The Biomonitoring Specialists

The Biomonitoring Specialists

EnviroScience, Inc.

The Biomonitoring Specialists

Who We Are…

• Established in 1989 as a Bioassay


• Rapidly grew into a comprehensive

ecological services firm

• 3 Major Divisions

• Bioassay

• Ecological

• Lake Management

• 55 Biologists / Environmental Scientists

based in Stow, Ohio serving clients


Bioassay Testing

Where did it come from?

• Stems from the

Clean Water Act

• NPDES permit


• State Agencies

• Individual NPDES

permit holders

What does it accomplish?

• Can address combinations of toxicants

• Address unknown substances

• Allows direct interaction with aquatic life

• Provides more comprehensive and

realistic picture of effects- a ‘real world’

measure of the impacts a discharge may

be having

Who can preform testing?

• State decides criteria for laboratory accreditation

• Must follow USEPA manuals

• Methods for Measuring the Acute Toxicity of Effluents to Freshwater and Marine

Organisms. 5 th Edition,USEPA,Office of Water, October 2002, EPA 821-R-02-013

• Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving

Waters to Freshwater Organisms. 4 th Edition,USEPA,Office of Water,October

2002,EPA 821-R-02-013

• Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving

Water to Marine and Estuarine Organisms, 3 rd Edition, EPA 821-R-02-014

• EnviroScience’s bioassay lab currently serves clients in

Ohio, Michigan, Pennsylvania, West Virginia, Indiana,

Illinois, New York, and Kentucky.

Common Toxicity Tests

• Chronic Toxicity

• Acute Toxicity

• Sediment Testing

• Elutriate Testing

• Product Testing

• Ceriodaphnia dubia

• Daphnia magna

• Pimphales promelas

• Hyalella azteca

• Chironomous dilutus

• Selenastrum


• Mysidopsis bahia

• Cyprinodon variegatus

• Menidia beryllina



Toxicity Testing Steps

• Obtain Samples

• Dilute Samples

• Add organisms

• Monitor daily/renew water/chemistry

• End test

• Data analysis

Obtaining Samples

• Grab vs Composite

• Containers

• Holding Time

• Temperature

• Chain of Custody

• Volume of Sample

• Determine dilution


• Dilution water

• UPS vs Lab water

• C.dubia vs P.promelas

• Acute vs Chronic


Test Initiation


• Chemical parameters

• DO

• pH

• Conductivity


• Hardness

• Alkalinity

• Temperature

• Ammonia (state


• Mortality

• Water Renewal

• Feeding

• Brood Size (water


Ending a Test

Acute Toxicity Tests

• Test Procedures

• 96 hours or less (species specific)

• Mortality is the measured endpoint

• Advantages

• Less expensive and time consuming

• Endpoint is easy to quantify

• Disadvantages

• Indicates only lethal concentrations

• Only the effects of fast acting chemicals are exhibited

Acute Acceptability Criteria

• Minimum control survival at least 90%

• Temperature maintained at 25 +/- 1°C

• Test organism age at initiation:

• 0-14 days for fish (State dependent)

• Less than 24 hours for daphnids

Acute Endpoints

• LC 50 (Lethal Concentration)

• Concentration of effluent that is lethal to 50

percent of the exposed organisms at a

specific time of observation

• Tu a (Acute Toxic Unit)

• Defined as 100÷LC 50

Chronic Toxicity Tests

• Test Procedures

• Typically 4-21 days

• Measures mortality, growth, and reproduction

• Advantages

• More sensitive than acute

• Assesses more parameters other than lethality

• Limitations

• More costly and time intensive than acute

Chronic Test Acceptability

• Minimum control survival 80%

• Minimum control dry weight (average):

• 0.25 mg for Fathead Minnows

• Minimum of 15 young (average) for control C. dubia

• Temperature maintained at 25 +/- 1°C

• Test organism age at initiation

Chronic Endpoints

• IC 25

• Inhibition Concentration- Concentration of effluent which has an

inhibitory effect on 25% of the test organisms for the monitored

effect, as compared to the control


• No Observable Effect Concentration- Highest concentration of

effluent tested which shows no statistically significant effect on

the organisms as compared to the control


• Lowest Observable Effect Concentration- Lowest concentration

of effluent tested which shows statistically significant effect on

the organisms as compared to the control

Chronic Endpoints

• % affected

• Typically used for ambient waters or one level


• ChV

• Chronic Value= square root of NOEC x LOEC

• Tu c

• Chronic Toxic Unit- Computed by

• 100÷ChV

• 100÷IC 25

• 100÷NOEC

Toxicity Identification/Reduction

(TIE/TRE) Evaluations

• Generally conducted in a phased iterative approach

• Does not necessarily involve extensive laboratory

evaluation and testing (the TIE part of the TRE)

• Variability of waste stream often poses significant

challenges, especially with low-level chronic toxicity

TRE Objectives

• Compare past biomonitoring and chemistry data to

look for patterns and indications as to the source of

toxicity in the final effluent

• Evaluate in-plant practices, processes, and chemical

usage in both process and treatment usage

• Identify the general class of toxicant(s) present and

then proceed to identify the specific toxicant(s) and


• Provide information, as needed, to support any

necessary treatment or process change

TRE Components

• Baseline Monitoring

• Chemical Optimization/ Operational


• Toxicity Identification Evaluation (TIE)

• Phase I Characterization

• Phase II Identification Analyses

• Phase III Confirmation Analyses

• Treatability Analyses

Operational Assessment

• Pre-laboratory phase of the TRE which

investigates possible toxicity sources

• Focuses on:

• Past analytical and bioassay data

• In-house chemical usage and treatment processes

• In-house practices and BMPs

• Combines in-plant knowledge of the facility and its

waste stream with an outside perspective to

identify possible sources of toxicity




Common Effluent Manipulation

Tests used in the TIE

• Baseline

• Aeration

• volatiles

• pH Adjustment

• Some metals

• Filtration

• Particle size

• Dissolved vs particulate


• C18 Solid Phase


• Nonpolar organics,


• Oxidant Reduction

• Chlorine, some metals

• Graduated pH

• Ammonia, some metals

• EDTA Chelation

• metals

Water Effect Ratio

• Used to derive site-specific limits for certain metals from

national and state aquatic life criteria that were developed

using laboratory toxicity data

• National aquatic life criteria for many metals may be

overprotective because most surface waters have greater

hardness and higher pH than the lab water that was used to

form the basis for the standard

The WER is derived by comparing toxicity differences

between site water and synthetic lab water for different lethal



Jessica Anderson

Martin Hilovsky

EnviroScience, Inc.


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