Environmental Stressors: Effects

on Aquatic Ecosystems
S. Marshall Adams
Environmental Sciences Division
Oak Ridge National Laboratory

Outline
Concepts of environmental stressors and stress

Assessing effects of stress using multiple response measures

Examples of Stress effects in aquatic systems

- stream polluted by multiple contaminants
- coastal estuary experiencing eutrophication
- ponds on Air Force Base affected by military activities

Relevance and Application

Definitions of Environmental Stress and Stressor

Stressor - Any environmental condition, situation, or factor that causes

a biological system to mobilize its resources and increase its energy
expenditure

Stress - A state produced by an environmental stressor which extends

the adaptive responses of an organism beyond its normal range, or
which disturbs normal functioning to such an extent that the chances
of survival are significantly reduced

1
 Healthy Ph
ys
iol
Increasing stress og
ica
lC
on
dit
ion
Stressed

Damaged
but Curable

Physiological Tolerance Limit

Irrepairable
Disease, Reproductive Impairment, Death
Incurable

Death
DISABILITY

Disease
Severely Damaged Zone 3
Moderately
Damaged Zone 2
Repair
Stressed Zone 1
Healthy
OK

Homeostasis Compensation Non-Compensation

IMPAIRMENT

Habitat Eutrophication
Availability Temperature
Hydrodynamic Regimes
Regimes Food
Varying Availability
Physiochemical
Factors Contaminants

S. Marshall Adams
Environmental Sciences Division
Oak Ridge National Laboratory
Integrated Response
A N INTEGRA TED FRA MEWORK FOR A SSESSING COA STA L
RESOURCE MA NA GEMENT ISSUES: EX A MINING CA USA L
RELA TIONSHIPS
GROWTHIN COMPLEX ENVIRONMENTS
REPRODUCTIVE
SUCCESS

POPULATION RESPONSE

2
 Environmental
Stressors
Direct Indirect

Biomolecular
Quality and Behavioral
Biochemical
quantity of food activities
Bioenergetic Behavioral
Physiological Organism
Pathological Metabolic (growth, repro)

Population

Community

Chemical Stressors Biochemical-Physiological

- contaminant exposure Responses
- water quality - organ dysfunction
- immune function
- genetic damage

Physicochemical
stressors Organ-Individual Responses
- hydrologic regimes - condition
- dissolved oxygen - growth
- behavior

Habitat
modification
Reproductive-population
- quantity
Responses
-quality
- steroid hormones
- egg size & quality
- abundance
- size distribution

Establishing Causality Between

Stressors and Biological Effects

Stressors

nutrients Land development-

Pollutants
sedimentation

Causal
Mechanisms?

Effects

3
 High sensitivity
& specificity

l a
logic
R
UL A
L EC

uno
MO

Imm
AL
IC L
OG ICA
Sublethal IO
L
OL
OG
YS TH
PH PA
TIC
Low ecological GE
ER
relevance OE
N
RE

BI
PR
O
DU

Low sensitivity
C

ON & specificity
TI

TI
VE

A
P UL
PO
TY
NI
M MU
CO

Ecologically
High ecological
Bioindicators
relevance relevant

4
 Downstream Gradient in Contaminants
Contaminant Concentration (PPM)

3.5
3
2.5
2
1.5
1
0.5
0
Site 1 PCBs-Sediment
Site 2
Site 3 PCBs-Fish
Site 4 Mercury-Water
REF
Stream Flow

5
 Spatial Trends in Benthic Macroinvertebrates

T o ta l T a x o n o m ic R ic h n e s s
40
Number of taxa/sample

30 Decreasing pollution

ce
en
fer
20

Re
10

0
EFK 24 EFK 23 EFK 14 B F K 7 .6

S ite

Spatial Pattern in Fish Divsrsity

Number of species

1990
20
Number of Species

15
nc e
Refere

10

0
EFK 24 EFK 23 EFK 14 BFK

6
 NUCLEOLUS

NUCLEUS

MITROCHONDRION

HEALTHY GILL CELL

MERCURY-EXPOSED GILL

7
 Spatial changes in Steroid Hormones

Sewage Treatment
Industrial Plant
Discharge

40
Industrial Discharge
Discharge
Sewage Outfall
Industrial
Industrial Discharge
Blood Steroid Hormones (ng/ml)

30
Estradiol
Testosterone

20

10
km =

0
1 2 3 4 Reference

stream flow

Genetic Diversity of Sunfish Population

35

30
Decreasing Contamination
Resistant Genotype
(% of populatiion)

25

20

15

10
References

0
3

F1

F2
K2

K1

K1

K0

RE

RE
EF

EF

EF

EF

Integrated Bioindicator Site Responses

in East Fork Poplar Creek

References

Contaminated
l Variate

C1
Third
Canonica

C2

C3
C4
te
ria

Firs
n ic n d

t Ca
Va

non
no eco

ical
al

Var
S

iate
Ca

8
 Isabel 9-18-03

Floyd 9-16-99

Sewage plume

9
 ts
utan
poll
ie nts-
Nutr

North
Carolina

Pam Pamlico Sound

lico
Rive
r

Cape
Hatteras

B
X
er
Riv
A u se
Ne
Atlantic Ocean

Affected site
X
Cape
Lookout

North
Reference site
sitena

Neuse River

0- 0.2 mg/L 0.2-2 2-4

4-6 mg/L 6-8

Lower Neuse/western Pamlico

10
 Hypoxic zone

Hypothesis: Fish sampled from areas in Pamlico Sound

with low dissolved oxygen are sublethally stressed
compared to fish from non-affected reference areas

Southern Flounder (ambush predator, carnivore)

Spot (omnivore, benthic forager)

11
A B

C D

Hematology-Organ Dysfunction Indicators

60 6

Spot
50 * 5
Leukocrit, Bun, Protein

40 4
Hematocrit

30 3
*
20 2

10 * 1

0 0
Reference Pamlico Sound
60 7
Leukocrit, Bun, Protein

50 Flounder 6
5
Hematocrit

40
4
30
3
20
2
10
*
1
0 0
Reference Pamlico Sound
*
Hematocrit BUN
leukocrit Serum protein

Condition indicators
700 2

600 Spot
*
Serum triglycerides (mg/dl)

1.5
500
Somatic indices

400
1
300
*
200
0.5
100
*
0 0
Reference Pamlico Sound

800 3
Serum triglycerides (mg/dl)

Flounder
Somatic indices

2.5
600
2

400 1.5

1
200
0.5

0 0
Reference Pamlico Sound
Serum lipid Spleno-somatic index
Liver-somatic index Condition factor

12
 Reproductive Indicators
1200
Plasma steroid hormones (pg/ml) 70
1000 Spot * 60

800 50

Percent
40
600
30
400
*
20
200 * * 10

0 0
Reference Pamlico Sound

1000
Plasma steroid hormones (pg/ml)

* 80
800
Flounder
60
600

Percent
40
400

200 20
*
0 0
Reference *
Pamlico Sound
testosterone (F) % ovaries developed
estradiol (F) % female estradiol non-detects

Integrated Health Responses

(discriminant analysis)
Indicators used in analysis +3
Spot
Immunological - 7
Reference
Hematology - 2 +2
Physiological - 4 Spot
Reproductive - 6 Pamlico
8.5
Second canonical variate

Condition - 5
+1
Total= 24

-1
Flounder
Flounder
-2 Reference
6.0 Pamlico

-3

-3 -2 -1 0 +1 +2 +3
First canonical variate

Gopher frog Gopher tortoise

Gopher frog range

Gopher tortoise range

13
 Sampling design & site selection
gopher frogs-Eglin AFB

High disturbance
Medium-low disturbance
Reference
B
C74 range

C
A
e
ng
ra
70 C-52 range
B-
Ref
Ref

Assessment of Military Impact

Sample site Explosive Ordinance/ Cratering/ Soil Burning/ Composite Miitary

residuals unit area unit area disturb. scorching score impact

C52N-1 3 3 3 3 2 14 High
C52N-2 3 3 3 3 1 13 High
C52N-3 3 3 3 2 2 12 High
C52C-1 1 1 2 2 1 7 Moderate
C52C-2 1 2 2 2 1 8 Moderate
C52C-3 1 1 2 2 2 8 Moderate
Reference-1 0 0 0 1 2 3 Low
Reference-2 0 0 0 1 1 2 Low
Reference-3 0 0 0 0 2 2 Low

Criteria for assessing military disturbance

3= moderate to high
2= low to moderate
1= none to low

Sampling design-Gopher frogs

¾9 “Treatment” areas
¾3 High disturbance (C52N, C74, C52CE) Tr 2
X
¾3 low disturbance (C52C, C52A, B70)
¾3 Reference X
X
¾2-6 Ponds per treatment
Transect 1 X X
C52A range X
low disturbance Pond X
Plots
C52N range
High disturbance Edge
Tr 3 X
Terrestrial
¾Habitat X
¾Each Pond
¾Morphology, water quality,
general substrate, vegetation, and
Reference military disturbance
¾Edge/TerrestrialBreeding Site
¾2-3 transects per pond
¾random plots within each
C52A range
community type
¾% cover plant layers; disturbance
metrics, no of burrows, etc.
¾Remote sensing 300 m home range

14
 Sampling devices deployed at Eglin
• Total sites with drift fences and traps - 40
• Total number of 30m drift fences with cages- 162
• Total feet of drift fencing- 5700
• Total number of traps at 40 sites- 940
• Number of traps at each site standardized
based on pond surface area

High disturbance site- C52 bombing range

300m home range

radii from pond

Low disturbance site-C52A range Undisturbed site-reference ponds

High activity sites-Eglin AFB

Drift fence with cages

fences + traps

B70 range fences + traps

C52N range

C74 Range

15
 Low activity and reference sites-Eglin AFB

C52A range
C52C range

Reference Reference

Construction and installation of sampling devices

Seep on C52N range

DriftDrift
fence with traps
fence
Drift fence
Trap
Trap

Wet/dry pond
Wet/dry pond on C52N range

Traps

New bomb crater New bomb crater

Oldbomb
Old bombcrater
crater

Old bomb crater

- DNA damage
Biomolecular
- glutathione
- malondialdehyde
- oxidative DNA damage
Physiological
Early warning, sensitivity, specificity

- transaminase enzy.
- creatinine
- serum electrolytes
- LDH Immunological
- bilirubin
- glucose - PHA test (cell mediated)
- BUN - bacterial killing (innate)
- AGAP - Corticosterone
- 24 total analysis
Reproductive
- Clutch size
- Egg size
- Steroid hormones
Population
- Hatching success - Relative abundance
- Vitellogenin - Age class distribution
- Size-freq distribution
- Sex ratio

Ecological Relevance

16
 Linking indicators to health and fitness

Hypothesis: Physiological indicators (bioindicators) can be used to assess

or predict the health and fitness of TES populations at military sites

• Canonical correlation analysis in conjunction with canonical variate

analysis used to predict and assess reproduction and population fitness
based on sets of measured bioindicator variables

Reproductive variables
Bioindicator variables NUCLEUS
Steroid hormones
correlate & Fecundity
Egg-oocyte quality
predict
Molecular Oocyte atresia
Biochemical Vitellogenin
Physiological
Immunological correlate and predict
Histopathological
Condition indices
correlate & Relative abundance
predict Age class distribution
Sex ratio
Size at age
Population variables

Organism-level responses provide a pivotal point through

which mechanistic understanding and the ecological
consequences of stress can be linked

S u b c e llu la r / T is s u e / P o p u la tio n C o m m u n ity

O rg a n is m
C e llu la r O rg an

RESPONSE
S E N S IT IV IT Y E C O L O G IC A L
RELEVANCE

B io m a r k e r s B io in d ic a t o r s

F o c u s o f F ie ld S tu d ie s
M in s /h o u rs D ays W e e k s /M o n th s Y e a rs B io lo g ic a l
g e n e ra tio n s
R E S P O N S E T IM E

Recovery Assessment
Ecological Risk Assessment
INJURY
Ecologically
EXPOSURE Community significant

Population
Sensitive fitness

Early Warning
Reproductive
Population
Histopath structure
Time scale
Physiological
Years
Months Immunological
Days/weeks Biochemical
Hours/days
Biomolecular
Minutes
SUB-ORGANISMAL INDIVIDUAL POPULATION--
COMMUNITY

Environmental
Stressors

17
 Biomarkers Bioindicators

Genetic
Mechanistic understanding

Biochemical

Histopathological
Immunological
Physiological

Populations
Reproductive

Communities

Increasing integration
Ecological significance

PREDATORS
Increasing Mechanistic Understanding
Increasing Ecosystem Integration

FISH PREY BASE

INVERTS

PRIMARY PRODUCTION

Physicochemical
Nutrients
Factors

Summary
Multiple indicators of stressare useful for:

• Early warning indicators of environmental problems

• Providing insights into causal mechanisms

linking stressors and biological effects

• Assessing cumulative and/or synergistic effects

of stress on organisms

• Identifying sources & causes of environmental

damage related to specific stressors

• Evaluating the effectiveness of environmental

cleanup (remedial) actions

18
 Establishing Causality Between
Stressors and Biological Effects

Stressors

Land development-
Pollutants nutrients
sedimentation

Multiple stress
responses

Establish causality

Effects

Environmental Stressors

Blood
X-Ray profile
Genetic Blood
damage profile Urine
Immune Histopath Profile Immune Endocrine
Enzymes

Environmental Medical
Diagnosis Diagnosis

19