protection Service, Ottawa, ON, Report EPS 1/RM/32, 131 pp.
  20. Environment Canada. 1997c. Biological Test Method: Test for Survival and Growth in Sediment Using the Freshwater Amphipod Hyalella azteca, Environmental Protection Service, Ottawa, ON, Report EPS 1/RM/33, 123 pp.
  21. Environment Canada. 1999a. Standard Operating Procedure for the Solid-Phase Toxicity Test Using Luminescent Bacteria (Vibrio fischeri), SOP No. IC50MS10.SOP, first draft, 18 p., prepared October 1999 by C. Buday. Pacific Environmental Science Centre, North Vancouver, BC.
  22. Environment Canada. 1999b. Guidance Document for the Application and Interpretation of Single- species Data from Environmental Toxicology Testing, Environmental Protection Service, Ottawa, ON, Report EPS 1/RM/34.
  23. Environment Canada. 2002. Biological Test Method: Solid-Phase Reference Method for Determining the Toxicity of Sediment Using Luminescent Bacteria (Vibrio fischeri), Environment Canada, Environmental Protection Series, EPS 1/RM/42.
  24. Environment Canada. 2003. Guidance Document on Statistical Methods for Environmental Toxicity Tests, Fifth Draft, March 2003, Environmental Protection Service, Ottawa, ON, Report EPS 1/RM/xx, in Preparation.
  25. Gaudet, I.D. 1998. Effect of Microtox Reagent Reconstitution Age on the Variability of Analytical Results from the Microtox Assay, Final Report to Western Canada Microtox Users Committee by Alberta Research Council, Edmonton, AB.
  26. Giesy, J.P., R.L. Graney, J.L. Newsted, C.J. Rosiu, A. Benda, R.G. Kreis, Jr., and F.J. Horvath. 1988. Comparison Of Three Sediment Bioassay Methods Using Detroit River Sediments. Environ. Toxicol. Chem. 7: 483 -498.
  27. Government of Canada. 2001. Disposal at Sea Regulations, SOR/2001-275 under the Canadian Environmental Protection Act, Canada Gazette Part II, 135 (17): 1655-1657, August 1, 2001, Ottawa, ON.
  28. ISO. 1993. Water quality -Determination of the Inhibitory Effect of Water samples on the Light Emission of Vibrio fischeri (Luminescent bacteria test). International Organization for Standardization. ISO/TC 47/SC 5N107 (Draft). 18 pp.
  29. Jacobs, M.W., J.J. Delfino, and G. Bitton. 1992. The Toxicity Of Sulfur To Microtox From Acetonitrile Extracts Of Contaminated Sediments. Environ. Toxicol. Chem. 11: 1137 - 1143.
  30. Johnson, B.T. 1998. Microtox Toxicity Test System -New Developments and Applications, p. 201- 218, Chapter 14 in: Microscale Testing in Aquatic Toxicology --Advances, Techniques, and Practice, P.G. Wells, K. Lee, and C. Blaise (eds.), CRC Press, New York, NY, USA. 679 pp.
  31. Kwan, K.K., and B.J. Dutka. 1995. Comparative Assessment Of Two Solid-Phase Toxicity Bioassays: The Direct Sediment Toxicity Testing Procedure (DSTTP) and The Microtox Solid-Phase Test (SPT). Bull. Environ. Contam. Toxicol. 55: 338 -346.
  32. McLeay, D., K. Doe, P. Jackman, P. Ross, C. Buday, G. van Aggelen, G. Elliott, W. Antoniolli, S. Trottier, J. Pickard, K. Kinnee, K. Lee, and G. Wohlgeschaffen. 2001. Interlaboratory Studies to Validate Environment Canada's New Reference Method for Determining the Toxicity of Sediment Using Luminescent Bacteria (Vibrio fischeri) in a Solid-Phase Test, Technical Report Prepared by McLeay Environmental Ltd. (Victoria, BC) and Environment Canada's Atlantic Environmental Science Centre (Moncton, NB) for the Method Development and Applications Section, Environment Canada, Ottawa, ON.
  33. Microbics. 1992. Detailed Solid-phase Test Protocol, p. 153-178 in Microtox Manual -A Toxicity Testing Handbook, Microbics Corporation, Carlsbad, CA, USA.
  34. Microbics. 1995a. Microtox Acute Toxicity Solid-phase Test, Microbics Corporation, Carlsbad, CA, 18 pp.
  35. Mueller, D.C., J.S. Bonner, S.J. McDonald, R.L. Autenrieth, K.C. Donnelly, K. Lee, K. Doe, J. Anderson. 2003. The use of toxicity bioassays to monitor the recovery of oiled wetland sediments. Environ. Toxicol. Chem. 22 (9): 1945-1955.
  36. NICMM (National Institute for Coastal and Marine Management/RIKZ). 1999. Standard Operating Procedures, Marine: Microtox Solid-phase (Vibrio fisheri) Sediment Toxicity Test, C.A. Schipper, R.M. Burgess, M.E. Schot, B.J. Kater, and J. Stronkhorst, Project SPECIE*BIO, RIKZ/AB-99.107x, Ver.1.1, June 1999, NICMM, Middelburg, The Netherlands, 23 pp.
  37. Porebski, L.M. and J.M. Osborne. 1998. The application of a Tiered Testing Approach to the Management of Dredged Sediments for Disposal at Sea in Canada, Chemistry and Ecology 14: 197-214.
  38. Porebski, L.M., K.G. Doe, B.A. Zajdlik, D. Lee, P. Pocklington, J. Osborne. 1999. Evaluating the techniques for a tiered testing approach to dredged sediment assessment -a study over a metal concentration gradient. Environ. Toxicol. Chem. 18 (11): 2600-2610
  39. Qureshi, A.A., K.W. Flood, S.R. Thompson, S.M. Janhurst, C.S. Innis, and D.A. Rokosh. 1982. Comparison of a Luminescent Bacterial Test with Other Bioassays for Determining Toxicity of Pure Compounds and Complex Effluents, p. 179-195 in J.G. Pearson, R.B. Foster, and W.E. Bishop, eds., Aquatic Toxicology and Hazard Assessment: Fifth Conference, ASTM STP 766, American Society for Testing and Materials, Philadelphia, PA., USA.
  40. Qureshi, A.A.,A.A. Bulich, and D.I. Isenberg. 1998. Microtox Toxicity Test Systems -Where Thet Stand Today, p. 185-199, Chapter 13 in: Microscale Testing in Aquatic Toxicology -- Advances, Techniques, and Practice, P.G. Wells, K. Lee, and C. Blaise (eds.), CRC Press, New York, NY, USA. 679 pp.
  41. Ringwood, A.H., M.E. DeLorenzo, P.E. Ross, and A.F. Holland. 1997. Interpretation of Microtox Solid-phase Toxicity Tests: the Effects of Sediment Composition. Environ. Toxicol. Chem. 16 (6): 1135-1140.
  42. Ross, P. and P.A. Leitman. 1995. Solid Phase Testing of Aquatic Sediments Using Vibrio fischeri: Test Design and Data Interpretation, p. 65-76, Chapter 6 in: Environmental Toxicology Assessment, M. Richardson (ed.), Taylor and Francis, London, UK.
  43. Ross, P. 1998. Role of Microbiotests in Contaminated Sediment Assessment Batteries, p. 549-556, Chapter 38 in: Microscale Testing in Aquatic Toxicology --Advances, Techniques, and Practice, P.G. Wells, K. Lee, and C. Blaise (eds.), CRC Press, New York, NY, USA.
  44. Ross, P., G.A. Burton, Jr., M. Greene, K. Ho, P. Meier, L. Sweet, A. Auwarter, A. Bispo, K. Doe, K. Erstfeld, S. Goudey, M. Goyvaerts, D. Henderson, M. Jourdain, M. Lenon, P. Pandard, A. Qureshi, C. Rowland, C. Schipper, W. Schreurs, S. Trottier, and G. van Aggelen. 1999. Interlaboratory Precision Study of a Whole Sediment Toxicity Test with the Bioluminescent Bacterium Vibrio fischeri. Environ. Toxicol. 14: 339-345.
  45. Schiewe M.H., E.G. Hawk, D.I. Actor, and M.M. Krahn. 1985. The Use Of Bacterial Bioluminescence Assay To Assess Toxicity of Contaminated Marine Sediments. Can. J. Fish. Aquat. Sci. 42: 1244 -1248.
  46. Sprague, J.B. and A. Fogels. 1977. Watch the Y in Bioassay, p. 107-118 in: Proceedings of the 3 rd Aquatic Toxicity Workshop, Halifax, NS, November 2-3, 1976, Environment Canada, Environmental Protection Service Technical Report No. EPS-5-AR-77-1.
  47. Tay, K.-L., K.G. Doe, S.J. Wade, J.D.A. Vaughan, R.E. Berrigan, and M.J. Moore. 1992. Sediment bioassessment in Halifax Harbour. Environ. Toxicol. Chem. 11: 1567-1581.
  48. Tay, K.L., K.G. Doe, A.J. MacDonald, and K. Lee. 1998. The Influence of Particle Size, Ammonia, and Sulfide on Toxicity of Dredged Materials for Ocean Disposal, p. 559-574, Chapter 39 in: Microscale Testing in Aquatic Toxicology --Advances, Techniques, and Practice, P.G. Wells, K. Lee, and C. Blaise (eds.), CRC Press, New York, NY, USA. 679 pp.
  49. True, C.J., and A.A. Heyward. 1990. Relationships between Microtox test results, extraction methods, and physical and chemical compositions of Marine sediment samples. Toxicol. Assess. 5: 29 -45
  50. Zajdlik, B.A., K.G. Doe, and L.M. Porebski. 2000. Report on Biological Toxicity Tests Using Pollution Gradient Studies -Sydney Harbour. Environment Canada, Marine Environment Division, Ottawa. Report EPS 3/AT/2. July, 2000. 104 pp.