gratitude), the Board concluded that it is appropriate that sponsors of
repellent efficacy research studies should be required to ensure that if
a subject is injured as a result of their participation, then the
subject will not have to assume the costs of medical care needed to
treat such injuries.

Charge to the Board

h. Please comment on any special considerations that should be addressed
in the informed consent materials provided people who are candidates to
become subjects in insect repellent efficacy research.

Board Response to the Charge

The general requirements for informed consent are outlined in 40 CFR
26.116 of the Agency’s final human studies rule.  A basic element in
seeking informed consent is that the subject should be told that the
study involved research and given an explanation of the purposes of the
research, the expected duration of the research, a description of the
procedures to be followed, and an identification of any procedures that
are experimental.

The informed consent should begin with a clear statement that this study
is research, and a statement as to whether the product being tested is
approved and marketed in the test formulation or still in the
experimental stage.  Information about the potential efficacy of the
product against the test insect should also be provided.

For the insect repellent studies, it is especially important to be very
clear about the experimental set-up, either the laboratory or the field,
and what the expectations are for the subject.  Because there appears to
be a tendency in these research studies to use “seasoned” subjects
(i.e., those who are in the scientific field or have participated in
these studies before), a detailed explanation of the procedures might
seem to be unnecessary to the investigator.

Nonetheless, the written details of the experimental procedure must be
sufficient to inform a potential subject who has never anticipated in
this type of study and to remind one who has done so.  A video,
PowerPoint presentation, or photographs might help the subject to
visualize what will occur to him/her during the study.  If it is a
laboratory study, it may help to have the subject place their arm into
the cage.  If the subject is expected to use an aspirator, training on
its use should occur prior to the beginning of the study.  A
demonstration of what a landing and probing feels like might be
appropriate because the dermal sensitivity of individuals will vary.

The length of time that the study would take should be clear in the
informed consent document, including whether the test would be repeated.
 The process for randomizing subjects to the test or experimental group
should be included.  In field studies that take the entire day, it might
be explained whether food would be provided to the subjects.

A clear discussion of the stopping rules should be included, especially
for the field studies.  For example, to discontinue participation, does
the subject raise their hand, return to the van, or find a study monitor
to express a desire to stop.

Another basic element of informed consent is a description of any
reasonably foreseeable risks or discomforts to the subjects.  Inclusion
of a Material Data Safety Sheet is not sufficient to adequately inform
the subject as far as all the inherent risks and benefits of study
participation.  For insect repellent studies, three types of risks are
reasonably foreseen.

The first is the risk of being bitten.  The informed consent document
should give an estimate of the potential number of bites that a subject
could receive in the control and experimental groups.  A statement that
the researcher strives for each subject to receive few to no bites is
not sufficient.  Additionally, should a subject have an allergic
reaction to the insect bites, medical procedures and remedies that would
be present should be clearly described in addition to any available
follow-up treatment (e.g. will subjects be given an antibiotic or
steroid cream for their bites?

The second risk is that of a sensitivity or allergic reaction to the
experimental product.  A synopsis of the animal studies and any human
data should be given to the subject along with a discussion of the
theoretical risk of a reaction occurring.  Emergency care procedures
should be presented for a subject who has a reaction to either the bite
itself or the repellent.

The third and most serious risk is the potential for acquisition of
vector-borne illness as a result of insect bites.  The severity of these
illnesses should be clearly explained, even if the researcher believes
the risk is minimal to non-existent due to either through the use of
disease-free insects in a laboratory study or the selection a
disease-free zone for the field study.  The risk of insect-borne
diseases might not occur to a subject who normally does not work in the
field.  The investigator should make sure that the subject clearly
understands both the risks of disease transmission and what symptoms to
look for with any potential insect-borne diseases.  One suggestion might
be to test the subject, either verbally or in writing, about their
understanding of the procedures and the risks.

Another basic element of the informed consent process is a clear and
complete description of any benefits to the subjects or others that are
reasonably expected to result from the research.  It should be very
clear in the informed consent document that there are likely to be no
direct benefits to the subject as a result of study participation.  The
only potential benefit is to society at large to have an arsenal of
insect repellents available.  Additionally payment for participation in
a research study can not be considered a benefit of the study.

Each research participant should be told the extent, if any, to which
confidentiality of the records identifying the subject should be
maintained.  The researcher should be careful to only include those
organizations that have jurisdiction over the study and might therefore
have the right to inspect the records.  It is equally important that
access to the records is limited to as few individuals as possible and
that strict confidentiality procedures be developed and are strictly
adhered to.

Since insect repellent studies should be classified as research
involving more than minimal risk, the subject should be clearly told if
the researcher will cover medical treatment if an injury occurs (this
issue was reviewed by the Board in more detail in response to question
g.) , including not only treatment during the research study but
long-term care, if needed (e.g. in those circumstances in which a study
subject contracts an insect-borne disease).  A phone number should be
provided to volunteers so that they can obtain additional information
about the risks and benefits of study participation, and their rights as
study subjects.

The subject should clearly be told that the study is voluntary and
refusal will not result in any loss of benefits or privileges.  For
studies that use only one control group, it is especially important for
a subject assigned to the control group to understand that they can
withdraw even if this withdrawal might invalidate the study.  It should
also be clear that the subject does not need to give a reason for
withdrawal from the study.  The consequences of a subject’s decision
to withdraw from the study should be addressed, including how it will
affect any payment for participation in the study.

Students or employees used as research subjects in this study are
considered “vulnerable subjects” because they might feel coerced
into participating either by their supervisor, thesis advisor, or even
fellow students/employees.  It should be clear that participation in
these types of studies is neither a condition of employment nor an
academic requirement for students.  An explanation of whom to contact if
the subject feels coerced should be provided.  This contact should not
be associated with the investigator and the subject should be guaranteed
anonymity.  Employees who report directly to the investigators or study
sponsors, and students of the investigator should be excluded from the
study.

The subjects should also be told that they will be informed if any new
information is found during the course of the study that might affect
the subject’s willingness to participate.  Additionally the subject
should be told that they will be informed if it is found that either a
test site, or a laboratory strain of insects used is discovered to have
a higher level of disease than previously thought.

The informed consent document should be written in a language
understandable to the subjects and the subjects should be informed about
any potential conflicts of interests that the researchers have.

HSRB Consensus and Rationale

The consensus of the HSRB was that informed consent should comply with
all of the requirements of 40 CFR 26.1116 of the Agency’s final human
studies rule.  To comply with the human studies rule, consent
information for pesticides studies must include: (a) detailed
information on the procedure (e.g., number of insect bites or landings
anticipated, nature apparatus or field context, length of time of
exposure); (b) a clear statement of the risks involved (e.g., discomfort
from bites, risk of vector-borne disease, medical consequences of the
disease, treatments available for the disease); (c) the voluntary nature
of participation (e.g., statements that eliminate the perception of
coercion for students or employees; specific instructions on how to
signal desire to withdraw from the study); (d) the fact that there was
no immediate direct benefit to the subject in participating as well as a
description of alternative available repellents; and (e) other steps
outlined above. In addition, informed consent information should be as
detailed for experienced subjects as for naïve subjects.

Charge to the Board

i. Does the HSRB recommend that the draft guideline be revised? If so,
please explain what aspects or sections might improve with revision.

Board Response to the Charge

The Board recommended that the Agency refer to Board responses to
previous questions to address revisions to the draft guideline.

Review of HSRB Protocol Criteria

Before the Board reviewed the presented proposed human studies research,
the Board developed science and ethics criteria as a guide for its
evaluation of such studies.  Reference to such criteria would be helpful
for the Agency, study investigators, and other members of the public to
understand the Board’s approach for the review of proposed human
studies.  The relative emphasis placed by the Board on each criterion
may be applied case-by-case and may vary with the nature of the chemical
product, study design, and participants.  Specific studies may also call
for additional criteria. A list of the science and ethics criteria
identified by the Board are provided below:

 Science Criteria

The criteria for the evaluation of the scientific quality of studies
involving human subjects was based on a series of questions which the
Board agreed needed to be addressed by the details provided in the study
protocol.

1) Is a valid scientific question addressed by the study?

2) Are existing data adequate to answer the scientific question?

3) Are new studies involving human subjects necessary to answer the
question?

4) What are the potential benefits of the study?

5) What is the likelihood that the benefits would be realized?

6) What are the risks? Are they serious or irreversible?

7) Is the purpose of the study clearly defined?

8) Are there specific objectives/ hypotheses?

9) Can the study as described achieve these objectives or test these
hypotheses?

10) What is the sample size and how is it derived?

11) What is the basis for the proposed dose levels and formulations in
the study?

12) Is there a plan allocating individuals to treatment?

13) Can the findings from this study be generalized beyond the study
sample?

14) Is there a justification for the selection of the target population?

15) Are participants representative of the population of concern? If
not, why not?

16) Are the inclusion/exclusion criteria appropriate?

17) Is the sample a vulnerable group?

18) Will the measurements be accurate and reliable?

19) Are measurements appropriate to the question being asked?

20) Are adequate quality assurance procedures described?

21) Can the data be statistically analyzed?

22) Is the statistical method appropriate to answer the question?

23) Are point estimates accompanied by measures of uncertainty?

24) Do laboratory conditions simulate real-world conditions?

25) Are field conditions representative of intended use?

26) Does the protocol include a stop rule plan, medical management plan,
and a safety monitor?

Ethics Criteria

The criteria for the ethical acceptability of environmental research
protocols involving human dosing and intentional exposure proposed by
the Board are grounded in the general criteria for IRB approval found in
Subpart K of the Agency’s human studies rule (40 CFR 26.1111 and
1116).  This approach is similar to that taken by the National Academy
of Sciences (2004) in formulating the criteria for scientific and
ethical acceptability (recommendation 5-1) and participant selection
(recommendation 5-2).

Scientific Validity and Social Value

One of the most important criteria for the ethical review of protocols
in fact is scientific.  The research design must be sound (i.e.,
scientifically valid) and the risks of the research must be reasonable
(or balanced) in relation to the importance of the knowledge that may
reasonably be expected to result.  Absent a sound research design, the
prospect of the research generating usable knowledge is severely
diminished.  Although the risks to research participants may be balanced
against anticipated benefits to these same subjects, much environmental
research (such as intentional exposure studies) will not offer any
direct benefit to the research subjects themselves  (See NAS
recommendation 3-1.).

The justifiable risks to which research subjects justifiably are exposed
should be directly proportional (i.e. reasonable or balanced) to the
importance of the knowledge expected to be gained.  In other words, the
information to be gained from the research study must be "worth
knowing".  The NAS took this approach in recognizing that scientific
accuracy alone is insufficient justification for exposing research
subjects to anything more than "no identifiable risk."   For example,
there must be a "reasonable certainty of no harm" to research subjects
if the only benefit of the research is to improve the scientific
accuracy of extrapolating animal to human data (NAS recommendation 4-1).
 As such, a research protocol needs to describe the benefits of the
knowledge that may be obtained so that the reasonableness of the risks
can be judged against the importance of that knowledge.  In addition to
improved scientific accuracy of risk assessment, such benefits may
include a more stringent regulatory standard, new public health measures
that could be adopted, or new products that may protect public health.

Federal regulations state that an IRB should not consider the possible
effects of the research on public policy when evaluating those research
risks that fall within the responsibility of the IRB (40 CFR 26.1111 (a)
(2)).  Nevertheless, the public policy implications of the knowledge
that may result from the research does affect the importance of that
information.  Further, the Human Studies Review Board is not limited by
Subpart K in evaluating the ethical acceptability of a proposed research
study.  As such, scientific accuracy alone may be an insufficient
justification for the importance of a research project.  The protocol
should address the potential benefit of improved scientific accuracy,
and to whom this benefit would accrue.  As recognized by the NAS in
recommendation 4-2, studies that may have a potential public health or
environmental benefit could involve a somewhat higher level of risk
while not causing any lasting harm to research subjects.

Minimizing Research Risk

The research should not expose any human subjects to unnecessary risk
(40 CFR 26.1111 (a) (1)).  This ethical principle has a number of
important corollaries.  First, the use of human subjects must be
absolutely necessary in order to answer an important scientific question
that could not otherwise be answered by using animal models.  In
addition, any intentional dosing studies can only be justified if
observational studies would neither answer the question nor be feasible.
 Admittedly, the judgment of feasibility may be ethically difficult
especially if the only consideration is time and expense.  Second, the
elimination of unnecessary risk means that there is no way to answer the
scientific question that involves less risk if human subjects are to be
used.  Third, the scientific protocol should involve no additional
exposure of study participants to risk unless absolutely necessary.  The
ethical responsibility for "using procedures already being performed on
the subjects" translates, in the environmental context, to studying
those situations in which human subjects are exposed to environmental
toxins as part of their usual activities without increasing their
exposure to those same toxins.  In addition to the ethical priority of
animal over human studies, there is an ethical priority for
observational research over intentional dosing research involving
environmental toxins if scientifically appropriate.  Whether a study
meets the scientific and ethical criteria necessary to justify the
exposure of human subjects to potential risk can only be evaluated in
the context of a given research protocol if the investigator and/or
sponsor specifically addresses alternative means of obtaining the
desired data.

Equitable Selection of Subjects

The selection of subjects should be equitable (40 CFR 26.1111 (a) (3)).
In practical terms, this means that the selection of subjects should
reflect the scientific purposes of the research and not the availability
of a particular population.  This ethical criterion may be especially
problematic in the context of environmental hazards research.  Often the
exposure to environmental hazards in the workplace or at home is greater
for those who are either socioeconomically or educationally
disadvantaged.  As such, subject selection based purely on scientific
design may be insufficient protection for the research subjects, with
additional safeguards.  The need for such safeguards must be assessed
within the specific context of a particular protocols based on an
in-depth knowledge of the community within which the research will take
place.  The ability to "minimize the possibility of coercion or undue
influence" (40 CFR 26.1116) may require the involvement of
representatives from the community from whom the research subjects will
be drawn and within which the research will take place.  A research
protocol also must include specific measures for assuring the equitable
selection of subjects, including recruitment practices, incentives
(financial or otherwise), impact on employment, and the possibility of
retaliation.  In addition, any incentive for participation (whether
financial or otherwise, such as time off from work) should not be
included in the analysis of risks and potential benefits of the
research.

Informed Consent

The information that is included in the informed consent process and
documentation should include all of the information found under the
general requirements for informed consent in 40 CFR 26.1116.  There are
a few specific features of the informed consent information that are
worth highlighting in the context of environmental research.  First, EPA
regulations do not allow for a waiver of either informed consent or the
written documentation of informed consent.  Second, the informed consent
information must include the identity of the pesticide and its mode of
action if the research involves intentional exposure of subjects to a
pesticide (40 CFR 26.1116 (e)).  Given the vulnerability of the research
subjects that are likely to be enrolled in environmental research (as
discussed above), the default position for any research on environmental
toxins (whether observational or intentional) should be that the risks
of any potential pesticide exposure be included in the informed consent
information.  However, if the risks of the toxins are not part of the
research, but instead are part of daily work life, this should be made
clear.  Third, the alternatives to research participation (40 CFR
26.1116(a)(4)) should include all steps that might minimize the risk of
exposure to environmental hazards, up to and including removing oneself
from that environment.  Fourth, as noted previously, the HSRB supports
the view that research subjects should receive needed medical care for
research related injuries at no cost to themselves (consistent with NAS
recommendation 5-5).  As such, the oft-used informed consent template
statement that “no program of compensation is available” would be
unacceptable in human dosing or pesticide exposure research.  The HSRB
acknowledges that the determination that any given injury may be
research-related might be difficult when the protocol combines
observational or interventional procedures with non-research related
exposure to environmental toxins.  Nevertheless, the principle of
providing medical care for research related injuries at no cost to
research subjects must be affirmed. Fifth, the voluntary nature of
participation must be carefully and explicitly described during the
consent process. Investigators, study sponsors and pesticide registrants
are obligated to ensure that neither employment status nor economic need
creates a coercive context for study participation. Finally, the process
and documentation of informed consent needs take into account special
circumstances that may arise in the context of any given research
setting, including language barriers, literacy, comprehension,
employment status, and the confidentiality of screening tests such as
for pregnancy.

Subject Safety

The research protocol must also discuss provisions for assuring the
safety of subjects enrolled in the research, both during and after the
research has been completed.  This obligation goes beyond simply
"monitoring the data collected" to include procedures for collecting
real-time exposure data to the environmental toxins during the research,
and procedures for intervening should the health of research subjects be
at risk from the environmental toxins (regardless of whether the
exposure is intentional or not).  The protocol should describe in detail
any procedures for reversing experimentally-induced harms.

IRB Approval

The HSRB believed that the ethical analysis of a research protocol
requires information concerning the potential risks to human subjects,
measures proposed to minimize risks, the nature and magnitude of all the
expected benefits, and to whom they may accrue, alternative means of
obtaining information comparable to what would be collected for the
proposed research, and the balance of risks and benefits of the proposed
research (40 CFR 26.1125(a)).  Further, the HSRB believes that an IRB is
unable to make the determinations required under 40 CFR 26.1111 absent
this information.  As such, the HSRB expects this information to be
found in the protocol submitted to the responsible IRB.  Although an IRB
may be able to gather this information from other sources, the lack of
this information in the protocol and the lack of a substantive
discussion of these issues in the IRB minutes would raise doubt about
the adequacy of the IRB review.

Insect Repellent Product Performance Efficacy Studies

Study EMD-003 from Carroll-Loye Biological Research

Charge to the Board

a. Does the proposed research described in study EMD-003 appear likely
to generate scientifically reliable data, useful for assessing the
efficacy of the repellent?

Board Response to the Charge

The protocol submitted for review by the HSRB outlined studies to
evaluate the efficacy of IR3535 as a tick repellent in human subjects.
The protocol described a laboratory study in which the movement of the
Western black-legged tick (Ixodes pacificus) up the forearm was to be
determined.  Studies in humans are required to assess the efficacy of
such repellents because laboratory animals differ in their
attractiveness to the pest, and therefore do not provide an accurate
assessment of efficacy in humans.  A more general protocol (CL-001),
which provided additional information relevant to study conduct, was
also submitted for review in combination with protocol EMD-003.

Overall, the protocol for EMD-003 was poorly prepared, with numerous
errors in referring to the nature of the material to be applied (as a
lotion, aerosol or spray) and the evaluation of repellency against
mosquitoes was indicated in the rationale provided for the study.
Furthermore, the protocol indicated that the dose to be applied was 1 mg
formulation/600 cm2, when in fact, the authors of the protocol intended
the applied dose to be 1 gram/600 cm2.   These mistakes were not
considered to be fatal errors in the protocol, but suggested a lack of
attention to the details of protocol preparation and review by the
investigators.  Staffers from the USEPA provided comments on the
numerous shortcomings of the proposed study, and the HSRB fully
concurred with these weaknesses.

IR3535 is commercially available, and there is a large amount of
toxicology data suggesting that it is a compound of low toxic potential.
 Therefore, human subjects are unlikely to be at risk of experiencing
adverse effects relative to exposure to the proposed formulations.

The HSRB recognized three major limitations to the protocol as submitted
to the HSRB for review.  These limitations include: (1) the lack of a
clear rationale underlying the conduct of the study; (2) the lack of
identification and characterization of the formulations to be tested and
(3) the scientific design of the study.  Of these issues, which are
discussed in more detail below, the design of the study was seen as the
most significant shortcoming of the proposed work.

With respect to the clear rationale for the conduct of the study, the
HSRB understood that all new formulations must be evaluated for
efficacy, and that such studies must be conducted in human subjects to
be valid.  However, the investigators failed to identify what was new
about the formulations being studied and failed to identify the
potential benefit of the formulations.  This shortcoming was considered
to be minor and could readily be addressed by providing such additional
information in the study protocol.

With respect to the formulations to be evaluated, the investigators
provided tables listing the percent of active ingredient along with
incipients used to formulate the spray, aerosol and lotion to be used in
the study.  However, there was no additional information regarding when
the formulations would be prepared relative to study execution, whether
the formulations would be characterized analytically to confirm active
ingredient composition, and whether the stability of the formulations
was to be determined.  This information is critical to the overall valid
execution of the study and could be remedied by providing such detail in
the protocol.

The major limitation with the scientific conduct of the study concerned
the study design and data collection.  In particular, the protocol
outlined a study using six test subjects for each product formulation,
with two additional subjects serving as a negative and a positive
control.  No information was provided to justify the group sizes used in
the research. Given the nature of these studies to evaluate tick
repellency, the HSRB considered that a test in which each subject served
as his own control (using one arm for the untreated, negative control
and one arm for the test) was a more appropriate design that would also
be more powerful statistically and more likely to generate reliable
results.

Additional questions were raised by the HSRB concerning how subjects
would be trained to accurately and consistently collect information
regarding the number of ticks crossing or repelled from the arm skin.
The protocol defined that a crossing is scored by the movement of a tick
by at least two centimeters toward the elbow starting from a line at the
wrist, and that subjects select a new tick from a pool of unused,
prescreened ticks every 15 minutes.  There was no information made
available to the Board as to how subjects were trained and qualified to
establish that they could collect accurate data on tick movement.  The
Board agreed that such information was important for establishing good
quality control of the data collection concerning repellency.

HSRB Consensus and Rationale

Overall, the HSRB concluded that there were numerous technical
deficiencies in protocol EMD-003, and the information provided in the
general protocol (CL-001) did not make up for the deficiencies in the
specific protocol.  Therefore, the Board concluded that the available
protocol did not warrant moving forward with the study.

Charge to the Board

b. Does the proposed research described in Study EMD-003 from
Carroll-Loye Biological Research appear to comport with the applicable
requirements of 40 CFR part 26, subparts K and L?

Board Response to the Charge

Background

The study proposed was to evaluate the efficacy of a compound known as
IR3535 as a tick repellent in human subjects. The study is to be
conducted by Carroll-Loye Biological Research, a private research
laboratory in Davis, California by using healthy volunteers and a
controlled laboratory environment. Two protocols were submitted for
review, a general protocol (CL-001) that provided considerable
background information about tests of insect repellency in general, and
the protocol for analysis of the efficacy of IR353 as a tick repellent
evaluated here.

For this protocol, the efficacy of IR3535 as a tick repellent would be
determined by placing Western black-legged ticks (Ixodes pacificus) on
IR3535-treated and –untreated forearms and measuring the speed and
distance that moving insects would penetrate into the treated area.

Strengths and Limitations

The Board concurred with the factual observations of the strengths and
weaknesses of the study, as detailed in the EPA’s Initial Ethics
Review (USEPA 2006b). This study, it was argued, would provide critical
data on the efficacy of IR3535 as a tick repellent. IR3535 is
commercially available and has been used as a repellent in Europe for
years with no evidence of toxicity, so the subjects enrolled in this
study were unlikely to be at increased risk of experiencing adverse side
effects upon exposure. The ticks used for the study also were bred and
raised in a laboratory environment and are considered to be
pathogen-free, minimizing the risk of vector-borne diseases.

The Board concluded, however, that given the deficiencies noted by the
Agency, the proposed research described in Protocol EMD-003 did not
comport with the applicable requirements of §40CFR26, particularly
subpart K. Carroll-Loye Biological Research and the IRB of record also
failed to obtain or to provide all of the documents necessary to be in
compliance with the requirements of §40CFR26, subpart M. The IRB, for
example, refused to release copies of the minutes documenting the
discussion of EMD-003 protocol, preventing the Board from evaluating
whether or not they considered fully the issues listed under the EPA’s
Final Human Studies Rule as part of their review.  Furthermore, the
protocol and supplementary documents submitted to the Board were poorly
written which, while not a fatal flaw in and of itself, should have
precluded IRB and HSRB review and approval.

The HSRB recognized several significant limitations to the protocol, as
submitted to the Board for review. There was, for example, lack of a
clear rationale justifying the conduct of the study as designed. The
Board’s concerns about this are discussed in detail with respect to
the companion protocol submitted by Carroll-Loye, EMD-004, but it was
felt that absent any clear scientific rationale for conducting such a
study, exposure of human subjects to the risks inherent in this protocol
would be unnecessary and unjustifiable.

Additional limitations of the study protocol provided to the Board can
be grouped into two broad categories: (1) concerns about equitable study
subject selection and recruitment; and (2) questions about whether or
not the documentation and process of study subject enrollment was
sufficient to meet prevailing standards of voluntary informed consent.

Subject Recruitment

The Board expressed concern about the potentially coercive nature of
study subject recruitment. Although the study is to be conducted by
Carroll-Loye Biological Research, a private research laboratory in
Davis, California, the Principal Investigator of the study and Co-Owner
of the research laboratory, Dr. Scott P. Carroll, also is an adjunct
faculty member of the Department of Entomology at the University of
California, Davis. As the majority of research participants will be
recruited from the University’s student population, including from Dr.
Carroll’s own department, the protocol and consent documents need to
be altered to define clearly the mechanisms in place to prevent any
coercive enrollment, as well as the additional concerns listed below.

Voluntary Informed Consent

The Board believed that the protocol and consent documents, as provided,
lacked sufficient information to ensure that all study participants were
adequately informed about the risks, benefits and alternatives to
participation in the study. It was unclear, for instance, that
participation in the study would have no direct benefit for volunteers
or that the study was being conducted solely for marketing research.
The major risks of participation in the study also needed to be more
clearly identified in the informed consent form and in supplementary
documents provided to study subjects. For example, one additional risk
that the study investigators may have failed to consider arises from the
plan to pre-screen female volunteers in order to exclude any subjects
who may be pregnant. In accordance with the newly promulgated provisions
in the EPA’s final human studies rule (§40CFR26.1701 - 26.1704),
minors and pregnant women are explicitly excluded from participation,
the latter being confirmed by requiring all female volunteers to undergo
a self-administered over-the-counter pregnancy test on the day of the
study. Because many of the volunteers are undergraduate or graduate
students at a nearby college, the unexpected revelation that a subject
may be pregnant could have a profound psychological or social impact;
Dr. Carroll also may have a professional relationship with these
students through his affiliation with the University. These risks should
be specifically addressed, and the Board recommended that a separate
consent document for female volunteers be prepared that addresses these
risks as well as the safeguards established by study investigators to
ensure that the results of over-the-counter pregnancy tests would be
kept private.

Study investigators also may wish to provide either a more detailed
explanation of the study protocol – including a detailed method for
manipulating the ticks used in the experiment and a clear description of
the study’s duration.

Finally, it was felt that the informed consent documents should be
re-written to: (a) comport with the reading and comprehension level of
the likely subject population; and (b) clarify the section on
compensation for research related injury.

HSRB Consensus and Rationale

The Board concurred with the initial assessment of the Agency that the
study submitted for review by the Board failed to meet the ethical
requirements established in the Agency’s final human studies rule
(§40CFR26).

The Board determined the proposed research described in this study did
not comport with the applicable requirements of §40CFR26, subparts K
and L. The study documents submitted for review also failed to comply
with the requirements of §40CFR26, subpart M.  However, the
deficiencies noted, while significant, were not irreparable.

Study EMD-004 from Carroll-Loye Biological Research

Charge to the Board

a. Did the proposed research described in Study EMD-004 from
Carroll-Loye Biological Research appear likely to generate
scientifically reliable data, useful for assessing the efficacy of a
test substance for repellent ticks?

Board Response to the Charge

Introduction

The Board began its review noting that this protocol addresses repelling
insects, not ticks.

The Board concluded that the proposed research should generate
scientifically useful data for assessing efficacy.  Protocol EMD-004
describes a test of the efficacy of 3-[N-butyl-N-acetyl]-aminopropionic
acid, ethyl ester (IR3535) to repel mosquitoes in field experiments.  It
describes the formulation and dose of the repellent and the number of
replications (6-10 for each formulation). The components of the three
formulations were provided by the Agency.  There would be one to two
untreated controls and one to two positive (DEET-treated) controls.  Two
locations would be used, in or adjacent to the Central Valley in
California and the Florida Keys. The experiment would be double-blinded.
The compound has a very low toxicity profile in animal tests and has
been used in Europe for over 20 years as a repellent for many years
without reports of adverse effects in humans.

General Scientific Criteria

The scientific question was stated (i.e., to test the efficacy of IR3535
in repelling mosquitoes).

It was not clear whether existing data were adequate to answer the
question.

It was not clear whether new studies involving human subjects were
necessary; however, if the repellency had never been tested with North
American mosquitoes, the tests may be necessary.

The potential benefits of the study were clear, i.e., that an effective
repellent would be available that would have either greater efficacy
and/or fewer drawbacks than what was currently approved.

It was likely that the benefits would be realized (i.e., efficacy as a
repellent) because there was a long positive history on this compound
from its European use.

The risks were not specifically noted.

The most likely relevant risk would be disease transmitted by the
mosquitoes, if the mosquitoes carried pathogens, and some mosquito-borne
diseases (e.g., West Nile virus-mediated disease) were serious. The
protocol did not indicate the likelihood of the mosquitoes in the two
test areas to be carriers of disease organisms that could be transmitted
to humans.  However, using the fewest number of untreated controls would
provide the least risk of disease to the participants. The protocol did
not indicate whether all the inert ingredients in the formulations are
GRAS compounds or have documented lack of toxicity at the exposure
levels anticipated.

Study Design Criteria

The purpose of the study was clearly defined (i.e., efficacy testing).

There were specific objectives/hypotheses (i.e., that IR3535 is an
effective repellent).

The study as described can test this hypothesis.

The sample size and how it was derived was not clear, but seems to have
been taken from the guidelines. The number of subjects listed in section
9.1.3 of the protocol listed potentially more subjects than in the table
in section 8.3.2. It was not clear if the stated number of subjects
would be repeated in both locations. The basis for the dose levels and
formulations were not provided. There were no controls with just the
formulation matrix without the repellent.

There was a plan allocating individuals to treatments.

The findings from this study can probably be generalized beyond the
study sample.

Participation Criteria

There was partial justification for the selection of the target
population.

The participants were representative of the population of concern.

The inclusion/exclusion criteria were appropriate.

The sample was not a vulnerable group.

Measurement Criteria

The measurements were expected to be accurate and reliable.

The measurements were appropriate to the question being asked.

Quality assurances issues did not appear to be addressed.

Statistical Analysis Criteria

The data should be able to be analyzed statistically if the efficacy
with time was the subject of the analysis and the comparisons are made
across time.  However, if there is only one untreated control (which
would be more protective against possible disease transmission), then
there would be difficulties with statistical analysis with comparisons
to the untreated control.

The statistical method seems to be appropriate.

Measures of uncertainty were not addressed.

Laboratory and Field Conditions

No laboratory experiments were proposed in this protocol, probably
because of the data already available due to the compound’s long
previous use.

The field conditions were representative of the intended use.

The protocol did not include a stop rule plan, medical management plan,
and a safety monitor.

HSRB Consensus and Rationale

It was not clear whether new studies involving human subjects were
necessary. If the repellency had never been tested with North American
mosquitoes, however, the tests were probably necessary. The potential
benefits of the study were clear, i.e., that an effective repellent
would be available that would have either greater efficacy and/or fewer
drawbacks than what was currently approved. However, empirical evidence
or procedures to determine risks to subjects (e.g., risks of contracting
a vector-borne disease) were not adequate. It was not clear if the
stated number of subjects would be repeated in both testing locations.
The basis for the dose levels and formulations were not provided. There
were no controls with just the formulation matrix without the repellent.
Therefore, the Board concluded that some of the more critical
deficiencies in information identified above would have to be adequately
addressed before this protocol could receive a positive recommendation.

Charge to the Board

b. Did the proposed research described in Study EMD-004 from
Carroll-Loye Biological Research appear to comport with the applicable
requirements of 40 CFR part 26, subparts K and L?

Brief Overview of the Study

The proposed study would evaluate the efficacy of three different skin
applied formulations of an already registered and marketed (in Europe)
insect repellent IR3535.  There would be two study sites, one located in
central California and the other located in the Florida Keys.  The test
compounds would be administered to a standardized skin surface area,
with a comparison to one positive control and one negative control.  The
subjects allocated to the intervention groups would be blinded to the
treatment.  The chosen outcome measures are "percent reduction in the
rate of alightments" and "complete protection time."  The protocol
stated that there would be 6 to 10 subjects per treatment group, with
one subject per control group.  However there was no discussion of
sample size justification.  As discussed below, the protocol lacked any
discussion of risks.

Ethics and Regulatory Compliance

Subpart K of the Agency’s final human studies rule requires that the
investigator submit to the EPA all information that pertains to the IRB
review of proposed research (40 CFR 26.1115a) as well as additional
information specified in 40 CFR 26.1125, if not already included in the
IRB documentation.  The information requested under 40 CFR 26.1125
includes a discussion of the potential risks to human subjects, the
measures proposed to minimize these risks, expected benefits if any and
to whom, alternative means to obtain comparable information, and the
balance of risk and benefits of the research.  In addition, subject
information sheets and approved written informed consent agreements
should be provided, along with any information about recruitment and the
presentation of this subject information.  Finally, the investigator
should provide copies of all correspondence with the IRB, including
official notification of IRB review and approval.

In the case of this protocol, the principal investigator made a request
to the reviewing IRB (Independent Investigational Review Board Inc.
located in Plantation, Florida) for the documents required under 40 CFR
26.1125.  The response from the IRB, dated May 12, 2006, did not include
the minutes of IRB meetings at which the protocol was discussed.  As a
result, the Board was unable to assess whether the IRB discussed or was
even aware of the controversial issues raised by this protocol.  The IRB
did provide templates of two different forms, the EPA Protocol Checklist
and Research Evaluation Form.  Although these forms were fairly
comprehensive, the Board was not provided with copies of the forms used
for the specific protocol review and thus cannot assess whether or not
the forms were used or the content of the IRB analysis and discussion.
The membership roster of the IRB was included.  Although the membership
was diverse and meets the regulatory requirements, there was no
scientific member that appears to have sufficient expertise in the
scientific issues involved in field testing of insect repellents to
assure that the IRB was qualified to make an adequate assessment of this
protocol.  The scientific and ethical assessment may have been adequate,
but the lack of IRB minutes made this determination impossible.  In
effect, the IRB response was to provide procedural documentation of the
IRB's compliance with 40 CFR 26.1115 but to withhold any substantive
documentation that this procedural compliance resulted in an adequate
ethical and scientific review of the submitted protocol.  As such, the
proposed research failed to meet the requirements of 40 CFR part 26,
subpart K.

The investigator, to his credit, remedied some of these deficiencies in
a supplementary document submitted to the EPA as part of the Board's
review.  This undated document was presumably written after the IRB
review.  In this document, the investigator addressed the potential
risks to human subjects, the measures proposed to minimize these risks,
the nature and magnitude of all expected benefits of the proposed
research and to whom they would accrue, the balance of risks and
benefits of the proposed research, and alternative means of obtaining
information comparable to what would be collected through the proposed
research.

Several observations are in order.  First, none of this material,
including the discussion of risks and benefits can be found in the
protocol submitted to the IRB.  The absence of this information in the
protocol further compounds the uncertainty created by the absence of
minutes  showing how the IRB made the determinations required under 40
CFR 26.1111.  The information about the potential risks to human
subjects, the measures proposed to minimize these risks, the nature and
magnitude of all expected benefits of the proposed research and to whom
they would accrue, and the balance of risks and benefits of the proposed
research should be part of the research protocol submitted for initial
IRB review.  Otherwise, the IRB lacked sufficient information to make an
appropriate assessment of the proposed research.  This was not to say
that the protocol would fail to meet the criteria for IRB approval, only
that there was no evidence that the IRB had sufficient information or
expertise to make these determinations.  Second, the protocol did
minimize the risk of vector-borne diseases by limiting the untreated
control group to  a single subject who was experienced in field biology
or entomology.  The risk was minimized further by using an outcome
measure that does not require biting, but rather preparatory activities
on the part of the mosquito after lighting on the subject followed by
aspiration and removal of the mosquito.  However, this approach raised
concerns about the scientific adequacy of the protocol design.  Third,
the investigator addressed the question of alternative means of
obtaining information by arguing that the protocol was designed in
compliance with previous EPA guidelines for registration of these
products.  The Board did not take a position on whether the protocol was
indeed in compliance with these previous guidelines.  However, there was
sufficient discussion by the Board of the new draft EPA guidance on
"Insect Repellent Product Performance Testing" to cast doubt on the
scientific adequacy and necessity of the approach taken in this
protocol.

Finally, the Board agreed with the ethical deficiencies noted by the EPA
(USEPA 2006c)   With the exception of amending the protocol to include
the applicability of additional standards of ethical conduct and the
process of informing appropriate regulatory authorities of any
amendments or deviations from the approved protocol, all of these
deficiencies related to the informed consent discussion and document.
These included a more accurate discussion of subject assignment, a more
extensive discussion of the risks (with specific information about the
risk of vector borne diseases), the correction of an important
typographical error in the pregnancy section, a clarification of the
section on compensation for research related injury, a clarification of
the lack of direct benefit to research subjects and additional
information under the heading of confidentiality.  The Board also
discussed the topic of pregnancy testing and whether there should be a
separate consent for such testing.  As many of the research subjects may
be in a professional relationship to the principal investigator (such as
graduate students or colleagues), a protocol and consent document needs
to discuss how the confidentiality of such pregnancy testing would be
protected.

HSRB Consensus and Rationale

The Board thus concluded that the proposed research described in Study
EMD-004 from Carroll-Loye Biological Research did not comport with the
applicable requirements of 40 CFR part 26, subpart K.  The proposed
research does comport with subpart L, as pregnant women and children
were excluded.  Although the ethical concerns identified by the Board
could be remedied, there were sufficient questions raised about the
adequacy of the research design to cast doubt on whether the proposed
research would meet the criteria for IRB approval found under 40 CFR
26.1111(a) (1).  In other words, absent a sound research design, any
exposure of human subjects to risk would be unnecessary and
unjustifiable.

Board Response to the Charge

Occupational Handler Exposure Monitoring Studies

Charge to the Board

The Agricultural Handlers Exposure Task Force (AHETF) had submitted
protocols for five pesticide exposure studies that are part of a larger
research program the AHETF is conducting.  The premise of the AHETF
research program is that data can be used generically by various
stakeholders (e.g., applicants, registrants, EPA, and others) for
calculating exposures for the occupational handlers of pesticides.  The
scope of the AHETF research program was very broad in that it intends to
address exposures related to many job functions in agriculture and also
to assess generally the impacts of various parameters on exposure (e.g.,
How do changes in the pounds of pesticide handled or acres treated
affect exposure levels?).  The protocols submitted for HSRB review
described studies to measure exposures for five specific scenarios.

The Agency believed these studies had the potential to improve EPA’s
ability to assess the risks of using pesticides because the data would
reflect current agricultural practices, equipment and techniques and
would allow for more refined exposure estimates.  Further, the
monitoring techniques to be used for these studies have been
standardized for use across the AHETF research program. These more
refined and reliable data would allow the Agency to estimate better how
worker exposure levels are affected by changes in various factors such
as the amount of active ingredient handled, type of application
equipment used, application rate used, volumes handled, and personal
protective equipment used.

	It should be noted, however, that the use of the data generated in this
study by the EPA and other stakeholders would depend upon the nature of
the results.  For example, the adequacy of the field or laboratory
quality control data may dictate that correction factors are applied to
adjust monitored exposure levels to account for losses from field
samplers or low performing analytical methods.

1. AHETF Closed System Mixing/ Loading of Liquids Protocol (AHE34)

a. Does the proposed research described in Study No. AHE34 from the
Agricultural Handlers Exposure Task Force appear likely to generate
scientifically reliable data, which would be useful, together with other
data, for assessing the potential levels of pesticide exposure received
by people when mixing, loading or applying a liquid pesticide with
closed systems? [Note:  In a few cases, corresponding application events
are also to be monitored; the same question applies to those elements of
the study.]

b. Does the proposed research described in Study No. AHE34 from the
Agricultural Handlers Exposure Task Force appear to comport with the
applicable requirements of 40 CFR part 26, subparts K and L?

2. AHETF Airblast Application to Trellis Crops in the West Protocol
(AHE36)

a. Does the proposed research described in Study No. AHE36 from the
Agricultural Handlers Exposure Task Force appear likely to generate
scientifically reliable data, which would be useful, together with other
data, for assessing the potential levels of pesticide exposure received
by people when making an airblast application of a pesticide to a
trellis crop under conditions found in the western United States? [Note:
 In a few cases, corresponding mixing/loading events are also to be
monitored; the same question applies to those elements of the study.]

b. Does the proposed research described in Study No. AHE36 from the
Agricultural Handlers Exposure Task Force appear to comport with the
applicable requirements of 40 CFR part 26, subparts K and L?

3. AHETF Airblast Application to Trellis Crops in the East Protocol
(AHE37)

a. Does the proposed research described in Study No. AHE37 from the
Agricultural Handlers Exposure Task Force appear likely to generate
scientifically reliable data, which would be useful, together with other
data, for assessing the potential levels of pesticide exposure received
by people when making an airblast application of a pesticide to a
trellis crop under conditions found in the eastern United States? [Note:
 In a few cases, corresponding mixing/loading events are also to be
monitored; the same question applies to those elements of the study.]

b. Does the proposed research described in Study No. AHE37 from the
Agricultural Handlers Exposure Task Force appear to comport with the
applicable requirements of 40 CFR part 26, subparts K and L?

4. AHETF Closed Cab Airblast Application to Orchards Protocol (AHE38)

a. Does the proposed research described in Study No. AHE38 from the
Agricultural Handlers Exposure Task Force appear likely to generate
scientifically reliable data, which would be useful, together with other
data, for assessing the potential levels of pesticide exposure received
by people when making an airblast application of a pesticide to orchard
crops? [Note:  In a few cases, corresponding mixing/loading events are
also to be monitored; the same question applies to those elements of the
study.]

b. Does the proposed research described in Study No. AHE38 from the
Agricultural Handlers Exposure Task Force appear to comport with the
applicable requirements of 40 CFR part 26, subparts K and L?

5. AHETF Fixed-Wing Aerial Application Protocol (AHE42)

a. Does the proposed research described in Study No. AHE42 from the
Agricultural Handlers Exposure Task Force appear likely to generate
scientifically reliable data, which would be useful, together with other
data, for assessing the potential levels of pesticide exposure received
by people making an aerial application of a pesticide from fixed-wing
aircraft? [Note:  In a few cases, corresponding mixing/loading events
are also to be monitored; the same question applies to those elements of
the study.]

b. Does the proposed research described in Study No. AHE42 from the
Agricultural Handlers Exposure Task Force appear to comport with the
applicable requirements of 40 CFR part 26, subparts K and L?

Board Response to the Charge

For the Board’s review of the agricultural handler protocols, the
Board decided to focus  its analysis addressing the common strengths,
limitations and overall conclusion of the five protocols.

Scientific Considerations

Study Overview

The pesticide handler exposure study protocols submitted to the HSRB
were part of a larger project that was initiated in December 2001 by the
Agricultural Handler Exposure Task Force (AHETF). The project will
produce a generic agricultural handler exposure database (AHED™).  EPA
and other regulatory agencies would use this database to calculate
pesticide handler exposures across a wide range of work conditions.  All
of the protocols follow a similar pattern. They involve personal
measurements of inhalation and dermal exposure among a group of workers
who conduct what is referred to as a “scenario”; that is, the study
participant would conduct a specified job task with specified equipment,
handling a particular product formulation that contains one of six
pesticides.

These studies can be referred to as scripted, so as to distinguish them
from purely observational studies. Workers are asked to conduct their
work activities under a set of scripted conditions similar, but maybe
not identical, to those they experience in their normal work activities.
The overall plan for the exposure database had been discussed with
regulators from EPA, California EPA, and Health Canada on a regular
basis. The presentation of these protocols to the HSRB is the first
independent scientific review of the task force project.

The task force had proposed 33 handler exposure scenarios, distinguished
by equipment type, work task, and pesticide formulation. The task force
has already conducted or initiated 14 exposure studies, and has
sponsored four studies. In addition to the five protocols presented to
the HSRB, the task force planned to conduct approximately 40 additional
studies over the next several years.

Each of the protocols focuses on one primary exposure scenario, but all
of the protocols include more than one scenario. The five protocols
reviewed by the HSRB are summarized in Table 1.

Table 1. Exposure Scenarios Included In The Five AHETF Protocols (N =
anticipated number of independent observations for a particular
scenario)

	AHE34	AHE 36	AHE37	AHE38	AHE42

Scenario

1	Closed system mix-load of a liquid	Open cab trellis crop airblast
application	Open cab trellis crop airblast application	Closed cab
orchard crop airblast application	Closed cockpit fixed-wing aircraft
application

N	10	5	5	8	7

Scenario

2	Open or closed cab airblast application	Closed cab trellis crop
airblast application	Closed cab trellis crop airblast application	Open
pour mix-load of a liquid	Open pour mix-load of a liquid

N	0-3	4	4	not specified

	0-3

Scenario

3	Closed cockpit fixed-wing aircraft application	Open pour mix-load of a
liquid 	Open pour mix-load of a wettable powder

Closed system mix-load of a liquid

N	0-3	0-4	4

0-3

Scenario

4	Closed cockpit rotary-wing aircraft application	Closed system mix-load
of a liquid

	N	0-3	0-4

	Total N	13	13	13	not specified	10

Chemical	Malathion 8

(80% a.i.)	Malathion 8

(80% a.i.)	Diazinon 50% WP	Carbaryl

4lb a.i./gal	Chlorothalonil

6lb a.i./gal

Location	CA	West (CA)	East (NY)	FL and GA	Pacific NW

The task force studies are using six different pesticides: three
organophosphorus insecticides (acephate, diazinon, malathion), one
carbamate insecticide (carbaryl), one organochlorine fungicide
(chlorothalonil), and one triazine herbicide (simazine). A description
of the selection criteria for these compounds was provided as a part of
the task force documentation package. These selection criteria did not
include the toxicity of the compounds, and toxicity was not discussed in
the Agency reviews of the protocols.  However, oral comments from task
force representatives at the June HSRB meeting indicated that the
formulations of these pesticides have been selected such that all fall
into EPA toxicity categories III or IV; i.e., relatively low toxicity
formulations. It was also stated that all workers in these studies wear
long-sleeve shirts, long-legged pants, as well as socks and shoes.
Protective equipment, such as chemical-resistant gloves and eye
protection, are provided to the workers if required by the pesticide
label.

The database to be developed from the task force studies is intended to
supersede an existing database – the Pesticide Handler Exposure
Database (PHED). This database was developed in the late 1980’s and
early 1990’s through the compilation of existing data. These data were
drawn from both registrant-sponsored studies and studies published in
the scientific literature. The studies included in PHED used a different
method for measuring dermal exposure. This method, known as the “patch
technique” (deposition coupons distributed over body regions attached
to the outer layer of clothing), has served as the standard method for
such studies since the 1960’s. When coupled with a hand rinse
technique, it provides an estimate of exposure to all body surfaces.

The documents submitted by the AHETF in support of the proposed exposure
studies consisted of the following:

Cover letter dated May 24, 2006

Analytical method validation reports for 5 of the 6 pesticides (missing
simazine)

List of 33 exposure scenarios

Description of selection requirements for surrogate compounds

32 standard operating procedure (SOP) documents

A generic field exposure monitoring protocol

5 exposure study protocols: AHE34, 36, 37, 38, and 42

IRB documents related to each protocol

In addition, the HSRB received an EPA review for each protocol, EPA,
Office of Pesticide Program guideline documents, and several general
documents on pesticide handler exposure. Finally, the AHETF provided
public comments (AHETF 2006) containing comments on the EPA review of
the five protocols.

Critique of Study

General Scientific Criteria

The primary aim of these studies is to generate personal measurement
data on pesticide handlers suitable for use in an agricultural handler
exposure database. The notion that such a generic database for pesticide
handlers can be developed is supported by substantial scientific
evidence. This evidence indicated that occupational pesticide exposure
in agriculture is largely process rather than chemical-dependent
(excluding chemicals with high volatility). Thus, if sufficient data can
be collected on the key variables that influence exposure, then a
database can be developed to estimate exposure for a wide range of
exposure scenarios. A major concern of the HSRB was that these protocols
included too many variables, and that, even when combined with the full
complement of studies proposed, the database would be inadequate for
meaningful exposure estimates.

The process that has guided the exposure database project had some
significant limitations. The database project has been developed over
the past 5 years by a pesticide industry task force with the input of
regulatory staff from EPA, California, and Canada. Such an ambitious
undertaking would have benefited from an initial independent scientific
peer review, particularly of the study design and statistical plan, as
the HSRB finds itself raising some fundamental questions mid-stream in
the project. Input from the labor community would also have enhanced the
project regarding procedures such as subject recruitment, selection of
pesticides to be used in individual studies, and informed consent. The
purpose of the project, after all, is to develop data to estimate worker
risks. It seems reasonable to give those who would be taking the risks
an opportunity to contribute to the design of the project.

In regard to justification for new human studies, the Agency currently
uses an existing generic pesticide handler exposure database, known as
PHED. It is recognized that new data have not been added to PHED in a
number of years, and that the existing data have a number of scientific
limitations.  However, the inadequacy of PHED was not documented in the
protocols. The Agency had not provided a compelling justification for
these new human studies in the materials provided.

Benefits of the study were not described in the protocols. However, the
AHETF comments (AHETF 2006) and the EPA review documents provided some
general information regarding the role of a handler exposure database in
EPA’s regulatory process.  It was not possible to determine the
likelihood that the benefits would be realized, since the protocols did
not include a description of the full database and how it would be used.

		Study Design Criteria

The purpose of these studies was clearly defined. The objective was to
collect high quality personal measurement data for use in a generic
exposure database. The protocols reviewed by the Board should be able to
produce such data.

Approximate sample sizes were presented in the protocols and discussed
in more detail in the Agency reviews of the protocols. Within each
protocol, the sample sizes for particular scenarios were quite small.
All of the protocols contained multiple scenarios, with sample size per
scenario ranging from 1-10 (see Table 1). The inability to define
exactly how many samples would be collected in each proposed study was
understandable, since the task force was attempting to take advantage of
‘real-world’ conditions. Weather, logistical challenges, and grower
decisions regarding pest management can all affect the number of workers
available for a given study. The effort to study exposures under
realistic conditions required expenditure of significant resources, and
was viewed by the HSRB as highly commendable. The HSRB understood that
these protocols should not be viewed as “stand-alone” studies, since
data from these studies would be combined with other similar studies.
Presumably all of the data collected in these five studies would be
allocated to one of the 33 exposure scenarios outlined by the task
force. Under these circumstances, there was insufficient information for
the HSRB to evaluate the adequacy of the sample size.

In regard to dose levels, participants would be handling varying amounts
of pesticides under variable exposure conditions. The HSRB presumed that
the conditions outlined in the protocols and reviewed by the Agency all
fall within parameters on the label. Actual dose during these studies
would likely be lower than normal, due to the wearing of a whole-body
cotton garment, and strict observance of label instructions.

Participation Criteria

Participants are referred to as ‘replicates’ both in the AHETF
protocols and in the Agency reviews. This term is problematic from a
scientific perspective, since it is used to refer to both a series of
independent observations (e.g., three persons doing the same thing one
time) and a series of repeated measures (e.g., one person doing the same
thing three times). This language needs to be altered such that
different terms are used for repeated measures on one person as compared
to observations on unique individuals. The AHE34 protocol, for example,
indicated that “ten different mixer/loader workers (or replicates)
will be monitored . . . each mixer/loader replicate should be performed
by a separate worker.”  This type of awkward description could be
eliminated through use of unambiguous terminology.

The protocols indicated that the participants would be “experienced
workers” recruited through their employers, but there was no detailed
description of the recruitment procedures, nor were there clear
inclusion/exclusion criteria other than age and pregnancy status. It was
hard to tell whether the workers who volunteer for a protocol exposure
study would be representative of the typical worker.  Random sampling
from a group of eligible workers would improve this aspect of the
protocols.

Measurement Criteria

AHETF investigators are using three different methods to measure skin
exposure: cotton garments (whole body dosimeters), hand rinse, and
face/neck wipes. The cotton garments should be able to capture pesticide
that would normally be deposited on skin.  However, no method for
preventing or monitoring garment breakthrough was presented. If
breakthrough occurs, the dermal exposure measurements would
underestimate true exposure. The hand rinse method and face/neck wipe
both measure the amount of material that can be removed from the skin at
the particular time of the sampling. This amount is some fraction of the
total material deposited on the skin, since some of the material would
have been absorbed into the skin. This method is likely to underestimate
the true exposure. Published laboratory and field studies have indicated
that the fraction of the amount deposited on skin that can be removed by
rinsing or wiping can be quite variable, depending on the nature of the
chemical, its formulation, skin characteristics, and the length of time
the chemical has been in contact with the skin.  In particular, the
face/neck wipe method may seriously underestimate exposure to these
surfaces. This method was not among the methods presented by the Agency
in its 875 guidelines (Occupational and Residential Exposure Test
Guidelines: OPPTS 875.1100 Dermal Exposure – Outdoor), and has not
been validated. The accuracy of these measurements could be improved
through the conduct of laboratory removal efficiency studies. The 875
guidelines do not require removal efficiency studies, but they do
indicate that investigators should address this concern. If such method
validation studies are contemplated, the HSRB recommended that they be
conducted as independent scientific studies published in the
peer-reviewed literature.

In summary, all of the methods for dermal exposure measurements have the
potential to underestimate exposure. The study investigators should
acknowledge this problem in the protocols, and explain what steps, if
any, they have taken to improve or verify the accuracy of the
measurements.

The protocols stated that hand rinse and wipe samples may be collected
multiple times during the work period (e.g., prior to eating, whenever a
worker would normally wash hands), and that this would vary from worker
to worker. The protocols did not explain how multiple measurements from
a single worker would be combined. They also did not discuss whether or
not samples across workers with different rinse/wipe regimens can be
considered comparable. For example, can the amount of pesticide
recovered in a single hand rinse from a worker at the end of the study
period be put in the same database as that from a worker who had four
hand rinses across the study period? The answer would seem to be
“no” from a sampling perspective, given the nature of dermal
absorption processes.

The quality assurance components of the protocols are of high quality.
There was substantial documentation regarding the reliability of
analytical methods available for each of the sampling media to be used.
There were detailed standard operating procedure documents for field and
laboratory quality assurance activities.

Statistical Analysis Critera

An inadequate statistical analysis plan was provided in the protocols.
The HSRB identified this deficiency as the most critical scientific
limitation for these protocols.  There was a need for a more
professional and comprehensive treatment of statistical issues in the
analysis of data, and in the design of individual protocols. Chief among
these issues was the question of statistical power. It is critical to
address the HSRB’s concern that the present design calls in most cases
for a single observation per experimental condition. In other words, it
appears that the present studies are intended to be parsed in terms of
formulation, container size, frequency of worker activity, equipment,
air temperature, wind speed, relative humidity, amount of cloud cover,
rainfall, crop, amount of material handled, rate of application, acreage
treated, and geographic location (along with other possible qualifiers).
As a result, the number of variables to be evaluated appears to approach
or even exceed the total number of subjects for a given scenario. One
may hope that some useful information might yet emerge from a properly
performed analysis of the full data set coming from studies involving
different chemicals, sites, and conditions. What is needed now, however,
is a cogent and thoughtful discussion, in the protocol, of just what can
be accomplished along these lines, and an explanation of how it can be
accomplished. Further thought may lead to the conclusion that the
current data-gathering plan is in fact overly optimistic in regard to
the issue of statistical power. It would then be essential to
restructure the plan and change the study design to ensure that the
enormous effort in this large and important project would not be wasted.

If the goal of these studies was to estimate the distribution of
exposures across a variety of application scenarios, it would be
important to include true repeated measures for at least some of these
scenarios to assess the extent of within-worker variability.

Laboratory and Field Conditions

The protocol states that the field conditions were selected to be
representative of real-world use of pesticides. In this regard,
participants are experienced workers, are allowed to wear their own
clothing, and conduct normal work activities in an actual agricultural
setting. Studies are distributed across the U.S. and across the year in
an attempt to develop a range of exposure conditions. While laudable in
scope, it is important to restate the concern that the large number of
variables included in these studies may prove extremely problematic for
analysis.

The issue of potential heat stress was discussed at length at the HSRB
public meeting. The HSRB concluded that the protocols should include
explicit criteria for halting a study due to heat stress risk.

The protocols stated that workers would be monitored “during a period
of time representative of a full day’s work”. The protocols also
indicated that monitoring times would  conform to a “typical”
workday.  However, none of the protocols defined the typical work period
for the specific tasks to be studied.  Instead, the protocols stated
that monitoring time “will involve work periods with a target of 4
hours.” This language suggested that even four hours of monitoring
might not be achieved in some cases, and there was no indication that
workdays as long as 8 or 9 hours would ever be monitored.  It was not
clear to the HSRB that these studies would necessarily reflect a full
workshift or a “typical” workday. Many factors can influence the
length of the workday, including weather conditions, and the need to
“get the job done” due to pest pressures or the stage of crop
development. Fatigue is an important factor that can affect exposure,
and usually inattention occurs at the end of the day. The HSRB
recommended that the protocols document the time of a typical workday
(or the range of these times) for each scenario, and that the monitoring
time be based on this information.

	Special Concerns Regarding Use of Diazinon in Protocol AHE37

Protocol AHE37 involves handling a wettable powder (50WP) formulation of
diazinon. Workers would be monitored during open cab trellis crop
airblast applications and open pour mixing-loading operations. The HSRB
was concerned that these practices are not consistent with current
Agency policy.  The Agency’s May 2004 interim registration eligibility
document (IRED) for diazinon stated that engineering controls are
required during handling.  The “IRED Facts for Diazinon” states,
“All application equipment must use lock and load engineering
controls. All wettable powder formulations must be packaged in
water-soluble bags. Closed cabs are required for all ground equipment,
except for applications to apples.” The IRED Executive Summary further
stated, “Occupational exposure to diazinon is of concern to the
Agency.  For agricultural uses of diazinon, most mixer/loader/applicator
risk scenarios currently exceed the Agency’s level of concern (i.e.,
MOEs are less than 100 for dermal exposure and MOEs are less than 300
for inhalation exposure).  Taking into consideration both the risks and
benefits of these uses, EPA has determined that most agricultural uses
may continue with the adoption of the following mitigation measures: . .
. engineering controls for mixers and loaders and closed cabs for
applicators for all application scenarios . . .” The above statements
indicate that open pouring of diazinon is not permitted, and that open
cab airblast applications are not permitted in trellis crops.

HSRB Consensus and Rationale

The five studies presented for HSRB review were components of a
large-scale exercise to create a contemporary database on occupational
exposure to agricultural pesticides. The undertaking is in itself likely
to be worthwhile in quantifying and improving our understanding of the
exposures of and risks to pesticide handlers. The potential benefits are
large and the risks appear to be relatively modest. However, the
materials supplied for HSRB review failed to deal adequately with risks
and benefits. None of these protocols can be properly evaluated in
regard to scientific validity because they lacked: (1) a developed
rationale documenting the need for new data; (2) a clear and appropriate
plan for the handling of the data (including its statistical analysis),
and (3) an explanation of the uses to which the data would be put and
adequate sample sizes and protocols for repeated measures to
appropriately estimate exposures within individuals and between
scenarios. These points need to be addressed, at least briefly, in each
specific protocol and, more fully, in a separate and new “governing
document” that is not simply a generic description of the planned
activities.

Additional validation studies are recommended to determine the extent to
which dermal exposure measurements may underestimate true exposure.
Laboratory-based removal efficiency studies or field-based biomonitoring
studies could be conducted to achieve this goal. Such studies should be
published in the peer-reviewed literature. Broader participation of the
scientific community and of parties with a direct interest in the
database project, such as the labor community, would likely improve the
quality of the database and enhance the credibility of its use in risk
assessments.

	The HSRB recommended that specific criteria for cessation due to heat
stress be included in these worker exposure protocols, and that the
protocols included a heat stress management plan.  In addition, the HSRB
recommended that the length of each study should be truly representative
of a full workday, and that each protocol should document the basis for
the proposed duration of the study.

	The HSRB was gratified to receive the Agency’s response to its query
regarding the use of diazinon in the AHE37. It is the understanding of
the HSRB that the Agency would inform the AHETF that it needs to
identify a pesticide other than diazinon in this protocol to evaluate
exposures associated with open pour activities and applications using
open cabs, and that the Agency would ensure that future protocols comply
with the most current risk mitigation measures specified in IREDs and
REDs.

Board Response to the Charge

Ethical Considerations

Background

These five studies are part of a series of studies that are to be
conducted by the AHETF, a coalition of 19 pesticide registrants that was
formed in December 2001 to share resources in the design, evaluation,
and development of a proprietary agricultural mixer/loader and
applicator exposure database for use in regulatory risk assessments.

The study protocols were designed by AHETF investigators after joint
discussions with the US EPA, Health Canada, and the California
Department of Pesticide Regulation, in accordance with the
recommendations of such guidance documents as: 1) US EPA, Occupational
and Residential Exposure Test Guidelines, Series 875.1000 through
875.1600 (1996); and 2) US EPA, Working Draft - Occupational and
Residential Exposure Test Guidelines, Series 875 Group-B,
Postapplication Exposure Monitoring Test Guidelines Version 5.4 (1998).
The supporting and supplementary study documents also assert compliance
with the Good Laboratory Practice (GLP) Standards established by the
1972 amendment to the Federal Insecticide, Fungicide, and Rodenticide
Act (FIFRA) (§40CFR160). Finally, these protocols were reviewed and
approved by the Western Institutional Review Board (WIRB) of Olympia,
Washington, prior to submission to the Agency.

The aims of these studies are to provide critical exposure information
for individuals who mix, load, and apply agricultural pesticides.
Agricultural producers (“growers”) would be recruited by the study
coordinators several months prior to initiation of the study; in
exchange for their participation in the study, each grower would
receive, free of charge, an amount of liquid pesticide equivalent to the
normal quantity mixed and loaded into closed-mixing systems and spray
rigs for the duration of the study (expected to be a single day).
Participating growers also may be asked to recruit other growers and
pesticide applicators into the research study.

Study investigators would recruit agricultural handlers on-site;
volunteers would receive $100/day for their participation in addition to
their regular pay. Voluntary informed consent would be solicited by
study investigators, and will be documented using a standardized
informed consent form. Because the study participants would be recruited
from a pool of experienced agricultural workers who routinely mix and
load liquid pesticides as part of their normal duties, the AHETF had
argued that participation in this study presents a negligible increase
in pesticide exposure risk to volunteers. In accordance with the newly
promulgated provisions in the EPA’s Final Human Studies Rule
(§40CFR26.1701 - 26.1704), minors and pregnant women are explicitly
excluded from participation, the latter being confirmed by requiring all
female volunteers to undergo a self-administered over-the-counter
pregnancy test on the day of the study.

Dermal exposure to pesticides would be ascertained through hand rinses
and face/neck wipes, as well as the use of long cotton underwear – as
a surrogate for skin – to be worn under the study participant’s
clothing. In addition to the long underwear, all participants would be
required to wear long sleeved shirts and long pants, shoes plus socks,
in accordance with accepted worker protection standards. Volunteers may
wear their own clothing provided they are freshly laundered;
alternatively, the AHETF would provide freshly laundered clothing. Any
personal protective equipment (PPE) that may also be required, such as
chemical resistant gloves and protective eyewear, will be provided. At
the conclusion of the four-hour study observation, the long underwear
would be removed and subjected to laboratory analyses to estimate
whole-body dermal pesticide exposure. Study participants would also be
asked to wear OSHA Versatile Samplers (OVS) outfitted with glass
filters, XAD-2 sorbent, and tygon tubes to measure inhalation exposure.
The tubes would be attached to the volunteer’s collars with the
openings positioned in their breathing zones. By using such
state-of-the-art monitoring techniques, the AHETF argues, this study
would provide critical exposure information for individuals who mix/load
liquid agricultural pesticides.

Strengths and Limitations

These studies would provide critical exposure information for
individuals who mix/load and apply agricultural pesticides. It is also
believed that the monitoring techniques proposed for these studies
represent the current state-of-the-art. However, the Agency also
recognized that use of the data resulting from this studies would take
careful scrutiny and may require a number of adjustments depending upon
the results. Finally, the overall design of these studies should be
considered in the context of the goals of the AHETF which are to develop
a broad-based database that can be generically used as a predictive tool
for estimating exposures to pesticide handlers and that the
interpretation of the results of these studies may or may not
necessitate the need for additional monitoring data.

The Board concurred with the factual observations of the strengths and
weaknesses of the studies, as detailed in the EPA’s Initial Ethics
Review of the AHETF Template Protocol and each individual study
protocol. The Board concluded that, given the deficiencies noted by the
EPA, the proposed research described in the AHETF Template Protocol and
each individual study protocol do not comport with the applicable
requirements of §40CFR26, subparts K and L. Furthermore, the AHETF and
WIRB failed to provide all of the documents necessary to be in
compliance with the requirements of §40CFR26, subpart M.

Although public comments from several members of the AHETF helped
assuage some of the Board’s concerns, the members of the HSRB believed
that further comments about this protocol were warranted. The comments
below are grouped into four broad categories: (1) whether the study was
designed to adequately minimize risk to study participants; (2) whether
the documentation and process of study subject enrollment was sufficient
to meet prevailing standards of voluntary informed consent; (3) whether
study participants would be adequately compensated in the event of a
study-related injury; and (4) whether appropriate alternatives to
participation are provided.

Minimization of Risks to Study Participants

This study proposes to measure dermal and inhalation exposure to liquid
pesticides by agricultural handlers who usually perform pesticide
mixing, loading, and application as part of their daily routine.
However, it was unclear to Board members, given the semi-scripted nature
of the protocol provided, as to whether or not study participants would
be exposed to greater quantities of these compounds than would normally
occur. Are the studies proposed purely observational in nature, or are
study investigators intervening by requesting that study participants
use different types and quantities of pesticide, or different mixing,
loading, and application methods, than they normally would? If the
latter is true, the assumption that this study represents a negligible
increase in pesticide exposure risk to volunteers may be unfounded.
Several Board members also expressed concern that the additional
requirements for donning and removing the equipment used to measure
pesticide exposure may inadvertently lengthen the participant’s normal
work day. If so, this should be clearly described during the consent
process, as should the question of whether the $100 paid for study
participation is expected, in whole or in part, to compensate for the
extension of the work day.

The protocol failed to detail the approach taken to ensure that
agricultural handlers are adequately trained in the proper mixing,
loading, and application of these compounds. Although pesticide mixing
instructions and Material Safety Data Sheets are made available to study
participants, given that many agricultural workers may not be fluent in
English (or may even be illiterate), a clear plan for ensuring that
volunteers are properly educated in minimizing their exposure to these
compounds should be included. Furthermore, study investigators may want
to make arrangements to provide volunteers with the results of the study
following completion.

One of the greatest risks to study participants is heat-related illness,
given that dermal exposure to pesticides will be determined by asking
volunteers to wear long underwear in addition to their normal protective
equipment (e.g., long sleeved shirts and long pants, and other
applicable protective gear). Although study coordinators are expected to
be vigilant for signs of heat-related illness among volunteers, in order
to minimize the risks posed to the study participants the protocol also
should include: a) explicit starting and stopping criteria based on a
quantifiable measure like ambient temperature or heat index; and b) a
clear description of the symptoms of heat-related illness in the
informed consent documents. There should also be a clear plan for
reporting any heat-related illness (or, for that matter, any other
adverse event) to the study investigators, Western IRB, and the EPA.

Because some of the study participants may be undocumented immigrants,
measures to ensure strict confidentiality should be developed. Many
undocumented workers, for example, may be loathe to report any adverse
study-related event requiring medical attention or hospitalization if
they believe that their illegal status will be reported to immigration
authorities. Alternatively, study investigators may wish to require
documentation of citizenship or immigration status as part of the
inclusion criteria for recruiting study participants. In addition,
because many pregnant day-laborers may fear job loss in the event that
their employer learns of their condition, extra care should be taken to
keep the results of over-the-counter pregnancy tests private.

Voluntary Informed Consent

Several Board members felt that the AHETF protocol, as provided, lacked
sufficient safeguards to ensure that all study participants were
adequately informed about the risks, benefits and alternatives to
participation in the study. For example, it was felt that the informed
consent documents provided were written at too high of a reading and
comprehension level. Given the sociodemographic characteristics of the
farm worker population, many of the study participants may have limited
education, may speak English as a second or even a third language, or
may even be illiterate. Study investigators should develop a clear
consent document which – in addition to including a more detailed
description of risks (including the risks of the pesticides being
handled) as described previously, as well as a clear distinction between
what comprises research versus normal activities – is written at a
lower grade-level and translated into the various languages likely to be
spoken by study participants. A brief oral test of comprehension should
also be developed, with volunteers required to demonstrate a clear
understanding of the purposes and the risks of the study prior to
enrollment.

The Board also expressed concern about the potentially coercive nature
of the study, given the potential for study participants to believe that
there is a direct relationship between study investigators and growers.
Absent additional safeguards, the “gift” of study pesticide to the
growers may contribute to undue influence on employees to participate in
the research. Western IRB, in its initial review of several of the AHETF
protocols, recommended that “extra care” be taken during the
recruitment and consent process to minimize coercion or undue influence
on study participants. However, no documentation was provided to the
HSRB as to how the AHETF addressed WIRB’s concern. For example, there
was no evidence to suggest that AHETF researchers solicited the help of
the farm-workers themselves or other community leaders to ensure that
study participants would not be covertly or overtly coerced into
participating in the study. The rights of participants to withdraw from
the study at any time also should be emphasized. It is unclear from the
informed consent or other study documents, for instance, as to whether
volunteers are entitled to receive monetary payment even if they chose
to withdraw during the course of the study. Although the Board was
reassured during the discussion that sufficient alternate work was
available, the protocol also failed to specify that workers would still
be paid for a day’s labor even if they refused to participate in the
research.

Compensation for Injury to Study Participants

The study protocol and informed consent documents state that: “If [a
study participant is] injured as a result of being in this study,
treatment will be available from a health professional at a nearby
medical facility.  The costs of such treatment will be covered by the
AHETF.  This does not cover any injuries resulting from [the
volunteer’s] normal activities.” Given the nature of the study
design, however, it is unclear whether a distinction between injuries
resulting from normal work activities versus participation in this study
can be made. Two of the symptoms of heat exhaustion, for example, are
dizziness and loss of coordination – will study coordinators be able
to distinguish between an accidental injury caused by clumsiness versus
an injury resulting from potentially-unrecognized symptoms of
heat-related illness? In light of these concerns, the Board recommends
that the AEHTF cover medical treatment for all participant illness and
injury occurring during the study period (i.e., the day of the test).

Alternatives to Participation

As noted above, the design of this study involves collaboration between
the researchers and growers in which the growers receive, free of
charge, a particular pesticide that they are required to apply to their
fields on the day of the study. That arrangement will lead in many
circumstances (except of the few coincidental instances when the grower
had already planned to use that chemical on that day) to a change in the
pesticide being applied by the grower.

The following question thus arises: What alternatives are offered to
agricultural handlers working for that grower who choose not to
participate in the study? One option is that they could be offered the
choice of applying that pesticide that day, but not needing to
participate in any other study procedures (such as wearing the long
underwear). Some members of the Board believed that if that is the only
alternative to participation, then this aspect of the study would not
comply with 40 C.F.R. Part 26, Subpart K. A primary purpose of the EPA
rule is to prevent a person from being intentionally exposed to a
pesticide without their voluntary informed consent. The EPA emphasized
this point when it promulgated the final version of its rule, commenting
that the term “research involving intentional exposure” covers
“any research on a substance, unless the subjects of the research
retain complete control over whether, when, and how they are exposed to
the substance.” 71 Fed. Reg. 6138, 6146 (2006).

Some members of the Board accordingly concluded that for agricultural
workers who had pre-existing expectations of earning money working for
the grower on the day of the study (either as employees or as
independent contractors with contractual expectations of working that
day), the protocol must provide them alternatives for earning that same
amount of money that do not require them to apply the pesticide used in
the study. Acceptable alternatives could include applying some other
pesticide they have in the past applied, performing some other task they
regularly perform, or being paid their expected earnings without needing
to work. Absent such alternatives, the protocol would appear to be
inappropriately coercing such persons into applying the study compound
or else losing the money they expected to earn that day.

HSRB Consensus and Rationale

The Board concurred with the initial assessment of the Agency that the
studies submitted for review failed to meet the ethical requirements
established in the 40CFR26.

The Board determined the proposed occupational handlers exposure studies
do not comport with the applicable requirements of 40CFR26, subparts K
and L. However, the deficiencies noted, while significant, were not
irreparable.

REFERENCES

AHETF 2006.  Comments to the Human Studies Review Board (HSRB) Regarding
Five Protocols Submitted for Review.  June 20, 2006.

NAS 2004.  Intentional Dosing Studies for EPA Regulatory Purposes:
Scientific and Ethical Issues. National Academy Press.

NBAC 2001.  Ethical and Policy Issues in Research Involving Human
Participants.  National Bioethics Advisory Commission.
www.bioethics.gov

Prentiss AM.  1937.  Chemical in War: A Treatise on Chemical Warfare.
McGraw-Hill Book Company, Inc.  New York.

USEPA 2006a.  Human Studies Review Board.  Weight of Evidence
Determination for Chloropicrin.  June 7, 2006

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USEPA 2006c. Ethics Review of Protocol for Human Study of Mosquito
Repellent Performance  June 9, 2006.

USGPO.  1949.  Trials of War Criminals before the Nuremberg Military
Tribunals under Control Council Law No. 10, Vol. 2, pp. 181-182.
Washington, D.C.

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