Document ID: EPA-HQ-OAR-2003-0065-0478
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2008-08-04T04:00Z

Comments	Action/Response	2nd comments 	Action / Response 2	Page #

211 (b) Research Group Reviewer

First paragraph, lines 1-2:

The sentence should be: "...and fetal growth resulted& no signs...".
Revised

	Not OK:  Still grammatically incorrect.

To be consistent with new interpretation, the sentence should read: 
“…and fetal growth resulted in signs of slight…”	See final draft
report	1-2

211 (b) Research Group QA/QC 

Reviewer

Compliance Statement:

Since it was the sponsor’s responsibility to maintain the method of
synthesis, fabrication, or derivation of the test fuel, and this has not
been completed, it must be included in the sponsor’s compliance
statement.	Added to the test substance section of the report.  We do not
consider this to be a compliance issue as it is now available.  	While
this is true, the fact remains that the study was performed without it,
and was not in compliance during its conduct.  The compliance statement
should state this.  	See final draft report	3-1

Page 3-5, Environmental Conditions:

These are the protocol-required ranges for temperature and humidity. The
actual measured ranges should be given.	The actual measured ranged were
added for the chambers.  The animals rooms were monitored to determine
that they were within the acceptable range, but values were not
recorded. 	  There must be some documentation that can account for the
temperatures and humidities in this room for this period of time.  It
must be included in the data.  	See final draft report	3-5

Most of the reviewers’ comments have been addressed.  But, because the
statistical analyses of the data were re-done after the reviewers
comments were received and some of the results changed, there are a few
additional areas that need attention.  

Inconsistent identification of the NOAEL for developmental toxicity

The NOAEL for developmental toxicity is described inconsistently as
10,000 and 20,000 mg/m3 in different parts of the report.  For example,
the final conclusion of the report (page 29) states, “Therefore, the
No Observable Adverse Effect Level for maternal and developmental
toxicity in this study was established at 10,000 mg/m3.”  However,
this conflicts with the conclusions of the Abstract and Summary, which
say that the NOAEL for maternal and developmental toxicity is 10,000 and
20,000 mg/m3, respectively.

	See final draft report

	NOAEL for maternal toxicity

Of note, both reviewers Goldsworthy and Schlesinger agreed with the
authors that the maternal toxicity NOAEL is 20,000 mg/m3.  The current
draft says that 10,000 mg/m3 is the maternal NOAEL.  This change in
conclusions about the maternal toxicity NOAEL is appropriate and
consistent with API and EPA comments.  No need to change anything here. 

	See final draft report

	Abstract

A sentence in the Abstract should be amended to indicate at what
exposure level maternal toxicity was observed.  The Abstract should
states, “There was evidence of slight maternal toxicity as indicated
by statistically significant decreased body weight changes at the GD
20-21 and GD 5-21 intervals.”  Suggest inserting the phrase “at
20,000 mg/m3” at the end of this sentence.  Otherwise, the reader
cannot know what exposure level(s) caused maternal toxicity until
reading the last sentence of the Abstract. 

	See final draft report

	Tables

The authors also reported decreasing linear trends in body weight, body
weight change, and food consumption.  These significant trend tests were
reported in the text, but not in any of the tables.  Usually, such
results are highlighted in the tables.  

Reviewer Goldsworthy asked for only critical results in tables.  The
revised version of the report has no tables at all.  All of the tables
are in the various appendices.  While the lack of tables in the main
report is inconsistent with the request of Reviewer Goldsworthy (and not
my first choice either), I am not sure it is necessary to change this.

	See final draft report

	Interpretation of Maternal Toxicity

The authors reported a statistically significant decrease in maternal
body weight gain at 20,000 mg/m3.  It is unusual for maternal toxicity
to manifest itself as weight loss on GD 20-21 and not earlier during the
exposure period.  Of interest, one animal at 20,000 mg/m3 had a delivery
in progress on GD21.  This dam (#757) lost 22 grams between GD20 and
GD21.  This weight loss suggests the dam had already delivered one or
more pups on GD21.  If it hadn’t been for this dam, the decrease in
maternal weight gain on GD 20-21 and 5-21 might not have been
statistically significant.  Another dam in the 10,000 group lost 28
grams on GD20-21.  The authors may want to take another look at the
individual animal data in their interpretation of maternal toxicity. 
Could the decreased weight gain on GD 20-21 have been an artifact of
early parturition (delivery) in one dam?  

	See final draft report

	Erroneous entries:

Title Page (vi, not v)

Approval Signatures (vii, not vi)

Personnel (viii, not vii)

Quality Assurance Statement (ix, not viii)

Experimental Design (3-7, not 3-6)

Mating (3-7, not 3-6)

Experimental Groups (3-7, not 3.6)

Administration etc. (3-7, not 3-6)

The Chamber (3-7, not 3-6)

The Test Atmosphere (3-8, not 3-7)

Experimental Evaluation (3-8, not 3-7)

Euthanasia and Cesarian Section (3-8, not 3-7)

Examination of Fetuses (3.9, not 3-7)

Tissue Preservation (3-9, not 3-8)

Records (3-9, not 3-8)

Statistical Analysis (3-9, not 3-8)

Gestation Food Consumption (4-2, not 4-1)

Visceral Observations (4-3, not 4-4)

Skeletal Observations (4-3, not 4-5)

Exposure and Chamber Conditions (4-4, not 4-6)

Discussion (4-5, not 4-7)

Protocol Exceptions (4-6, not 4-8)

List of Abbreviations (4-7, not 5-1)

Statistical Symbols and Abbreviations (4-7, not 5-1)

Uterine Implantation Data Abbreviations (4-8, not 5-2)

Fetal External and Visceral Examination Abbreviations (4-8, not 5-2)

References (5-1, not 6-1)

Table 4-1 (4-4, not 4-5)	See final draft report	Table of Contents, page
2 of 262:

Table 4-1 (4-4, not 4-5)	See final draft report

	Page 10, 5th paragraph:  delete underline for the word, “in” and
“p<”	See final draft report	10

Page 11, 1st sentence:  grammatical correction: …”and fetal growth
resulted in signs of slight…”	See final draft report	11

Page 14, Supplier:  Please correct to:  Chevron Research and Technology
Center	See final draft report	14

Page 20, Administration of Test Substance and Exposure Schedule:  two
grammatical/typographical errors:  “…was at six hours…” and
“cequilibration”	See final draft report	20

Page 22, Statistical Analysis, 2nd paragraph:  grammatical: 
“Continuous data were…”	See final draft report	22

Page 23, grammatical:  “The following data were…”	See final draft
report	23

Page 26, Gestation Food Consumption:   “These trends in the gestation
body weight data are considered a reflection of the trends in the food
consumption data.	See final draft report	26

Page 27, Fetal Observations:  please revise:  “…were statistically
significantly increased in the 10,000 mg/m3 group when compared…”
See final draft report	27

Page 27, External Observations:  typographical:  “statistical”

Red material around placenta:  how is this a fetal observation rather
than a uterine (i.e., maternal) observation	See final draft report	27

Page 93, H-2:  extraneous underline	See final draft report	93; H-2

Page 4-5 indicates the NOAEL for both Maternal and Developmental
Toxicity at 10,000 mg/m3, while the Summary indicates NOAELs of 10,000
mg/m3 for Maternal Toxicity and 20, 000 mg/m3 for Developmental
Toxicity, and the Abstract only indicates a NOAEL for Maternal Toxicity
of 10,000 mg/m3.  These should agree. 

	See final draft report	4-5

Appendix H:  It appears that several findings were seen in the original
report that are not included in this second draft.  These are Individual
Cartilagious Structural Variations, Sternebrae Anlage, Hypoplastic; Rib
Anlage, Hypoplastic Thoracic;  Vertebrae Anlage Thoracic Centra
hypoplastic and Thoracic Arch Hypoplastic.  I know that several findings
are grouped for statistical purposes, but in Appendix H, all of these
findings should be there.  As well, one of the Ossification findings,
Thoracic Centra Hypoplasia appears in the summary under Individual
Cartilaginous variations.  

	See final draft report	Appendix H

Appendix H, page 1:  The incidences of ‘Total Fetuses with Skeletal
Variations’ and ‘Total litters with Skeletal Variation’ appear to
have changed.  Since the individual data reported has not changed, it is
not clear why these incidences would change.  As well, during the
original QA audit these original incidences were verified with the
reported data and were seemingly correct.  

1:  The incidences of ‘Total Fetuses with Skeletal Variations’ and
‘Total litters with Skeletal Variation’ appear to have changed. 
Since the individual data reported has not changed, it is not clear why
these incidences would change.  As well, during the original QA audit
these original incidences were verified with the reported data and were
seemingly correct.  

	See final draft report	Appendix H, page 1

211(b) Toxicology Research Group

Ethanol Rat Developmental Toxicity Study Report 

Reviewer Checklist 2 – Additional Comments

				  PAGE  1 	                                                         
                                  DATE \@ "M/d/yyyy"  7/31/2008