EP A-HSRB-21-1
Dr. Jennifer Orme-Zavaleta
EPA Science Advisor
Office of the Science Advisor
1200 Pennsylvania Avenue, NW
Washington, DC 20460
Subject: January 26, 2021 EPA Human Studies Review Board Meeting Report
Dear Dr. Orme-Zavaleta,
The United States Environmental Protection Agency (EPA) requested that the Human Studies
Review Board (HSRB) provide scientific and ethics review of a study protocol involving human
participants. On January 26, 2021, the HSRB considered a study protocol for field evaluation of
two topically-applied insect repellent products containing IR3535. Briefly, the goal of the
proposed study is to determine the efficacy and duration of protection of two skin-applied
products preventing mosquito landings on human hosts.
The HSRB's responses to the charge questions presented at the meeting on January 26, 2021
along with detailed rationale and recommendations for their conclusions are provided in the
enclosed final meeting report.
Signed,
Jennifer Cavallari, ScD, CIH
Chair, EPA Human Studies Review Board

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INTRODUCTION
On January 26, 2021, the United States Environmental Protection Agency (EPA or Agency)
Human Studies Review Board (HSRB or Board) met to address the scientific and ethical charge
questions related to a study protocol "Field evaluation of two topically applied insect repellent
products containing IR3535 against mosquitoes in Louisiana." In accordance with 40 CFR
26.1601, EPA sought HSRB review of the study protocol.
REVIEW PROCESS
The Board conducted a public meeting on January 26, 2021. Advance notice of the meeting was
published in the Federal Register as "Human Studies Review Board; Notification of a Public
Meeting" (EPA, FRL- 10017-40-ORD). This Final Report of the meeting describes the HSRB's
discussion, recommendations, rationale and consensus in response to the charge questions on
ethical and scientific aspects of the completed and proposed research.
The Agency staff presented their review of the scientific and ethical aspects of the proposed
research, with each presentation followed by clarifying questions from the Board. The HSRB
solicited public comments and next proceeded to address the charge questions under
consideration. The Board discussed the science and ethics charge questions and developed a
consensus response to each question. For each of the charge questions, the Chair called for the
Board to vote to confirm concurrence on a summary statement reflecting the Board's response.
For their evaluation and discussion, the Board considered materials presented at the meeting,
study reports, related materials and documents provided by the study sponsors, the Agency's
science and ethics reviews of the study, as well as oral comments from Agency staff and the
investigators during the HSRB meeting discussions. A comprehensive list of background
documents is available https://www.epa.gov/osa/january-26-2021-meeting-human-studies-
review-board.

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Protocol for Field Evaluation of Two Topically Applied Insect Repellent Products
Containing IR3535 against Mosquitoes in Louisiana
Charge to the Board- Science:
Is the protocol "Field evaluation of two topically applied insect repellent products containing
IR3535 against mosquitoes in Louisiana" likely to generate scientifically reliable data, useful for
estimating the amount of time each of the products tested repels mosquitoes?
HSRB Response:
The protocol "Field evaluation of two topically applied insect repellent products containing
IR3535 against mosquitoes in Louisiana" is likely to generate scientifically reliable data, useful
for estimating the amount of time each of the products tested repels mosquitoes provided that the
comments and recommendations provided by the EPA and HSRB are adequately addressed.
Science review
The HRB reviewed the protocol and related documentation for the field evaluation of two skin-
applied insect repellent products containing IR3535. The data collected in this study will be used
to support product registration for AKIVA 20 lotion and wipe formulations of skin-applied
repellent products containing IR3535. This study is designed to determine the median complete
protection time (CPT). This study is sponsored by LivFul, Inc. and will be conducted by the
London School of Hygiene and Tropical Medicine's ARCTEC and Louisiana State University
(LSU).
Field-testing of these products has been conducted previously at two test sites. The HSRB
reviewed the data generated to support this registration under a different protocol in October
2019. For this study, the proposed site will be selected from locations in Louisiana to replace
data from a previous study at one Florida site where data were insufficient to support protection
time for more than three hours due to low landing pressure from predominantly Culex species.
Product registration requires data from two distinct sites. Data resulting from this study will be
combined with existing data that support a CPT of 14 hours for the lotion and of 13 hours for the

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wipe. The rationale for this testing is to collect data to show the efficacy of these products at
repelling mosquitoes to support registration of these products by the EPA.
While the submission included two versions of a protocol (one dated 17 April 2020 and one
dated 2 May 2020), both the EPA and the HSRB review pertain to the 17 April 2020 protocol
version. EPA has provided a thorough scientific review of the protocol and supporting
documents and made a number of recommendations that the HRSB agrees with.
Briefly, the field testing will be conducted in the following manner:
Control subjects will monitor ambient landing pressure by exposing their untreated leg for up to
5 minutes prior to each exposure period. Adequate landing pressure is defined as five landings
within five minutes or less on both control subjects.
Immediately following the control subject landing pressure assessment, treated subjects will
expose their treated limbs for five minutes every 30 minutes, beginning 2 hours post product
application. Once the first confirmed landing has occurred within a five minute time interval (a
single landing followed by a second landing within 30 minutes of the first recorded landing), the
time point of repellent failure and period of complete protection will be recorded for that
particular subject.
The test will be stopped for: safety reasons; more than four non-consecutive exposure periods or
more than three consecutive exposure periods are missed due to bad weather, rain or wind speed
>10mph; landing pressure is below acceptable levels for more than four non-consecutive
exposure periods or more than three consecutive exposure periods; the end of field test period is
reached (maximum duration reached or sufficient number of subjects experience FCL); at the
Study Director's discretion for any reason.
We agree with the EPA recommendations as presented in their scientific review and report and
would like to emphasize the importance of the following recommendations: revising the study
objective and purpose with more detailed working; replacing references to bites with references
to landings; adding details on randomization assignments; improving the site monitoring plan
(including updates from the Local Health Department); assuring the presence of 3 species of
mosquitos at the testing site; clarifying how the volume of the test substance will be applied;

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clarifying skipped periods. CPT and landing pressure, delayed exposure periods; inclusion of
additional data elements on field data forms; explicit criteria for subject withdrawal and
replacement; clarification of statistical methods including Kaplan Meier Survival curves; and
clarification with respect to the power analysis and statistical design.
Recommendations:
HSRB would like EPA to consider the following additional recommendations in a revised
protocol.
Field site selection
We strongly support EPA's requirement that the test area be assessed for activity of all 3
mosquito species of interest prior to conducting the field testing. Failing to do so would seem to
increase the likelihood of having less than sufficient data at the end of the study. HRSB concurs
that a more detailed plan for site monitoring needs to be included in the protocol. In particular,
we recommend that a 'trap type' be added to the description of the methods for site monitoring.
The Study Director will be relying on information collected from the local health departments
and mosquito control districts (MCD). In addition to specifying how the Study Director will
coordinate with local these local agencies, please consider adding (as appendices) the SOPs for
the MCDs of the testing locations/sites to further clarify the methods of site surveillance.
Dose application
In addition to the aforementioned need for detail on who will apply the test substance to subjects,
HSRB further recommends that for consistency in application, it is best for one person to apply
this dose. However, HRSB recognizes that this may cause interference with consistent times of
application as a single individual may require time to work with each participant. HRSB
recommendation is then for the researchers to ensure as best as possible, consistency in the
application method for all participants.
Monitoring weather impacts

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The protocol indicates that the study will be stopped for certain conditions of rain and wind.
Study personnel should add to protocol that every effort will be made to check the weather
conditions ahead of time and only schedule the field study in good weather.
Alternate study designs
While not part of the required review, the HSRB noted that the May version of the Protocol
mentions a differing design of staggering time of 2 hours and 8 hours over two days. The HRSB
wants to recognize some concerns if this were to be considered: 1) there needs to be consistency
in how these studies are performed, 2) for the staggered individuals at 8 hours inside the
building, the repellent on the skin is not subjected to as many weather events, which may or may
not affect performance. In addition, EPA has mentioned that if for the 8 hours staggered
individuals, mosquito landing occurred immediately upon entering field, then an earlier CPT
could not be established. The HRSB agrees with this additional concern.
Statistical Review
With respect to the statistical aspects of the proposed study, the sample size for this study was
appropriately computed using EPA guidance and calculations. There will be 13 individuals
enrolled in the study with 2 untreated controls. Five alternates will be included for each product
under study for a total of 20 participants per product or 40 total participants. The participants will
be randomized to treatment or control within the males and within the females. Thus there will
be one female control and one male control. Kaplan-Meier survival functions are appropriate to
estimate the mCPT and 95% confidence intervals.
Recommendations
•	HSRB recommends additional clarification of the randomization scheme. Specifically,
detail to further clarify how the participants will be randomized to treatment or control is
suggested. For example, with regard to sex, it is unclear how it will be ensured that one
male and one female will be assigned as untreated controls
•	The end of field test period is indicated as occurring when "maximum duration reached
or sufficient number of subjects experience FCL". The stopping rule for the 'sufficient

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number of subjects experience FCL' should be further clarified to indicate 'at least 50%
of subjects experience FCL'.
• Clarification is needed with respect to how withdrawn participant data will be used in the
statistical analysis. The protocol indicates that data from participants that have withdraw
from the study will be used in the statistical analysis unless the participant requests that
their data is not used. In its recommendations, "EPA recommends that a withdrawn
subject whose data are right censored should not be replaced." The HSRB recommends to
explicitly state that the data from participants that withdraw will be considered right
censored in the statistical analyses. The definition of right-censoring should be clarified
in the protocol.
Charge to the Board - Ethics:
Is the research likely to meet the applicable requirements of 40 CFR part 26, subparts
K and L?
Response:
The research proposed in the protocol "Field evaluation of two topically applied insect repellent
products containing IR3535 against mosquitoes in Louisiana." and related documents is likely to
meet the applicable requirements of 40 CFR part 26, subparts K and L, if the recommendations
made by the EPA and HSRB are adequately addressed.
Ethics review:
The EPA has reviewed the aforementioned protocol for a field testing at a single site of two
formulations of skin-applied skin repellents containing IR3535 from both scientific and ethics
perspectives. This study is designed to determine the median complete protection time (CPT) for
AKIVA 20 lotion and wipe formulations of a skin-applied repellent containing IR3535. The
repellents will be tested against mosquitoes in the field. Ethical aspects of the proposed research
are assessed in terms of the standards defined by 40 CFR 26 subparts K and L.
The data collected in the study will be used to support product registration. The research has
societal value because people are at risk of contracting mosquito-borne diseases, and the data

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supporting currently registered skin-applied repellents do not show the efficacy of these
formulations of products containing IR3535. The rationale for this testing is to collect data to
show the efficacy of these products at repelling mosquitoes. As intended, the data resulting from
this proposed study will be used to support registration of one or more of these skin-applied
repellents.
The study team has implemented strategies and safeguards to minimize the risks of participants.
Overall, the study activities are informed by prior research in this field that has been previously
approved by the EPA and HSRB. The study products have been tested on human subjects in a
similar study in Florida, the dose and exposure levels of IR3535 for this study are well below
dermal toxicity levels and testing day safety procedures are thorough and well-documented.
The HRSB agrees that the testing periods should correspond with prior CPT findings for both the
lotion (14 hours) and the wipe (13 hours) rather than the proposed 16 hours in the protocol. This
will reduce unnecessary exposure to both the product and mosquitoes.
The HSRB agrees with the EPA recommendation that site landing pressure should be assessed
and confirmed prior to field testing and that the project team should coordinate with local health
officials to ascertain the presence of vector-borne illness at planned testing sites.
The HSRB agrees with increasing recruitment allowance to account for all potential participant
discontinuation or withdrawal scenarios.
With respect to subject selection and recruitment, the strategy for equitable selection of subjects
is adequate. The inclusion/exclusion criteria should be updated to reflect the suggestions put
forth in the EPA review. Specifically, further rationale needs to be given for the age range of
recruitment; specifically, if they agree with the EPA and will increase the age range beyond 55,
or their justification for keeping it at the proposed 18-55.
Recommendations:
COVID-19 precautions

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EPA has asked for COVID19 precautions to be included: wearing of masks and social
distancing, and in general adherence to any CDC, local or state requirements. The HRSB
discussed the following additional recommendations or considerations:
•	Testing for SARS-CoV-2 prior to participation, testing of study team members who will
be present on site can be considered. In addition, the HSRB recommends the
incorporation of screening questions and/or temperature checks as relevant to identify
study team members or participants at risk of infection or transmission.
•	Data forms can include where the subject is to be located for each study period in the
field, and where randomization is being used to place the participants at the site (i.e.,
seating layout of all subjects). This would facilitate contact tracing, if necessary.
•	The protocol should also discuss what steps will be taken to address participants who fail
to abide by social distancing and mask wearing provisions.
•	The consent form and recruitment script should also note that subjects will be required to
abide by mask wearing and social distancing requirements while participating in the
study.
•	The HSRB strongly supports EPA's comments concerning the issue of transportation to
and from the test site, which is only vaguely discussed in the protocol and not in the
consent form at all. Clarification is needed with respect to the options and/or
expectations of subjects in terms of driving (e.g., can they drive their own vehicle to the
test site if they prefer).
•	We recommend adding follow-up contacts post-study to ask about possible symptoms or
diagnoses. Furthermore, the collection of alternate contact information for someone who
can reach the study participant if contact tracing is necessary.
Prevention of heat stress
Further clarification on what participants will wear (light clothes vs. Tyvek) during testing
periods is required; otherwise, the study forms need to be harmonized and reflect the final
proposed design. The HRSB recommends that no Tyvek suits be used; it is our understanding
that Tyvek suits have not been utilized in other studies and present a heat stress risk. Due to heat
related concerns, add a question and exclusion criteria for potential volunteers concerning

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previous history of heat related injury (e.g. heat exhaustion, heat stroke, etc). Individuals that
have experienced this in the past are more prone to future events. Excluding them from the
volunteer pool could reduce overall risk to subjects. Furthermore, the HRSB recommends that
fans be used in the enclosure for the comfort of the participants.
Consent form modifications
The consent form, as submitted, is inadequate and should include the following information as
identified by the EPA:
•	More information about the study sponsor
•	More information about the pre-field-testing activities and requirements: attractiveness
test; aspirator training and verification; COVID precautions; relative location of test sites
and transportation thereto; and previously mentioned changes to compensation.
•	The HSRB recommends that the risk section be constructed in a table form so that it's
easier for participants to read, understand, and assess potential risks of involvement. If
risks are supported by prior research or literature, then such phrases as "slight risk"
should be further quantified (e.g., "less than 1% of people have X reaction") if possible.
The HSRB noted a number of inconsistencies across the protocol and supporting documents that
should be reconciled.
•	This include the factors that subjects are expected to refrain from the day before testing.
The consent form and protocol, and recruitment events should all consistently mention all
factors (i.e., spicy foods, not smoking etc.).
•	Likewise, a net over the head and neck is mentioned in some areas is inconsistently
presented across the materials. The protocol states that subjects will wear a head net and
gloves during the study. The HSRB suggests clarifying whether these are disposable
product and if new sets are provided to each subject.
•	In several places the protocol refers to spray, lotion or wipe but it appears in others that
only a lotion or wipe is to be used (there are also inconsistent references to 2 or 3
products). This should be corrected, as also noted by EPA.

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In the Protocol it is stated that the "Purity" of the test product is 20%. The expectation is
that the concentration of the AI in the product is 20% but that the purity (i.e.,
concentration relative to contaminants, degradants) of the AI is actually much higher.
This should be clarified.
In various parts of the protocol it is stated that subjects who are allergic to any ingredients
of the test material will be excluded (e.g., Protocol p. 7 emphasis added) but, aside from
the AI, the ingredients of the test material are never discussed. So how is this potential
allergy to be evaluated? If the check will be just against allergy to the AI, then that
should be how the statement is given.
It is stated on page 13 of the protocol that the results of the pregnancy test will not be
recorded yet elsewhere it is stated that negative results will be recorded (protocol, p. 21).
The protocol should be clarified for consistency.
Regarding the recruitment script "If you have a known allergy to insect bites, or have
participated in an insect repellent study in the last 3 months or insect repellent study in
the past 72 hours you will not be able to participate in the study. " The wording is unclear
and doesn't distinguish between the 3 month and 72 hour requirements. This is explained
better elsewhere. "If you are an employee, manager, or spouse of employees of Louisiana
State Univeristy and OR of the study sponsor, LivFul, you will not be able to participate
in the study." Note also that University is mis-spelled. EPA's final comment on the
"Survey Consent Form", final item "Signatures" is incomplete.
It is unclear if the study team has a detailed procedure for guaranteeing that participants
can use the aspirator with proficiency. For example, are a certain number of successful
aspirations required before including someone in the study? It is unclear with the
proficiency is tested or confirmed in the field.
HSRB recommends clarifying whether participants and alternates be paid for shortened
field-testing days (whether truncated by weather or inadequate landing pressure).
While, the justification for the inclusion of English-speaking participants only (written
materials are in English; study team speaks only English) appears acceptable. It is noted,
though, that the fact that there are no direct benefits to participants is not generally
considered justification for limiting study participation to people who speak a specific

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•	It is unclear whether participants who sustain "open cuts, scrapes, or skin problems"
during the course of a field-test be withdrawn from the study. As this is an exclusion
criterion, checking or querying for this before and during test days may be considered.
•	The HSRB recommends clarifying the role of the Medical Monitor in assessing adverse
events; it appears that the Study Director will have a large role in assessing and
interpreting adverse events but does not have a medical/clinical background.

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