Page 1

UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

 WASHINGTON, D.C. 20460 

OFFICE OF CHEMICAL SAFETY AND 

POLLUTION PREVENTION

 September 29, 2015 

MEMORANDUM 

SUBJECT: Ethics Review of Completed Study entitled, “Laboratory Evaluation of Bite 

Protection from Repellent-Impregnated Clothing for the United States 

Military”

FROM: Maureen Lydon, Human Research Ethics Review Officer

Office of the Director (on Detail)

Office of Pesticide Programs 

TO: Marietta Echeverria, Chief, Invertebrate-Vertebrate Branch 1

Registration Division

Office of Pesticide Programs

REF: Bernier, U.; Staeben, J.; Hummel, R. (2015) Laboratory Evaluation of Bite 

Protection from Repellent-Impregnated Clothing for the United States 

Military. Unpublished document prepared by the United States Department 

of Agriculture – Agricultural Research Service, Center for Medical, 

Agricultural, and Veterinary Entomology. July 30, 2015. 285 p. (MRID 

49684002) (D429130)

I have reviewed available information concerning the ethical conduct of the 

research study entitled, “Laboratory Evaluation of Bite Protection from RepellentImpregnated Clothing for the United States Military.” If the research is determined to be 

scientifically acceptable, I find no barrier in regulation to the U.S. Environmental 

Protection Agency’s (EPA’s) reliance on this study in actions under the Federal Insecticide, 

Fungicide, or Rodenticide Act (FIFRA) or the Federal Food, Drug, and Cosmetic Act 

(FFDCA). The Human Studies Review Board (HSRB) will be asked to comment on this 

study.

Completeness of Submission

All requirements of §26.1303 were satisfactorily addressed in the completion of this 

study as noted in the checklist in Attachment 1.
Page 2

Background and Ethics-related Chronology

The purpose of this study was to determine the bite protection level of the 

etofenprox-treated U.S. military Fire Resistant Army Combat Uniforms (FRACUs) treated 

initially at an application rate of approximately 0.9% (wt/wt), and to assess bite protection 

performance prior to washing (0x) and after washing 20 times (20x), 50 times (50x), and 75 

times (75x). Among the insecticide-treated uniform types that the U.S. military personnel 

use, the more open weave construction of the FRACU uniform makes this uniform the 

most difficult to prevent bite-through. It has relatively low efficacy compared to other 

military uniform types; as a result, it was selected as the worse-case scenario for treatment 

of military uniforms.

The protocol for this study was approved by the overseeing institutional review 

board, the Western Institutional Review Board (WIRB), and submitted to EPA for review. 

The protocol and EPA’s review, dated March 20, 2014, were discussed in a public meeting 

by the Human Studies Review Board (HSRB) on April 9, 2014. As documented in the 

final HSRB meeting report, dated June 25, 2014, the HSRB concluded, with respect to 

ethics, that “the protocol submitted for review, if modified in accordance with EPA...and 

HSRB recommendations, is likely to meet the applicable requirements of 40 CFR 26, 

subparts K and L.”

In follow-up to the April 9, 2014 HSRB meeting, the protocol, self-certification 

questionnaire, telephone screening script, consent form and advertisement were revised to 

address comments, including EPA and HSRB comments described in Attachment 2, and 

submitted to the Western Institutional Review Board (WIRB) for review and approval prior 

to initiating the study. The last WIRB approval for revisions was issued on July 20, 2015. 

The WIRB approvals which occurred from March 2015 through July 2015 and 

impacted the ethical conduct of the study are listed below, along with pertinent discussions 

with the U.S. Environmental Protection Agency (EPA): 

Chronology of WIRB Approvals

a) 3/19/15 – The study director submitted revisions to the protocol, subject selfcertification questionnaire, telephone screening script and consent form to the 

WIRB for review and approval. These revisions addressed EPA and HSRB 

comments.

b) 4/7/15 – WIRB approved the revised protocol dated 3/19/15, subject selfcertification questionnaire, telephone screening script, and consent form.

c) 5/14/15 – The U.S. Department of Agriculture’s Agricultural Research Service

(USDA/ARS) discussed with EPA’s Office of Pesticide Programs (OPP) their 

proposal to include 75 x wash cycle specimens in the study consistent with the rest 

of the protocol. 

Inclusion of 75x wash cycle:
Page 3

The inclusion of the 75x wash cycle specimens in this study, consistent with the 

protocol, resulted in not having to test 8 new subjects with controls in the future. 

The additional test time for each subject was 15 minutes for mosquito species 

Aedes aegypti (1 hour and 15 minute total test time) and 15 minutes for Anopheles

Albimanus (1 hour and 15 minute total test time). As proposed to EPA, the extra 

two tests would increase compensation from $200 to $250 and would be reflected in 

the revised consent form and advertisement. 

d) 5/21/15 – EPA approved the changes described above. The revised protocol, 

including the revised consent form and advertisement, with changes incorporated as 

noted above, were submitted to the WIRB for review and approval. Revisions 

submitted to the WIRB also addressed recommendations from the Quality 

Assurance (QA) advisor for Good Laboratory Practices (GLPs).

e) 5/28/15 – Using an expedited review process, the WIRB approved changes 

submitted on 5/21/15.

The May 21st protocol, which includes the WIRB-approved changes, is in Appendix 

I to the completed study submitted to the HSRB for review. 

f) 7/13/15 – The study director submitted two protocol amendments, described on 

pages 5-6 of this memo, to the WIRB, along with corresponding language revisions 

for the protocol and consent form. 

g) 7/20/15 – Using an expedited review process, the WIRB approved the revised 

protocol dated 7/13/15 and consent form reflecting the amendments.

The required forms requesting the aforementioned changes and the WIRB approval 

certificates are provided in separate files for the HSRB entitled “WIRB correspondence.”

Subject Recruitment 

A printed advertisement in the Gainesville Sun (local newspaper) and on bulletin 

boards in the University of Florida buildings recruited subjects from the general population 

in Gainesville, Florida. The advertisement briefly described the testing and financial 

compensation for participation. Subjects were compensated for participating in the initial 

consent meeting and for each set of sleeves worn in the testing paradigm. Subjects 

received $250 for completion of testing with the full set of 10 pairs of sleeves. The 

advertisement provided a phone number where interested respondents were instructed to 

leave a message. The study director reviewed the messages and called respondents, who 

underwent preliminary screening via telephone to determine if they met the inclusion/ 

exclusion criteria. (The study discusses preliminary screening on page 19 of the study.) 

None of the test subjects had a relationship with the study director or any test personnel.

Subject Inclusion/Exclusion Criteria

The following inclusion/exclusion criteria were used for subject selection:
Page 4

1. Must be between 18 – 62 years old.

2. Must speak and read English.

3. Children (under the age of 18) and pregnant or lactating women were excluded.

4. People in poor health or physical condition were excluded.

5. People hypersensitive to or phobic of mosquito bites were excluded.

6. People known to be sensitive to the test material, pesticides or other chemical 

products were excluded.

7. People with open cuts, scrapes or skin conditions (e.g. psoriasis or eczema) on their 

hands or forearms were excluded.

8. People with latex sensitivity or allergy were to be offered nitrile gloves.

9. People with a relationship to the study director or sponsor (students or employees of 

the study director or sponsor) were excluded.

Exclusion of Pregnant Women

Potential female subjects completed the home pregnancy test (HPT) in a private 

setting at the mosquito testing lab at the Center for Medical, Agricultural, and Veterinary 

Entomology (CMAVE). After self-reading the results, these participants were asked if they 

would like to continue as a participant (irrespective of results). Those requesting to 

continue were asked to show the test results to a female staff member for verification of 

negative results. The results were recorded in the raw data, but will be kept confidential.

Random Selection of Subjects

A list of at least 20 qualified potential subjects was created after the telephone 

screening process. Eight test subjects were chosen at random from the list and enrolled for 

testing. The remaining qualified subjects were considered to be alternates. During the 

course of the study, a ninth subject was enrolled to supplant one of the original eight 

subjects. After completion of testing with the initial four female subjects, inspection of the 

data revealed that the non-treated control mosquito rate for both mosquito species was 

extremely low for one female subject. Rather than retest this subject against both species, 

data were acquired from a fifth female subject, selected from the list of alternates, to satisfy 

minimum testing requirements. Due to one alternate being enrolled, there were nine 

subjects, including four male and five females. None of the subjects withdrew from the 

study.

Informed Consent 

The study director and each subject held consent meetings; a representative of the 

study sponsor attended in order to pay subjects for participating in the consent meeting. 

Respondents were asked to complete self-certification forms attesting that their responses 

given during the telephone pre-screening were true and accurate. The study director 

provided a detailed explanation of study procedures and the consent form, outlining risks of 

participating in the study. The participants were informed of how many bites they were 

likely to obtain and what symptoms of arthropod-borne reactions they should be alert for 
Page 5

after participating in the study. (The representative of the study sponsor only commented 

during the study director’s explanation if she believed a particular point needed to be 

added.) There was consistency in information delivered in each consent meeting. After the 

detailed explanation of the procedures, the study director escorted the subject to the 

conference room where the video of the testing process was set up; a member of the study 

director’s trained staff played the video of the testing process. (The study staff members 

were the same trained individuals who helped to implement the study.) The candidate was 

allowed to ask questions at any time, including afterwards. After the video was viewed and 

questions were answered, the study director recalled the sponsor’s representative to provide 

payment for participating in the consent meeting. The subject then left with their 

compensation for attending the consent meeting and their unsigned consent form.

The process allowed sufficient opportunity for subjects to consider whether or not 

to participate before signing the consent form. On the day of testing, subjects entered the 

laboratory with their signed consent form and verbally confirmed their intent to engage in 

the study. The signed consent form was then accepted by the study director. Two copies 

were made of the consent form and one copy was provided to the subject. (The second 

copy, with the subject’s name and signature blacked out, was inserted into the study 

notebook for documentation that the subject provided consent.)

Protocol Amendments

The study protocol, dated May 21, 2015, was amended twice after it was approved 

by the Western Institutional Review Board (WIRB) on May 28, 2015. The modifications 

did not negatively affect participants’ rights, health or safety. As reflected in the “WIRB 

Correspondence” files provided to the HSRB, the protocol amendments were approved by 

the Western Institutional Review Board (WIRB) on July 20, 2015, using an expedited 

review process, and are summarized as follows:

Amendment 1:

• Approve additional subject to be tested to replace Subject 3 for a total of 9 subjects.

• Reason: Additional testing of subject was to ensure that data was collected with 

mosquitoes behaving avidly for each subject. After completion of testing for 

Subject 3, inspection of the data set revealed that the non-treated control mosquito 

bite rate was extremely low for this subject. Data from a 9th subject was to be used 

to satisfy testing requirements. 

• Additional Ethics-related Information: Consistent with the compensation section of 

the protocol, Subject 3 was compensated the full amount of $250.

Amendment 2:

• Allow Subject 4 to be retested against a mosquito species because it was known that 

a set of data was collected with mosquitoes that had been incorrectly maintained.
Page 6

• Reason: Additional testing of subject was to ensure that data were collected with 

mosquitoes behaving avidly for each subject. Subject 4 had one set of sleeves with 

very low bite through amounts for controls. It was determined that this was the 

result of mosquitoes that had been incorrectly maintained and therefore did not 

respond as avidly. The subject was willing to be retested, and the results from the 

retest were used in place of the original results, thereby allowing analysis of data 

based on mosquitoes that were not compromised.

• Additional Ethics-related Information: Subject 4 was compensated at the rate of 

$125 for retesting 5 pairs of sleeves and therefore received an additional $125 

above the $250 for the original tests. The same precautions were observed for the 

retest as for the original test. There was a break of about three weeks between the 

original test and the retest.

The amendments did not negatively affect participants’ rights, health or safety.

WIRB Approvals in Response to Amendments 

Additional Language Approved for Protocol

On July 20, 2015, the WIRB approved additional language for the protocol in 

response to the amendments. The latest WIRB-approved clean version of the protocol is 

provided in a separate electronic file for the HSRB; the electronic file is dated July 20, 

2015. (For the HSRB’s convenience, the same protocol, in track changes format, with the 

WIRB-approved changes highlighted, is also provided and dated July 13, 2015.) 

The additional language approved by the WIRB for the revised protocol includes the 

following:

1) In section 3.6, under “Summary of Experimental Design” the following statement 

was added: “Should it be determined that tests have been conducted with mosquitoes 

that are not behaving properly (e.g. very low control biting rates), then a subject may 

be asked to repeat the test set.”

2) In section 8.1.1, under “Recruitment,” the following statement was added: “In 

the event that a subject repeats a set of sleeves, then they will be compensated at 

the rate of $25 per pair.”

3) In section 8.4, under “Summary of Human Subject Test Procedure,” the 

following statement was added: “Should a situation arise where mosquito bite 

through rates are very low for a test set, and the cause of this [is] even suspected to be 

due to unhealthy mosquitoes, then a subject may be asked to repeat tests with an entire 

set of sleeves, against the mosquito species in question.”
Page 7

(On page 24 in the red-line/strike-out version of the revised protocol, a fourth sentence is 

highlighted. This statement is not new to the protocol; instead, it was simply moved from a 

different location.)

Additional Language Approved for Consent Form

Related to the protocol amendments, on July 20, 2015, the WIRB approved similar 

revisions to the consent form. The most recent version, with the July 20th WIRB approval 

stamp, is provided in a separate file for the HSRB. The additional language approved by 

the WIRB for the revised consent form includes the following:

1) At the end of the study procedures section, language was added which reads: “In the 

event that we discover that the mosquito bite rates for a set of tests is too low, we may 

ask you to repeat the tests on another day. If you agree, we will ask you to test the 

control sleeves and 4 sets of treated sleeves. This additional testing will take about 2 

hours.”

2) Under costs and payment, the following clarification was added: “If you are 

repeating a set of tests, you will be paid at the same rate of $25 per pair.”

Application Rate Amendment

There was another change in implementing the protocol that EPA recommends the 

study director report to the WIRB. The original protocol stated that etofenprox-treated 

U.S. military Fire Resistant Army Combat Uniforms (FRACUs) would be treated initially 

at an application rate of 1% wt/wt, and not at a rate of approximately 0.9% (wt/wt), as 

indicated in the final study. This change did not negatively impact participants’ rights, 

health or safety, but should be reported to the WIRB.

Follow-up action by EPA: 

EPA will ask the study director and sponsor to ensure that all amendments are 

reported to and approved by the WIRB in future studies.

Protocol Deviations 

The study documents eight protocol deviations, on pages 228-230, which occurred 

during 2015. The study director consulted with the WIRB with regard to their guidance on 

reporting deviations. The WIRB confirmed for the study director that the nature of these 

deviations did not require prompt reporting to the WIRB within five days; protocol 

deviations which require prompt reporting include those that harmed a subject or placed a 

subject at risk of harm and protocol deviations made without prior IRB approval to 

eliminate an immediate hazard to a subject. (If you look at the second page of the WIRB 

Certificates of Approval provided to the HSRB in the separate files entitled “WIRB 

Correspondence,” item # 7 outlines the information that must be reported to the WIRB 

within five days.)
Page 8

The eight protocol deviations which occurred during 2015 are outlined below:

Deviation 1

• A sponsor representative attended consent interviews to pay subject candidates in 

order to streamline the interview process.

Additional Ethics-related Information for HSRB:

The language of the protocol stipulated that the study director would perform the 

consent meeting and did not mention that a sponsor’s representative might sit in in 

order to pay subjects for participating in the consent meeting. 

The sponsor representative honored the confidentiality of the consent process. No 

information about the subject’s identity or participation in the study was disclosed 

by the sponsor representative as a result of her attending the consent interview.

Also, the consent form indicated that information from the study would be given to 

the sponsor.

Related to the consent meeting, the protocol did not explicitly state that the study 

director’s trained staff would play the video of the testing process or respond, along 

with the study director, to subjects’ questions after they watched the video; this

occurred during the study. Section 8.1.6 of the protocol stated, “During the consent 

meeting, respondents will be given a detailed explanation of the procedures of the 

study and be asked to watch a movie of the testing process.”

Follow-up action by EPA:

Deviation 1 did not negatively impact the rights of the subjects or the informed 

consent process. However, EPA will request that the study director and sponsor 

adhere closely to the specifics of the consent meeting and consent process, as 

explicitly outlined in approved protocols, in future studies.

Deviation 2

• Chain of custody letters were not shipped with fabric samples from Warmkraft, Inc. 

and Natick Soldier Research, Development and Engineering Center (NSRDEC).

Deviation 3

• NSRDEC provided chemical analysis information to CMAVE prior to the biteprotection assay. The reason was to determine whether fabric swatches were 

treated in accordance with the label rate.

Deviation 4

• Some mosquito test populations were outside of the range required by the protocol 

due to inaccurate estimation of the number of females collected for testing.

Deviation 5
Page 9

• Sleeves from the coat fabric were placed on the left arm and sleeves from the 

trouser fabric were placed on the right arm. This was an oversight; previous 

protocols consistently had coat fabric placed on the right arm and trouser fabric 

places on the left arm.

Deviation 6

• The number of blood-fed mosquitoes per each set of tested mosquitoes was 

determined by counting the number of blood-fed mosquitoes twice instead of in 

triplicate. This was an oversight. As a result, counting ceased once the same 

number of blood-fed mosquitoes was determined twice.

Deviation 7

• Human subject number codes were not preceded by a gender code (M or F) as 

stated on page 27 of the protocol. However, the gender of the participants was still 

noted using the gender code (M or F) in a separate column on the data sheet.

Deviation 8

• Bite protection data were not analyzed using a generalized linear model (GLiM) 

with a log link. GLiM-based confidence intervals are inappropriate when there is 

subject-to-subject variation. A t-distribution confidence interval was used instead 

because it provided more accurate confidence intervals.

The deviations, listed above, did not negatively affect participants’ rights or their 

health or safety, and did not adversely impact the scientific integrity of the study. 

Regulatory and Statutory Standards 

The following provisions of 40 CFR 26 Subpart Q, as amended, define the 

applicable ethical standards, which read in pertinent part: 

§26.1703: Except as provided in §26.1706, EPA must not rely on data from 

any research subject to this subpart involving intentional exposure of any 

human subject who is a pregnant woman (and therefore her fetus), a nursing 

woman, or a child. 

§26.1705: Except as provided in §26.1706, EPA must not rely on data from 

any research subject to this section unless EPA determines that the research 

was conducted in substantial compliance with all applicable provisions of 

subparts A through L of this part. . . . 

In addition, §12(a)(2)(P) of the Federal Insecticide, Fungicide and Rodenticide Act 

(FIFRA) applies. This passage reads: 

In general, [i]t shall be unlawful for any person . . . to use any pesticide in 

tests on human beings unless such human beings (i) are fully informed of 

the nature and purposes of the test and of any physical and mental health 
Page 10

consequences which are reasonably foreseeable therefrom, and (ii) freely 

volunteer to participate in the test. 

Findings 

Prior HSRB and EPA Review

Because this study was initiated after April 7, 2006, prior submission of the 

protocol and supporting materials to EPA was required by 40 CFR §26.1125. The 

requirements of 40 CFR §26.1125 for prior submission of the protocol to EPA and of 

§26.1606 for HSRB review of the protocol were satisfied. The study protocol was approved 

by the Western Institutional Review Board (WIRB) prior to submittal to EPA. The HSRB 

discussed the protocol at its April 9, 2014 meeting, and concurred with EPA’s assessment 

that the protocol, if revised as suggested by the Agency and the HSRB, would meet the 

applicable requirements of 40 CFR part 26, subparts K and L. 

Responsiveness to HSRB and EPA reviews

EPA’s and the HSRB’s ethics comments on the protocol were addressed before the 

research was conducted. Please see Attachment 2 for details. 

Prohibition of research involving intentional exposure of pregnant or nursing 

women or of children 

All enrolled subjects were at least 18 years old and there were no female subjects. 

The prohibition in 40 CFR §26.1703 of research involving intentional exposure of pregnant 

or nursing women or of children under age 18 was satisfied. 

Substantial compliance with 40 CFR 26 subparts A through L 

40 CFR §26.1705 requires that EPA not rely on data from any research subject to 

this section unless EPA determines that the research was conducted in substantial 

compliance with all applicable provisions of subparts A through L of this part. Within this 

range, only subparts K and L are directly applicable to the conduct of third-party research

such as this. The study documents substantial compliance with subparts K and L. 

Compliance with 40 CFR 26 subpart M 

As documented in Attachment 1 to this review, the central requirements of 40 CFR 

26 subpart M, §26.1303 to document the ethical conduct of the research were satisfactorily 

addressed. 

Compliance with FIFRA §12(a)(2)(P) 

The requirement of FIFRA §12(a)(2)(P) that human subjects of research be “fully 

informed of the nature and purposes of the test and of any physical and mental health 
Page 11

consequences reasonably foreseeable therefrom,” and “freely volunteer to participate in the 

test,” was met for this study. 

EPA Conclusion 

The study entitled, “Laboratory Evaluation of Bite Protection from RepellentImpregnated Clothing for the United States Military,” reports research conducted in 

substantial compliance with the applicable requirements of 40 CFR 26, subparts K and L. 

The conduct of the study met all applicable ethical standards for the protection of human 

subjects of research, and requirements for documentation of ethical conduct of the research 

were satisfied. 

For future studies, EPA will recommend to the study director and sponsor that they 

closely adhere to the details of the consent process as described in the approved protocol 

and ensure that all amendments are reported to the WIRB. 

If this study is determined to be scientifically valid and relevant, there is no 

regulatory barrier to EPA reliance on it in actions under FIFRA or §408 of FFDCA. 

Attachment 1: §26.1303 completeness check

Attachment 2: Responsiveness to EPA and HSRB Comments on Protocol
Page 12

Attachment 1

§ 26.1303 Checklist for Completeness of Reports of Human Research Submitted for EPA Review

Any person who submits to EPA data derived from human research covered by this subpart shall provide at the time of submission 

information concerning the ethical conduct of such research. To the extent available to the submitter and not previously provided to 

EPA, such information should include:

Requirement Y/N Comments/Page References

(a) Copies of all of the records relevant to the research specified by §26.1115(a) to be prepared 

and maintained by an IRB

§1115(a)(1): Copies of 

• all research proposals reviewed, 

• scientific evaluations, if any, that accompany the proposals, 

• approved sample consent documents, 

• progress reports submitted by investigators, and reports of injuries to 

subjects.

Y

n/a

Y

n/a

§1115(a)(2): Minutes of IRB meetings which shall be in sufficient detail to show 

• attendance at the meetings; 

• actions taken by the IRB; 

• the vote on these actions including the number of members voting

for, against, and abstaining; 

• the basis for requiring changes in or disapproving research; 

• a written summary of the discussion of controverted issues and their 

resolution.

Y Separately provided to HSRB 

members.

§1115(a)(3): Records of continuing review activities. Y

§1115(a)(4): Copies of all correspondence between the IRB and the 

investigators. Y

WIRB correspondence which 

occurred in follow-up to the April 

9, 2014 HSRB meeting was 

provided in a separate file to the

HSRB.

§1115(a)(5): 

• A list of IRB members identified by name; earned degrees; 

representative capacity; indications of experience such as board 

certifications, licenses, etc., sufficient to describe each member’s chief 

anticipated contributions to IRB deliberations; 

• any employment or other relationship between each member and the 

institution

Y

Y

Previously provided to HSRB 

members with the protocol. 

Updated information was included 

in a separate file provided to 

HSRB members with the 

completed study.

§1115(a)(6): Written procedures for the IRB in the same detail as described in § 

26.1108(a) and § 26.1108(b). Y Previously provided to EPA with the protocol.

§1115(a)(7): Statements of significant new findings provided to subjects, as 

required by § 26.1116(b)(5). n/a

Pls. 

see

note

Note: As discussed in the study 

and EPA review, one subject had 

to be retested. This does not 

constitute a significant new finding 

from the entire study for all 

subjects; it does constitute a 

finding which needed to be 

provided to one subject.

(b) Copies of all of the records 

relevant to the information identified 

in §26 1125(a) (f)

§1125(a)

A discussion of:

(1) The potential risks to human subjects; Y Discussed in consent form and p. 

39-40 of revised protocol.

(2) The measures proposed to minimize risks to the human subjects; Y Discussed in consent form and p. 

39-40 of revised protocol.

(3): The nature and magnitude of all expected benefits of such research, 

and to whom they would accrue; Y Discussed in consent form and p. 41 of revised protocol.

(4) Alternative means of obtaining information comparable to what would 

be collected through the proposed research; and Y Discussed in consent form and p. 42 of revised protocol.

(5) The balance of risks and benefits of the proposed research. Y Discussed on p. 41 of revised 

protocol.

§1125(b): All information for subjects and written informed consent agreements 

as originally provided to the IRB, and as approved by the IRB. Y Provided in separate file to EPA and HSRB

§1125(c): Information about how subjects will be recruited, including any 

advertisements proposed to be used. Y

Pages 18-19 of study, Appendix 1 

and pages 19-20 of revised 

protocol.
Page 13

Requirement Y/N Comments/Page References

§1125(d): A description of the circumstances and methods proposed for 

presenting information to potential human subjects for the purpose of obtaining 

their informed consent.

Y

Page 20 of study, revised consent 

form, and p. 21 of revised 

protocol.

§1125(e): All correspondence between the IRB and the investigators or 

sponsors.

Y

Separate file provided to HSRB 

with requests sent to WIRB and 

certificates of approval from 

WIRB.

§1125(f): Official notification to the sponsor or investigator, in accordance with 

the requirements of this subpart, that research involving human subjects has 

been reviewed and approved by an IRB.

Y

(c) Copies of sample records used to document informed consent as specified by 

§26.1117, but not identifying any subjects of the research Y Revised consent form approved by WIRB was provided to HSRB.

(d) If any of the information listed in paragraphs (a) through (c) of this section is not 

provided, the person shall describe the efforts made to obtain the information. n/a
Page 14

Attachment 2 - Responsiveness to EPA’s and HSRB’s Ethics Comments

Comments from EPA and HSRB Has comment been addressed?

1. In section 8.6 of the protocol, entitled 

“Detailed Stepwise Test Procedure,” insert an 

explanation of how, when, and by whom 

subjects’ hands and arms will be inspected for 

cuts or other skin conditions. 

Yes. The following statement was added to 

step 3 of section 8.6 in the protocol: “The 

laboratory technician or study PI will request 

the subject wash their hands with soap and 

water and then inspect the arms of the subject 

for cuts and scrapes.”

2. In section 8.6 of the protocol, entitled 

“Detailed Stepwise Test Procedure,” insert an 

explanation of how the pregnancy testing for 

female subjects will be handled, clarifying 

where in the sequence of stepwise procedures 

the pregnancy testing will occur. 

Yes, in section 8.6, the following statement 

was added to step 3: “If the participant is 

female, they will be required to take a 

pregnancy test as following the procedures of 

section 8.1.4.”

3. Please clarify which member of the research 

team will verify pregnancy test results of 

female subjects who desire to remain in the 

study after taking the pregnancy test.

Yes, section 8.1.4 of the protocol was revised 

to read in part, “Those requesting to continue 

will be asked to show the test results to a 

female staff member for verification of 

negative results.”

4. Please revise the first sentence in section 11.0 

of the protocol, titled “Benefits and to Whom 

Benefits Accrue,” as follows: “While there are 

no direct benefits to the subjects participating 

in this research study beyond a small 

compensation for their time, there are...” The 

proposed payment to subjects is considered 

compensation for lost time and inconvenience, 

not a benefit of participating in the research. 

This study provides no direct benefits to 

subjects.

Yes, the reference to compensation in this 

sentence was deleted from the protocol as 

requested. The revised sentence reads, 

“While there are no direct benefits to the 

subjects participating in this research study, 

there are indirect benefits to both the subjects 

and society.”

5. In section 8.1.2 of the protocol, titled 

“Inclusion/Exclusion Criteria,” consider 

expanding exclusion #6 as follows: “Exclusion 

of people known to be sensitive to the test 

material, pesticides, or other chemical 

products.”

Yes, exclusion criteria #6 was expanded as 

requested.

6. In section 8.1.2 of the protocol, titled 

“Inclusion/Exclusion Criteria”, add an 

exclusion for people with open cuts or scrapes 

or skin conditions such as psoriasis or eczema 

on their hands or forearms. Cut, scrapes or 

other skin conditions might increase the risk of 

skin reactions or sensitivity during the testing. 

Also, add this exclusion to the “Restrictions” 

section of the consent form. 

Yes, the requested exclusion was added as 

exclusion #7. The exclusion for people with 

cuts, scrapes or skin conditions was also 

added to the restrictions section of the

consent form.

7. In the “Study Procedures” section of the 

consent form, insert text similar to what 

appears below as #2, and adjust the numbering 

of the subsequent procedures accordingly: “2. 

Yes, the requested language was added to the 

consent form and the steps which followed in 

the study procedures section were 

renumbered accordingly.
Page 15

It is important that you NOT be in this study if 

you are pregnant. So, before the testing 

begins, each female volunteer will be asked to 

go to a private area and will be given a home 

pregnancy test kit. A female researcher will be 

able to explain how to use it and answer 

questions. If you are a female, you will be 

asked to take the test in a private restroom. If, 

after taking the test, you still wish to 

participate in the study, you will be asked to 

show the result of the test to a female 

laboratory technician so that she can verify 

that you are not pregnant. If you withdraw 

from the study after taking the pregnancy test, 

you will not be asked to share the result of the 

test with anyone.”

8. Although a question about allergy/sensitivity 

to latex is listed on the enrollment 

questionnaire, latex allergy/sensitivity does 

not appear as an exclusion criterion in the 

recruitment interview script or consent 

document. Latex allergy/sensitivity should be 

included in all statements of exclusion criteria. 

Alternatively, the protocol could be modified 

to use nitrile gloves instead of latex.

Yes, the language, “People with latex 

sensitivity or allergy will be offered nitrile 

gloves” was added to inclusion/exclusion 

criteria in the protocol and the restrictions list 

in the consent form.

9. The protocol currently calls for test sleeves to 

be made in a single size, despite the likely 

variation in arm size among participants and 

34 different uniform sizes available to military 

personnel. The single sleeve size may both 

skew the results of the testing across 

participants of various arm sizes and it may 

also cause embarrassment to participants who 

attempt to put on sleeves that do not fit easily. 

The Board recommended that researchers 

include a question about shirt sleeve size in 

the recruitment interview and have test sleeves 

available in different sizes.

Yes, as discussed on p. 20 of the study, a

range of sleeve sizes, designated small, 

medium and large, were constructed prior to 

study initiation. The fabric dimensions were 

altered by one-half inch increments, as 

needed, to yield sewn sleeves that were snug 

on the forearms of subjects. Due to the 

ability to alter the sleeves as noted, it was 

unnecessary to include a question about shirt 

sleeve size in the recruitment interview.

10. Currently, stopping rules depend on 

researchers’ judgment on key criteria of 

subjects’ attractiveness and sensitivity, and 

mosquitoes’ biting pressure. The Board 

recommended that the investigators articulate 

objective measures for stopping criteria.

Yes, stopping rules in the protocol were 

expanded in response to this comment. The 

revised stopping rules in section 14 of the 

protocol read as follows: “The study will be 

stopped if the test site becomes unsafe for any 

reason, biting pressure falls below threshold 

needed (<50% of the mosquitoes in a cage 

contact the fabric worn by a subject), biting 

pressure rises too high for subject comfort 

(expressed verbally by subject) or safety, 

subject asks to withdraw irrespective of the 

point they are in the study, subject is 

unattractive to target species (<50% of 

mosquitoes land on fabric surface during test 
Page 16

interval), subject exhibits hypersensitivity to 

insect bites during test (large areas of 

swelling generally over 0.5 cm per bite), 

subject exhibits sensitivity to the test 

materials during the test (redness, swelling or 

other skin reaction), study is terminated or 

discontinued.”

11. The Board noted that clarification is needed in 

the consent form about a nurse being on call 

but not present at the study site.

The study director’s interpretation of the 

comment was that it wasn’t clear on the 

consent form that the nurse was on call; 

therefore, language was added to indicate 

this. Page 39 of the original and revised 

protocol included language indicating that the 

nurse would be on call. The original consent 

form submitted to the HSRB with the 

protocol did not include a reference on the 

first page that a nurse would be on call. This 

language was added to the consent form, 

along with the name and telephone number of 

the on-call nurse. 

12. Currently, partial (per sleeve) compensation is 

provided to participants who leave the study 

early for any reason. The HSRB recommended 

that, to prevent coercion or to compensate for 

dropping out or being excluded by the 

Principal Investigator (PI), full compensation 

be provided to all participants.

With respect to payment of subjects and 

prevention of coercion, the study director 

stated that he tried to ensure that the study did 

not include any factors that could lead to 

coercion. The payment system was 

established for fair compensation of

participants for the time spent on the study 

and to pay subjects for each set of sleeves on 

which they initiated testing; the approved 

consent form states, “If you begin the study 

but do not complete it, you will be paid for 

each pair of sleeves at a rate of $25 per pair”. 

If a subject was to start a study and withdraw 

for any reason, they would still be 

compensated for any sleeves tested, 

regardless of whether or not they finished the 

15 minute test period. This represented a 

compromise to compensate participants, 

regardless of their ability to complete a full 

set of sleeves. In fairness to other 

participants who completed testing on all sets 

of sleeves, a participant who did not complete 

a full set of sleeves had less time invested, 

and therefore did not receive the same level 

of compensation as a participant who 

completed all sets. 

13. Language in the consent form indicating that 

laboratory mosquitoes pose no risk of 

transmitting disease needs to be modified. To 

address the unlikely possibility that any of the 

test mosquitoes is found to carry a disease, 

investigators should articulate a standard 

Yes, the language in the protocol under “risk 

of exposure to disease” was revised in 

response to this comment. This section 

currently reads, “There are risks of getting 

sick when you are bitten by mosquitos in the 

outdoors. The test mosquitos for this study 
Page 17

method and message for notifying participants 

of exposure.

have been grown in the laboratory and should 

not have had a chance to carry disease. In the 

unlikely event that they are found to carry 

disease, you will be notified immediately and 

will receive appropriate treatment at the 

hospital.”

14. It is recommended that researchers complete a 

course in human subject protections within 

three years of study initiation and completion. 

Depending on when the study occurs, some 

investigators may exceed this recommended 

time limit.

Yes, the study director and his research staff 

completed a number of courses, including 

two University of Florida courses (HIPAA 

for Researchers) and an IRB training course. 

The latter course is every 3 years. They also 

completed NIH training. They updated their 

training earlier this year per the 

recommendation of the HSRB.