Biological test method for determining toxicity of sediment using luminescent bacteria: chapter 11
Appendices
- Biological Test Methods and Supporting Guidance Documents Published by Environment Canada’s Method Development & Applications Section
- Members of the Inter-Governmental Environmental Toxicity Group (as of April 2002)
- Environment Canada Regional and Headquarters Offices
- Members of the Scientific Advisory Group
- Procedural Variations for Solid-Phase Sediment Toxicity Tests Using Luminescent Bacteria, as Described in Canadian, United States, and Netherlands Methodology Documents
- Bench Sheet for Sediment Toxicity Tests Using V. fischeri
Appendix A: Biological Test Methods and Supporting Guidance Documents Published by Environment Canada’s Method Development and Applications SectionFootnote 18
| Title of Biological Test Method or Guidance Document | Report Number | Publication Date | Applicable Amendments |
| Acute Lethality Test Using Rainbow Trout | EPS 1/RM/9 | July 1990 | May 1996 |
| Acute Lethality Test Using Threespine Stickleback (Gasterosteus aculeatus) | EPS 1/RM/10 | July 1990 | March 2000 |
| Acute Lethality Test Using Daphnia spp. | EPS 1/RM/11 | July 1990 | May 1996 |
| Test of Reproduction and Survival Using the Cladoceran Ceriodaphnia dubia | EPS 1/RM/21 | February 1992 | November 1997 |
| Test of Larval Growth and Survival Using Fathead Minnows | EPS 1/RM/22 | February 1992 | November 1997 |
| Toxicity Test Using Luminescent Bacteria (Photobacterium phosphoreum) | EPS 1/RM/24 | November 1992 | -- |
| Growth Inhibition Test Using a Freshwater Alga Selenastrum capricornutum | EPS 1/RM/25 | November 1992 | November 1997 |
| Acute Test for Sediment Toxicity Using Marine or Estuarine Amphipods | EPS 1/RM/26 | December 1992 | October 1998 |
| Fertilization Assay Using Echinoids (Sea Urchins and Sand Dollars) | EPS 1/RM/27 | December 1992 | November 1997 |
| Toxicity Tests Using Early Life Stages of Salmonid Fish (Rainbow Trout, Coho Salmon, or Atlantic Salmon) | EPS 1/RM/28 1st Edition |
December 1992 | January 1995 |
| Toxicity Tests Using Early Life Stages of Salmonid Fish (Rainbow Trout) | EPS 1/RM/28 2nd Edition |
July 1998 | -- |
| Test for Survival and Growth in Sediment Using the Larvae of Freshwater Midges (Chironomus tentans or Chironomus riparius) | EPS 1/RM/32 | December 1997 | -- |
| Test for Survival and Growth in Sediment Using the Freshwater Amphipod Hyalella azteca | EPS 1/RM/33 | December 1997 | -- |
| Test for Measuring the Inhibition of Growth Using the Freshwater Macrophyte, Lemna minor | EPS 1/RM/37 | March 1999 | -- |
| Title of Biological Test Method or Guidance Document | Report Number | Publication Date | Applicable Amendments |
| Reference Method for Determining Acute Lethality of Effluents to Rainbow Trout | EPS 1/RM/13 1st Edition |
July 1990 | May 1996, December 2000 |
| Reference Method for Determining Acute Lethality of Effluents to Rainbow Trout | EPS 1/RM/13 2nd Edition |
December 2000 | -- |
| Reference Method for Determining Acute Lethality of Effluents to Daphnia magna | EPS 1/RM/14 1st Edition |
July 1990 | May 1996, December 2000 |
| Reference Method for Determining Acute Lethality of Effluents to Daphnia magna | EPS 1/RM/14 2nd Edition |
December 2000 | -- |
| Reference Method for Determining Acute Lethality of Sediment to Marine or Estuarine Amphipods | EPS 1/RM/35 | December 1998 | -- |
| Title of Biological Test Method or Guidance Document | Report Number | Publication Date | Applicable Amendments |
| Guidance Document on Control of Toxicity Test Precision Using Reference Toxicants | EPS 1/RM/12 | August 1990 | -- |
| Guidance Document on Collection and Preparation of Sediment for Physicochemical Characterization and Biological Testing | EPS 1/RM/29 | December 1994 | -- |
| Guidance Document on Measurement of Toxicity Test Precision Using Control Sediments Spiked with a Reference Toxicant | EPS 1/RM/30 | September 1995 | -- |
| Guidance Document on Application and Interpretation of Single-Species Tests in Environmental Toxicology | EPS 1/RM/34 | December 1999 | -- |
Appendix B: Members of the Inter-Governmental Environmental Toxicity Group (as of April, 2002)
Federal, Environment Canada
C. Blaise
Centre St. Laurent
Montreal, Quebec
M. Bombardier
Environmental Technology Centre
Ottawa, Ontario
U. Borgmann
National Water Research Institute
Burlington, Ontario
J. Bruno
Pacific Environmental Science Centre
North Vancouver, British Columbia
C. Buday
Pacific Environmental Science Centre
North Vancouver, British Columbia
K. Doe
Atlantic Environmental Science Centre
Moncton, New Brunswick
G. Elliott
Environmental Protection Service
Edmonton, Alberta
F. Gagné
Centre St. Laurent
Montreal, Quebec
M. Harwood
Environmental Protection Service
Montreal, Quebec
D. Hughes
Atlantic Environmental Science Centre
Moncton, New Brunswick
P. Jackman
Atlantic Environmental Science Centre
Moncton, New Brunswick
N. Kruper
Environmental Protection Service
Edmonton, Alberta
M. Linssen
Pacific Environmental Science Centre
North Vancouver, British Columbia
D. MacGregor
Environmental Technology Centre
Ottawa, Ontario
D. Moul
Pacific Environmental Science Centre
North Vancouver, British Columbia
W.R. Parker
Environmental Protection Service
Fredericton, New Brunswick
L. Porebski
Marine Environment Branch
Hull, Quebec
D. Rodrique
Environmental Technology Advancement Directorate
Hull, Quebec
R. Scroggins
Environmental Technology Centre
Ottawa, Ontario
A. Steenkamer
Environmental Technology Centre
Ottawa, Ontario
T. Steeves
Atlantic Environmental Science Centre
Moncton, New Brunswick
G. van Aggelen (Chairperson)
Pacific Environmental Science Centre
North Vancouver, British Columbia
R. Watts
Pacific Environmental Science Centre
North Vancouver, British Columbia
P. Wells
Environmental Conservation Service
Dartmouth, Nova Scotia
Federal, Fisheries & Oceans Canada
R. Roy
Institut Maurice Lamontagne
Mont-Joli, Quebec
Provincial
C. Bastien
Ministère de l’Environnement du Québec
Ste. Foy, Quebec
B. Bayer
Manitoba Environment
Winnipeg, Manitoba
D. Bedard
Ontario Ministry of Environment
Rexdale, Ontario
M. Mueller
Ontario Ministry of Environment
Rexdale, Ontario
D. Poirier
Ontario Ministry of Environment
Rexdale, Ontario
J. Schroeder
Ontario Ministry of Environment
Rexdale, Ontario
Appendix C: Environment Canada Regional and Headquarters Offices
Headquarters
351 St. Joseph Boulevard
Place Vincent Massey
Hull, Quebec
K1A 0H3
Atlantic Region
15th Floor, Queen Square
45 Alderney Drive
Dartmouth, Nova Scotia
B2Y 2N6
Quebec Region
105 McGill Street,
8th Floor
Montreal, Quebec
H2Y 2E7
Ontario Region
4905 Dufferin St., 2nd Floor
Downsview, Ontario
M3H 5T4
Prairie and Northern Region
Room 210, Twin Atria No. 2
4999 - 98 Avenue
Edmonton, Alberta
T6B 2X3
Pacific and Yukon Region
224 West Esplanade Street
North Vancouver, British Columbia
V7M 3H7
Appendix D: Members of the Scientific Advisory Group
SAG Members
Mr. Craig Buday
Environment Canada
Pacific Environmental Science Centre
2645 Dollarton Highway
North Vancouver, BC V7H 1B1
Phone: (604) 924-2514
Fax:(604) 924-2554
e-mail: craig.buday@ec.gc.ca
Dr. Allen Burton, Jr.
Institute for Environmental Quality
Wright State University
Dayton, OH 45435, USA
Phone: (937) 775-4995
Fax: (937) 775-4997
e-mail: aburton@wright.edu
Mr. Ken Doe
Environment Canada
Toxicology Laboratory
Environmental Quality Section, ECB
Atlantic Environmental Science Centre
P.O . Box 23005
Moncton, NB E1A 6S8
Phone: (506) 851-3486
Fax:(506) 851-6608
e-mail: Ken.Doe@ec.gc.ca
Dr. Kay Ho
Atlantic Ecology Division
U .S. Environmental Protection Agency
27 Tarzwell Drive
Narragansett, RI 02882, USA
Phone: (401) 782-3196
Fax: (401) 782-3030
e-mail: ho.kay@epamail.epa.gov
Ms. Paula Jackman
Environment Canada
Toxicology Laboratory
Environmental Quality Section, ECB
Atlantic Environmental Science Centre
P.O. Box 23005
Moncton, NB E1A 6S8
Phone: (506) 851-2886
Fax: (506) 851-2907
e-mail: Paula.Jackman@ec.gc.ca
Mr. Jim Osborne
Environment Canada
Marine Environment Branch
P lace Vincent Massey, 12 th Floor
351 St. Joseph Boulevard
Hull, P Q K1A 0H3
Phone: (819) 953-2265
Fax: (819) 953-0913
e-mail: Jim.Osborne@ec.gc.ca
Ms. Linda Porebski
Environment Canada
Marine Environment Branch
Place Vincent Massey, 12th Floor
351 St. Joseph Boulevard
Hull, PQ K1A 0H3
Phone: (819) 953-4341
Fax: (819) 953-0913
e-mail: Linda.Porebski@ec.gc.ca
Dr. Philippe Ross
Environmental Science and Engineering Division
Colorado School of Mines
Golden, CO 80401-1887, USA
Phone: (303) 273-3473
Fax: (303) 273-3413
e-mail: pross@mines.edu
Mr. Paul Topping
Environment Canada
Marine Environment Branch
Place Vincent Massey, 12th Floor
351 St. Joseph Boulevard
Hull, PQ K1A 0H3
Phone: (819) 953-0663
Fax: (819) 953-0913
e-mail: Paul.Topping@ec.gc.ca
Mr. Sylvain Trottier
Environment Canada
Centre Saint-Laurent
105 McGill Street
Montreal, PQ H2Y 2E7
Phone: (514) 496-7104
Fax: (514) 496-7143
e-mail: sylvain.trottier@ec.gc.ca
Scientific Authority
Mr. Rick Scroggins
Environment Canada
Method Development and Applications Section
Environmental Technology Centre
3439 River Road South
Ottawa, ON K1A 0H3
Phone: (613) 990-8569
Fax: (613) 990-0173
e-mail: scroggins.richard@etc.ec.gc.ca
Consultants
Dr. Don McLeay
McLeay Environmental Ltd.
2999 Spring Bay Road
Victoria, BC V8N 5S4
Phone: (250) 472-2608
Fax: (250) 472-2609
e-mail: mcleayenvir@islandnet.com
Mr. Gary Wohlgeschaffen
Marine Environmental Sciences
Bedford Institute of Oceanography
P.O. Box 1006
Dartmouth, NS B2Y 4A2
Phone: (902) 426-2479
Fax: (902) 426-6695
e-mail: WohlgeschaffenG@mar.dfo-mpo.gc.ca
Appendix E: Procedural Variations for Solid-Phase Sediment Toxicity Tests Using Luminescent Bacteria, as Described in Canadian, United States, and Netherlands Methodology Documents
Source documents are listed here chronologically, by originating agency rather than by author(s).
EC 1992 represents EC (Environment Canada), “Biological Test Method: Toxicity Test Using Luminescent Bacteria (Photobacterium phosphoreum)”, Conservation and Protection, Ottawa, ON, Report EPS 1/RM/24, 61 p. (1992).
MICROBICS 1992 represents Microbics, “Detailed Solid-Phase Test Protocol”, p. 153-178, in: Microtox Manual--A Toxicity Testing Handbook, Microbics Corporation, Carlsbad, CA (1992).
MICROBICS 1995a represents Microbics, “Microtox Acute Toxicity Solid-Phase Test”, Microbics Corporation, Carlsbad, CA, 18 p. (1995a).
ASTM 1995 represents ASTM (American Society for Testing and Materials), “Standard Guide for Conducting Sediment Toxicity Tests with Luminescent Bacteria”, Draft No. 8, ASTM, Philadelphia, PA (1995).
EC 1996a represents EC (Environment Canada), “Procedure for Conducting a Microtox Solid-Phase Test”, Toxicology Section, Environmental Quality Laboratories, Environmental Conservation Branch, Moncton, NB, Standard Operating Procedure #43, 10 p., Prepared January 1996 by G. Wohlgeschaffen. Atlantic Environmental Science Centre, Moncton, NB (1996a).
AZUR 1997 represents AZUR, “Basic Solid-Phase Test”, AZUR Environmental, Carlsbad, CA, Draft 6-11-97, 4 p. (1997).
AZUR 1998a represents AZUR, “Basic Solid-Phase Test (Basic SPT)”, AZUR Environmental, Carlsbad, CA, 16 p. (1998).
AZUR 1998b represents AZUR, “Solid-Phase Test (SPT)”, AZUR Environmental, Carlsbad, CA, 19 p. (1998).
EC 1999a represents EC (Environment Canada), “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 (1999a).
NICMM 1999 represents NICMM (National Institute for Coastal and Marine Management/RIKZ), “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, 23 p, NICMM, Middelburg, The Netherlands (1999).
| Document | Sample Type |
Collection Procedure |
Transport Temp (°C) |
Container | Field Replicates? |
| EC 1992 | soil, sediment, sludge | follow guidance in ASTM (1991) and EC (1994) | 1-7 | glass or TeflonTM-coated are preferred; polyethylene, polypropylene acceptable; 500 mL to 1 L | NI |
| MICROBICS 1992 | soil, sediment | NITable note a.1 | NI | new borosilicate TFE-lined caps | NI |
| MICROBICS 1995a | soil, sediment | NI | NI | new borosilicate Teflon-lined caps | NI |
| ASTM 1995 | sediment | follow ASTM Standard Guides E 1367, E 1383, and E 1391 | 4 ± 2 | newTable note b borosilicate TFE-lined caps | yesTable note c |
| EC 1996a | soil, sediment | NI | NI | NI | NI |
| AZUR 1997 | soil, sediment | NI | NI | NI | NI |
| AZUR 1998a | soil, sediment | NI | NI | NI | NI |
| AZUR 1998b | soil, sediment | NI | NI | NI | NI |
| EC 1999a | soil, sediment | NI | NI | 50 mL centrifuge tube | NI |
| NICMM 1999 | sediment | NI | NI | NI | NI |
| Document | Airspace? | Temp (°C) | Light/Dark | Maximum Time |
| EC 1992 | no | 4 ± 2 | dark | 6 weeks |
| MICROBICS 1992 | no | 2-8 | NI | 48 h if possible |
| MICROBICS 1995a | no | fridge | NI | 48 h if possible |
| ASTM 1995 | no | 4 ± 2 | dark | 2-8 weeks |
| EC 1996a | NITable note a.2 | NI | NI | NI |
| AZUR 1997 | NI | NI | NI | NI |
| AZUR 1998a | NI | NI | NI | NI |
| AZUR 1998b | NI | NI | NI | NI |
| EC 1999a | no | 4 ± 2 | dark | 6 weeks |
| NICMM 1999 | NI | 4 | dark | < 4 weeks |
| Document | Sieve? | Remove Large Organisms? |
Remove Large Particles? |
Homogenize Sample? |
Sediment Characterization |
| EC 1992 | NITable note a.3 | NI | NI | mix thoroughly | colour |
| MICROBICS 1992 | NI | NI | yes(≥1 mm) | yes | pH |
| MICROBICS 1995a | NI | NI | NI | yes (1-3 min) |
pH |
| ASTM 1995 | no | yes | yes(>5 mm) | yes | organic carbon or total volatile solids, particle size distribution, ammonia, percent water, pH |
| EC 1996a | NI | NI | NI | yes | NI |
| AZUR 1997 | NI | NI | NI | yes (1-3 min) |
NI |
| AZUR 1998a | NI | NI | NI | mix thoroughly | NI |
| AZUR 1998b | NI | NI | NI | mix thoroughly | NI |
| EC 1999a | no | yes | yes | mix thoroughly | NI |
| NICMM 1999 | NI | NI | NI | yes | for porewater:Table note b.1 pH, salinity, sulphide, ammonia |
| Document | Amount (g) | Replicates | Weighing Precision (g) | Temperature (°C) |
Time |
| EC 1992 | NITable note a.4 | NI | NI | NI | NI |
| MICROBICS 1992 | 5 | NI | NI | NI | NI |
| MICROBICS 1995a | 5 ± 0.2 | 3 | ± 0.01 | 100 ± 5 | 24 h |
| ASTM 1995 | a few | NI | NI | ~100 | overnight |
| EC 1996a | 3-5 | 3 | ± 0.01 | 60-100 | 48 h |
| AZUR 1997 | NI | NI | NI | NI | NI |
| AZUR 1998a | NI | NI | NI | NI | NI |
| AZUR 1998b | NI | NI | NI | NI | NI |
| EC 1999a | 5 ± 0.2 | 3 | ± 0.01 | 100 ± 5 | 24 h |
| NICMM 1999 | ~ 5 | 3 | NI | 100 | 24 h |
| Document | Subsample Weight (g) |
Discard Pore Water? |
Type of Diluent |
Volume of Diluent (mL) |
Container for Primary Dilution |
Container for Test Concentrations |
| EC 1992 | 0.4 | yes (centrifuge) | Solid-Phase Diluent | 4.0 | NATable note a.5 | SPT tubeTable note b.2 |
| MICROBICS 1992 | 0.3 | yes (centrifuge) | Solid-Phase Diluent | 3.0 | NATable note a.5 | SPT tubeTable note b.2,Table note c.1 |
| MICROBICS 1995a | 7.00 | optional | NaCl or NaNO3 Diluent | 35 | new 50 mL glass or plastic | SPT tubeTable note b.2,Table note c.1 |
| ASTM 1995 | 7 ± 0.35Table note d | no | 3.5% NaCl or 3.02% NaNO3Table note e |
35 | new, clean borosilicate glassTable note f | new, clean borosilicate glass tubeTable note f |
| EC 1996a | 7 ± 0.05 | NITable note g | Solid-Phase Diluent | 35 | new 50 mL disposable polypropylene | SPT tubeTable note b.2,Table note c.1 |
| AZUR 1997 | 7.00 | NI | Diluent or reference seawater | 35 | ~50 mL beaker | cuvetteTable note h |
| AZUR 1998a | 7.00 | NI | Solid-Phase Diluent | 35 | ~50 mL beaker | cuvetteTable note h |
| AZUR 1998b | 7.00 | NI | Solid-Phase Diluent | 35 | ~50 mL beaker | SPT tubeTable note b.2,Table note c.1 |
| EC 1999a | 0.3 | yes (centrifuge) | 3.5% NaCl | 3.0 | NATable note a.5 | SPT tubeTable note b.2,Table note i |
| NICMM 1999 | 7.0 | NI | Solid-Phase Diluent | 35 | serum tube | SPT tubeTable note b.2,Table note c.1 |
| Document | pH Adjustment? |
Mixing Device |
Vortex Depth |
Mixing Time (min) |
| EC 1992 | NITable note a.6 | shake and use pipette | NATable note b.3 | NI (mix well) |
| MICROBICS 1992 | NITable note c.2 | shake and use pipette | NA | NI (completely dissociate and suspend sample) |
| MICROBICS 1995a | NI | magnetic plate, TeflonTM stir bar | half the height of liquid level | 10 |
| ASTM 1995 | noTable note d.1 | magnetic plate, Teflon stir bar | half the height of liquid level | 20Table note e.1 |
| EC 1996a | NI | magnetic stirrer | 1/3 the height of liquid level | 10 |
| AZUR 1997 | NI | magnetic plate, Teflon stir bar | half the height | 10 of liquid level |
| AZUR 1998a | NI | magnetic plate, Teflon stir bar | half the height of liquid level | 10 |
| AZUR 1998b | NI | magnetic plate, Teflon stir bar | half the height of liquid level | 10 |
| EC 1999a | no | shake and use pipette | NA | NI (completely dissociate and suspend sample) |
| NICMM 1999 | NI | magnetic plate, Teflon stir bar | half the height of liquid level | 10, or until suspension is homogeneous |
| Document Factor | Dilution | Number of Test Concentrations | Volume Transferred from Primary Dilution to Tube with Maximum Test Concentation (mL) | Number of Times to Mix with Pipette to Resuspend Particles | Volume Transferred Serially from Maximum to Lowest Concentration (mL) |
| EC 1992 | 1:2 | 9 | NATable note a.7 | NITable note b.4 | 2.0 |
| MICROBICS 1992 | 1:2 | 12Table note c.3 | NA | 5-6× | 1.5 |
| MICROBICS 1995a | 1:2 | 13Table note c.3 | 1.5Table note d.2 | NI | 1.5 |
| ASTM 1995 | 1:2Table note c.3 | 12Table note c.3 | 1.5Table note e.2 | several | 1.5 |
| EC 1996a | 1:2 | 12 | 1.5Table note d.2,Table note e.2 | 3-4× | 1.5 |
| AZUR 1997 | 1:2 | 9 | 2.0Table note f.1 | thoroughly | 1.0 |
| AZUR 1998a | 1:2 | 9 | 2.0Table note f.1,Table note g.1 | mix | 1.0 |
| AZUR 1998b | 1:2 | 13 | 1.5Table note d.2,Table note g.1 | mix | 1.5 |
| EC 1999a | 1:2 | 10 | 1.5Table note e.2 | 5-6× | 1.5 |
| NICMM 1999 | 1:2 | 13 | 1.5 | 10× | 1.5 |
| Document | Maximum Test ConcentrationTable note a.8 |
Number of Control SolutionsTable note b.5 | Duplicates of Each Test Concentration? | Initial (I0) Light Readings? | Containers Inoculated with ReagentTable note c.4 |
| EC 1992 | 10% | 1 | optional | no | SPT tubes |
| MICROBICS 1992 | 98 684 mg/L or 9.868% | 3 | optional | no | SPT tubes |
| MICROBICS 1995a | 19.737% | 4 | yes | no | SPT tubes |
| ASTM 1995 | 10% | 3 | no | no | glass tubes |
| EC 1996a | 197 368 mg/L | 3 | no | no | SPT tubes |
| AZUR 1997 | 9.9% or 99 000 mg/L | 1 | optionalTable note d.3 | yesTable note e.3 | cuvettesTable note f.2 |
| AZUR 1998a | 99 000 mg/L | 1 | optionalTable note d.3 | yesTable note e.3 | cuvettesTable note f.2 |
| AZUR 1998b | 9.9% or 99.01 mg/LTable note g.2 | 2 | no | no | SPT tubes |
| EC 1999a | 9.868% | 3 | no | no | SPT tubes |
| NICMM 1999 | 19.737% | 4 | yes | no | SPT tubes |
| Document | Equilibration Temperature (°C) | Equilibration Time (min) |
| EC 1992 | 15 ± 0.3 | NI |
| MICROBICS 1992 | 15 | 10 |
| MICROBICS 1995a | 15 | 10 |
| ASTM 1995 | 15 ± 0.5 | NI |
| EC 1996a | 15 | 10 |
| AZUR 1997 | NITable note a.9 | 5 |
| AZUR 1998a | NITable note a.9 | 5 |
| AZUR 1998b | 15 | 5 |
| EC 1999a | room temperature (~20) | 5 |
| NICMM 1999 | 15 | 10 |
| Document | Species | Strain | Source | Storage Temperature (°C)Table note a.10 |
| EC 1992 | P. phosphoreumTable note b.6 | NRRL B-11177Table note c.5 | Microbics | -20 |
| MICROBICS 1992 | P. phosphoreumTable note b.6 | NI | Microbics | NI |
| MICROBICS 1995a | NITable note d.4 | NI | Microbics | NI |
| ASTM 1995 | V. fischeri | NRRL B-11177Table note c.5 | NI | -20 |
| EC 1996a | P. phosphoreumTable note b.6 | NI | NI | NI |
| AZUR 1997 | NI | NI | AZURTable note e.4 | NI |
| AZUR 1998a | NI | NI | AZUR | NI |
| AZUR 1998b | NI | NI | AZUR | NI |
| EC 1999a | V. fischeri | NRRL B-11177Table note c.5 | AZUR | -20 |
| NICMM 1999 | V. fischeri | NI | AZUR | NI |
| Document | Source of Reconstitution SolutionTable note a.11 |
Temp (°C) |
Usable Life | Swirl Vial | Mixing Reconstituted Bacteria |
| EC 1992 | Microbics | 3-5 | 2 h | 3- 4× | 20× with 0.5 mL pipette |
| MICROBICS 1992 | Microbics | NITable note b.7 | 2 h | 3-4× | 20× with 0.5 mL pipette |
| MICROBICS 1995a | Microbics | NITable note b.7 | NI | NI | 10× with 0.5 mL pipette |
| ASTM 1995 | water | 5.5 ± 1 | NI | a few times | 20× with 0.5 mL pipette |
| EC 1996a | AZUR | NITable note b.7 | NI | 3× | 10× with 0.5 mL pipette |
| AZUR 1997 | AZUR | NITable note b.7 | 3 h | NI | 10× with 0.5 mL pipette |
| AZUR 1998a | AZUR | NITable note b.7 | 3 h | 3-4× | 10× with 0.5 mL pipette |
| AZUR 1998b | AZUR | NITable note b.7 | 3 h | 3-4× | 10× with 0.5 mL pipette |
| EC 1999a | lab-distilled water | NITable note b.7 | 3-4 h | 3-4× | 20× with 0.5 mL pipette |
| NICMM 1999 | AZUR | NITable note b.7 | 4 h | noTable note c.6 | 20× with 1 mL pipette |
| Document | Volume of Inoculum (µL) | Mixing Procedure | Times Mixed | Incubation Time (min) | Incubation Temperature (°C) | Insert Filter Columns?Table note a.12 |
| EC 1992 | NITable note b.8 | NI | NI | 20 | 15 ± 0.3 | yes |
| MICROBICS 1992 | 20 | 1.5 mL pipette | 5-6× | 20 | 15 | yes |
| MICROBICS 1995a | 20 | shake, then 1.5 mL pipette | NI | 20 | 15 | yes |
| ASTM 1995 | 20 | shake or vortex | NI | 20 | 15 ± 0.5 | yes |
| EC 1996a | 20 | 1.5 mL pipette | 2× | 20 | 15 | yes |
| AZUR 1997 | 10Table note c.7 | shake | NI | 30Table note c.7 | NITable note d.5 | noTable note e.5 |
| AZUR 1998a | 10Table note c.7 | shake | NI | 30Table note c.7 | NITable note d.5 | noTable note e.5 |
| AZUR 1998b | 20 | 1.5 mL pipette | NI | 20 | 15 | yes |
| EC 1999a | 20 | 1.5 mL pipette | 5-6× | 20 temperature (~20) | room | yes |
| NICMM 1999 | 20 | manual | NI | 20 | 15 | yes |
| Document | Analyzer | Volume Transferred (µL) | Cuvette Type | Stabilization ConditionsTable note a.13 | Reading Cuvette |
| EC 1992 | Microtox Model 2055, Model 500 | 500 | glassTable note b.9 | 5 min, 15 ±0.3°C |
manually inserted |
| MICROBICS 1992 | Microtox Model 500 | 500 | glassTable note b.9 | 5 minTable note c.8 | manually inserted |
| MICROBICS 1995a | Model 500 | 500 | glassTable note b.9 | 10 minTable note c.8 | manually inserted |
| ASTM 1995 | photometer | 500 | glassTable note b.9 | 10 min, 15 ± 0.5°C |
photometer instruction manual |
| EC 1996a | Model 500 | 500 | glassTable note b.9 | NITable note c.8,Table note d.6,Table note e.6 | manually inserted |
| AZUR 1997 | NITable note d.6,Table note f.3 | 500Table note g.3 | NITable note d.6,Table note h.1 | NITable note c.8,Table note d.6,Table note i.1 | manually inserted |
| AZUR 1998a | Model 500 | 500Table note g.3,Table note j | NITable note d.6,Table note h.1 | NITable note c.8,Table note d.6,Table note i.1 | manually inserted |
| AZUR 1998b | Model 500 | 500 | NITable note d.6,Table note h.1 | 10 minTable note c.8 | manually inserted |
| EC 1999a | Microtox Model 500 | 500 | glassTable note b.9 | 5 minTable note c.8 | manually inserted |
| NICMM 1999 | Microtox Model 500 | 500 | glassTable note b.9 | NITable note c.8 | manually inserted |
| Document | Use and Properties of Reference Sediment | Use to Correct Sample Values?Table note a.14 |
| EC 1992 | must useTable note b.10; parallel test; should be similar in particle sizes and organic/inorganic content | NITable note c.9 |
| MICROBICS 1992 | optional/desirable; “control” samples from area useful in d istinguishing toxicity | NI |
| MICROBICS 1995a | desirable; match with test material for particle size, organic and mo isture content | optional; analytical data “reduced” versus reference |
| ASTM 1995 | recommended; important to match with test material for particle size | optionalTable note d.7; analytical data “reduced” versus reference |
| EC 1996a | optional | NI |
| AZUR 1997 | recommended; match with test material for particle size, organic and moisture content | noTable note e.7 |
| AZU R 1998a | recommended; match with test material for particle size, organic and moisture content | noTable note e.7 |
| AZUR 1998b | recommended; match with test material for particle size, organic and moisture content | NI |
| EC 1999a | recommended; “control” samples from area with similar particle size useful in distinguishing toxicity | no |
| NICMM 1999 | correction made for the measured silt and clay fraction of sample using an equation derived from own data and from Ringwood et al. (1997) | use equation to correct EC50 |
| Document | Date Entry | Method of Analysis | EndpointTable note a.15 (Units) |
Guidelines for Judging Toxicity?Table note b.11 |
| EC 1992 | software data capture | manual or software graphingTable note c.10 | IC50 (% t:v) | no |
| MICROBICS 1992 | software data capture | manual or software graphing | ECxx (mg/L) | no |
| MICROBICS 1995a | software data capture | manual or software graphing | ECxx (% wt:v) | no |
| ASTM 1995 | manual | manual calculations and graphs | ICxx (% wt:v) | yesTable note d.8 |
| EC 1996a | software data capture | software graphing | ICxx (mg/L) | no |
| AZUR 1997 | software data capture | software graphing | EC50 (% wt:v) or EC50 (mg/L) |
yesTable note e.8 |
| AZUR 1998a | NITable note f.4,Table note g.4 | software graphing | IC50 (mg/L) | yesTable note h.2 |
| AZUR 1998b | NITable note f.4,Table note g.4 | NITable note c.10,Table note f.4 | NI | no |
| EC 1999a | software data capture | software graphing | IC50 (% wt:v) | yesTable note i.2 |
| NICMM 1999 | software data capture | manual or software graphing | ECxx (% wt:v) | no |
| Document | Normalize for Moisture? | Correct for Colour? | Correct for Turbidity? |
| EC 1992 | NITable note a.16 | NI | NI |
| MICROBICS 1992 | no | yesTable note b.12 | yesTable note b.12 |
| MICROBICS 1995a | yesTable note c.11 | yesTable note d.9 | yesTable note d.9 |
| ASTM 1995 | optional | optionalTable note d.9,Table note e.9 | NI |
| EC 1996a | yes | yes | yes |
| AZUR 1997 | NI | noTable note f.5 | noTable note g.5 |
| AZUR 1998a | NI | noTable note f.5 | noTable note g.5 |
| AZUR 1998b | NI | noTable note f.5 | noTable note g.5 |
| EC 1999a | yesTable note c.11 | no | no |
| NICMM 1999 | yesTable note c.11 | NI | NI |
| Document | Reference Toxicant | Control Sediment? | Requirement for Valid Sediment Toxicity Test |
| EC 1992 | phenol, zinc, potassium dichromate, sodium lauryl sulphate | yesTable note a.17 | valid numerical estimate of ICp based on concentrations showing light inhibition both greater than, and less than, the inhibition at the ICp |
| MICROBICS 1992 | phenol, zinc | NITable note b.13,Table note c.12 | NITable note b.13 |
| MICROBICS 1995a | NITable note b.13 | NITable note b.13 | if Gamma for each test concentration is >1, a repeat test is required using <7 g in primary dilution |
| ASTM 1995 | phenol, zinc | yes (may be the reference sediment) | if SD of the light reading for the three control solutions exceeds ~12% of the mean, the test should be considered unacceptableTable note d.10 |
| EC 1996a | NITable note b.13 | NITable note b.13 | NITable note b.13 |
| AZUR 1997 | NITable note b.13 | NITable note b.13 | NITable note b.13,Table note e.10 |
| AZUR 1998a | NITable note b.13 | NITable note b.13 | NITable note b.13,Table note e.10 |
| AZUR 1998b | NITable note b.13 | NITable note b.13 | NITable note b.13 |
| EC 1999a | HS-6 standard sediment | Roberts Bank “clean” sediment | IC50s for “clean” sediment and HS-6 within warning limits (± 2 SD) of respective control charts |
| NICMM 1999 | zinc | yesTable note a.17 | porewater sulphide, <16.5 mg/L; porewater pH, 6.5-7.5 (± 0.5), porewater salinity, >8‰; EC50 for reference toxicant (ZnSO4) within ± 3 SD of control chart; EC50 for control sediment, <1000 Toxic Units |
Appendix F: Bench Sheet for Sediment Toxicity Tests Using V. fischeri
| Project: | |||||||
| Test Method: | Analyst: | ||||||
| Reagent Lot: | Expiry Date: | ||||||
| Date of Test | Filename and/or Sample | Date Collected in Field |
Date Received in Lab |
Temp (°C) on Arrival |
Reagent Recon TimeTable note a.18 |
Test Start Time | NotesTable note b.14 |
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