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UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

WASHINGTON D.C., 20460 

OFFICE OF CHEMICAL SAFETY AND 

POLLUTION PREVENTION 

March 20, 2014 

MEMORANDUM

SUBJECT: Science and Ethics Review of USDA Protocol for Laboratory Evaluation of Bite 

Protection from Repellent-treated Clothing for the United States Military 

FROM: Kevin J. Sweeney, Senior Entomologist 

 Registration Division 

 Office of Pesticide Programs 

Kelly Sherman, Human Research Ethics Review Officer 

 Office of the Director 

 Office of Pesticide Programs 

TO: Deborah McCall, Chief, Insecticides Branch 

 Registration Division 

 Office of Pesticide Programs 

REF: Bernier, U. (2014) Laboratory Evaluation of Bite Protection from RepellentImpregnated Clothing for the United States Military. Unpublished document 

prepared by United States Department of Agriculture – Agricultural Research 

Service, Center for Medical, Agricultural and Veterinary Entomology. March 7, 

2014. 210 p. 

We have reviewed the referenced protocol for a laboratory test of mosquito repellentimpregnated clothing for the United States Military from both scientific and ethics perspectives. 

This review assesses the scientific aspects of the proposed research for a special efficacy study to 

assess etofenprox-treated U.S. Military uniforms in terms of the recommendations of the EPA 

and of the EPA Human Studies Review Board. Ethical aspects of the proposed research are 

assessed in terms of the standards defined by 40 CFR 26 subparts K and L and the 

recommendations of the EPA Human Studies Review Board. 
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A. Completeness of Protocol Submission 

The submitted protocol was reviewed for completeness against the required elements 

listed in 40 CFR §26.1125. EPA’s checklist is appended to this review as Attachment 5. All 

elements of required documentation are provided in the submitted protocol package. 

Volume 1 of USDA’s submitted package includes the following documents—all 

considered in this review: 

 WIRB-approved protocol dated 2/28/14 (pp. 5-192) 

 WIRB-approved informed consent form (approved 3/04/14) (pp. 193-203) 

 WIRB-approved recruiting advertisement (p. 204) 

 WIRB-approved telephone script for receiving phone calls in response to 

advertisement (p. 205) 

Volume 2 of USDA’s submitted package includes documentation of communications 

with WIRB, as well as copies of CVs and ethics training certifications for the principal 

investigator and staff. 

Volume 3 of USDA’s submitted package provides statistics analysis for the proposed 

study. 

B. Summary Assessment of Ethical Aspects of the Proposed Research 

Here is a summary of our observations about the ethical aspects of the proposed protocol. 

Supporting details are in the attachment. 

1. Societal Value of Proposed Research: This study is designed to determine the bite 

protection level of etofenprox-treated U.S. Military Fire Resistant Army Combat 

Uniforms (FRACUs) treated initially at an application rate of 1% wt/wt, and to assess the 

bite protection performance after 0x, 20x, and 50x washes. The results of this research 

will allow for determination of whether etofenprox-treated FRACUs meet the 

Department of Defense’s specifications for minimum bite protection level. The research 

has societal value because U.S. military personnel serving domestically and abroad are at 

risk of contracting insect-transmitted diseases. Currently, permethrin is the only pesticide 

that is EPA registered for treatment of military uniforms. Due to global issues regarding 

disease vector resistance to pyrethroids such as permethrin, other insecticide candidates 

must be considered. This protocol provides a method for evaluating the bite protection of 

fabrics that are treated or impregnated with substances that repel or reduce arthropod 

bites. 

 

2. Subject Selection: Ten adult subjects will be recruited from the general population in 

Gainesville, Florida (8 initially assigned for participation, plus two alternates). 

Participants will self-identify in response to a printed advertisement in a local newspaper 

(the Gainesville Sun) or posted on bulletin boards in University of Florida buildings. 

Callers responding to the advertisement will be screened, scheduled for informed consent 
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meetings, and enrolled. While it is possible that people who respond to the advertisement 

are different in some unknowable ways from those who do not respond, there is no reason 

to think that respondents in the Gainesville, Florida area are not typical of people who 

would respond to these types of advertisements in other areas of the United States. 

3. Risks to Subjects: Risks to subjects include the risk of exposure to biting mosquitoes; 

the risk of exposure to disease vectors; the risk of exposure to the test material; risks 

related to receiving an unexpected result on a pregnancy test; and the risk of a loss of 

confidentiality related to the pregnancy test and subjects’ participation in the study. Risks 

are minimized in the protocol by excluding candidates known to be hypersensitive to or 

phobic of mosquito bites; using disease-free colony-raised mosquitoes; excluding 

candidates known to be sensitive to the test material; applying clear stopping rules; and 

by incorporating procedures to keep the subjects’ identities and results of pregnancy 

testing private, and to permit discrete withdrawal. All practical steps to minimize subject 

risks have been taken, and the remaining risks have a low probability of occurrence. 

4. Benefits: This research offers no direct benefits to subjects, but may provide indirect 

benefits to subjects and society by providing data that could be used by EPA to register a 

novel repellent treatment for military clothing, thereby allowing for better protection of 

American military forces from nuisance bites and bites that lead to arthropod-borne 

diseases. The results may also lead to better protection of U.S. civilians both domestically 

and while they travel abroad because novel repellents may be incorporated into civilian 

clothing as a potentially better alternative to the repellent-treated clothing already 

available on the market. 

5. Risk/Benefit Balance: No practical opportunities to further reduce risk to subjects while 

maintaining the robustness of the scientific design have been overlooked. The residual 

risk to subjects is very low, and reasonable in light of the potential benefits of the data to 

society. 

6. Independent Ethics Review: The Western Institutional Review Board (WIRB) has 

reviewed and approved the protocol, informed consent form, and recruitment materials. 

WIRB is independent of the investigators and sponsors. Satisfactory documentation of 

WIRB procedures and membership is on file with the Agency and has been provided to 

the HSRB members with the background materials for this protocol. 

7. Informed Consent: The protocol contains a complete and satisfactory description of the 

process by which potential subjects will be recruited and informed, and the process for 

seeking their consent to participate. A copy of the WIRB-approved consent documents 

meeting all requirements of 40 CFR §§26.1116 and 26.1117 is included in the proposal. 

8. Respect for Subjects: Subject-identifying information will be recorded only once; all 

subsequent data records and reports will refer to individual subjects only by an arbitrary 

code. Provision is made for discrete handling of the pregnancy testing that is required of 

female subjects on the day of testing. Candidates and subjects will be informed that they 

are free to decline to participate or to withdraw at any time for any reason. Subjects who 
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withdraw will be compensated for time spent up to the point of withdrawal. Medical care 

for research-related injuries will be provided by the sponsor Landis International at no 

cost to the subjects. 

C. Compliance with Applicable Ethical Standards 

This is a protocol for third-party research involving intentional exposure of human 

subjects to a pesticide, with the intention of submitting the resulting data to EPA under the 

pesticide laws. Thus the primary ethical standards applicable to this proposal are 40 CFR 26, 

Subparts K and L. In addition, the requirements of FIFRA §12(a)(2)(P) for fully informed, fully 

voluntary consent of subjects apply. A point-by-point evaluation of how this protocol addresses 

the requirements of 40 CFR 26 Subparts K and L and the criteria recommended by the HSRB is 

appended as Attachment 1. 

EPA Ethics Comments

Before the research is conducted, the documents should be revised as follows and 

resubmitted for review by the approving IRB: 

 In section 8.6 of the protocol (V1:33), titled “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. Language similar to what appears on page 4 of the informed consent 

(V1:196) form would suffice. 

 In section 8.6 of the protocol (V1:33), titled “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. Language 

similar to what appears on page 3 of the informed consent form (V1:195) or in section 8.1.4 

of the protocol (V1:24) would suffice. 

 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. The protocol and 

consent form are inconsistent on this point. Section 8.1.3 of the protocol says that the result 

must be presented “to a staff member” (V1:24), and section 8.1.4 of the protocol states that 

the result must be shown to “the study director or an appropriate designee for verification of 

negative results” (V1:24). The consent form states that if, after taking the test, a subject still 

wishes to participate, she must show the result to a female laboratory technician. (V1:195) A 

female member of the research team would be the best person to verify negative pregnancy 

test results. 

 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, 
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not a benefit of participating in the research. This study provides no direct benefits to 

subjects. (V1:45) 

 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” (V1:23) 

 In section 8.1.2 of the protocol, titled “Inclusion/Exclusion Criteria” (V1:23), 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 (V1:195). 

 In the “Study Procedures” section of the consent form (V1:196), insert text similar to what 

appears below as #2, and adjust the numbering of the subsequent procedures accordingly: 

“2. 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.” 

40 CFR 26 Subpart L, at §26.1703, as amended effective August 22, 2006, provides in 

pertinent part: 

EPA shall not rely on data from any research involving intentional exposure of 

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

woman, or a child. 

This protocol requires that subjects be at least 18 years old and excludes female subjects who are 

pregnant or lactating. Thus §26.1703 would not forbid EPA to rely on a study executed 

according to this protocol. 

D. Summary Assessment of Scientific Aspects of the Proposed Research 

 “The objective of this proposed study is to determine the bite protection level of etofenprox 

treated U.S. Military Fire Resistant Army Combat Uniforms (FRACUs) treated initially at an 

application rate of 1% etofenprox (weight/weight), and to assess the bite protection performance 

after 0x, 20x, and 50x washes. This is a non-guideline study; therefore, it is not designed to fulfill 

the requirements of a specific OCSPP (formerly OPPTS) Guideline. This is considered a special 

study. This study will be conducted in accordance with EPA, FIFRA (Federal Insecticide, 
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Fungicide and Rodenticide Act), Good Laboratory Practice Standards (GLP); 40 CFR, Part 160 

(October 1989).” (V1:16). 

The basic experimental unit in this study is a sleeve test. Each test involves a subject 

exposing a (unwashed treated, untreated, washed treated) fabric-sleeved arm into a cage of one 

species of mosquito for 15 minutes per hour for up to eight hours (See Figure 8.2.1, V1:26). The 

data obtained from each 15 minutes test with each experimental unit will be counts of the 

number of bloodfed female mosquitoes and the total number of female mosquitoes in each test 

cage. The observed bite-through proportion (or ‘rate’) is the proportion of bloodfed female 

mosquitoes to the total number of mosquitoes in each test cage, which will be expressed as 

percent bite protection. Etofenprox treatment bite through rates will be corrected using Abbott’s 

formula for ‘background’ bite through rates in the control (untreated fabric sleeve). To increase 

testing precision, each subject will serve as their own treatment and control. Therefore, the 

experimental design consists of 4 groups tested in the following order per mosquito species. The 

test groups are: 

 1 test with an untreated FRACU fabric-sleeve, which serves as the control. 

 1 test with treated unwashed (0x) FRACU fabric. 

 1 test with treated washed (20x) FRACU fabric. 

 1 test with treated washed (50x) FRACU fabric. 

FRACU fabric from coats (shirts/blouses) and trousers will be tested as described in the tables 

below (3.5.3, and 3.5.4). Each subject (8 subjects) will test each group once per species for a 

total 8 replicates per group per species, resulting in 16 replicates per fabric group for this 

experiment as shown in Table 3.5.2. 

The widely accepted method of evaluating the efficacy of insecticide treated clothing includes 

laboratory aging of this treated clothing by laundering through standardized wash cycles per the 

American Association of Textile Chemists and Colorists (AATC) laundering protocol (See 

Appendix E on V1:171). Testing will be conducted with treated and untreated clothing prior to 

laundering (0x wash cycle) and at the 20x and 50x wash cycles. This protocol is similar to the 

protocol accepted as Standard Operating Procedure by the U.S. Military to evaluate their 

repellent treated combat uniforms. 

The unit of measure for determination of the repellent effects in this proposed experiment (% 

bite protection based on the proportion of bloodfed to total mosquitoes in a cage) differs from 

skin applied repellent evaluations where the “Landing with Intent to Bite” measure is used and 

efficacy is measured as Complete Protection Time. A detailed justification for the proposed test 

system is presented in the protocol (V1:16). In brief, the repellent effect created by skin applied 

repellents is instantaneous and non-toxic, whereas mosquitoes exposed to the treated clothing 

must remain in contact with the treated cloth for a longer time period to illicit a repellent effect. 

The resulting repellent effect is usually a toxic effect that results in ‘excito-repellency’ or 

incapacitation due to exposure to the fast acting insecticide. The proposed specifications for 

testing success are shown in the table below, which is taken from the table on p.12 of 210 in 

Volume 1 and page 3 of 21 in Volume 3. These specifications are in-line with current U.S. 

Military specifications for treated uniforms. 
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Bite Protection Specifications (%) 

Uniform 0x wash cycle 20x wash cycle 50x wash cycle 

Army FRACUs (test 

material in the proposed 

protocol) 

85% 80% 70% 

This protocol also proposes to evaluate the repellent effect (% bite protection) of treated clothing 

using only two mosquito species - unlike skin applied repellent studies conducted under field 

conditions where three species are evaluated. In the proposed study representative species from 

the genus Anopheles (malaria vector) and genus Aedes (vector of dengue and yellow fever) will 

be evaluated. A mosquito species from the genus Culex (vector of West Nile virus or St. Louis 

encephalitis) will not be tested. Justification for exclusion of the third species is not mentioned 

with referral to current military test specifications instead. 

The study director included a justification for sample size in Table 3.5.1 (V1:19). These tables 

show that the overall bite protection value for a treated fabric is dependent on control bite 

through rates and the percentile of bite protection. Precision improves with the number of 

subjects based on the width of the 95% confidence interval for the 80th and 95th percentiles with 

control bite through of 50% and 20%. However, this precision does not improve very much 

beyond 8 test subjects. Precision can decline if percent control bite through rates decline or are 

low, therefore, as proposed by the study director, mosquitoes will be screened for their 

‘aggressiveness’ to ensure that control bite through rates are substantial. The testing paradigm 

including replication is described in Tables 3.5.2, 3.5.3, and 3.5.4 (V1:20). 

The objective of the data analysis is to estimate the mean level of bite protection and associated 

95% confidence intervals for different ‘treatments’ [i.e. different combinations of fabric types 

(coats and trousers), number of washes, and mosquito species]. The numbers of bloodfed and 

total female mosquitoes found with treated and control fabric (clothing) for each subject will be 

analyzed as binomial distributed data in a generalized linear model (GLiM) using a log link, 

generalized estimating equations or a mixed effect GLiM. This is largely dependent on the 

‘subject term’, which may be treated as a fixed or random effect to adjust for within-subject 

differences (V1:22). Volume 3 includes a very detailed explanation of the analysis and 

justification for the study design. Alternate subject data appear to be treated together with 

existing subject data in the event of a dropout but the protocol is silent on this aspect of data 

analysis. The analysis does not take into account what happens if less than 8 subjects complete 

the study. 

1. Study design: 

Replicate subjects will be used in this study to evaluate bite protection for a U.S. military 

clothing fabric (FRACU) treated with insecticide/repellent (etofenprox). A fabric’s “bite 

protection” is a measure of the relative level to which a treated fabric prevents bites 

compared to the untreated control. As described in §8.5 (V1:30) of the protocol the 

observed bite protection for a subject is calculated using the subject’s bite-through rates 

for the treated fabric and for a corresponding untreated/unwashed control fabric. (Each 

subject serves as their own control.) The purpose of the control is to compensate for 

influences related to the subject’s individual attraction level, the general host-seeking 
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response of the test mosquito population, and to correct for bite-through rate of the 

untreated fabric. The treatment and control values for a subject are then used in Abbott’s 

formula to calculate the observed bite protection level of the sleeve for that subject. 

Treated clothing sets will be evaluated at the specific standardized wash intervals: 

unwashed (0x), 20x washes, and 50x washes. Separate fabric specimens for each wash 

interval are tested, similar to that described in U.S. military GL/PD specifications 

(Appendix C, pg. 34 and Appendix D, pg. 36). Two species of mosquitoes, Aedes 

aegypti and Anopheles albimanus, will be tested separately. Eight subjects (preferably 

the same eight subjects) will be used to test each fabric and mosquito species 

combination. Two alternate subjects will also be recruited. The exposure time to 

mosquitoes at each test interval (once per hour) for control and treated replicates is 15 

minutes per arm. A summary of the experimental design and testing paradigm for each 

mosquito species are shown below (Vol 1:18-20): 

Table 3.5.2 Experimental Design 

Fabric and Treatment Condition1

Number of 

Fabric 

Specimens 

Number of 

Subjects 

Number of 

Species2

Total Replicates 

per Fabric 

Type 

Coat Untreated Unwashed Control3 1 8 2 16 

Coat Treated Washed 50x 1 8 2 16 

Coat Treated Washed 20x 1 8 2 16 

Coat Treated Unwashed (0x) 1 8 2 16 

Trouser Untreated Unwashed 

Control3 1 8 2 16 

Trouser Treated Washed 50x 1 8 2 16 

Trouser Treated Washed 20x 1 8 2 16 

Trouser Treated Unwashed (0x) 1 8 2 16 

1 Fabric treatment conditions are either untreated and unwashed (Control) or treated and unwashed (0x), 

treated and washed 20 times (20x) or treated and washed 50 times (50x). 2The test species are Aedes aegypti or Anopheles albimanus. 3Each subject serves as their own control for the bite protection calculation. 

 Table 3.5.3 Testing Paradigm using Aedes aegypti

Subject Right Arm Subject Left Arm 

Test 

Set1 Treatment Condition Specimen 

Designation 

Treatment Condition Specimen 

Designation 

1 Coat Untreated Unwashed 

Control2

Sleeve 1 Trouser Untreated Unwashed 

Control2

Sleeve 2 

2 Coat Treated Washed 50x Sleeve 3 Trouser Treated Washed 50x Sleeve 4 

3 Coat Treated Washed 20x Sleeve 5 Trouser Treated Washed 20x Sleeve 6 

4 Coat Treated Unwashed (0x) Sleeve 7 Trouser Treated Unwashed (0x) Sleeve 8 

*

Each subject will have both their right arm and left arm tested simultaneously and complete Test 

Set 1-4 for Aedes aegypti. Each subject will have a break between test sets when new cages are being 

 filled with mosquitoes. All cages will be washed after all test sets for each participant are completed. 

 1 Each test set runs for 15 minutes. 2Each subject serves as their own control for the bite protection calculation. 

 
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Table 3.5.4 Testing Paradigm using Anopheles albimanus 

Subject Right Arm Subject Left Arm 

Test 

Set1

Treatment Condition Specimen 

Designation 

Treatment Condition Specimen 

Designation 

5 Coat Untreated Unwashed 

Control2

Sleeve 9 Trouser Untreated Unwashed 

Control2

Sleeve 10 

6 Coat Treated Washed 50x Sleeve 11 Trouser Treated Washed 50x Sleeve 12 

7 Coat Treated Washed 20x Sleeve 13 Trouser Treated Washed 20x Sleeve 14 

8 Coat Treated Unwashed (0x) Sleeve 15 Trouser Treated Unwashed (0x) Sleeve 16 *

Each subject will have both their right arm and left arm tested simultaneously and complete Test 

 Set 5-8 for Anopheles albimanus. Each subject will have a break between test sets when new cages are 

being filled with mosquitoes. All cages will be washed after all test sets for each participant are completed. 1 Each test set runs for 15 minutes. 

2

Each subject serves as their own control for the bite protection calculation.

Laboratory-reared 6-11 day old adult mosquitoes will be used for the bite protection assay. 

Mosquito species have differing behavior and levels of ‘aggressiveness’, therefore, adult 

female mosquitoes of two of the more aggressive and anthropophilic species will be tested. 

One of these selected species will be Aedes aegypti, a vector of yellow fever and dengue 

fever that is found heavily in Asia and South America. The second species will be 

Anopheles albimanus, a tropical mosquito that is a highly aggressive biter, one of the most 

tolerant species when tested with topical skin repellents, and is a competent vector for 

malaria transmission. The mosquito colonies are reared at the Center for Medical, 

Agricultural, and Veterinary Entomology in a USDA facility in Gainesville, Florida. The 

colonies used have been maintained since being established in 1952 while in Orlando for 

Ae. aegypti, and in El Salvador in 1974 for An. albimanus. There have been period 

introductions of wild type species. Mosquitoes from a colony will respond on the whole 

more aggressively to attractant stimuli than strains reared from freshly collected wild-types 

(V1:27).

“The test cages are approximately 59,000 cm3 in volume and each will contain 175 to 225 

female mosquitoes (density of ~1 mosquito/300 cm3

). Female mosquitoes will be 

preselected from stock cages by using a specially designed draw box that uses odors from 

the hand of a laboratory staff person to attract mosquitoes upwind in to a trap (Fig. 8.3.1). 

The trap containing the mosquitoes will then be transferred to the test cage for subsequent 

testing by subjects (Fig 8.3.2, V1:28).” 

2. Statistical design: 

 The proposed research will include eight human subjects and two alternates. As 

described, the subjects will serve as their own treatments and controls. Each fabric type 

will be replicated 16 times, 8x for each species. An analysis of the impact of the number 

of replications on the number of subjects was conducted as described below in Table 

3.5.1 (V1:19) from the protocol. The effects of control bite through rates (20% and 50%) 

as well as bite protection (80th and 95th percentile) were evaluated in this analysis using 

1000 simulated datasets for each number of subjects. (Volume 3 includes the complete 

explanation of this approach, the simulations, and related justifications on pp. 5-12) 
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These simulated data show that reliable data sets can be collected with 8 subjects and that 

increasing the number of subjects slightly increases precision, but this increase in 

precision does not provide enough information to justify a larger experiment. 

Table 3.5.1 Impact of the Number of Replications on the Number of Subjects 

True bite-through rate for 

control fabric (θC) 50% 20% 

True bite protection for 

treated fabric (βT)

1 80% 95% 80% 95% 

Number of Subjects Expected half-width of a 95% confidence interval 

for % bite protection2 

3 5.2% 2.7% 8.8% 4.5% 

4 4.5% 2.3% 7.5% 3.8% 

5 4.0% 2.0% 6.7% 3.4% 

6 3.7% 1.9% 6.0% 3.0% 

7 3.4% 1.7% 5.6% 2.8% 

8 3.2% 1.6% 5.2% 2.6% 

9 3.0% 1.5% 4.9% 2.4% 

10 2.8% 1.4% 4.7% 2.3% 

15 2.3% 1.2% 3.8% 1.9% 

20 2.0% 1.0% 3.3% 1.3% 

1

Bite incidence for treated fabric is calculated from bite protection as θT = θC(1 - βT/100) 2

Average half-width from 1,000 simulated datasets. Each dataset consisted of S subjects testing a pair 

of fabrics (control and treated). For each pair the total number of mosquitoes (M) was a Poisson 

(200) random variable, and the number of bloodfed mosquitoes was simulated as a binomial (θ,M) 

random variable. Subject-subject differences were simulated by adding a subject-specific normal 

(0,0.3) random variable to the logit of the true incidence for both control and treatment fabrics. For 

each simulated dataset a binomial generalized linear model was fit to the data using the GENMOD 

procedure in SAS. The model specified fixed effects for both subject and test material and used a 

log link. Bite protection confidence intervals were then obtained by back-transforming the 

confidence intervals for the contrast log(θT)-log(θC). 

The primary objective of the data analysis is to estimate the overall (or ‘mean’) level of bite 

protection and associated 95% confidence interval for different ‘treatments’ (i.e., different 

combinations of fabric type, number of washes, and mosquito species). Subject-specific 

bite protection values will be calculated for each treatment using Abbott’s formula as 

described in §8.5. These values will be averaged over all subjects to obtain mean observed 

bite protection values that can be used as a check on any model-based bite protection 

estimates. 

The numbers of bloodfed and total female mosquitoes found with treated and control fabric 

for each subject will be analyzed as binomial distributed data in a generalized linear model 

(GLiM) using a log link. A subject term will be added as a fixed effect in the model to 

adjust for subject-subject differences. (Alternatively, subjects could be treated as a random 

effect and the within-subject correlation accommodated using either generalized estimating 

equations or a mixed effect GLiM. The decision on how to analyze the collected data needs 

to be finalized.) Use of the log link makes it possible to obtain an estimate and confidence 
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interval for the ratio of the treatment and control bite-through rates. The estimates and 

confidence intervals for percent bite protection are obtained from the relationship: 

Percent Bite Protection = [1 – (treatment rate) / (control rate)] × 100% 

The GLiM model-based bite protection estimates could be obtained by analyzing multiple 

models each with just a single treatment group and the matched control group. However, it 

may also be of interest to compare the bite protections of different types of treated fabric, 

number of washes, or mosquito species. In this case, it would be necessary to include all of 

the treatments (and species) of interest in the same model. Because the GLiM uses a log 

link, hypothesis tests concerning ratios of bite protection can be formulated as linear 

contrasts in the GLiM (V1:21-22) (V3:3-5). 

3. How and to what will human subjects be exposed? 

 Subjects will be exposed to test material and two species of caged mosquitoes in the 

laboratory. Each subject will have etofenprox treated sleeves placed on both forearms and 

both arms will be exposed to caged mosquitoes for 15 minutes per hour [The step-wise 

procedure is described in detail in §8.4 (V1:28-30) and §8.6 (V1:33-42)]. This period of 

time allows mosquitoes to land, probe, and bloodfeed. The greatest number of bites is 

expected to be received during the first set of tests with the untreated, unwashed control 

sleeves. Subsequent tests will involve treated sleeves and it is expected that far fewer 

bites will be received by the test subjects. 

4. Endpoints and Measures: 

Efficacy will be measured as percent bite protection. The proposed study will estimate the 

overall (or ‘mean’) level of bite protection and associated 95% confidence interval for 

different ‘treatments’ (i.e., different combinations of fabric type, number of washes, and 

mosquito species). Subject-specific bite protection values will be calculated for each 

treatment using Abbott’s formula as described in §8.5 based on exposure to mosquitoes 

during a 15 minutes bioassay every hour for up to 8 hours. These values will be averaged 

over all subjects to obtain mean observed bite protection values that can be used as a check 

on any model-based bite protection estimates. 

E. Compliance with Applicable Scientific Standards

 This protocol adequately addresses the following elements according to applicable 

scientific standards: 

 Prerequisite acute toxicity research to characterize toxicological profile of the 

formulation and calculate margin of exposure (MOE). 

 Experimental design 

 Pre-training of subjects 
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EPA Science Comments

The following elements in the protocol require revision before the research goes forward: 

 Provide justification for testing two vector mosquito species instead of three (V1: 27). 

 The statistical analysis must be finalized. Two approaches to GLiM use are discussed in the 

present version (V1: 21-22). “Recommendations for Data Analysis” expands on this 

discussion in V3: 13. The protocol should be amended to include the selected analysis before 

the study is executed. 

 The protocol proposes only two alternate subjects. Consider recruiting more than two 

alternates in case more than two subjects withdraw midway through the study. 

 Describe how the data will be analyzed if the number of test subjects at the end of the test is 

less than eight. 

 Please add more details to the protocol about what will happen if a subject withdraws 

midway through the study and an alternate is brought into the study as a replacement. Will 

data from the subject who withdraws be discarded, or used alone, or used in combination 

with data from the alternate subject? Will an alternate who replaces an original subject 

complete all eight pairs of sleeves, or only the pairs of sleeves that were not completed by the 

original subject? If the latter, that person must also be tested with an untreated sleeve against 

each mosquito species to establish their baseline. The statistical plan for analyzing the data 

will need to take into account how alternate subjects will be handled. There may be statistical 

implications of combining partial data sets from different subjects. 

 The subjects and alternates need to be randomly selected from a larger pool of qualified 

potential subjects. The protocol states that the first 20 respondents to the advertisement will 

undergo a preliminary telephone screening. Please continue screening respondents to the 

advertisement until you have at least 20 qualified potential subjects. Then, randomly select 

the 8 subjects and 2 or more alternates from the pool of qualified potential subjects. The 

random selection may be accomplished by drawing names from a hat or through the use of a 

randomizer program such as http://randomizer.org. 

 Section 8.1.5 of the protocol states that “it is desired that random selection will yield a nearly 

equal distribution of male and female candidates...” (V1:24) However, it is possible that 

random selection will yield an unequal distribution of males and females. Please revise the 

protocol to specify exactly what will happen if there is unequal distribution or if only one sex 

is represented. Will you continue to randomly select from the pool of eligible subjects until 

you have an equal number of males and females? Or until you have at least one subject of 

each sex? 

 Due to dermal observations resembling skin irritation in a 28-day dermal toxicity study 

conducted with technical etofenprox on rabbits, the etofenprox registrant, Mitsui Chemicals, 

will soon be conducting a product-specific 28-day dermal toxicity study in rabbits with 
Page 13 of 36 

etofenprox-treated fabric. The new study will test the same fabric that subjects will be 

exposed to in the proposed efficacy study under review here. Do not initiate this research 

until the results of the product specific fabric study have been submitted to and 

reviewed by EPA. 

 Present values for 70% true bite protection for treated fabric in the protocol in Table 3.5.1 to 

illustrate the change in the 95% confidence interval with the number of subjects as this value 

was selected as the performance standard for FRACU fabric washed 50x (V1: 12). The 

change in the width of the 95% confidence interval is addressed in Volume 3 for the 70% 

level of bite protection performance in Tables 4, 5 and 6 on pp. 10-12. 

 Data compilation and processing. The protocol provides a sample data sheet that is to be 

used for every 15 minute sleeve test (V1:190 and 192). However, little detail is provided in 

the protocol on how the data from these sheets will be compiled and processed before entry 

into Excel, JMP, or SAS and how the resulting records will be handled and archived (§9.0, 

V1:43). 

 Amend the protocol to identify the Quality Assurance Unit (V1:15). 

 

 Attachments: 

1. Summary Review of Protocol (dated 2/28/14) 

2. §26.1111 Criteria for IRB approval of research 

3. §26.1116 General requirements for informed consent 

4. §26.1117 Documentation of informed consent 

5. §26.1125 Criteria for Completeness of Proposals for Human Research
Attachment 1 

Page 14 of 36 

EPA Protocol Review 

Title: Laboratory Evaluation of Bite Protection from Repellent-Impregnated Clothing for the 

United States Military 

Date: February 28, 2014 

Principal Investigator and any sub-investigators: Ulrich R. Bernier, Ph.D.

 

Participating Laboratory: 

United States Department of Agriculture-Agricultural Research Service

Center for Medical, Agricultural, and Veterinary Entomology 

1600 SW 23rd Drive 

Gainesville, FL 32608 

Sponsor: 

United States Department of Agriculture-Agricultural Research Service 

Center for Medical, Agricultural, and Veterinary Entomology 

1600 SW 23rd Drive 

Gainesville, FL 32608 

IRB: 

Western Institutional Review Boards 

1019 39th Avenue, SE Suite 120 

Puyallup, WA 98374-2115 

1. Societal Value of Proposed Research 

(a) What is the stated purpose of the proposed research? 

In this study, military uniform fabric will be treated with a pesticide called etofenprox. 

Treated fabric will be compared to untreated fabric to determine if the treatment can 

decrease mosquito bites to human skin that is covered by the sleeve. Specifically, this 

study will determine the bite protection level of etofenprox treated U.S. Military Fire 

Resistant Army Combat Uniforms (FRACUs) treated initially at an application rate of 

1% wt/wt, and to assess the bite protection performance after 0x, 20x, and 50x washes 

against two species mosquitoes (Aedes aegypti and Anopheles albimanus). (V1:161)

 

1 This pagination convention is used throughout this review. “V1” refers USDA Volume 1, “V2” refers to USDA 

Volume 2, etc. Entries after the colon are page references; many page images bear more than one page number. In 

Volume 1, the cited page number is from the expression “Page n of 210” found at the bottom right-hand corner. 

Volume 2 page references are from the expression “Page n of 580” found at the bottom right-hand corner. Volume 3 

page references are from the expression “Page n of 25” found at the bottom center of the page. 
Attachment 1 

Page 15 of 36 

(b) What research question does it address? Why is this question important? 

Would the research fill an important gap in understanding? 

The purpose of this study is to develop a standardized protocol that can be used to evaluate 

the bite protection of fabrics that are treated or impregnated with substances that repel or 

reduce arthropod bites to determine if they provide sufficient protection against 

mosquitoes. Due to global issues regarding mosquito resistance to ester-pyrethroids, other 

insecticide candidates must be considered. Thus, comparisons of new insecticide/repellent 

candidates should be evaluated in products where performance has already been 

determined with a known standard, i.e. permethrin. 

A standardized protocol will enable the EPA to receive consistent and scientifically reliable 

data for new repellent clothing treatments. The bite protection data will provide 

information about: 1) the relative level to which bites are received through the fabric with 

the repellent treatment compared to bites received through the untreated control fabric; 2) 

whether the fabric is in the “plateau” region of bite protection for the repellent-treated 

surface, such that application of additional chemical does not improve bite protection 

performance; 3) the relative bite protection capability of one fabric construction and 

composition vs. another fabric construction and composition; and 4) the bite protection 

efficacy of a new product for EPA registration. Because these data are acquired in a 

laboratory setting, there are fewer associated risks than determining where the optima lie 

than using wild-type mosquitoes in a field setting. Bites are measured in these studies by 

the presence of a blood meal in the abdomen of the female mosquito (V1:9). 

(c) How would the study be used by EPA? 

EPA will consider the study to satisfy product specific efficacy data requirements and 

acceptable label claims for repellent efficacy for the test material. 

(d) Could the research question be answered with existing data? If so, how? If not, 

why not? 

EPA requires product-specific efficacy data to support its registration. No previous 

testing of this product against mosquitoes under the proposed use pattern has been 

conducted. 

(e) Could the question be answered without newly exposing human subjects? If so, 

how? If not, why not? 

Human subjects are required because they represent the target system for the test 

material, and sufficiently reliable non-human models for repellency testing have not been 

developed. 

 
Attachment 1 

Page 16 of 36 

2. Study Design 

(a) What is the scientific objective of the study? If there is an explicit hypothesis, what 

is it? 

“The objective of this study is to determine the bite protection level of etofenprox treated 

U.S. military Fire Resistant Army Combat Uniforms (FRACUs) treated initially at an 

application rate of 1% wt/wt, and to assess the bite protection performance after 0x, 20x, 

and 50x washes.” (V1:16)

(b) Can the study as proposed achieve that objective or test this hypothesis? 

The objective cited may be achieved by the study as proposed if the protocol is revised 

and amended to explain, in more detail, the following items noted on pages 12-13 of this 

review. 

2.1 Statistical Design 

(a) What is the rationale for the choice of sample size? 

 

 The rationale for the sample size appears on pp. 18-19 of 210 (§ 3.5) in Volume 1. 

The researcher’s justification for sample size is based on the argument that the 

precision of the overall (mean) bite protection value for a treated fabric will depend 

on the true bite through rate for the control fabric and the true level of bite protection. 

This justification is illustrated in Table 3.5.1 on p. 19 of 210 in Volume 1. Based on 

the width of the 95% confidence intervals in this table: 1) the experimental variability 

decreases as control treatment bite through rates increases; 2) as true bite protection 

increase, the width of the 95% interval decreases; and 3) the precision of overall bite 

protection value does not appear to increase very much once the number of subjects 

in the experiment exceeds 8. 

(b) What negative and positive controls are proposed? Are proposed controls 

appropriate for the study design and statistical analysis plan? 

 Each subject will serve as their own treatment and negative control for each test set as 

described on p. 20 of 210 in Volume 1 in Tables 3.5.2, 3.5.3, and 3.5.4. The controls 

are appropriate to calculate the overall bite protection because percent bite protection 

will be calculated by counting bloodfed female mosquitoes in the treatments and 

comparing them to the untreated control. Both arms will serve as a control treatment 

replicate, one for coat fabric and the other for trouser fabric. 

(c) How is the study blinded? 

 The study is not blinded. Untreated sleeves will be tested first followed by 50x, 20x 

and 0x fabrics. 

 
Attachment 1 

Page 17 of 36 

(d) What is the plan for allocating individuals to treatment or control groups? 

All subjects will be assigned to control and treatment groups as described on p. 20 of 

210 in Volume 1. 

The subjects and alternates need to be randomly selected from a larger pool of 

qualified potential subjects. The protocol states that the first 20 respondents to the 

advertisement will undergo a preliminary telephone screening. Please continue 

screening respondents to the advertisement until you have at least 16 qualified 

potential subjects. Then, randomly select the 8 subjects and 2 or more alternates from 

the list of 16+ qualified potential subjects. The random selection can be accomplished 

by drawing names from a hat or through the use of a randomizer program such as 

http://randomizer.org. 

(e) Can the data be statistically analyzed? 

 Yes. “Treated clothing sets will be evaluated at the specific standardized wash 

intervals: unwashed (0x), 20x washes, and 50x washes. Separate fabric specimens for 

each wash interval are tested, similar to that described in U.S. military GL/PD 

specifications (Appendix C, pg. 34 and Appendix D, pg. 36). Two species of 

mosquitoes, Aedes aegypti and Anopheles albimanus, will be tested separately. Eight 

subjects will be tested using each fabric and mosquito species combination” (V1:19). 

Each subject will serve as their own treatment and control. The experimental design 

together with the testing paradigms for Ae. aegypti and An. albimanus, respectively,

are illustrated in tables 3.4.2, 3.5.3, and 3.5.4 on p. 20 of 210 in Volume 1. 

(f) What is the plan for statistical analysis of the data? 

“The primary objective of the data analysis is to estimate the overall (or ‘mean’) level 

 of bite protection and associated 95% confidence interval for different ‘treatments’ 

 (i.e., different combinations of fabric type, number of washes, and mosquito species). 

 Subject-specific bite protection values will be calculated for each treatment using 

 Abbott’s formula as described in Section 8.5. These values will be averaged over all 

 subjects to obtain mean observed bite protection values that can be used as a check on 

 any model-based bite protection estimates. 

“The numbers of bloodfed and total female mosquitoes found with treated and control 

 fabric for each subject will be analyzed as binomial distributed data in a generalized 

 linear model (GLiM) using a log link. A subject term will be added as a fixed effect in 

 the model to adjust for subject-subject differences. (Alternatively, subjects could be 

 treated as a random effect and the within-subject correlation accommodated using 

 either generalized estimating equations or a mixed effect GLiM.) Use of the log link 

 makes it possible to obtain an estimate and confidence interval for the ratio of the 

 treatment and control bite-through rates. The estimates and confidence intervals for 

 percent bite protection are obtained from the relationship: 
Attachment 1 

Page 18 of 36 

“Percent Bite Protection = [1 – (treatment rate) / (control rate)] × 100% 

 “The GLiM model-based bite protection estimates could be obtained by analyzing 

multiple models each with just a single treatment group and the matched control group. 

However, it may also be of interest to compare the bite protections of different types of 

treated fabric, number of washes, or mosquito species. In this case, it would be 

necessary to include all of the treatments (and species) of interest in the same model. 

Since the GLiM uses a log link, hypothesis tests concerning ratios of bite protection can 

be formulated as linear contrasts in the GLiM” (V1:21-22). A complete explanation 

and related justifications can be found in Volume 3. 

(g) Are proposed statistical methods appropriate to answer the research question? 

 The analysis will provide the overall bite protection values for each treatment group 

and the controls. As proposed, the analysis addresses mean values and associated 

uncertainties. 

(h) Does the proposed design have adequate statistical power to definitively answer 

the research question? 

Statistical power is adequately described in §3.5 of Volume 1 and is illustrated in 

Table 3.5.1. An in-depth analysis is presented in Volume 3. As expected the width 

of the 95% confidence interval decreases as the number of subjects increases but the 

relative improvement or ‘gain’ from increasing the number of subjects decreases as 

the number of subjects increase. The proposed sample size of 8 subjects represents a 

reasonable compromise between decreasing confidence interval width and limiting 

unnecessary human experimentation. 

2.2 How and to what will human subjects be exposed? 

Subjects will be exposed to test material and mosquitoes in the laboratory. The test 

material will be cut out of treated military trousers or coats and sewn into “sleeves” at the 

USDA-ARS testing facility. The test material’s active ingredient, etofenprox, has a low 

acute and chronic risk profile (§3.3 and §3.4, V1:17-18), and the inert ingredients are 

classified by the Agency as safe for this use. The test material has been tested in animals 

for acute toxicity. Subjects with known allergic reactions (§8.1.2) are excluded from 

participation in the test. 

Subjects will be exposed to laboratory reared populations of mosquitoes free of 

mosquito-borne pathogens in the laboratory (V1:27, 44). Subjects with known allergic 

reactions to mosquito bites will be excluded from research participation (V1:23). 

 

(a) What is the rationale for the choice of test material and formulation? 

 Efficacy data to satisfy product performance requirements and to support label claims 

for this product are required by EPA for registration. EPA requires submission of 
Attachment 1 

Page 19 of 36 

product performance data for all products claiming efficacy against public health 

pests.

(b) What is the rationale for the choice of dose/exposure levels and the staging of 

dose administration? 

 There is no dosimetry phase in this experiment. Test sample sleeves contain 1% 

etofenprox (wt/wt). As explained in Volume 1 on page 11 of 210 in the ‘Background’ 

section of this protocol, the registrant performs the dosimetry experiments before 

efficacy testing is conducted to determine: 1) the ‘plateau region’ at which no 

additional bite protection is gained from additional compound and 2) the point at 

which the bite protection begins to decrease substantially, typically this done by 

treating fabric with a range of concentrations to determine the dose-response curves. 

The range of concentrations for these preliminary tests is selected to be within an 

acceptable range based on the toxicity profile of the insecticide/repellent compound to 

minimize risk to the individual wearing the fabric. 

(c) What duration of exposure is proposed? 

The exposure period is eight 15-minute periods (2 hours total) for both arms of each 

subject.

2.3 Endpoints and Measures 

 (a) What endpoints will be measured? Are they appropriate to the question(s) being 

 asked? 

 Endpoints/Measures for efficacy evaluation: 

 Number of bloodfed and total number of females mosquitoes in each test. The 

proportion of bloodfed/total will be calculated and expressed as a percentage 

value. This calculation will be performed for untreated control sleeves and 

treated sleeves (0x, 20x, and 50x washes). 

 For each test set, the treatment % bite values will be corrected to account for 

the bite through values in the untreated control using Abbott’s Formula. 

 The overall % bite protection will be calculated and expressed as a mean 

value for each treatment: 0x, 20x, 50x washes for coats and trousers. 

 The endpoints are appropriate to the questions being asked and address uncertainty 

 associated with the samples size, between subject variation, % bite values, and the 

 overall bite protection value. 

 The data form for each 15 minutes sleeve test is presented in Appendix I on page 

 190 of 210. A completed sample data sheet is shown in Appendix J on page 192 

 of 210. 
Attachment 1 

Page 20 of 36 

(b) What steps are proposed to ensure measurements are accurate and reliable? 

 Standard Operating Procedures (SOPs) will be in place that must meet Good 

Laboratory Practices requirements. 

 Laboratory technicians will assist subjects with placing the test sleeves on 

their arms and excluding all exposed skin from mosquito exposure. 

 Laboratory technicians will assist subjects with insertion and removal of their 

arms in/from the cages. 

 Laboratory technicians and the study director will track test sleeve samples 

and closely monitor the testing. 

 Alternate subjects will be enrolled to ensure adequate sample size. 

 Counts of bloodfed mosquitoes and the total number of mosquitoes in the cage 

will be determined by a research technician. 

 The test sleeve samples will be assayed by the Analytical Unit (p. 15 of 210) 

and the amount of etofenprox reported (pp. 31-32 of 210) as a surface 

concentration of etofenprox in units of mg/cm2

, which is commonly done for 

treated fabrics. 

 (c) What QA methods are proposed? 

 As explained in Volume 1, §7.0 on p. 22 of 210 a separate, professional Quality 

Assurance Unit (QAU) will inspect the study: “Quality assurance of this study will be 

carried out in accordance with Good Laboratory Practice (GLP) Standards 40 CFR 160. 

Written reports of all findings from the Quality Assurance Officer will be provided to 

the study director and management. Any part of the study found by the Quality 

Assurance Officer to be likely to affect the integrity of the study will be brought the 

attention of the study director. A statement signed by the Quality Assurance Officer 

listing the phases inspected, inspection dates, and dates reported to the study director 

and management will be included in the final report. All deviations and amendments 

will be recorded and reported as per GLP guidelines.

 Additionally, fabric samples will be retained indefinitely for further analysis and 

 verification as requested by NSRDEC for quality control. 

 The quality assurance unit of the analytical laboratory will provide the study director 

 and the study director’s management with relevant data, process, and report audits to 

 meet Environmental Protection Agency GLP requirements.”

 (d) How will uncertainty be addressed? Will point estimates be accompanied by 

 measures of uncertainty? 

Uncertainty is addressed in the experimental design and selection of the number of 

subjects as described in §3.5. The statistical analysis also discusses uncertainty in 

§6.0. Volume 3 provides any in-depth discussion of uncertainty associated with the 

experimental design and data to be generated and analyzed. The objective of the data 

analysis is to estimate the mean level of bite protection and associated 95% 
Attachment 1 

Page 21 of 36 

confidence intervals for different ‘treatments’ [i.e. different combinations of fabric 

types (coats and trousers), number of washes, and mosquito species]. The numbers of 

bloodfed and total female mosquitoes found with treated and control fabric (clothing) 

for each subject will be analyzed as binomial distributed data in a generalized linear 

model (GLiM) using a log link, generalized estimating equations or a mixed effect 

GLiM. This is largely dependent on the ‘subject term’, which may be treated as a 

fixed or random effect to adjust for within-subject differences (V1:22). Volume 3 

includes a detailed explanation of the statistical analysis. 

3. Subject Selection 

3.1 Representativeness of Sample 

(a) What is the population of concern? 

The population of concern is U.S. military personnel who would wear FRACUs treated 

with etofenprox. 

 (b) From what populations will subjects be recruited? 

Subjects will be recruited from the general population in Gainesville, Florida through the 

use of an advertisement placed in a local newspaper and on bulletin boards in University 

of Florida campus buildings. 

 (c) Are expected participants representative of the population of concern? 

 If not, why not? 

Since Gainesville, Florida is a university community and given that the advertisements 

will be placed within campus buildings, the population from which subjects will be 

recruited is likely younger than the general U.S. population, but may mirror the 

population of concern – U.S. military personnel – in terms of over-representation of 

younger individuals. 

 (d) Can the findings from the proposed study be generalized beyond the study sample? 

Yes. 

3.2 Equitable Selection of Subjects 

(a) What are the inclusion/exclusion criteria? Are they complete and appropriate?

The inclusion/exclusion criteria are complete and appropriate except that 

“sensitivities to pesticides or other chemical products” and “people with cuts, scrapes 

or skin conditions such as psoriasis or eczema on their arms or hands” should be 

added as additional exclusions. 
Attachment 1 

Page 22 of 36 

“Inclusion/Exclusion Criteria 

1. Must be between the ages of 18-62 years old. 

2. Must be able to speak and read English. 

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

4. Exclusion of people in poor health or physical condition. 

5. People who are hypersensitive to or phobic of mosquito bites will be excluded. 

6. Exclusion of people known to be sensitive to the test material. 

7. Exclusion of people with a relationship to the study director or sponsor (students 

or employees of the study director or sponsor).” (V1:23) 

(b) What, if any, is the relationship between the investigator and the subjects? 

None. People with a relationship to the study director or sponsor (students or 

employees of the study director or sponsor) are excluded from becoming subjects. 

 (c) Are any potential subjects from a vulnerable population? 

No. 

(d) What process is proposed for recruiting and informing potential subjects? 

“Subjects will be recruited from the general population in Gainesville, Florida 

through a printed advertisement placed in the Gainesville Sun (local newspaper) and 

on bulletin boards in University of Florida buildings. The advertisement (Appendix F) 

will contain a brief description of the testing and financial compensation for 

participation. Subjects will be compensated $20 for participating in the initial 

research consent meeting and $25 for each set of sleeves in the testing paradigm. 

Subjects that are pregnant (see below) will not be allowed to participate but will be 

paid $25 for screening process. Subjects will receive $200 for completion of testing 

with the full set of 8 pairs of sleeves. The advertisement will provide a phone number 

where interested respondents can leave a message. The messages will be reviewed by 

the study director. Respondents will be called by the study director and undergo 

preliminary screening via telephone interview to determine if they meet the inclusion 

and exclusion criteria. Individuals will be recruited from the community and are not 

from any particular employer or agency, and will have no relationship to the study 

director.” (V1:23) 

(e) If any subjects are potentially subject to coercion or undue influence, what 

specific safeguards are proposed to protect their rights and welfare? 

Subjects will be recruited through advertisements in local newspapers. There will be 

no connection or communication between the researchers and the potential subjects’ 

employers, which minimizes the potential for coercion or undue influence. In 
Attachment 1 

Page 23 of 36 

addition, students or employees of the study director or sponsor are excluded from 

participation. 

3.3 Remuneration of Subjects

(a) What remuneration, if any, is proposed for the subjects? 

“Subjects will be compensated $20 for participating in the initial research consent 

meeting and $25 for each set of sleeves in the testing paradigm. Subjects that are 

pregnant (see below) will not be allowed to participate but will be paid $25 for 

completing the screening process. Subjects will receive $200 for completion of testing 

with the full set of 8 pairs of sleeves.” (V1:23) Subjects that begin the study but do 

not complete it will be paid for each pair of sleeves at a rate of $25 per pair. (V1:201) 

(b) Is proposed remuneration so high as to be an undue inducement? 

No. 

(c) Is proposed remuneration so low that it will only be attractive to economically 

disadvantaged subjects? 

No. 

(d) How and when would subjects be paid? 

Payment will be made in cash before subjects leave the test facility. 

4. Risks to Subjects 

4.1 Risk characterization 

(a) Have all appropriate prerequisite studies been performed? What do they show 

about the hazards of the test material? 

Etofenprox is an EPA-registered pesticide with an essentially complete supporting 

toxicity database. It has been tested extensively in animals and is of low toxicity by 

all routes of exposure. The acute dermal LD50 of etofenprox is greater than 2,100 

mg/kg body weight. Etofenprox is not a skin sensitizer. 

The treated fabric that will be placed on the forearm of subjects will contain at most 

105.9 mg of etofenprox (for 0x washes), which results in a maximum dose of 211.8 

mg/subject. Assuming an adult weighs 70kg, the maximum etofenprox dose from 

exposure to the treated fabric equals 3.02mg/kg per subject, assuming that there is 

100% absorption through the skin, resulting in an estimated MOE = 695. This MOE 

does not exceed EPA’s level of concern (MOE= 100) (V1:18). 
Attachment 1 

Page 24 of 36 

 Results from toxicity testing: 

 A primary eye irritation study on rabbits showed that etofenprox is a minimal 

irritant to the eyes. 

 A dermal sensitization study in Guinea pigs (Buehler method) showed that 

etofenprox is not a contact sensitizer. 

 A primary skin irritation study in rabbits study showed that etofenprox is 

minimally irritating to the skin. 

 The single dose acute dermal LD50 of the etofenprox is >2,100 mg/kg in rabbits. 

 The acute oral LD50 of etofenprox is >5,000 mg/kg in dogs. 

Due to dermal observations resembling skin irritation in a 28-day dermal toxicity 

study with rabbits, the etofenprox registrant, Mitsui Chemicals, will soon be 

conducting a 28-day dermal toxicity study in rabbits with etofenprox-treated fabric. 

Do not initiate this research until the results of that study have been submitted to and 

reviewed by EPA. 

 (b) What is the nature of the risks to subjects of the proposed research? 

Risks to subjects include the risk of exposure to biting mosquitoes; the risk of 

exposure to disease vectors; the risk of exposure to the test material; risks related to 

receiving an unexpected result on a pregnancy test; and the risk of a loss of 

confidentiality. 

(c) How do proposed dose/exposure levels compare to the established NOAELs for 

the test material? 

A dose/endpoint was not chosen for a dermal risk assessment since no systemic 

toxicity was observed at the limit dose (1000 mg/kg/day) following repeated dermal 

exposure to rabbits for 28 days to etofenprox technical. However, in the same study a 

dermal LOAEL was identified that equaled 400mg/kg/day based on an increased 

incidence of dermal observations (scabbing, crusting, desquamation, and exfoliation) 

together with histopathological changes that included diffuse epidermal hyperplasia 

in both sexes of rabbits. Based on a dermal absorption study in rats with etofenprox 

technical, very little etofenprox is absorbed through skin (7% of the applied dose) and 

metabolism is rapid with nearly all etofenprox excreted in feces and urine within 24 

hours after application to the skin of test animals. Taken together, subjects in this 

study will be exposed to very little etofenprox when the treated fabric is placed on the 

skin during testing with the maximum dose of 3.02 mg/kg per subject being much 

lower than the systemic NOAEL >1000 mg/kg/day for the dermal route of exposure 

and more than 100x less than the dermal LOAEL observed in rabbits. 

Due to dermal observations resembling skin irritation in a 28-day dermal toxicity 

study conducted with technical etofenprox on rabbits, the etofenprox registrant, 

Mitsui Chemicals, will soon be conducting a product-specific 28-day dermal toxicity 

study in rabbits with etofenprox-treated fabric. The new study will test the same 

fabric that subjects will be exposed to in the proposed efficacy study under review 
Attachment 1 

Page 25 of 36 

here. Do not initiate this research until the results of the product specific fabric study 

have been submitted to and reviewed by EPA. 

(d) What is the probability of each risk associated with the research? How was this 

probability estimated? 

No numerical probability is estimated, but risks have a low probability of occurrence. 

Risks are minimized in the protocol by excluding candidates known to be 

hypersensitive to or phobic of mosquito bites; using disease-free colony-raised 

mosquitoes; excluding candidates known to be sensitive to the test material; applying 

clear stopping rules; and by incorporating procedures to keep the results of pregnancy 

testing private and permit discrete withdrawal. 

4.2 Risk minimization 

(a) What specific steps are proposed to minimize risks to subjects? 

Risks from exposure to biting mosquitoes. 

 Candidates who are hypersensitive to or phobic of mosquito bites are 

excluded (V1:23, 195) 

 Subjects are alerted in the consent form to the possibility of experiencing a 

skin reaction to mosquito bites, and are advised to inform the study director, 

laboratory technician, or on-call nurse if they believe they are having a 

reaction (V1:199) 

 Over-the-counter topical steroid cream to relieve itching will be available for 

use by subjects (V1:199) 

 A nurse familiar with the protocol will be on-call to provide advice or 

assistance in the event that a subject experiences a reaction (V1:199) 

Risks of exposure to disease vectors. 

“Since these mosquitoes are colony reared and not exposed to disease agents, the 

risk of contracting an infectious disease from bites is negligible. Mosquitoes from 

colony have been reared through immature stages and delivered to the laboratory 

testing room just prior to pupal eclosion. Mosquito pupae are received from 

colony and allowed to eclose into cages in the laboratory. For those mosquitoes 

to transmit pathogens such as dengue fever or malaria, they would need to 

escape, bite an infected human and then return back to the cage for selection in 

tests. Even if a mosquito escapes, the probability that it could return into a cage 

and be used in testing is negligible. There is also an incubation period during 

which the disease must be in the mosquito in order to transmit to another 

organism. Additionally, all mosquitoes that are contained in the cages are not 

allowed to feed on any organisms and thus do not have the opportunity to be 

infected with pathogens prior to testing. The mosquito colonies are reared at the 

Center for Medical, Agricultural, and Veterinary Entomology in a USDA facility 

in Gainesville, Florida using a membrane feeding system and bovine blood. The 

citrated bovine blood is pathogen free (certified sterile by the supplier). The 
Attachment 1 

Page 26 of 36 

mosquitoes will not have been fed on any humans prior to the study. The colonies 

used have been maintained in this manner since being established in 1952 while 

in Orlando for Ae. aegypti, and in El Salvador in 1974 for An. albimanus.” 

(V1:27) 

Risks from exposure to the test material.

 Candidates who are known to be sensitive to the test material are excluded. 

(V1:23) It is also recommended that the protocol exclude subjects who are 

sensitive to pesticides or other chemical products. 

 It is recommended that the protocol exclude subjects with cuts, scrapes, or 

skin conditions such as psoriasis or eczema on their arms or hands. These 

conditions could increase the possibility of a reaction to test material. 

Risks of stress and loss of confidentiality related to pregnancy testing

 The protocol provides for discrete handling of the pregnancy testing that is 

required of female subjects on the day of the study. 

 Female subjects self-administer the pregnancy test in a private restroom. 

(V1:34, 195)

 After completing the test, each female subject is asked if she would like to 

continue in the study. If her answer is no, then no further questions are asked; 

she will not be asked to share the result with anyone. If her answer is yes, the 

result of pregnancy test will be verified by only one member of the research 

(EPA has recommended that the protocol specify that it be a female member 

of the research team). (V1:34, 195)

 For females who proceed with the testing, the result of the pregnancy test is 

recorded in the raw data and kept confidential. (V1:34) 

 (b) What stopping rules are proposed in the protocol? 

“The study will be stopped if the test site becomes unsafe for any reason, biting 

pressure falls below threshold needed, biting pressure rises too high for subject 

comfort or safety, subject asks to withdraw, subject is unattractive to target species, 

subject exhibits hypersensitivity to insect bites during test, subject exhibits sensitivity 

to the test materials during the test.” (V1:46) 

(c) How does the protocol provide for medical management of potential illness or 

injury to subjects? 

“On the day of the testing, there will be a nurse on call who has read the protocol 

and discussed the research with the study director to assist if needed. UF Health 

Shands Hospital is located 0.5 miles from the testing facility. Medical care for 

research-related injuries will be provided at no cost to the subjects.” (V1:43) 

“If you are injured or become ill during the study, tell the study director, laboratory 

technician, or on call nurse immediately. The study director will obtain emergency 

medical treatment for you, if necessary. If your illness or injury is a direct result of 

being in this study, the sponsor of this research will cover the costs of any necessary 
Attachment 1 

Page 27 of 36 

medical treatment that is not covered by your insurance or the insurance of a third 

party under which you are covered. 

“If, after participating in the study, you believe you have become ill as a direct result 

of your participation in the study, please call the study director, Ulrich Bernier 

Ph.D., at (352) 871-8469 (24 hours) or 352-374-5917 or Wendy L. Morrison, RN, 

BSN at 352-339-2179 (24 hours).” (V1:202) 

(e) How does the protocol provide for safety monitoring? 

Subjects are clearly and repeatedly informed that they may remove themselves for 

any reason from the study at any time. All subjects are asked to immediately tell the 

study director, laboratory technician, or on-call nurse if they believe they are 

experiencing a reaction or feel ill during the study. The consent form also states that 

if, after participating in the study, a subject believes he or she has become ill as a 

result of their participation in the study, they should contact the Study Director or the 

on-call nurse anytime, 24-hours a day. Their telephone numbers are provided. 

On the day of testing, a nurse who has read the protocol and discussed the research 

with the study director will be on call. (V1:43) 

(f) How does the protocol provide for post-exposure monitoring or follow-up? Is it 

of long enough duration to discover adverse events which might occur? 

The consent form states: “If, after participating in the study, you believe you have 

become ill as a direct result of your participation in the study, please call the study 

director, Ulrich Bernier Ph.D., at (352) 871-8469 (24 hours) or 352-374-5917 or 

Wendy L. Morrison, RN, BSN at 352-339-2179 (24 hours).” There is no time limit 

given. (V1:202) 

(g) How and by whom will medical care for research-related injuries to subjects be 

paid for? 

 “If your illness or injury is a direct result of being in this study, the sponsor of this 

research will cover the costs of any necessary medical treatment that is not covered 

by your insurance or the insurance of a third party under which you are covered.” 

(V1:202) 

5. Benefits 

(a) What benefits of the proposed research, if any, would accrue to individual subjects? 

There are no direct benefits to subjects.

 
Attachment 1 

Page 28 of 36 

(b) What benefits to society are anticipated from the information likely to be gained 

through the research? 

“While there are no direct benefits to the subjects participating in this research study 

beyond a small compensation for their time, there are indirect benefits to both the 

subjects and society. First, the data collected in this study will be used establish the level 

at which insecticide/repellent treated uniforms prevent mosquito bites. Data generated 

from this study may be used for U.S. EPA registration of a novel insecticide/repellent 

treatment for military clothing. 

“It is expected that this laboratory data can be translated to bite protection of covered 

areas of the body for individuals in the field. Because not all individual biting arthropods 

carry disease agents, a reduction (not necessarily complete elimination) of biting 

pressure is expected to also minimize the risk of disease transmission and subsequently 

reduce overall arthropod-borne disease rates. Reduction of disease rates is dependent 

upon the infectivity rates of the insects in a given area, the overall biting pressure, and 

use compliance. Through factory treatment, we can ensure a high level of compliance for 

use of treated military clothing which should significantly reduce overall arthropodborne disease among military populations, particularly when used in conjunction with 

other prevention methods, such as topical repellents and mosquito netting. 

“These studies are conducted to better protect American military forces from nuisance 

bites and from bites that lead to arthropod-borne diseases. The results of these studies 

may also lead to better protection of U.S. civilians both domestically and while they 

travel abroad because novel repellents can be incorporated into civilian clothing as a 

potentially better alternative to the permethrin-treated clothing already available on the 

market.” (V1:45) 

(c) How would societal benefits be distributed? Who would benefit from the proposed 

research? 

One beneficiary will likely be the sponsor who is seeking EPA-registration for 

etofenprox-treated clothing. Indirect beneficiaries would include the U.S. military and 

civilians who may benefit from wearing etofenprox-treated clothing. 

(d) What is the likelihood that each identified societal benefits would be realized? 

The testing is likely to demonstrate that the formulation is effective at providing the level 

of bite protection sought by the U.S. military, which could result in better protection of 

the U.S. military from mosquito bites and perhaps better protection of U.S. civilians 

through the use of a new type of repellent-treated clothing. 

 
Attachment 1 

Page 29 of 36 

6. Risk/Benefit Balance 

(a) How do the risks to subjects weigh against the anticipated benefits of the research, 

to subjects or to society? 

“Risks have been minimized in terms of disease (through use of colony mosquitoes in a 

laboratory setting only) and with respect to numbers of replicates that a subject will be 

testing (one control and 3 treated specimens). In this study, the biting of subjects will be 

incurred voluntarily to help develop a new product that will provide optimized protection 

against mosquito bites in a field setting where the risk of disease is a bona fide threat.” 

(V1:45) 

The protocol systematically reduces risks to subjects without reducing the robustness of 

the scientific design. No reasonable opportunities to further reduce subject risk have 

been overlooked. The resulting residual risk to subjects is very low. The potential 

benefits from availability of a wider variety of effective repellent-treated clothing are 

likely to be realized, and make the residual risks to subjects in this proposed research 

reasonable. 

7. Independent Ethics Review 

(a) What IRB reviewed the proposed research? 

Western Institutional Review Board 

(b) Is this IRB independent of the investigators and sponsors of the research? Yes 

(c) Is this IRB registered with OHRP? Yes 

(d) Is this IRB accredited? If so, by whom? 

WIRB has full AAHRPP accredditation.

(e) Does this IRB hold a Federal-Wide Assurance from OHRP? 

Yes. 

(f) Are complete records of the IRB review as required by 40 CFR 26.1125 provided? 

Yes. 

(e) What standard(s) of ethical conduct would govern the work? 

This is a protocol for third-party research involving what EPA has interpreted to be 

intentional exposure of human subjects to a pesticide. The study is being conducted with 

the intention of submitting the resulting data to EPA under the Federal Insecticide 
Attachment 1 

Page 30 of 36 

Fungicide and Rodenticide Act (FIFRA). Thus, the primary ethical standards applicable 

to this proposal are 40 CFR 26, Subparts K and L. In addition, the requirements of 

FIFRA §12(a)(2)(P) for fully informed, fully voluntary consent of subjects apply. 

8. Informed Consent 

(a) Will informed consent be obtained from each prospective subject? 

Yes. 

(b) Will informed consent be appropriately documented, consistent with the 

requirements of 40 CFR 26.1117? 

Yes. 

(c) Do the informed consent materials meet the requirements of 40 CFR 26.1116, 

including adequate characterization of the risks and discomforts to subjects from 

participation in the research, the potential benefits to the subject or others, and the 

right to withdraw from the research? 

Yes. 

(d) What is the literacy rate in English or other languages among the intended research 

subjects? 

Ability to speak and read English is a requirement for participation. 

(e) What measures are proposed to overcome language differences, if any, between 

investigators and subjects? 

N/A

(f) What measures are proposed to ensure subject comprehension of risks and 

discomforts? 

Frequent opportunities to ask questions during the consent process. 

(g) What specific procedure will be followed to inform prospective subjects and to seek 

and obtain their consent? 

 “The consent meeting will be held privately between the respondent and the study 

director....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. They 

will be allowed sufficient opportunity to consider whether or not to participate before 

signing the consent form. The participants will be informed of how many bites they are 

likely to obtain and what symptoms of arthropod-borne reactions they should be alert for 
Attachment 1 

Page 31 of 36 

after participation in the study. Participants will be provided a detailed consent form 

(Appendix H) outlining the risks of participating in the study.” (V1:25) 

(h) What measures are proposed to ensure fully voluntary participation and to avoid 

coercion or undue influence? 

Candidates are offered repeated opportunities to decide not to participate; participants are 

offered repeated opportunities to withdraw. Exclusion factors rule out participation by 

employees or students of the Study Director. Recruitment of alternate subjects reduces 

the likelihood that subjects might be reluctant to withdraw. 

9. Respect for Subjects 

(a) How will information about prospective and enrolled subjects be managed to 

ensure their privacy? 

Subject-identifying information will be recorded only once; all subsequent data records 

and reports will refer to individual subjects only by an arbitrary code. “To protect your 

privacy, we will give you an identification number for this study and we will record and 

report all data under that number. We will keep only one record linking your name to this 

number, and we will store it away from other data. We will not identify you by name or in 

any other way in the study report.” (V1:201) 

Provision is made for discrete handling of the pregnancy testing that is required of female 

subjects on the day of testing. “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.” (V1:195) 

(b) How will subjects be informed of their freedom to withdraw from the research at 

any time without penalty? 

The informed consent form states: “Your participation in this study is voluntary. You may 

decide not to be in the study or you may leave the study at any time. Your decision will 

not result in any penalty or loss of benefits to which you are entitled.” (V1:202)

“If you cannot keep your arms in the cage for the entire 15 minutes, you may remove 

them at any time.” (V1:199)

(c) How will subjects who decline to participate or who withdraw from the research be 

dealt with? 

Subjects who decide not to participate after completing the informed consent meeting 

will be paid $20. Subjects who decide not to participate after taking a pregnancy test will 
Attachment 1 

Page 32 of 36 

be paid $25. Subjects who withdraw from the research will be paid for the number of 

pairs of sleeves that they completed at the rate of $25 per pair. (V1:201) 
Attachment 2 

Page 33 of 36 

§ 26.1111 Criteria for IRB approval of research 

USDA Protocol for Bite Protection of Etofenprox-treated Fabric 

Criterion Y/N Comment/Page Reference

(a)(1)(i) Risks to subjects are minimized by using procedures which are consistent with 

sound research design and which do not unnecessarily expose subjects to risk. 

Y 

(a)(1)(ii) Risks to subjects are minimized, whenever appropriate, by using procedures 

already being performed on the subjects for diagnostic or treatment purposes. 

N/A 

(a)(2) Risks to subjects are reasonable in relation to anticipated benefits, if any, to 

subjects, and the importance of the knowledge that may reasonably be expected to 

result. In evaluating risks and benefits, the IRB should consider only those risks and 

benefits that may result from the research (as distinguished from risks and benefits 

subjects would receive even if not participating in the research). The IRB should not 

consider possible long-range effects of applying knowledge gained in the research (for 

example, the possible effects of the research on public policy) as among those 

research risks that fall within the purview of its responsibility. 

Y 

(a)(3) Selection of subjects is equitable, taking into account the purposes of the 

research and the setting in which it will be conducted, and being particularly cognizant 

of the special problems of research involving vulnerable populations, such as 

prisoners, mentally disabled persons, or economically or educationally disadvantaged 

persons. 

Y 

(a)(4) Informed consent will be sought from each prospective subject or the subject’s 

legally authorized representative, in accordance with, and to the extent required by 

§26.1116. 

Y 

(a)(5) Informed consent will be appropriately documented, in accordance with, and to 

the extent required by §26.1117. 

Y 

(a)(6) When appropriate, the research plan makes adequate provision for monitoring 

the data collected to ensure the safety of subjects. 

Y 

(a)(7) When appropriate, there are adequate provisions to protect the privacy of 

subjects and to maintain the confidentiality of data. 

Y 

(b) When some or all of the subjects are likely to be vulnerable to coercion or undue 

influence, additional safeguards have been included in the study to protect the rights 

and welfare of these subjects. 

N/A 
Attachment 3 

Page 34 of 36 

§26.1116 General requirements for informed consent 

USDA Protocol for Bite Protection of Etofenprox-treated Fabric 

Criterion Y/N Comment/Page Reference

No investigator may involve a human being as a subject in research covered by this 

subpart unless the investigator has obtained the legally effective informed consent of 

the subject or the subject’s legally authorized representative 

Y 

An investigator shall seek such consent only under circumstances that provide the 

prospective subject or the representative sufficient opportunity to consider whether or 

not to participate and that minimize the possibility of coercion or undue influence 

Y 

The information that is given to the subject or the representative shall be in language 

understandable to the subject or the representative 

Y 

No informed consent, whether oral or written, may include any exculpatory language 

through which the subject or the representative is made to waive or appear to waive 

any of the subject’s legal rights, or releases or appears to release the investigator, the 

sponsor, the institution or its agents from liability for negligence 

Y 

(a) In seeking informed consent the following information 

shall be provided to each subject 

(1) A statement that the study involves research, an explanation of the 

purposes of the research and the expected duration of the subject’s 

participation, a description of the procedures to be followed, and 

identification of any procedures which are experimental 

Y 

(2) A description of any reasonably foreseeable risks or discomforts to the 

subject 

Y 

(3) A description of any benefits to the subject or to others which may 

reasonably be expected from the research 

Y 

(4) A disclosure of appropriate alternative procedures or courses of 

treatment, if any, that might be advantageous to the subject 

Y 

(5) A statement describing the extent, if any, to which confidentiality of 

records identifying the subject will be maintained 

Y 

(6) For research involving more than minimal risk, an explanation as to 

whether any compensation and an explanation as to whether any medical 

treatments are available if injury occurs and, if so, what they consist of, or 

where further information may be obtained 

Y 

(7) An explanation of whom to contact for answers to pertinent questions 

about the research and research subjects’ rights, and whom to contact in 

the event of a research-related injury to the subject 

Y 

(8) A statement that participation is voluntary, refusal to participate will 

involve no penalty or loss of benefits to which the subject is otherwise 

entitled, and the subject may discontinue participation at any time without 

penalty or loss of benefits to which the subject is otherwise entitled 

Y 

(b) When appropriate, one or more of the following elements of information shall also be provided to each subject 

(1) A statement that the particular treatment or procedure may involve 

risks to the subject (or to the embryo or fetus, if the subject may become 

pregnant) which are currently unforeseeable 

Y 

(2) Anticipated circumstances under which the subject’s participation may 

be terminated by the investigator without regard to the subject’s consent 

Y 

(3) Any additional costs to the subject that may result from participation in 

the research 

Y 

(4) The consequences of a subject’s decision to withdraw from the 

research and procedures for orderly termination of participation by the 

subject 

Y 

(5) A statement that significant new findings developed during the course 

of the research which may relate to the subject’s willingness to continue 

participation will be provided to the subject 

N/A 

(6) The approximate number of subjects involved in the study Y 

(e) If the research involves intentional exposure of subjects to a pesticide, the subjects 

of the research must be informed of the identity of the pesticide and the nature of its 

pesticidal function. 

Y 
Attachment 4 

Page 35 of 36 

§26.1117 Documentation of informed consent 

USDA Protocol for Bite Protection of Etofenprox-treated Fabric 

Criterion Y/N Comment/Page Reference

(a) Informed consent shall be documented by the use of a written consent form 

approved by the IRB and signed by the subject or the subject’s legally authorized 

representative. A copy shall be given to the person signing the form. 

Y Consent form pp. 193-203 

(b)(1) The consent form may be a written consent document that embodies the 

elements of informed consent required by §26.1116. This form may be read to the 

subject or the subject’s legally authorized representative, but in any event, the 

investigator shall give either the subject or the representative adequate opportunity to 

read it before it is signed; or 

Y Consent form meets requirements of 

§26.1116; procedure described in 

protocol §8.1.6 provides adequate 

opportunity to read it before it is 

signed. 

(b)(2) The consent form may be a short form written consent document stating that the 

elements of informed consent required by §26.1116 have been presented orally to the 

subject or the subject’s legally authorized representative. When this method is used, 

there shall be a witness to the oral presentation. Also, the IRB shall approve a written 

summary of what is to be said to the subject or the representative. Only the short form 

itself is to be signed by the subject or the representative. However, the witness shall 

sign both the short form and a copy of the summary, and the person actually obtaining 

consent shall sign a copy of the summary. A copy of the summary shall be given to the 

subject or the representative, in addition to a copy of the short form. 

N/A 
Attachment 5 

Page 36 of 36 

40 CFR 26.1125 Submission of proposed human research for EPA review

USDA Protocol for Bite Protection of Etofenprox-treated Fabric 

Any person or institution who intends to conduct or sponsor human research covered by §26.1101(a) shall, after receiving 

approval from all appropriate IRBs, submit to EPA prior to initiating such research all information relevant to the proposed 

research specified by §26.1115(a), and the following additional information, to the extent not already included: 

Requirement Y/N Comments/Page Refs 

The following Information, to the 

Extent not already included: 

§1125(a) 

a discussion of: 

(1) The potential risks to human subjects Y V1:43-44 

(2) The measures proposed to minimize risks to the human subjects; Y V1:43-44 

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

research, and to whom they would accrue Y V1:45 

(4) Alternative means of obtaining information comparable to what 

would be collected through the proposed research; and Y none 

(5) The balance of risks and benefits of the proposed research. Y V1:45 

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

agreements as originally provided to the IRB, and as approved by the IRB. Y V1:25 

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

advertisements proposed to be used. Y V1:23, 204 

§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 V1:25 

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

sponsors. 

Y Volume 2 

§1125(f): Official notification to the sponsor or investigator. . . that research 

involving human subjects has been reviewed and approved by an IRB. Y V1:208-210 

all information relevant to the proposed research 

specified by § 26.1115(a) 

(1) Copies of 

 all research proposals reviewed by the IRB, 

 scientific evaluations, if any, that accompanied the proposals 

reviewed by the IRB, 

 approved sample consent documents, 

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

subjects. 

Y 

n/a 

Y 

n/a 

Volume 2 

V1:193-203 

(2) Minutes of IRB meetings . . . 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 

Y 

Y 

n/a 

n/a 

Separately provided to HSRB 

members 

(3) Records of continuing review activities. n/a 

(4) Copies of all correspondence between the IRB and the investigators. Y Volume 2 

(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, for example, full-time employee, a member of 

governing panel or board, stockholder, paid or unpaid consultant. 

Y 

Y 

V2:22 

(6) Written procedures for the IRB in the same detail as described in 

§26.1108(a) and §26.1108(b). N Separately provided to HSRB members 

(7) Statements of significant new findings provided to subjects, as required by 

§26.1116(b)(5). n/a n/a for protocols