SEDIMENT HEALTH QUOTIENT: APPLICATION … HEALTH QUOTIENT: APPLICATION DOCUMENT FOR FRESHWATER SYSTEMS Prepared for: Golder Grundteknik kb Gata 12 …

SEDIMENT HEALTH QUOTIENT:

APPLICATION DOCUMENT FOR

FRESHWATER SYSTEMS

Prepared for:

Golder Grundteknik kb

Gata 12

416 64 Göteborg, Sweden

Sweden

Prepared by:

HydroQual Laboratories Ltd.

#3, 6125 12th Street SW

Calgary, AB

Canada T2H 2K1

February 2000

Sediment Health Assessment - Application Document Freshwater

Written by SG on 1998/07/09 HydroQual Laboratories Ltd. Version 2.0 Revised by SG/JFH on 2000/02/14 i File: sediment health quotient(bil6).doc

Executive Summary

The sediment health quotient is an approach for assessing the health and ecological

potential of sediments. The quotient is based on a physical, chemical, and biological

characterization and rating of sediment conditions. The approach is flexible, permits

comparisons amongst sediments with widely different properties and contaminant levels,

and it can be adapted to site specific conditions.

The rationale and development of the quotient are documented in this report along with

sample handling, assessment methods, and quality assurance practices. Standardized

reporting formats have also been developed for compiling and presenting the findings.

An interpretive guide is included for the reporting formats and how to apply the results to

site specific conditions. This guide was written for freshwater sediments. The same

approach can be taken for marine sediments but with different test species.


Written by SG on 1998/07/09 HydroQual Laboratories Ltd. Version 2.0 Revised by SG/JFH on 2000/02/14 ii File: sediment health quotient(bil6).doc

Table Of Contents

Executive Summary ................................................................................... i

Table of Contents ...................................................................................... ii

List of Tables ........................................................................................... iii

List of Figures ........................................................................................... iii

1.0 Sediment Health Quotient .......................................................................... 1

1.1 Background .................................................................................... 1

1.2 Sediment Health Quotient .............................................................. 2

2.0 Assessment Methods................................................................................. 4

2.1 Sediment Handling Procedures ...................................................... 4

2.2 Biological Assessment.................................................................... 5

2.2.1 Microbes ............................................................................. 5

2.2.2 Plants.................................................................................. 7

2.2.3 Invertebrates....................................................................... 8

2.2.4 Sediment Community Processes ........................................ 9

2.2.5 Genotoxicity Testing ........................................................... 10

2.3 Quality Assurance Practices........................................................... 11

3.0 Reporting Formats ..................................................................................... 12

4.0 Interpretive Guide ...................................................................................... 12

5.0 General References................................................................................... 14

6.0 Tables ........................................................................................... 15

7.0 Figures ........................................................................................... 16

8.0 Appendices ........................................................................................... 17

8.1 Examples of Data Summary Tables


Written by SG on 1998/07/09 HydroQual Laboratories Ltd. Version 2.0 Revised by SG/JFH on 2000/02/14 iii File: sediment health quotient(bil6).doc

List Of Tables

1. Summary of Test Methods

2. Criteria for Ranking Test Results

3. Test Results Summary

4. Data Summary Tables

List Of Figures

1. Flow Chart for Processing Test Sediments


Written by SG on 1998/07/09 HydroQual Laboratories Ltd. Version 2.0 Revised by SG/JFH on 2000/02/14 1 File: sediment health quotient(bil6).doc

1.0 Sediment Health Quotient

The sediment health quotient is an approach for assessing the health and ecological

potential of contaminated sediments. The quotient is based on physical, chemical and

biological information on the sediments and porewater. The level of impairment is

ranked relative to what is defined as a non-toxic, or a healthy state and condition of the

sediment. The focus of the sediment health quotient is ecological. It can be adapted to

site specific conditions to address specific questions and information requirements in

both freshwater and marine systems..

The rationale for the development of the sediment health quotient is presented in this

section. Specific handling and test methods are briefly described in the next section. A

guide for interpreting and applying the findings is also provided and the data reporting

formats are appended.

1.1 Background

Conventional approaches to contaminated site assessments are well documented and

will not be reviewed in detail here. They typically involve a review of existing information

on the site, some chemical analyses, and interpretation of the findings based on

published data. More recently, biological testing has been incorporated into

assessments to compliment chemical analyses and to confirm that no bioactive

substances have been overlooked (due diligence and final validation). However, this has

involved limited testing with usually one or two different species.

Sediments are complex systems and factors affecting the availability of contaminants

are not well understood. Reliance on chemical specific criteria alone is limiting because

the approach does not address:

• potential interactive effects (synergistic, additive, etc.),

• factors affecting bioavailability (exposure pathways),



• effects of background conditions,

• unknown toxic constituents and conditions, and

• ecosystem structure and function (the biological component).

There are potentially billions of permutations and combinations of the basic building

blocks of organic compounds (carbon, hydrogen, oxygen, nitrogen, etc.). Over five

hundred thousand new compounds are registered every year and there are currently

over fourteen million known and registered compounds. Chemical criteria have been

established for only a few hundred compounds. Hence, the potential for missing

chemicals of real concern in site assessments is both real and large.

Chemical criteria are also based on the presence of the material alone and do not

provide consideration for interactive effects with other compounds. These interactions

can be additive, synergistic (greater than the sum of either), or even negative. Further,

chemical criteria do not provide information on how sediment conditions affect

bioavailability and the structure and function of a viable ecosystem.

No life forms may be present in sediments with conditions considered ideal for

supporting a viable ecosystem. This would be considered a system with high ecological

potential even though there are no life forms present. Alternatively, conditions may not

be suitable for supporting growth of one or more trophic levels. This impairment may not

be a result of chemical toxicity but could arise from the absence of nutrients, organic

matter, or sediment conditions unfavorable for supporting growth of microbes, plants

and other trophic levels (physical factors). This system would have a low ecological

potential. These issues are of particular relevance in assessing areas that are absent of

life within an impacted site. Do you reclaim or rehabilitate areas with naturally low

ecological potentials to conditions considered more ecologically self-sustaining?

The sediment health quotient provides a means to integrate chemical, physical and

biological information into an overall rating for comparative assessment purposes. It

provides a tool for dealing with naturally versus chemically impaired systems. The



quotient provides a tool to establish realistic, site specific clean-up criteria targeted at

protecting ecological integrity (ecosystem structure and function).

1.2 Sediment Health Quotient

Sediment health is defined in terms of abiotic and biotic properties and how these relate

to the existing ecological state and future potential. Abiotic factors include physical and

chemical conditions such as sediment porewater pH, electrical conductivity (salts),

particle size distribution (sand, silt, clay), porewater ammonium, total organic carbon

(TOC) and total Kjeldahl nitrogen (TKN) levels, colour and odour. Additional parameters

can be included in the initial characterization as required (metals and acid volatile

sulfides for example).

Most of these parameters are gross measures of the quality of the physical and

chemical environment. Organisms must respond to changes in the quality of their

environment in order to survive. The physiological tolerances of most living things are

also well defined. However, frequent and large exceedences of their physiological limits

can be stressful and ultimately lethal.

There are two biotic components. The first is an assessment of indigenous microbial,

invertebrate and plant populations (what’s present in the sample). The microbes are

plated out onto a nutrient agar and allowed to grow. The presence of invertebrates and

plants is noted on the project sheet during sample sign-in.

The second component involves measurement of two types of responses. The first

involves community processes and second is the response of individual organisms

exposed to the sample and sample extracts. A multi-trophic level approach is taken in

both cases. The absence of life forms may not be a result of chemical or physical

conditions or lack of food. In this respect, living organisms are employed as detectors of

environmental quality. The test battery is composed of organisms representative of

major trophic levels and could include:



• microbes

⇒ bacterial luminescence

⇒ bacterial growth

⇒ enumeration of fungal and bacterial populations

⇒ sediment oxygen demand (community process)

• plants

⇒ inhibition of plant growth in water and methanol extracts (root, algal,

duckweed)

⇒ presence/absence of plants in the sample as received

• invertebrates

⇒ survival or sediment dwelling organisms (chironomids, amphipods, or

Lumbriculus)

⇒ zooplankton survival in water and methanol extracts (i.e. the waterflea

Daphnia magna)

⇒ presence/absence of invertebrates in the sample as received

• vertebrates

⇒ fish survival (fathead minnow or other species)

• other - depends on information needs (some examples are listed below)

⇒ amphibian development and survival

⇒ invertebrate reproduction

⇒ microbial genetic diversity

⇒ endocrine disrupting compounds

⇒ mixed function oxidase induction

⇒ in vitro testing with cell cultures

⇒ genotoxcity (AMES, SOS, Mutatox)

⇒ mammalian and avian studies

HydroQual Laboratories Ltd. has developed a large and diverse battery of screening test

methods for assessing site specific issues (freshwater and marine). Most of the species

are representative of major trophic levels in aquatic and sediment systems. Plants and

microbes convert chemical energy and light (plants) into biomass, and they also serve



as primary food sources for invertebrates. Microbes and invertebrates come into

intimate contact with the sediments and sediment-bound contaminants. Hence,

exposure is through direct contact and from water column.

Invertebrates are preyed upon by other invertebrates and fish. They provide a pathway

for contaminants from the sediments to the water column and other life forms (birds and

mammals).

The response of the test organism is proportional to the level of stress; in other words

dose dependent. The intensity of the response is factored into the rating system for the

health quotient. The test battery is not limited to the organisms and measurements

defined above and alternate tests can be included to address site specific conditions.

Further, this document is for freshwater sediments and the same approach can be taken

for marine systems with ecologically similar species.

Invertebrate species native to the area under investigation can be added to the test

battery. Alternatively, some species can be removed if considered inappropriate or

insensitive. However, it is important to include one or more standard test organisms in

each trophic level as benchmarks to relate the findings to other sites and conditions.

A major benefit of employing different life forms in a test battery is that different species

have different sensitivities to different compounds and conditions. Hence, effects are

less likely to be missed with a test battery. The approach also permits resolution of the

sensitivity of ecosystem components to different contaminants and conditions. Other

applications of the sediment health quotient include:

• assess ecological risk,

• map or delineate impacted areas (isopleths of ecological impairment),

• basis for selection of remedial management options, and

• tool to assess effectiveness of remediation and validation of treatments (confirm

ecosystem structure and function adequately restored to acceptable conditions).



2.0 Methods

This document does not cover sampling procedures. The sample collected in the field

must be representative of the site under investigation. The number of samples and or

composites is determined by the study objectives.

2.1 Sediment Handling Procedures

Tests are conducted on the sediments as received, porewater (if available) and on water

and methanol extracts (Figure 1). Observations are made on the colour, texture and

odour of the sediment with a simplified key. Colour is recorded as either pale or dark

yellow, brown, red, olive, green, gray or clear. Texture is evaluated as percent

sand/silt/clay by hydrometer. Odour is rated as either strong or mild hydrocarbon,

organic, chemical, or none. Measurements for total organic carbon and Kjeldahl nitrogen

provide some information on the nutritional value of the sediment (organic substrate

available). Additional chemical and physical analyses may be included as required.

The sediments are homogenized with hand mixing. A 100 g sample is removed and

centrifuged at 15,000 x g for 20 minutes. The pore water volume is recorded and the

supernatant decanted and archived for testing. The solids are removed, dried at 105oC

overnight, and reweighed to get the moisture content. The dry solids are homogenized

and 20 g samples separately extracted with 80 mL of reagent grade methanol and

deionized water. The extracts are shaken to disperse the solids. The colour, odour, pH

and conductance of the porewater and/or aqueous extract are measured. All sediments

and extracts are stored at 5oC in darkness until tested.

Aqueous extracts may not be done on samples with high porewater volumes (>30%). In

these instances, the porewater is tested directly.



The water and methanol solvents permit differential extraction of potential contaminants

based on their physical and chemical properties. Methanol extracts are tested at a level

known to have no adverse effects on the test organism (NOEC or no observed effect

concentration).

2.2 Biological Test Methods

The test descriptions and results are grouped by major trophic level. Tests are

performed on the sediments as received (solid phase) and on the porewater or aqueous

extract, and the methanol extract.

The results from each test are rated from 1 to 5. Higher values are assigned to lower

responses or no effect (non-toxic condition). Toxic responses are assigned lower

values. These ratings are then carried forward to a trophic level assessment, then finally

an overall sediment health quotient.

2.2.1 Microbes

Microbes are an integral component of sediment systems. They play vital roles in the

degradation of organic mater, the cycling of organic nutrients and metals, and serve as

an important food source for many invertebrates. The microbial tests included bacterial

luminescence, bacterial growth inhibition and enumeration of sediment bacteria and

fungi. Additional tests can be included if required such as sediment oxygen demand,

assessment of genetic diversity, and enumeration of microbial community structure.

The bacterial luminescence test is based on light output by the marine bacterium Vibrio

fischeri (Environment Canada, 1992). Substances that are toxic or stressful will reduce

light output. Hence, light output is related to sample strength or toxicity. The aqueous

samples are tested at 91% full strength; the slight dilution resulting from the addition of a

salt solution to osmotically adjust the sample (marine bacterium requires salt). The

methanol extracts are tested at 5%, which is the highest concentration of methanol

tolerated by the bacterium (NOEC or no observed effect concentration for methanol).



Light levels are measured at 15oC with a Microtox Model 500 Unit. The results are

expressed as a percentage of light output in the controls (water or 5% methanol) and

then ranked (Table 2).

Sediment bacterial populations are enumerated with a most probable number (MPN)

method (Carter, 1993). A one hundred fold dilution of the porewater or aqueous extract

is prepared with growth media (trypticase soy broth). This working solution is dispensed

into three wells of a 96 well microplate with a further ten fold dilution. The solutions in

these wells are then diluted one in ten six times in adjacent wells with media.

The bacterial density is rated from 1 to 5, depending on the extent of growth. A value of

5 is assigned to aqueous extracts of sediment with a large bacterial population (>106

bacteria/mL); lower values are assigned to extracts with less growth (fewer bacteria).

Fungal counts are performed on the porewater or water extract. A 0.1 mL aliquot of

porewater is plated onto a Rose-Bengal medium containing streptomycin (to prevent

bacterial growth). This medium is acidic, therefore favoring the growth of fungi. The

number and characteristics of colonies growing on the plates are recorded after seven

days of incubation at 25 ± 2oC.

The fungal counts are ranked 1 to 5, based on the number and diversity of colonies, as

well as the overall coverage of the plate. No growth is assigned a value of one. A few

colonies of the same type, with coverage of less than 15% is assigned two. Plates

ranked three have numerous colonies of the same type, with relative coverage of 15%

to 40%. A four is given to plates with numerous colonies of the same type with 40% to

75% coverage. Plates with greater than 75% coverage by numerous different types of

colonies are ranked five (Table 2). These high-ranking plates contained high densities of

fast growing, diverse fungi.

The bacterial growth test is done on the porewater or water extract and the methanol

extract. The methanol extract is tested at 5% (v/v) and the aqueous extracts at 80%



(v/v). All tests are done in 96 well microplates with three replicates per sample. The

results are expressed as a percent of control growth based on the absorbance of a

tetrazoleum dye (forms a dark blue diformazan pigment when enzymatically reduced).

2.2.2 Plants

The plant growth tests were done with a terrestrial plant, a floating microphyte and a

unicellular green alga. The floating microphyte and alga are both common aquatic plants

found in most freshwater systems.

Germination and root elongation in a terrestrial plant are rapid and sensitive methods for

detecting the presence of potentially harmful substances present in the sediments. The

root is composed of a population of cells that are reproducing enlarging. The cells in the

root also provide a conduit for absorption and transport of water and nutrients. Although

simple, germination and root growth can provide insight on a number of important

processes affecting plant establishment and performance. The results from the seed

test can also be extrapolated to assessing potential effects on shoreline vegetation.

Hence, the species for this trophic level cover all major compartments in aquatic

systems (sediment, water column, and shoreline).

Root elongation tests are conducted following the procedure of Greene et al. (1989).

Lettuce seed was chosen and is a standard plant species widely used in research and

for assessment of contaminated sites. Lettuce was selected based on size, sensitivity

and speed of response (germination and root growth). Ten seeds are placed on a

Whatman No. 3 filter paper in a 10 cm plastic Petri dish moistened with 4 mL of the

porewater or aqueous extract or the methanol extract (1% methanol - the highest

methanol concentration that has no adverse effect on germination and root growth).

The dishes are capped with lids, sealed with Parafilm and incubated at 23 ± 2oC in

darkness for five days. Seeds with root tips emerging or with a split seed coat are

considered germinated. Lettuce root lengths (hypocotyl) are measured from the root tip



to the base of the shoot (epicotyl). The transition between the root and shoot of lettuce

seeds is clearly defined by a sharp bend. The results are expressed as a mean percent

of controls before ranking (Table 2).

Duckweed is a common, fast growing plant found in ponds worldwide. This plant is fed

upon by fish and birds hence a pathway for contaminants from sediments to higher

trophic levels. The test is done with 10 mL volumes; the methanol extracts are diluted to

10% for testing. The solutions are placed into a 40 mL clear plastic cups. One three

frond plant is placed into each cup and growth is scored after a seven day incubation at

4000 ± 400 lux; 25 ± 2oC. Growth is based on the increase in frond number relative to

controls and ranked one to five (one representing toxicity and five representing no

toxicity; Table 2).

The algal growth inhibition test is done with the unicellular green alga Selenastrum

capricornutum (formerly Monoraphidium, Environment Canada, 1992). This species is

common to many freshwater lakes and ponds throughout North America and Europe.

The tests are performed on the porewater and methanol extracts supplemented with

nutrients. The methanol extracts are diluted to 0.1% with deionized water prior to testing

(this is the highest concentration of methanol that has no effect on algal growth). The

tests are conducted in 96 well microplates. Growth is assessed after a three day

incubation period under continuous light (4000 ± 400 lux; 25 ± 2oC) by an increase in

cell numbers (particle counts or turbidity).

Any substance or condition that is stressful will inhibit or retard growth, resulting in a

lower final cell density. Increases in final cell densities over the controls may result from

the presence of nutrients or other essential trace substances in the samples. The results

are expressed as a percent of controls, then ranked from 1 to 5. A high value (4-5)

indicates optimal growth, values less than three indicate growth inhibition (Table 2).

2.2.3 Invertebrates



The invertebrate test species include the freshwater microcrustacean Daphnia magna,

and sediment organisms such as Hyalella azteca, Chironomus tentans, and Lumbriculus

sp. Other invertebrates can be included in the test battery as required. The water flea,

Daphnia magna is found throughout the world in freshwater systems. It is a standard

test species and it’s inclusion provides a means to benchmark effects.

Tests with the sediment dwelling organisms are done on the sediment, porewater or

aqueous extract and the methanol extract (tested at 5%, NOEC for methanol). The tests

are conducted with 20 mL sample volumes in 40 mL plastic containers. The tests on the

sediment are done with laboratory dilution water. Five organisms are placed into each

container, and mortality scored after four days incubation at 25oC with an 8 hour dark

and 16 hour light photoperiod (intensity at the water surface of 800 lux).

The tests with Daphnia magna are done in 40 mL cups under the same conditions. The

results are expressed as a percent of control survival and ranked from one to five (Table

2).

2.2.4 Vertebrate Testing

Fish are an integral part of the aquatic systems. They feed on plants and invertebrates

and are preyed upon by other fish, birds and mammals. The tests are done with larval

fathead minnows. This is a standard test species and allows benchmarking of effects.

The larval stage is also considered the most sensitive of fish development. The tests are

done under the same conditions as the invertebrate tests.

2.2.5 Other Testing Endpoints

Other tests can be included in the sediment health quotient as required. They may

include tests for mutagens, bioaccumulation, biomagnification, genetic diversity, bio-

diversity, community processes, hormonally active substances, or whatever is of interest

from an ecological or human health perspective. The method would be modified to

permit ranking from 1 to 5.



2.3 Quality Assurance

A number of quality assurance procedures are incorporated into the study design.

These procedures are in addition to those routinely followed as part of HydroQual’s

Quality System. Specific procedures include the use of positive controls (reference

toxicants), negative controls and replicates.

Reference toxicants are positive controls for assessing the health, condition and

sensitivities of the test populations. When test organisms are exposed to the reference

toxicant, the test result or response must fall within predefined limits. Values outside

these limits can indicate a change in the sensitivity of the organism or change in test

conditions.

Reference toxicants also provide a means to benchmark test results for comparison to

other studies and species. The reference toxicant test results are expressed as the

concentration required giving a 50% change in the response measured, relative to

controls (IC50, inhibitory concentration; EC50, effective concentration; LC50, lethal

concentration).

A negative control is a treatment that does not have an effect on the test organism (a

baseline or laboratory control). The response in the negative controls must not exceed a

predefined response level for a test to be considered valid. It should be noted that the

test data are all generally expressed as a percent of controls (normalized relative to the

test controls).

Duplicate tests are performed on every tenth sample, and for some tests multiple

replicates are set up on all samples (seedling emergence and fungal counts). The

accuracy and precision of the testing procedures are monitored by including extra

replicates within a test and repeating tests on selected sediments.



The last element in the quality system is the reporting of data. All data are independently

reviewed and verified by the Quality Assurance Unit.

3.0 Reporting Formats

The results are compiled in a formatted Excel workbook. The workbook contains six

worksheets with the following information:

1. physical and chemical characteristics

2. sediment test data

3. porewater test data

4. methanol extract test data

5. summary data sheet

6. sediment health ratings

The test data are entered into the appropriate worksheet and the results ranked (scale

of 1 to 5). The rankings are summarized in one sheet. The rank for each trophic level

and overall sediment health quotient are then summarized in the final sheet.

The first four worksheets are appended to the test report. The summary data sheet and

sediment health ratings are tables in the report (assigned numbers 3 and 4

respectively).

Copies of all worksheets are appended.

4.0 Interpretive Guide

The Sediment Health Quotient tests were designed to provide a measure of the

biological health and condition of sediment. The results must be interpreted in light of

other test results and sample information. It is more important to seek consistent, logical

trends in the data, rather than absolutes. Relative effects and patterns in the toxicity



tests must also be interpreted in light of sample location and any chemical data that may

be available. For example, sediment organisms are not only sensitive to major

contaminant types, but can be affected by sediment pH, texture, organic content, and

the particle size distribution.

Consistent results must be obtained amongst the tests within each trophic level. For

example, sediments with high numbers of bacteria and fungi should be non-toxic to

bacterial luminescence. The survival and growth results for sediment organisms should

be consistent with different species. Differences may arise due to factors affecting water

availability in the tests on the solid phase (sediment).

All test results are normalized on a scale from 1 to 5. Five is non-toxic or the response

was not different from controls. A lower value is assigned to sediments or extracts that

are toxic. The following approach is recommended for interpreting the results based on

the overall sediment health quotient.

• sediment health quotient of 4 to 5: healthy sediment with high ecological potential

⇒ no further assessment required

• sediment health quotient of 2 to 3: slight but significant impairment

⇒ review trophic level ratings

• one sensitive level

∗ review species ratings

• one species overly sensitive

• could be species specific effects; review

physical and chemical data in light of species

tolerance limits

• all species equally sensitive

• review physical and chemical data for potentially

adverse factor(s) common to all species

• all levels equally sensitive



∗ review physical and chemical data for potentially adverse

factor(s) common to all species

• sediment health quotient of 1: impaired with low ecological potential

⇒ review physical and chemical data for potentially adverse factor(s) common to

all species across trophic levels; sediments with these ratings will have either

high contaminant levels or physical and chemical properties outside of the

physiological tolerances of the test organisms

The patterns in the test data can provide insight on the nature of what is causing the

effect. An adverse response or low rating with one or two species may occur and can

often be explained in light of species specific tolerances. Effects detected amongst all

species within one trophic level but not in other levels can indicate the presence of very

specific types of contaminants (for example, phytotoxins that differentially affect plants).

Effects detected across trophic levels indicate the presence of combinations of

contaminants or unsuitable sediment conditions. These types of results require further

assessment based on available physical and chemical data and information about the

site.

The level of effort required to interpret the findings is determined by the overall sediment

quotient, followed by the pattern in trophic level ratings, and then by effects on individual

species. The final interpretation must include relevant information on the physical and

chemical properties of the sediment in light of an appropriate background condition, i.e.

a control site.



Sample Calculation: The following example is provided for illustrative purposes. The results are grouped by tests done on sediments, porewater and methanol extracts. Each result is ranked or scored from 1 to 5 based on the criteria in Table 2. The averages for each trophic level within each group (sediments, porewater and methanol extracts) are then calculated (right column). An overall trophic level rank is derived followed by a sediment health quotient or rating (bottom of table). Test Result Rank Average Sediments • Invertebrates: Lumbriculus 100% 5 = 5 Porewater • Microbes = (3+4+5+3) / 4 = 4 Bacterial counts 105 3 Bacterial luminescence 67 4 SOS-Chromotest 5 5 Fungal counts 3 3 • Plants = (5+5+4) / 3 = 5 Algal growth 84% 5

Root elongation: (lettuce) 89% 5 Duckweed growth 73% 4 • Invertebrates : Lumbriculus survival 100% 5 = (5+5+4) / 3 = 5 Chironomus survival 100% 5 Daphnia survival 75% 4 • Vertebrates Fathead Minnow survival 100% 5 = 5 Methanol Extracts • Microbes = (5+5) / 2 = 5 Bacterial luminescence 90 5 SOS-Chromotest 5 5 • Plants = (4+3+5+5) / 4 = 4 Algal growth 75% 4 Root elongation: (lettuce) 58% 3 • Invertebrates : Lumbriculus survival 100% 5 = (5+5+4) / 3 = 5 Chironomus survival 100% 5 Daphnia survival 75% 4 • Vertebrates Fathead Minnow survival 100% 5 = 5



Average ranks per trophic level for each group of tests are summarized below.

Trophic Sediment Porewater Methanol Trophic Level Rank Level (untreated) Extract (from above) Microbes 4 5 = (4+5) / 2 = 4.5 Plants 5 4 = (5+4) / 2 = 4.5 Invertebrates 5 5 5 = (5+5+5) / 3 = 5 Vertebrates 5 5 = (5+5) / 2 = 5 A sediment health quotient for the sample is the average of the Trophic Level scores

Sediment health quotient (SHQ) = (4.5+4.5+5+5) / 4 = 5



4.0 General References

American Society of Testing and Materials. 1996a. Standard Guide for Conducting a

Laboratory Soils Toxicity Test with Lumbricid Earthworm Eisenia foetida. E1676-95.

ASTM vol. 11.05

American Society of Testing and Materials. 1996b. Standard Practice for Conducting

Early Seedling Growth Tests. E1598-94 vol. 11.05.

Burton, G., 1992. Sediment Toxicity Assessment. Lewis Publishers, Chelsea, MI.

Environment Canada. 1999. Biological Test Method: Test for Measuring the Inhibition of

Growth Using the Freshwater Macrophyte, Lemna minor, EPS1/RM/37.

Environment Canada. 1992a. Biological Test Method: Growth Inhibition Test Using the

Freshwater Alga Selenastrum capricornutum, EPS1/RM/25.

Environment Canada. 1997. Biological Test Method: Test for Survival and Growth in

Sediment Using the Freshwater Amphipod Hyalella azteca, EPS1/RM/33.

Environment Canada. 1992a. Biological Test Method: Test for Survival and Growth in

Sediment Using the Larvae of Freshwater Midges, EPS1/RM/32.

Environment Canada. 1992b. Biological Test Method: Toxicity Test using Luminescent

Bacteria (Vibrio fischeri), EPS 1/RM/24.

Carter, M.R., 1993. Soil Sampling and Methods of Analysis. Lewis Publishers, Boca

Raton.

Gorsuch, J., W. Lower, M. Lewis, W. Wang, 1991. Plants for Toxicity Assessment:

Second Volume. American Society for Testing and Materials, Philadelphia.



Greene, J.C., C.L. Bartels, W.J. Warren-Hicks, B.R. Parkhurst, G.L. Linder, S.A.

Peterson, and W.E. Miller. 1989. Protocol for Short Term Toxicity Screening of

Hazardous Waste Sites, EPA/3-88-029.

Landis, W., Yu, M., 1999. Introduction to Environmental Toxicology. Lewis Publishers,

Boca Raton.

Legault, R., C. Blaise, D. Rokosh, and R. Chong-Kit, 1994. Comparative Assessment of

the SOS-Chromotest Kit and the Mutatox Test with the Salmonella Plate Incorporation

(Ames Test) and Fluctuation Tests for Screening Genotoxic Agents. Environmental

Toxicology and Water Quality: An International Journal, Vol. 9 (1994) 45-57. John

Wiley & Sons, Inc.

Newman, M., 1998. Fundamentals of Ecotoxicology. Ann Arbor Press, Chelsea, MI.

Organization for Economic Cooperation and Development. OECD Guidelines for the

Testing of Chemicals, 1993. Method 203. Terrestrial Plants Growth Test. Paris.



6.0 Tables



Table 1. Summary of Test Methods

Trophic Level Test Reference

Microbes bacterial luminescence Environment Canada, 1992b total heterotrophic bacteria Carter, 1993 fungal enumerations Carter, 1993

Plants algal growth Environment Canada, 1992a Duckweed growth HydroQual Laboratories Ltd. root elongation Greene et al., 1989

Invertebrates Sediment organisms HydroQual Laboratories Ltd. Daphnia survival HydroQual Laboratories Ltd.

Vertebrates Fathead minnow survival HydroQual Laboratories Ltd.

Human Health SOS-Chromotest HydroQual Laboratories Ltd.



Table 2. Ranking Criteria for Test Results

Test Endpointnon toxic marginal mild moderate toxic

5 4 3 2 1

sediment organism survival survival >80% 60-80% 40-59% 20-39% <19%

bacterial luminescence reduction in light production >80% 60-80% 40-59% 20-39% <19%total heterotrophic bacteria growth in highest dilution of extract >106 106 105 104 103

fungal enumerations coverage of plate >75% 40-75% 15-40% <15% nonenumber and diversity of colonies high same type same type same type

SOS-chromotest Induction –S9 <2 2-3.9 4-5.9 6-7.9 >8SOS chromotest Induction +S9 <1.5 1.5-2 2.1-2.5 2.6-3 >3algal growth cell number based on optical density >80% 60-80% 40-59% 20-39% <19%duckweed survival frond growth >80% 60-80% 40-59% 20-39% <19%root elongation root length (compared to controls) >80% 60-80% 40-59% 20-39% <19%

daphnid survival survival >80% 60-80% 40-59% 20-39% <19%

fathead minnow survival survival >80% 60-80% 40-59% 20-39% <19%

Rank



7.0 Figures



Figure 1. Flow Chart for Processing Test Sediments

Sediment

Solid Phase Aqueous Extract Methanol Extract

bacterial enumeration bacterial luminescencesediment organism survival fungal enumeration SOS chromotest

bacterial luminescence fathead minnow survivalduckweed survival algal growthdaphnid survival daphnid survival

SOS choromotestalgal growth

fathead minnow suvival

sediment organism survivalsediment organism survival



8.0 Appendices

Data Compilation Tables and Reporting Formats

The data collected on each batch of samples are reported in seven separate tables. The

test data are reported on Tables A to D (bench sheets) and the resultssummarized in

Tables 1 and 2.

Table

1. Sediment Health Ratings and Quotient

2. Test Results Summary

a. Physical and Chemical Characterization of Test Sediment

b. Sediment Test Data

c. Porewater Test Data

d. Methanol Extract Test Data

Documents

SEDIMENT HEALTH QUOTIENT: APPLICATION … HEALTH QUOTIENT: APPLICATION DOCUMENT FOR FRESHWATER SYSTEMS Prepared for: Golder Grundteknik kb Gata 12 …