Course Title: Ecotoxicology

Course Code: ESRM 4101

Chapter 05

Environmental Risk Assessment

Dr. Tanmoy Roy Tusher

Associate Professor
Dept. of ESRM, MBSTU
What is Risk?
✓ Risk is defined as the probability of an adverse effect in an
organism, system, or population caused under specific
circumstances by exposure to a hazardous agent (stressor).

Probability of
Severity of adverse
ecological effect(s) ✕
Risk = occurrence of adverse
ecological effect(s)

✓ Risk does not exist unless two conditions are satisfied:

1) The stressor has the inherent ability to cause one or more
adverse effects, and
2) The stressor co-occurs with or contacts an ecological
component long enough and at sufficient intensity to
elicit the identified adverse effect.
Risk Assessment
✓ Risk assessment is the systematic scientific evaluation of
potential adverse effects resulting from the exposures to
hazardous agent(s).

✓ It is also defined as the process of assessing the risks

associated with each identified hazard, so that
appropriate control measures can be implemented based
on the likelihood of occurrence and the severity of
potential consequences.

✓ The risk assessment process includes four steps:

1) Hazard identification
2) Hazard characterization
3) Exposure assessment
4) Risk characterization
Steps Involved in Risk Assessment
Steps Involved in Risk Assessment
✓ Hazard identification:

• It is the first step in risk assessment, consisting of the

determination of substance(s) of concern and the
identification of the adverse effects the substance may
have inherently on target systems under certain
conditions of exposure.

• Examples of health hazards include either acute (e.g.,

skin and eye irritation, lethal effects) or chronic (e.g.,
carcinogenicity, effect on reproductive system, effect on
nervous system, effect on organs).

• Examples of ecological hazards include mortality

(acute) or reduced growth and reproduction (chronic) to
representative species.
Risk Assessment (Hazard Identification)
Effects of Jute Ecofilm (JE) Leachate
on Reproduction of C. elegans :
Days JCBB Leachate Control
(2%) (No Leachate)
Plate Plate Plate Plate Plate Plate a
1 2 3 1 2 3
Day 1 29 29 28 57 47 56

Day 2 39 65 97 80 72 68

Day 3 31 38 26 22 32 38

Day 4 61 55 15 13 22 14

Day 5 0 0 0 5 7 9

Day 6 0 0 0 2 0 0 No acute toxicity

on the reproduction of C. elegans
Total 160 187 166 179 180 185 was observed
(small letter ‘a’ indicates p >0.05)
Steps Involved in Risk Assessment
✓ Hazard characterization:

• It is the second step in the process of risk assessment,

consisting of the qualitative and, wherever possible,
quantitative description of the nature of the hazard
associated with the substance(s).

• Examples include
mechanisms of
action involved,
dose-response and
dose-effect
relationships.
 Risk Assessment (Hazard Characterization)
Effects of Jute Ecofilm (JE) Leachate on Radish Seed Germination
and Seedling Growth:
JE-High JE-Low Control
(2% Leachate) (0.2% Leachate) (No Leachate)

JE-High JE-Low
(2% Leachate) (0.2% Leachate)

120
a
Germination rate (%)

100 ab b
80
60
40
20
0
JE-High JE-Low Control
Seedling morphology
Steps Involved in Risk Assessment
✓ Exposure assessment:
• It is the third step in risk assessment, consisting of a
qualitative and quantitative determination of the
concentration of a substance (including its derivatives) in a
given environment as well as the estimation of its rate of
intake in target organisms.

• For example, finding the concentration of aflatoxin in peanut

butter and determining the dose an ‘average’ person would
receive.

• Exposure assessment may involve considering the duration,

frequency, and concentration of exposure, including the
bioavailability of the substance.

• Estimates of exposure are obtained by determining the

sources, pathways, and rates of the movement of the
substance in the environment.
Steps Involved in Risk Assessment
✓ Risk characterization:

• Risk characterization is the integration of information on

hazard, exposure, and dose-response to provide an
estimate of the likelihood that any of the identified
adverse effect(s) will occur.

• It consists of a qualitative estimate in terms of low,

moderate, high, and/or quantitative or numerical
estimation of the probability of occurrence and the
severity of an adverse effect (known or potential).

• It helps decision-makers to choose appropriate risk

management strategies by providing information on the
probability of occurrence of adverse effect, its severity,
and the exposed population.
Risk Assessment Matrix
Types of Risk Assessment

Risk
assessment

Human health Ecological risk

risk assessment assessment

Health related risks Ecological risks at sites

(Carcinogenic and non-carcinogenic)
Human Health Risk Assessment
✓ Human health risk assessment is a process intended to
estimate the risk to a population from exposure to a
substance of concern.

✓ The process considers:

• the type and composition of the substance,
• it's potential to harm,
• the way in which people may be exposed (such as through
direct exposure, inhalation of air or food and water
consumption), and
• how long people are exposed and how much they might be
exposed to.

✓ The human health risk assessment process also includes

hazard identification, hazard characterization (dose-
response relationship), exposure assessment, and risk
characterization.
Human Health Risk Assessment
• Hazard identification:

WOE = Weight-of-the-evidence
 Human Health Risk Assessment
• Dose-response relationship:

BMD = Benchmark dose

Human Health Risk Assessment
• Dose-response relationship:

For arsenic = 15 (mg/kg-day)-1

Human Health Risk Assessment
• Exposure assessment:
Human Health Risk Assessment
• Exposure assessment:

Chronic daily intake:

Where, C is the concentration of Cd in water (e.g., mg/L)

IR is the ingestion rate (L/day)
EF is the exposure frequency (days/year)
ED is the exposure duration (years)
AT is the average time (For non-cancer: AT = ED × 365 days;
For cancer risk: AT = 70 years × 365 days)
BW is the body weight (kg; different for adults and children)
Human Health Risk Assessment
• Risk characterization:

Where, HQ is the hazard quotient (no unit)

CDI is the chronic daily intake (mg/kg-day)
RfD is the oral reference dose (mg/kg-day)

➢ For multiple pollutants, the hazard index (HI) is calculated by summing the
Hazard Quotient (HQ) values of each pollutant.

➢ HQ or HI <1 indicates no non-carcinogenic risks

➢ HQ or HI >1 indicates potential non-carcinogenic risks
 Human Health Risk Assessment
• Risk characterization:

CR = CDI ✕ SFo

Where, CR is the carcinogenic risk (no unit)

CDI is the chronic daily intake (mg/kg-day)
SFo is the oral slope factor (mg/kg-day)-1

➢ CR value >10-4 indicates cancer risks to the both adult and children
 Human Health
Risk Assessment
Download PDF
Human Health Risk Assessment
Values of the variables for the estimation of human health risk exposure to water
Human Health Risk Assessment
Plots of non-carcinogenic risk for adults

Plots of non-carcinogenic risk for children

Procedure of human health risk assessment

based on the Monte Carlo simulation
Ecological Risk Assessment
✓ Ecological risk assessment (ERA) is a process for evaluating how
likely it is that the environment (or ecosystem) might be impacted
as a result of exposure to one or more environmental stressors
(e.g., chemicals, land-use change, and invasive species).

✓ ERA can be used to identify vulnerable and valued resources as

well as link human activities to their potential effects.

✓ The results of ERA provide a basis for comparing different

management options, enabling decision-makers and the public to
make better informed decisions about the management of
ecological resources.

✓ All ERA plans must:

• determine the scope and boundaries of the assessment,
• select ecological entities for the focus of the assessment, and
• ensure that the product of the assessment will support
environmental decision making.
Steps in Ecological Risk Assessment Process

Screening-level ecological risk

assessment

e.g., Contaminated
land by chemicals

Baseline ecological risk assessment

➢ Data quality objective (DQO) process:
Clarify objectives, define appropriate
types of data to collect; Specify tolerable
levels of potential errors
Steps in Ecological Risk Assessment Process
✓ Step 1: Screening-level problem formulation and toxicity evaluation

• Problem formulation- information is gathered about the site

• Site visit- investigators visit the site
• Toxicity evaluation- review of scientific literature to determine at
what levels the chemicals present will have the adverse effects

Site visit

Literature
Chemicals review for
toxicity
evaluation

✓ Step 2: Screening-level exposure estimate and risk calculation

• Estimate exposures- calculating how much plants and animals

are exposed to chemicals at the site
• Risk calculations- calculation of hazard quotients (HQ)- the
comparison of the levels of chemical contamination at the site to
levels that are known to cause harm
Steps in Ecological Risk Assessment Process
✓ Step 3: Problem formulation
• Refining contaminants of potential ecological concern- deciding
whether or not chemicals found at the site should be considered
further in determining potential ecological risk
• Contaminant fate and transport- what happens to the chemicals
(biologically, chemically, and physically) and how they move among
plants and animals
• Mechanisms of toxicity- what are the harmful effects on plants and
animals
• Ecosystems potentially at risk- habitats; plants and animals present;
sensitivity by plants and animals to chemicals; water bodies present
• Exposure pathways- how the chemicals pass through the food web
from the source of contamination
• Assessment endpoints- the characteristic(s) of a plant or animal that
can be measured in terms of harmful effects
• Conceptual site model- collection of exposure pathways
• Risk questions- using various lines of evidence, answer the question
"does chemical X cause harmful effects in plant or animal Y?"
Steps in Ecological Risk Assessment Process

Genotoxicity

Neurotoxicity Carcinogenicity

Cytotoxicity
(apoptoosis/ Toxicity Ocular
toxicity
necrosis) endpoints

Reproductive
&
Immunotoxicity
developmental
toxicity

Oxidative Dermal
stress toxicity
Steps in Ecological Risk Assessment Process
✓ Step 4: Study design and data quality objective process

• Work plan and sampling plan- a written description of how the

investigation will proceed at the site
• Measurement endpoints- what is measured (i.e., reproduction,
mortality, growth, etc.) in relationship to the assessment
endpoints
• Study design- what type of testing will be done to evaluate the
potential ecological risk at the site
• Data quality objectives and statistical considerations- a series of
planning steps to insure the quality, type, and quantity of data will
be adequate and defendable

✓ Step 5: Verification of field sampling plan

• Determining if the site conditions still allow the investigation to
proceed as described in Step 4 or if new conditions require
modifications of the work plan
Steps in Ecological Risk Assessment Process
✓ Step 6: Site investigation and data analysis

• Site investigation- the work as described in the work plan is

carried out
• Data analysis- the information gathered is analyzed according to
the work plan and statistical design

✓ Step 7: Risk characterization

• Risk estimation and characterization- combining the results of

the studies performed to produce an estimate of the ecological
risk and describe that risk in terms of extent, future potential for
risk, how long might contamination remain, and what are the
prospects of natural recovery if no action is taken

✓ Step 8: Risk management

• Decisions are made concerning what future actions, if any, are to

be undertaken
Toxicity Testing Methods
✓ Aquatic toxicity: Aquatic toxicity in general refers to the effects of a
chemical on organisms living in water and is determined with
organisms representing the three trophic levels:
• Algae or plants, representing "primary producers”,
• Invertebrates (e.g., crustaceans such as Daphnia spp., Artemia
spp.), representing "primary consumers/secondary producers”,
and
• Vertebrates (usually fish), representing "secondary consumers”.
Toxicity Testing Methods
✓ Terrestrial toxicity: Terrestrial toxicity refers to the effects of a
chemical on land-dwelling organisms and is determined using
species that represent various ecological roles and trophic levels:
• Plants, representing primary producers, which are crucial for
photosynthesis and ecosystem productivity,
• Invertebrates (e.g., earthworms, springtails), representing
primary/secondary consumers and important indicators of soil
health, and
• Vertebrates (e.g., rodents, birds), representing higher-level
consumers, often used in evaluating long-term or systemic
effects.

Exposure to
cypermethrin
(insecticide)
Toxicity Testing Methods some of these

Plant and Animal Species Used in Standard Toxicity Tests

Some Examples of Ecotoxicity Tests
✓ Earthworm, acute toxicity test: The purpose of this test is to
determine the effects of a substance on the mortality of earthworm.
• The initial screening test (filter paper contact test) involves
exposing earthworms to test substances on moist filter paper in
order to identify potentially toxic chemicals to earthworms in soil.
• Five or more treatment levels in a geometric series and, at least,
ten replicates (one worm per vial) for each treatment should be
used.
• Tests are done in the dark and for a period of 48 hours (or for a
longer period in case of chronic toxicity).
• The artificial soil test gives toxicity data more representative of
natural exposure of earthworms to chemicals.
• It involves keeping earthworms in samples of a precisely defined
artificial soil.
Some Examples of Ecotoxicity Tests
✓ Earthworm, acute toxicity test:
Some Examples of Ecotoxicity Tests
✓ Growth inhibition test in Alga and Cyanobacteria: The purpose of this
test is to determine the effects of a substance on the growth of
freshwater microalgae and/or cyanobacteria.
• Exponentially growing test organisms are exposed to the test
substance in batch cultures over a period of normally 72 hours.
• The system response is the reduction of growth in a series of algal
cultures exposed to, at least, five concentrations of a test
substance. Three replicates at each test concentration should be
used.
• The response is evaluated as a function of the exposure
concentration in comparison with the average growth of control
cultures.
• Growth and growth inhibition are quantified from measurements of
the algal biomass as a function of time.
Some Examples of Ecotoxicity Tests
✓ Growth inhibition test in Alga and Cyanobacteria:

Microalgae
Raphidocelis
subcapitata
Some Examples of Ecotoxicity Tests
✓ Terrestrial plants, seed germination test:
• Seeds are exposed to the test substance under controlled
conditions (can be done in Petri dish), and their germination
response is evaluated over a typical period of 5 to 14 days,
depending on the species.
• Germination is generally defined by the emergence of a 2 mm
radicle, which is observed and recorded as a primary indicator of
successful germination.
• Endpoints measured include germination percentage, mean
germination time (MGT), root/shoot length, and visual signs of
phytotoxicity (e.g., discoloration, malformation).
• The assay can be performed either as a dose-response test using
multiple concentrations or as a limit test at a single concentration,
depending on study objectives.
Some Examples of Ecotoxicity Tests
✓ Terrestrial plants, seed germination test:
Some Examples of Ecotoxicity Tests
✓ Terrestrial plants, growth test:
• Seeds are placed in contact with soil treated with the test
substance and evaluated for effects following usually 14 to 21 days
after 50% emergence of the seedlings in the control group.
• Endpoints measured are visual assessment of seedling
emergence, biomass measurements, shoot height, and the visible
detrimental effects on different parts of the plant.
• The test can be conducted in order to determine the dose-
response curve, or at a single concentration/rate as a limit test,
according to the aim of the study.
• An appropriate statistical analysis is used to obtain effective
concentration ECx or effective application rate ERx for the most
sensitive parameter(s) of interest.
• Also, the no observed effect concentration (NOEC) and lowest
observed effect concentration (LOEC) can be calculated in this
test.
Some Examples of Ecotoxicity Tests
✓ Terrestrial plants, growth test:

Experiment
50% seedling ends for fast
emergence in growing plants
Sowing control? Observation Observation

Application Observation

0 7 14 21 Days
Assessment of Whole Effluent Toxicity Short note

✓ Whole Effluent Toxicity (WET) testing evaluates the combined toxic

effect of all chemical, physical, and biological components present
in a wastewater (effluent) sample on living organisms.

✓ It is used to assess whether the effluent as a whole may cause acute

or chronic toxicity to organisms, especially to aquatic life.

✓ Test organisms typically include representatives of three trophic

levels: Fish, Invertebrates, and Algae.

✓ The endpoints include survival, growth, reproduction, biomass or

cell density (for algae).

✓ WET tests are conducted under standardized laboratory conditions,

and results are expressed in terms of: NOEC, LOEC, IC₅₀, LC₅₀, EC₅₀,
and Toxic Units (TU): TU = 100 / EC₅₀.

✓ WET is widely used in regulatory compliance, effluent permitting,

and environmental monitoring to protect receiving waters from
harmful discharges.
Assessment of Whole Effluent Toxicity
Thanks to All