Systematic Review Supplemental File: Data Quality Evaluation of Environmental Toxicology Studies for Asbestos


PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
xvEPA
March 2020
United States	Office of Chemical Safety and
Environmental Protection Agency	Pollution Prevention
Draft Risk Evaluation for Asbestos
Systematic Review Supplemental File:
Data Quality Evaluation of Ecological Hazard Studies
March 2020

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
HERO
ID
621276
3080106
3093600
3093856
3584231
3585046
Table of Contents
Data Type
Reference
Chronic (>21 days); Aquatic;
Plants
Chronic (>21 days); Aquatic;
Plants
Trivedi, A. K.,Ahmad, I.,Musthapa, M. S.,Ansari, F. A.. 2007. Environmental
contamination of chrysotile asbestos and its toxic effects on antioxidative system
of Lemna gibba. Archives of Environmental Contamination and Toxicology
52:355-362
Trivedi, A. K.,Ahmad, I.,Musthapa, M. S.,Ansari, F. A.,Rahman, Q.. 2004. En-
vironmental contamination of chrysotile asbestos and its toxic effects on growth
and physiological and biochemical parameters of Lemna gibba. Archives of
Environmental Contamination and Toxicology 47:281-289
Chronic (>21 days); Aquatic; In-
vertebrates
Chronic (>21 days); Aquatic; In-
vertebrates
Chronic (>21 days); Aquatic; Fish
Chronic (>21 days); Aquatic; Fish
Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. UPTAKE OF
CHRYSOTILE ASBESTOS FIBERS ALTERS GROWTH AND REPRODUC-
TION OF ASIATIC CLAMS. Canadian Journal of Fisheries and Aquatic Sci-
ences 43:43-52
Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. SEASONAL BEHAVIORAL
AND GROWTH CHANGES OF JUVENILE CORBICULA-FLUMINEA EX-
POSED TO CHRYSOTILE ASBESTOS. Water Research 20:1243-1250
Belanger, S. E.,Schurr, K., Allen, D. J.,Gohara, A. F.. 1986. Effects of chrysotile
asbestos on coho salmon and green sunfish: evidence of behavioral and patho-
logical stress. Environmental Research 39:74-85
Belanger, S. E.,Cherry, D. S.,Cairns, J.. 1990. FUNCTIONAL AND PATHO-
LOGICAL IMPAIRMENT OF JAPANESE MEDAKA (ORYZIAS-LATIPES)
BY LONG-TERM ASBESTOS EXPOSURE. Aquatic Toxicology 17:133-154
12
15
19

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
Study Citation: Trivedi, A. K.,Ahmad, I.,Musthapa, M. S.,Ansari, F. A.. 2007. Environmental contamination of chrysotile asbestos and its toxic effects
on antioxidative system of Lemna gibba. Archives of Environmental Contamination and Toxicology 52:355-362
Data Type: Chronic (>21 days); Aquatic; Plants
Hero ID: 621276
Domain
Metric
Rating1
MWF* Score
Comments^
Domain 1: Test Substance
Metric 1:
Metric 2:
Test Substance Identity
Test Substance Source
Metric 3: Test Substance Purity
High
Low
Low
X 2 2 The test substance was identified definitively.
X 1 3 Although the test material source not defined there
is no indication this impacted the results of the
study.
X 1 3 Although the purity was not reported, there is no
indication that this had an effect on the results.
Domain 2: Test Design
Metric 4: Negative Controls
Medium
Metric 5: Negative Control Response
Metric 6: Randomized Allocation
Medium
Low
x 2
x 1
x 1
Although aquatic plants used as controls were cul-
tured in nutrient mediumwithout chrysotile fiber,
they were from the third generation of plants ob-
tained from a natural habitat in an aquatic body
that was contaminated with asbestos. There are un-
certainties (e.g., due to epigenetics) around how the
initial exposure to asbestos at the parent generation
would affect the plants from the third generation.
There were minor uncertainties or limitations re-
garding the biological responses of the negative con-
trol group(s).
Researchers did not report how organisms were al-
located to study group.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Unacceptable x 2
tion
The test organisms were cultured in a media contain-
ing asbestos, while also being exposed at a rate that
is reported in terms of exposure per frond. The au-
thors did not provide sufficient detail about the test
organisms or exposure regime (ex. how many fronds
per plant? Does excess test media applied to fronds
enter the test suspension?) to allow the reviewer to
confirm the scientific validity of this study.
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page
Study Citation: Trivedi, A. K.,Ahmad, I.,Musthapa, M. S.,Ansari, F. A.. 2007. Environmental contamination of chrysotile asbestos and its toxic effects
on antioxidative system of Lemna gibba. Archives of Environmental Contamination and Toxicology 52:355-362
Data Type: Chronic (>21 days); Aquatic; Plants
Hero ID: 621276
Domain Metric Rating^ MWF* Score Comments^
Metric 8: Consistency of Exposure Administration
Unacceptable x 1
Metric 9: Measurement of Test Substance Concentra- N/A
tion
Metric 10: Exposure Duration and Frequency
High
Metric 11: Number of Exposure Groups/Spacing of Ex- Low
posure Levels
Metric 12: Testing at or Below Solubility Limit N/A
x 1
x 1
N/A
N/A
There were serious flaws in how the aquatic plants
were exposed to asbestos. The authors described
the test media as containing asbestos, while describ-
ing the exposure of asbestos to the fronds. This led
the reviewer to question the source of the effects ob-
served in this study and whether it was due to as-
bestos in the media or the asbestos applied to the
frond. In addition, the lack of detail about the pro-
cedure used to apply asbestos to the fronds meant
that the exposure cannot be adequately understood
from the information provided in this study.
Exposure concentrations to fronds in the plants were
not measured due to the insoluble nature of asbestos
fibers.
Experiments were carried out for a test duration of
28 days. EPA determined this to be acceptable.
There were no Justifications provided for the selec-
tion of the test concentrations.
Test media was left in suspension because asbestos
is an insoluble particle. .
Domain 4: Test Organism
Metric 13: Test Organism Characteristics Medium x 2
Metric 14: Acclimitization and Pretreatment Conditions Unacceptable x 1
Metric 15: Number of Organisms and Replicates per High
Group
Metric 16: Adequacy of Test Conditions
x 1
Unacceptable x 1
4 There are minor reservations or uncertainties about
the source of test organisms.
4 Plants were cultured in a media containing asbestos
which may interfere with the ability of the authors to
adequately quantify the effects of the test material.
1 The numbers of test organisms and replicates were
reported and sufficient to characterize toxicological
effects.
4 Plants were cultured in a media containing asbestos
in addition to having suspensions containing as-
bestos applied to their fronds.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Low
x 2
The results were sufficiently reported, but uncertain-
ties regarding the exposure led the reviewer to ques-
tion the applicability of the results.
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page
Study Citation: Trivedi, A. K.,Ahmad, I.,Musthapa, M. S.,Ansari, F. A.. 2007. Environmental contamination of chrysotile asbestos and its toxic effects
on antioxidative system of Lemna gibba. Archives of Environmental Contamination and Toxicology 52:355-362
Data Type: Chronic (>21 days); Aquatic; Plants
Hero ID: 621276
Domain Metric Rating^ MWF* Score Comments^
Metric 18: Consistency of Outcome Assessment Medium x 1 2 There were incomplete reporting of minor details of
outcome assessment protocol execution, but these
uncertainties or limitations are unlikely to have sub-
stantial impact on results.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and Medium
Procedures
Metric 20: Outcomes Unrelated to Exposure High
x 2
x 1
The study reported minor differences among the
study groups with respect to environmental con-
ditions or other non-treatment-related factors, but
these are unlikely to have a substantial impact on
results.
There were no differences among groups that could
influence the outcome assessment.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
High x 1 1
Unacceptable x 2 8
Metric 23: Explanation of Unexpected Outcomes
High
x 1
Statistical methods were clearly described and ap-
propriate for dataset(s).
Results were reported in terms of asbestos applied
to each frond, but there were critical details lack-
ing about the characteristics of the test organisms,
particularly regarding the number of fronds /plant.
There were no unexpected outcomes, or unexpected
outcomes were satisfactorily explained.
Overall Quality Determination^ Unacceptable 4.0
Extracted No
** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, five of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
* MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
* The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / ^ MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
Study Citation: Trivedi, A. K.,Ahmad, I.,Musthapa, M. S.,Ansari, F. A.,Rahman, Q.. 2004. Environmental contamination of chrysotile asbestos and
its toxic effects on growth and physiological and biochemical parameters of Lemna gibba. Archives of Environmental Contamination
and Toxicology 47:281-289
Data Type: Chronic (>21 days); Aquatic; Plants
Hero ID: 3080106
Domain
Metric
Rating1
MWF* Score
Comments' t
Domain 1: Test Substance
Metric 1:
Metric 2:
Test Substance Identity
Test Substance Source
Metric 3: Test Substance Purity
High
Low
Low
X 2 2 The test substance was identified definitively.
X 1 3 Although the test material source not defined there
is no indication this impacted the results of the
study.
X 1 3 Although the purity was not reported, there is no
indication that this had an effect on the results.
Domain 2: Test Design
Metric 4: Negative Controls
Medium
Metric 5: Negative Control Response
Metric 6: Randomized Allocation
Medium
Low
x 2
x 1
x 1
Although aquatic plants used as controls were cul-
tured in nutrient mediumwithout chrysotile fiber,
they were from the third generation of plants ob-
tained from a natural habitat in an aquatic body
that was contaminated with asbestos. There are un-
certainties (e.g., due to epigenetics) around how the
initial exposure to asbestos at the parent generation
would affect the plants from the third generation.
There were minor uncertainties or limitations re-
garding the biological responses of the negative con-
trol group(s).
Researchers did not report how organisms were al-
located to study group.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- Unacceptable x 2
tion
The test organisms were cultured in a media contain-
ing asbestos, while also being exposed at a rate that
is reported in terms of exposure per frond. The au-
thors did not provide sufficient detail about the test
organisms or exposure regime (ex. how many fronds
per plant? Does excess test media applied to fronds
enter the test suspension?) to allow the reviewer to
confirm the scientific validity of this study.
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page
Study Citation: Trivedi, A. K.,Ahmad, I.,Musthapa, M. S.,Ansari, F. A.,Rahman, Q.. 2004. Environmental contamination of chrysotile asbestos and
its toxic effects on growth and physiological and biochemical parameters of Lemna gibba. Archives of Environmental Contamination
and Toxicology 47:28f-289
Data Type: Chronic (>21 days); Aquatic; Plants
Hero ID: 3080106
Domain Metric Rating^ MWF* Score Comments"^
Metric 8: Consistency of Exposure Administration
Unacceptable x 1
Metric 9: Measurement of Test Substance Concentra- N/A
tion
Metric 10: Exposure Duration and Frequency
High
Metric 11: Number of Exposure Groups/Spacing of Ex- Low
posure Levels
Metric 12: Testing at or Below Solubility Limit N/A
x 1
x 1
N/A
N/A
There were serious flaws in how the aquatic plants
were exposed to asbestos. The authors described
the test media as containing asbestos, while describ-
ing the exposure of asbestos to the fronds. This led
the reviewer to question the source of the effects ob-
served in this study and whether it was due to as-
bestos in the media or the asbestos applied to the
frond. In addition, the lack of detail about the pro-
cedure used to apply asbestos to the fronds meant
that the exposure cannot be adequately understood
from the information provided in this study.
Exposure concentrations to fronds in the plants were
not measured due to the insoluble nature of asbestos
fibers.
Experiments were carried out for a test duration of
28 days. EPA determined this to be acceptable.
There were no Justifications provided for the selec-
tion of the test concentrations.
Test media was left in suspension because asbestos
is an insoluble particle.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics Medium x 2 4
Metric 14: Acclimitization and Pretreatment Conditions Unacceptable x 1 4
Metric 15: Number of Organisms and Replicates per High
Group
x 1
Metric 16: Adequacy of Test Conditions
Unacceptable x 1
There are minor reservations or uncertainties about
the source of test organisms.
Plants were cultured in a media containing asbestos
which may interfere with the ability of the authors to
adequately quantify the effects of the test material.
The numbers of test organisms and replicates were
reported and sufficient to characterize toxicological
effects.
Plants were cultured in a media containing asbestos
in addition to having suspensions containing as-
bestos applied to their fronds.
Domain 5: Outcome Assessment
Continued on next page

ties regarding the exposure led the reviewer to ques-

tion the applicability of the results.
Metric 18:
Consistency of Outcome Assessment
Medium
X
1
2
There were incomplete reporting of minor details of

outcome assessment protocol execution, but these

uncertainties or limitations are unlikely to have sub-

stantial impact on results.
Domain 6: Confounding / Variable Control

Metric 19:
Confounding Variables in Test Design and
Medium
X
2
4
The study reported minor differences among the

Procedures

study groups with respect to environmental con-

ditions or other non-treatment-related factors, but

these are unlikely to have a substantial impact on

results.
Metric 20:
Outcomes Unrelated to Exposure
High
X
1
1
There were no differences among groups that could

influence the outcome assessment.
Domain 7: Data Presentation and Analysis

Metric 21:
Statistical Methods
High
X
1
1
Statistical methods were clearly described and ap-

propriate for dataset(s).
Metric 22:
Reporting of Data
Unacceptable
X
2
8
Results were reported in terms of asbestos applied

to each frond, but there were critical details lack-

ing about the characteristics of the test organisms,

particularly regarding the number of fronds /plant.
Metric 23:
Explanation of Unexpected Outcomes
High
X
1
1
There were no unexpected outcomes, or unexpected

outcomes were satisfactorily explained.
Overall Quality Determination"'"
Unacceptable

4.0

Extracted

Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. .. continued from previous page
Study Citation: Trivedi, A. K.,Ahmad, I.,Musthapa, M. S.,Ansari, F. A.,Rahman, Q.. 2004. Environmental contamination of chrysotile asbestos and
its toxic effects on growth and physiological and biochemical parameters of Lemna gibba. Archives of Environmental Contamination
and Toxicology 47:281-289
Data Type:	Chronic (>21 days); Aquatic; Plants
Hero ID:	3080106
Domain	Metric	Rating"^" MWF* Score	Comments^
** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4),
EPA will determine the study to be unacceptable. In this case, five of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score
is presented solely to increase transparency.
* MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
$ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / ^ MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed
out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. UPTAKE OF CHRYSOTILE ASBESTOS FIBERS ALTERS GROWTH AND
REPRODUCTION OF ASIATIC CLAMS. Canadian Journal of Fisheries and Aquatic Sciences 43:43-52
Data Type: Chronic (>21 days); Aquatic; Invertebrates
Hero ID: 3093600
Domain Metric Rating^ MWF* Score Comments^
Domain 1: Test Substance
Metric 1:
Metric 2:
Test Substance Identity
Test Substance Source
Metric 3: Test Substance Purity
High X 2 2 Grade 5 chrysotile asbestos mined ore was used.
Low X 1 3 The study authors did not report the specific com-
mercial supplier or batch/lot # used to obtain the
test substance.
Low X 1 3 The study authors mentioned "Grade 5 chrysotile
asbestos" was used but did not define what the
"Grade 5" represents.
Domain 2: Test Design
Metric
4:
Negative Controls
High
X
2
2
The study authors used an appropriate concurrent
negative control groups for all experiments,.
Metric
5:
Negative Control Response
High
X
1
1
The biological responses of the negative control
groups were reported and had acceptable variations.
Metric
6:
Randomized Allocation
Low
X
1
3
Study authors obtained clams from New River, Vir-
ginia and transported these clams to their lab at
Virginia Tech. There were no discussions about sep-
arating these clams into formal randomized groups.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
High
x 2
Low
x 1
The experimental system and methods for prepara-
tion of test media were described in adequate de-
tail and appropriately accounted for the physical-
chemical properties of the test substance. Specifi-
cally, the exposure system used for the asbestos ex-
periments kept the asbestos fibers in suspension by
employing a raised plexiglass tray above a stir bar.
Difficulties with measuring asbestos accurately
posed challenges in consistent administration of test
substance. Study authors used nominal concen-
trations of asbestos in their experiments and men-
tioned that the detection limits for all concentra-
tions ranged from 1.79E4 to 6.91E4 fibers. However,
they tested concentrations up to 10E8. This issue is
an inherent challenge to asbestos, a difficult to test
chemical.
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . continued from previous page
Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. UPTAKE OF CHRYSOTILE ASBESTOS FIBERS ALTERS GROWTH AND
REPRODUCTION OF ASIATIC CLAMS. Canadian Journal of Fisheries and Aquatic Sciences 43:43-52
Chronic (>21 days); Aquatic; Invertebrates
3093600
Study Citation:
Data Type:
Hero ID:
Domain
Metric
Rating"!" MWF* Score
Comments'^
Metric 9: Measurement of Test Substance Concentra- N/A
tion
Metric 10: Exposure Duration and Frequency
High x 1
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
x 1
Metric 12: Testing at or Below Solubility Limit
N/A
N/A Due to the physical chemical properties and diffi-
cult to test nature of asbestos, the nominal values
of asbestos are highly variable, i.e., the effect con-
centrations reported in this study may misrepresent
the actual effect concentrations. Consequently, this
metric is not applicable.
1 For the 96-hr and 30 day experiments, the duration
of exposures and exposure frequency were reported
and appropriate.
1 The number of exposure groups and spacing of expo-
sure levels were justified and adequate to observe the
short-term and long-term effects of asbestos effects
in Corbicula.
N/A Asbestos fibers are insoluble in water and organic
solvents. The study authors cannot test the asbestos
fibers at or below the solubility limit.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
High
x 2
Metric 14: Acclimitization and Pretreatment Conditions High x 1
Metric 15: Number of Organisms and Replicates per Medium x 1
Group
The Corbicula test organism was adequately de-
scribed and obtained from a reliable source. Cor-
bicula was an appropriate test organism to evalu-
ate the environmental effects posed by asbestos to
aquatic invertebrates for the following four reasons:
1) it resides in every major river system in the U.S
and, therefore, is likely to be found within several
chrysotile-contaminated waterways; 2) mollusks are
known to accumulate asbestos; 3) clams are easy to
collect and observe in laboratory environments; and
4) known effects of other toxicants on clams allow
comparisons.
Clams were acclimatized to laboratory conditions for
1-2 weeks prior to experiments and all pretreatment
conditions were the same for control and exposed
populations
Minor uncertainties or limitations were identified re-
garding the number of test organisms and replicates
used for each experiments. These uncertainties are
unlikely to have a substantial impact on the test re-
sults.
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. UPTAKE OF CHRYSOTILE ASBESTOS FIBERS ALTERS GROWTH AND
REPRODUCTION OF ASIATIC CLAMS. Canadian Journal of Fisheries and Aquatic Sciences 43:43-52
Data Type: Chronic (>21 days); Aquatic; Invertebrates
Hero ID: 3093600
Domain Metric Rating^ MWF* Score Comments^
Metric 16: Adequacy of Test Conditions High X 1 1 Clams were adequately housed and fed in a specially
designed exposure system that employed a raised
plexiglass tray above a stir bar that was used to keep
the asbestos fibers in suspension.
Domain 5: Outcome Assessment
Metric 17; Outcome Assessment Methodology High X 2 2 The outcome assessment methodology addressed the
intended biological effects.
Metric 18: Consistency of Outcome Assessment High x 1 1 Details of the outcome assessment protocol were
reported and outcomes were assessed consistently
across study groups (e.g., at the same time after ini-
tial exposure) using the same protocol in all study
groups.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High x 2
Procedures
Metric 20: Outcomes Unrelated to Exposure
High
x 1
There were no reported differences among the study
groups in environmental conditions or other factors
that could influence the outcome assessment.
Mortality of adult claims were not observed in the
96-hours experiments and not statistically signifi-
cant in the 30-days experiments.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
High x 1
High x 2
High x 1
Kruskal-Wallis test used was adequate for test ob-
jectives. Statistical methods were clearly described
and appropriate for datasets.
Data for exposure-related findings were presented
for each treatment and control group and were ad-
equate to determine values for the endpoint(s) of
interest.
All unexpected outcomes were satisfactorily ex-
plained.
Overall Quality Determination"'"
High
1.3
Extracted
Yes

Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. .. continued from previous page
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. UPTAKE OF CHRYSOTILE ASBESTOS FIBERS ALTERS GROWTH AND
REPRODUCTION OF ASIATIC CLAMS. Canadian Journal of Fisheries and Aquatic Sciences 43:43-52
Data Type:	Chronic (>21 days); Aquatic; Invertebrates
Hero ID:	3093600
Domain	Metric	Rating^ MWF* Score	Comments^
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.

-------
PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. SEASONAL BEHAVIORAL AND GROWTH CHANGES OF JUVENILE
CORBICULA-FLUMINEA EXPOSED TO CHRYSOTILE ASBESTOS. Water Research 20:1243-1250
Data Type: Chronic (>21 days); Aquatic; Invertebrates
Hero ID: 3093856
Domain Metric Rating^ MWF* Score Comments^
Domain 1: Test Substance
Metric
1:
Test
Substance
Identity
High
X
2
2
Chrysotile asbestos
Metric
2:
Test
Substance
Source
Low
X
1
3
Source of asbestos not specified
Metric
3:
Test
Substance
Purity
High
X
1
1
Test is conducted with a fiber; Asbestos fiber stocks
used in exposures were prepared by lightly milling
400 mg of asbestos, followed by sonicating 500 ml of
a 0.060mgl -j chrysotile stock for 2h with a Fisher ul-
trasonic cleaner to eliminate large blocks and cleav-
age fragments. Micrographs were taken of the first
15-25 fibers encountered and subsequently measured
for length, width and aspect ratio.
Domain 2: Test Design
Metric 4
Metric 5
Metric 6
Negative Controls
Negative Control Response
Randomized Allocation
High
High
Medium
x 2
x 1
x 1
randomization procedure not specified, no evidence
that this affected the results of the study
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara-
tion
Metric 8: Consistency of Exposure Administration
High
High
x 2
x 1
Asbestos fiber stocks used in exposures were pre-
pared by lightly milling 400 mg of asbestos, followed
by sonicating 500 ml of a 0.060mgl -j chrysotile stock
for 2h with a Fisher ultrasonic cleaner to eliminate
large blocks and cleavage fragments. Suspension
of asbestos fibers was maintained through magnetic
stirring.
Clams were exposed to 0, 102, 104, 105, 106 and
108 fibers -1 m chrysotile asbestos, aquaria situated
above a magnetic stirrer that kept asbestos in sus-
pension.
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. SEASONAL BEHAVIORAL AND GROWTH CHANGES OF JUVENILE
CORBICULA-FLUMINEA EXPOSED TO CHRYSOTILE ASBESTOS. Water Research 20:1243-1250
Data Type: Chronic (>21 days); Aquatic; Invertebrates
Hero ID: 3093856
Domain Metric Rating^ MWF* Score Comments^
Metric 9: Measurement of Test Substance Concentra- High
tion
Metric 10: Exposure Duration and Frequency High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit N/A
x 2
x 1
x 1
Asbestos fiber concentrations in water were deter-
mined by the TEM method described above except
that water samples were directly filtered onto Nude-
pore filters. Background and blanks were processed
simultaneously. Measured asbestos concentration
for 0, 102, 104, 105, 106 and 108 fibers/L were below
detection at 0, 104, 5.7 x 105, 1.3 x 107 and2.1 " 108
fibers/L, respectively.
30-day exposure
N/A Insoluble fiber maintained in a suspension
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
Medium x 2
Metric 14: Acclimitization and Pretreatment Conditions High x 1
Metric 15: Number of Organisms and Replicates per High x 1
Group
Metric 16: Adequacy of Test Conditions High x 1
Juvenile Corbicula (5.2-8.6 mm shell length) were
collected from the New River, Va, by dip net adja-
cent to an industrial pumphouse station (Celanese
Fibers Corp., Narrows, Va). It was uncertain if the
collection site was polluted, but the controls showed
no ill effects, or accumulated fibers so it was assumed
that this collection site was appropriate.
Juvenile clams were sorted from adults and sed-
iment in the field and returned to Virginia Tech
where they were acclimated to constant tempera-
ture ( 20"C)laboratory conditions for 7 days in 40-1.
aquaria.
10 clams/group
Groups of 10 clams were placed in a raised plexiglass
platform of 315 cm2 surface area in each tank.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology
Metric 18: Consistency of Outcome Assessment
High
High
x 2
x 1
Domain 6: Confounding / Variable Control
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . continued from previous page
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns J, J. R.. 1986. SEASONAL BEHAVIORAL AND GROWTH CHANGES OF JUVENILE
CORBICULA-FLUMINEA EXPOSED TO CHRYSOTILE ASBESTOS. Water Research 20:1243-1250
Data Type:	Chronic (>21 days); Aquatic; Invertebrates
Hero ID:	3093856
Domain
Metric
Rating"!" MWF* Score
Comments'^
Metric 19: Confounding Variables in Test Design and High x 2 2
Procedures
Metric 20: Outcomes Unrelated to Exposure	High x 1	1
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods
High
Metric 22: Reporting of Data
Metric 23: Explanation of Unexpected Outcomes
High
High
x 1
x 2
x 1
Nonparametric statistical techniques were applied in
allanalyses. The one-way analysis of variance rank-
analogue,the Kruskal-Wallis Test, was used for one-
way layout data. If significant differences were indi-
cated ( = 0.05), a rank-like Least Significant Differ-
ences Procedure was used to determine the relation-
ships betweengroups. In cases of two sample data
(e.g. planimetricanalysis of gill tissue), Wilcoxon's
Rank Sum Test was usedto test differences between
groups
Overall Quality Determination"'"
High
1.1
Extracted
Yes


*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
( 4	if any metric is Unacceptable
Overall rating =
I V. (Metric Score,- x MWF,') I V ..
'3
J]. (Metric Score; x MWF;) / J] . MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating,
ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
Study Citation: Belanger, S. E.,Schurr, K.,Allen, D. J.,Gohara, A. F.. 1986. Effects of chrysotile asbestos on coho salmon and green sunfish: evidence
of behavioral and pathological stress. Environmental Research 39:74-85
Data Type: Chronic (>21 days); Aquatic; Fish
Hero ID: 3584231
Domain
Metric
Rating"!" MWF* Score
Comments^ f
Domain 1: Test Substance
Metric 1:
Metric 2:
Test Substance Identity
Test Substance Source
Metric 3: Test Substance Purity
High X 2 2 Asbestos was in the form of mined chrysotile.
Medium X 1 2 Asbestos used in this study was a gift from a major
asbestos producer.
Low X 1 3 Purity and/or grade of test substance were not re-
ported. The test chemical was in the form of mined
chrysotile.
Domain 2: Test Design
Metric 4: Negative Controls
Metric 5: Negative Control Response
Metric 6: Randomized Allocation
High X 2 2 Study authors reported using an appropriate con-
current negative control group.
High X 1 1 The biological responses of the negative control
group(s) were adequate (e.g., mortality of control
fish "20 percent in the chronic tests).
High X 1 1 The study reported that organisms were randomly
allocated into study groups.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- High x 2
tion
Metric 8: Consistency of Exposure Administration
High
Metric 9: Measurement of Test Substance Concentra- N/A
tion
Metric 10: Exposure Duration and Frequency
High
x 1
x 1
2 The experimental system and methods for prepara-
tion of test media were described in adequate de-
tail and appropriately accounted for the physical-
chemical properties of the test substance.
1 Test organisms were consistently dosed with as-
bestos (i.e., only once at the beginning of the ex-
periment).
N/A Nominal values are highly uncertain due to the na-
ture of the test substance. As a result, the effect
concentrations reported in this study may misrepre-
sent the actual effect concentrations.
1 Test organisms were dosed with asbestos only once
at the beginning of the experiment. This is sufficient
because asbestos fibers are insoluble and the possi-
bility of the fibers degrading during the experiment
is low. The length of exposure was adequate for the
objectives of the experiments.
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page
Study Citation: Belanger, S. E.,Schurr, K.,Allen, D. J.,Gohara, A. F.. 1986. Effects of chrysotile asbestos on coho salmon and green sunfish: evidence
of behavioral and pathological stress. Environmental Research 39:74-85
Data Type: Chronic (>21 days); Aquatic; Fish
Hero ID: 3584231
Domain Metric Rating^ MWF* Score Comments^
Metric 11: Number of Exposure Groups/Spacing of Ex-
posure Levels
Metric 12: Testing at or Below Solubility Limit
High X 1 1 Two levels of exposure were used (i.e., 1.5E6 and
3E6 fibers/liter). These concentrations are similar
to concentrations found in many aquatic environ-
ments at the time of the study.
N/A N/A Asbestos fibers are insoluble in water and organic
solvents. Nominal values are highly uncertain due to
the nature of the test substance. The effect concen-
trations reported in these studies may misrepresent
the actual effect concentrations.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
High x 2
Metric 14: Acclimitization and Pretreatment Conditions High x 1
Metric 15: Number of Organisms and Replicates per Medium x 1
Group
Metric 16: Adequacy of Test Conditions
High x 1
This study was designed to evaluate the effects of
chrysotile asbestos on recentlyhatched coho salmon
larvae (Oncorhynchus kisutch) and juvenile green-
sunfish (Lepomis cyanellus). These species and
life stages were chosen due tothe importance of
salmonids (e.g., coho) in the Great Lakes ecosys-
tem and theprobable susceptibility of young fish to
asbestos intoxication. The test organisms were ade-
quately described and were obtained from a reliable
source.
Fish were allowed to acclimate for 5 days at room
temperature (20.0 + 2.0"C) and were randomly di-
vided into six groups of 20 fish each. The test or-
ganisms were acclimatized to test conditions and all
pretreatment conditions were the same for control
and exposed populations, such that the only differ-
ence was exposure to test substance.
The numbers of test organisms and replicates were
sufficient to characterize toxicological effects, but
minor uncertainties or limitations were identified re-
garding the number of test organisms and/or repli-
cates that are unlikely to have a substantial impact
on results.
Organism housing, environmental conditions, food,
water, and nutrients were conducive to maintenance
of health and biomass loading was appropriate.
Domain 5: Outcome Assessment
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page

Study Citation: Belanger, S. E.,Schurr, K.,Allen, D. J.,Gohara, A. F.. 1986. Effects of chrysotile asbestos on coho salmon and green sunfish: evidence
of behavioral and pathological stress. Environmental Research 39:74-85
Data Type: Chronic (>21 days); Aquatic; Fish
Hero ID: 3584231
Domain Metric Rating^
MWF*
Score
Comments^
Metric 17: Outcome Assessment Methodology High
Metric 18: Consistency of Outcome Assessment Medium
x 2
x 1
2
2
The outcome assessment methodology addressed or
reported the intended outcome(s) of interest and was
sensitive for the outcomes(s) of interest.
Details of the outcome assessment protocol were re-
ported but the outcomes were not assessed consis-
tently across study. The experiments with higher
concentrations of asbestos occurred for a lesser du-
ration compare to the experiments with lower con-
centrations of asbestos.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High
Procedures
Metric 20: Outcomes Unrelated to Exposure Medium
x 2
x 1
2
2
There were no reported differences among the study
groups in environmental conditions or other factors
that could influence the outcome assessment.
Data on attrition and/or outcomes unrelated to ex-
posure were not reported for each study group, but
this deficiency is not likely to have a substantial im-
pact on results.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods High
Metric 22: Reporting of Data High
Metric 23: Explanation of Unexpected Outcomes High
x 1
x 2
x 1
1
2
1
Statistical methods were clearly described and ap-
propriate for dataset(s).
Data for exposure-related findings were presented
for each treatment and control group and were ade-
quate to determine values for the endpoint(s) of in-
terest. Negative findings were reported qualitatively
or quantitatively.
There were no unexpected outcomes, or unexpected
outcomes were satisfactorily explained.
Overall Quality Determination"'" High

1.2

Extracted Yes
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. .. continued from previous page
Study Citation: Belanger, S. E.,Schurr, K.,Allen, D. J.,Gohara, A. F.. 1986. Effects of chrysotile asbestos on coho salmon and green sunfish: evidence
of behavioral and pathological stress. Environmental Research 39:74-85
Data Type:	Chronic (>21 days); Aquatic; Fish
Hero ID:	3584231
Domain	Metric	Rating^ MWF* Score	Comments^
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
Overall rating =
^ (Metric Score; x MWF;) / ^ MWFj
if any metric is Unacceptable
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns, J.. 1990. FUNCTIONAL AND PATHOLOGICAL IMPAIRMENT OF JAPANESE MEDAKA
(ORYZIAS-LATIPES) BY LONG-TERM ASBESTOS EXPOSURE. Aquatic Toxicology 17:133-154
Data Type: Chronic (>21 days); Aquatic; Fish
Hero ID: 3585046
Domain
Metric
Rating"!" MWF* Score
Comments^ f
Domain 1: Test Substance
Metric 1: Test Substance Identity
Metric 2: Test Substance Source
Metric 3: Test Substance Purity
Medium x 2
Low x 1
Low x 1
Study authors mentioned "Grade 5 chrysotile as-
bestos" but did not define what the "Grade 5"
means.
Study authors did not report the specific commer-
cial supplier or batch/lot # used to obtain the test
substance. In addition, they only used nominal con-
centrations of asbestos in their experiments.
Purity and/or grade of test substance were not re-
ported.
Domain 2: Test Design
Metric 4: Negative Controls
Metric 5: Negative Control Response
Metric 6: Randomized Allocation
High x 2
High x 1
Medium x 1
Study authors reported using an appropriate concur-
rent negative control group (i.e., all conditions equal
except chemical exposure).
The biological responses of the negative control
group(s) were adequate (e.g., mortality of control
fish "20 percent in the chronic tests).
The study reported methods of allocation of organ-
isms to study groups, but there were minor limita-
tions in the allocation method.
Domain 3: Exposure Characterization
Metric 7: Experimental System/Test Media Prepara- High x 2
tion
Metric 8: Consistency of Exposure Administration
High
x 1
The experimental system and methods for prepara-
tion of test media were described in adequate de-
tail and appropriately accounted for the physical-
chemical properties of the test substance. Water
and asbestos were completely changed every other
week and loading (wet weight of fish per liter) did
not exceed 0.33 g/1. Analyses of asbestos concen-
trations were performed before and after one water
exchange every 4 weeks for 4 months of exposures,
and 1 month of recovery following exposure (n = 20
for each concentration).
Details of exposure administration were reported
and exposures were administered consistently across
study groups.
Continued on next page . . .

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns, J.. 1990. FUNCTIONAL AND PATHOLOGICAL IMPAIRMENT OF JAPANESE MEDAKA
(ORYZIAS-LATIPES) BY LONG-TERM ASBESTOS EXPOSURE. Aquatic Toxicology 17:133-154
Data Type: Chronic (>21 days); Aquatic; Fish
Hero ID: 3585046
Domain Metric Rating^ MWF* Score Comments^
Metric 9: Measurement of Test Substance Concentra- N/A
tion
Metric 10: Exposure Duration and Frequency
High
Metric 11: Number of Exposure Groups/Spacing of Ex- High
posure Levels
Metric 12: Testing at or Below Solubility Limit
N/A
N/A Nominal values are highly uncertain due to the na-
ture of the test substance. As a result, the effect
concentrations reported in this study may misrepre-
sent the actual effect concentrations.
X 1 1 The duration of exposure and/or exposure frequency
were reported and appropriate for the study type
and/or outcome(s) of interest.
X 1 1 The number of exposure groups and spacing of ex-
posure levels were justified by study authors and ad-
equate to address the purpose of the study
N/A Asbestos fibers are insoluble in water and organic
solvents. Nominal values are highly uncertain due to
the nature of the test substance. The effect concen-
trations reported in these studies may misrepresent
the actual effect concentrations.
Domain 4: Test Organism
Metric 13: Test Organism Characteristics
High
Metric 16: Adequacy of Test Conditions
x 2
Metric 14: Acclimitization and Pretreatment Conditions High x 1
Metric 15: Number of Organisms and Replicates per High x 1
Group
High x 1
The test organisms were adequately described and
were obtained from a reliable source. The test
species, strain, sex, age, size, life stage, and/or em-
bryonic stage of the test organisms reported and
appropriate for the evaluation of the specific out-
come^) of interest
The test organisms were acclimatized to test condi-
tions and all pretreatment conditions were the same
for control and exposed populations, such that the
only difference was exposure to test substance.
The numbers of test organisms and replicates were
reported and sufficient to characterize toxicological
effects.
Organism housing, environmental conditions, food,
water, and nutrients were conducive to maintenance
of health and biomass loading was appropriate.
Domain 5: Outcome Assessment
Metric 17: Outcome Assessment Methodology High X 2 2 The outcome assessment methodology addressed or
reported the intended outcome(s) of interest and was
sensitive for the outcomes(s) of interest.
Continued on next page

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PEER REVIEW DRAFT - DO NOT CITE OR QUOTE
. . . continued from previous page
Study Citation: Belanger, S. E.,Cherry, D. S.,Cairns, J.. 1990. FUNCTIONAL AND PATHOLOGICAL IMPAIRMENT OF JAPANESE MEDAKA
(ORYZIAS-LATIPES) BY LONG-TERM ASBESTOS EXPOSURE. Aquatic Toxicology 17:133-154
Data Type:	Chronic (>21 days); Aquatic; Fish
Hero ID:	3585046
Domain	Metric	Rating^ MWF* Score	Comments^
Metric 18: Consistency of Outcome Assessment	High	x 1	1 Details of the outcome assessment protocol were
reported and outcomes were assessed consistently
across study groups (e.g., at the same time after ini-
tial exposure) using the same protocol in all study
groups.
Domain 6: Confounding / Variable Control
Metric 19: Confounding Variables in Test Design and High x 2
Procedures
Metric 20: Outcomes Unrelated to Exposure
Medium x 1
There were no reported differences among the study
groups in environmental conditions or other factors
that could influence the outcome assessment.
Data on attrition and/or outcomes unrelated to ex-
posure were not reported for each study group, but
this deficiency is not likely to have a substantial im-
pact on results.
Domain 7: Data Presentation and Analysis
Metric 21: Statistical Methods	High	X 1	1 Statistical methods were clearly described and ap-
propriate for dataset(s) (e.g., ANOVA).
Metric 22: Reporting of Data	High	x 2	2 Data for exposure-related findings were presented
for each treatment and control group and were ade-
quate to determine values for the endpoint(s) of in-
terest. Negative findings were reported qualitatively
or quantitatively.
Metric 23: Explanation of Unexpected Outcomes	High	X 1	1 There were no unexpected outcomes, or unexpected
outcomes were satisfactorily explained.
Overall Quality Determination^	High	1.3
Extracted	Yes
*	MWF = Metric Weighting Factor
t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value.
*	The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High.
if any metric is Unacceptable
Overall rating =
J]. (Metric Score; x MWF;) / ^ MWFj
(round to the nearest tenth) otherwise
where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is
crossed out and an arrow points to the new rating.
^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments.

-------