UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

WASHINGTON D.C., 20460

OFFICE OF CHEMICAL SAFETY AND 

POLLUTION PREVENTION

 December 15, 2015

MEMORANDUM

SUBJECT: Ethics Review of Davis et al Research on Flea Collars with TCVP

FROM: Maureen Lydon, Human Studies Ethics Review Officer

Office of the Director

Office of Pesticide Programs

TO: Dana Vogel, Director

Wade Britton, Environmental Health Scientist

Health Effects Division

Office of Pesticide Programs

REF: M. Keith Davis, J. Scott Boone, John E. Moran, John W. Tyler, Janice E. 

Chambers, Assessing intermittent pesticide exposure from flea control collars 

containing the organophosphorous insecticide tetrachlorvinphos, Journal of 

Exposure Science and Environmental Epidemiology (2008) 18, 564-570.

I have reviewed the referenced article on two studies which assess intermittent pesticide 

exposure from flea control collars containing the organophosphorous insecticide 

tetrachlorvinphos (TCVP). 

As described in the article, “Because TCVP has been used in flea collars, the amount of

exposure to TCVP that could occur in children and adults from the use of a TCVP-containing 

collar on a pet dog was assessed. A long study (about 4 months) was conducted first to determine 

the time course of transferable residue peak and dissipation as assessed by transferable residues 

of TCVP from the fur of dogs to white cotton gloves used to rub the dogs.” Study 1, conducted 

in 1998, included dog plasma cholinesterase measurements as well. “This was followed by a 

shorter study conducted over 3 weeks to include human biomonitoring of the TCVP metabolite 

2,4,5-trichloromandelic acid (TCMA) in urine of children and adults. TCVP residues transferred 

to tee shirts worn by children by contact with the dog were also quantified to determine whether 

tee shirts might serve as a surrogate of exposure.” As with the first study, study 2, conducted in 

2002, also assessed transferable residues of TCVP from the fur of dogs to white cotton gloves 

used to rub the dogs. In summary, four types of data were collected addressing: 1) transferable 

residues of TCVP from the fur of dogs to white cotton gloves: 2) TCVP residues on tee shirts 

worn by children; 3) the levels of the metabolite TCMA in urine from children and adults; and 4) 

dog plasma cholinesterase activity
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Studies 1 and 2 are systematic investigations designed to develop or contribute to 

generalizable knowledge and so meet the regulatory definition of research. Through collection 

of urine and tee shirt samples, study 2 obtained data about individuals and so meets the 

regulatory definition of human subject research. Although the families involved in the studies 

already used flea collars, the researchers bought and provided specific flea collars to the 

participating families and asked that their dogs wear the flea collars during the studies. As a 

result, the research constitutes intentional exposure. 

EPA’s Office of Pesticide Programs (OPP) wants to rely only on one sub-set of the data 

generated from this research, specifically the data on transferable residues of TCVP from the fur 

of dogs to white cotton gloves used to rub the dogs. The generation of this data did not involve 

children. However, the data was collected as part of broader research which involved children as 

study participants when they wore tee shirts and provided urine samples. As a result, specific 

federal regulations come into play before EPA can potentially rely on the TCVP glove residue 

data. 

40 CFR Subpart Q, §26.1703, prohibits EPA from relying on data from any research 

subject to this subpart involving intentional exposure of any human subject who is a pregnant 

woman (and therefore her fetus), a nursing woman, or a child. §26.1706 provides an exception. 

Under 40 CFR §26.1706, EPA can only rely on this research if it is crucial to making a decision 

to impose a more stringent regulatory restriction than could be justified without the data. If 

EPA’s Office of Pesticide Programs (OPP) decides to rely on the TCVP glove residue data,

under 40 CFR §26.1706, OPP must first complete three required steps. EPA must obtain the 

views of the Human Studies Review Board, provide an opportunity for public comment, and 

publish a full explanation of its decision to rely on the data, including a thorough discussion of 

the ethical deficiencies of the underlying research and the full rationale for finding that EPA met 

the standard in 40 CFR §26.1706 (c) (i.e., that the research is essential to a more stringent 

regulatory action to improve protection of public health). 

I defer to others for a full review of the scientific validity of the TCVP glove residue data

on which EPA wishes to rely. If the data were determined not to have scientific validity, it 

would also not be ethically acceptable and EPA could not use it. If EPA decides to rely on the 

TCVP residue data on gloves, and completes the required activities noted above, I find that there 

is no regulatory barrier to EPA relying on this research in actions taken under FIFRA or §408 of 

FFDCA. The rest of this memorandum and attachment(s) provide OPP’s ethics review of the 

research. 

I. Summary Characteristics of the Research

A. Summary of Studies Described in Article

The published research article is entitled “Assessing intermittent pesticide exposure from 

flea control collars containing the organophosphorous insecticide tetrachlorvinphos” by M. Keith 

Davis, J. Scott Boone, John E. Moran, John W. Tyler, and Janice E. Chambers. The Davis et al 

article discusses two studies which collected data useful for assessing the amount of exposure to 

tetrachlorvinphos (TCVP) that could occur in children and adults from the use of a TCVPcontaining collar on a pet dog. The studies were conducted in Oktibbeha County, Mississippi, 
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with 55 volunteer households who routinely used flea control products on their pet dogs. In each 

study, Hartz Control Ultimate Flea Collars F 14.55% TCVP were purchased from a local 

department store. 

“Study 1 was a longer term, 112-day study conducted from September to December 1998

because the collar was recommended by the manufacturer for 6-month use. Twenty-three dogs 

from different households were used, and petting samples from fur and dog plasma 

cholinesterase (ChE) measurements were obtained.” Study 1 “was conducted first to determine 

the time course of transferable residue peak and dissipation as assessed by transferable residues 

of TCVP from the fur of dogs to white cotton gloves used to rub the dogs” at three locations: 

near the base of the tail, at the neck with the collar removed, and at the neck with the collar in 

place. “Dogs were petted in a marked 10 x 4 inch area with clean, white, cotton gloves for a 

continuous 5-minute period.” Samplers were students enrolled at the College of Veterinary 

Medicine, Mississippi State University. For study 1, rubbing samples were obtained prior to

collar placement and at 4 hours, and 3, 7, 14, 28, 56, 84, and 112 days post-collar application. 

Study 2 was a shorter, 21-day study conducted in September and October 2002 which 

included human biomonitoring of the TCVP metabolite 2, 4, 5-trichloromandelic acid (TCMA) 

in urine of children and adults. Study 2 involved 44 volunteer human subjects including one 

child and one adult who were selected from each of the 22 families who participated in study 2. 

Study 2 involved 22 pet dogs from different households and quantified the residues of TCVP 

from: 1) glove/rubbing samples; 2) tee shirt samples; and 3) levels of the metabolite TCMA in 

the urine of children and adults.

1. Glove/Rubbing Samples: Under one component of the study, “dogs were petted in a 

marked 10 x 4 inch area with clean, white, cotton gloves for a continuous 5-minute 

period.” For study 2, rubbing samples were obtained prior to collar placement (day 0) 

and again at 5 and 12 days following collar placement. 

“Samplers utilized during both studies were students enrolled in the veterinary 

medicine curriculum at the College of Veterinary Medicine, Mississippi State 

University.” 

2. Tee Shirt Samples: Under the second component of the study, child participants were 

supplied with a new, laundered, white cotton tee shirt to wear on the day before the 

treatment and on each of days 7–11 after the collars were placed on the dogs; the tee 

shirts were worn by children for about 4 hours at selected times after pesticide 

application. 

3. Urine Samples: The first morning urine samples were collected from the child 

wearing the tee shirt and from one adult in the same household on the day prior to the 

treatment and then again on each of days 8–12 post-collar placement. 

B. EPA Reliance on Glove Residue Data and Reason for Presentation to HSRB

In order to improve the protection of public health, EPA would like to rely only on the 

TCVP glove residue data from study 1 and study 2 in its risk assessment and to potentially 
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impose a more stringent regulatory restriction than the Agency could do otherwise without the 

data. Study 2 also involved the collection of data based on tee shirts worn by children and urine 

samples from children and adults participating in the study. With the exception of the dog

plasma cholinesterase measurement, all of the data collected constitutes exposure assessment 

research and EPA cannot reasonably separate the different types of data in these studies as 

different types of human research. Because study 2 encompassed more than TCVP glove residue 

data and involved children wearing tee shirts and providing urine samples, study 2 constitutes 

research involving intentional exposure of children. For that reason, even though OPP does not 

wish to rely on the data involving children, OPP is submitting the research to the Human Studies 

Review Board for their review.

From a science perspective, OPP is interested in the Board’s perspective on the scientific 

validity of the TCVP glove residue data, which is the only data on which EPA intends to rely. 

From an ethics standpoint, OPP is interested in the HSRB’s perspective on two topics: 

1) Does the HSRB have any comments on EPA’s determination that the samplers were 

not human subjects?

2) Does the HSRB have any comments on the ethical conduct of the research? 

C. Support from EPA Grants

The acknowledgements at the end of the research article note that the research was 

supported by grants from the US EPA’s Science to Achieve Results (STAR) grant program 

(specifically grant numbers R825170 and R828017). I contacted EPA’s Office of Research and 

Development (ORD), the Office of the Science Advisor (OSA) to determine if ORD had 

documentation associated with either or both grants. ORD provided the documentation in 

Attachment 1 related to ORD’s review of grant proposal R828017. (ORD reviewed the grant 

proposal at the time because it was proposed for partial funding by EPA and involved human 

research. Documentation for the other grant, R825170, is not available because it did not require 

ORD review.) The ORD file presents several documents, including the abstract of the proposed 

research entitled, “Assessing Levels of Intermittent Exposure of Children to Flea Control 

Insecticides from the Fur of Dogs.” Page 11 of the abstract cites the research objectives and 

describes “3 specific aims, each of which will involve a correlation of residues from the rubbing 

procedure with cotton gloves (the technique we are using at present), residues from tee shirts 

worn by a child in the household, and urinary metabolites of this child and an adult in the 

household: 1. To determine the exposure of children to permethrin resulting from residues from 

the fur of dogs from a permethrin spot treatment; 2. To determine the exposure of children to 

TCVP resulting from residues from the fur of dogs treated with a TCVP flea collar; and 3. To 

determine the exposure of children to chlorpyrifos resulting from residues from the fur of dogs 

treated with a chlorpyrifos flea collar.” The published article by Davis et al describes the two 

studies which support aim #2, noted above, related to the TCVP flea collars. The primary 

investigator for the two studies, Dr. Janice Chambers from Mississippi State University, 

confirmed that the TCVP research described in the ORD file relates to the two studies discussed 

in the Davis et al article. For that reason, pertinent information from the ORD file is referenced 

throughout this ethics review. The ORD documentation in Attachment 1 includes:

1) The Mississippi State University (MSU) Statement of “Assurance of Compliance with EPA 

Regulations for Protection of Human Research Subjects”; 
Page 5 of 18

2) Institutional Review Board (IRB) statement of institution and investigator compliance with 40 

CFR 26; 

3) Certification of IRB Approval and Institutional Endorsement signed by MSU’s Vice President 

for Research, the MSU IRB Chairman at the time and the MSU project investigator Janice 

Chambers; 

4) List of MSU IRB members at the time of the research; 

5) Draft minor’s assent form; 

6) Draft consent form for adult human subjects (also referred to as the authorization for 

participation in the research project on flea collars); 

7) 16-page abstract describing the research along with 3.5 pages of additional references; and 

8) ORD’s comments on the draft consent forms. 

The file and Attachment 1 also includes an email from ORD’s then Assistant Director for EPA’s 

National Center for Environmental Research (NCER) identifying ORD’s comments on the

consent forms. The primary investigator (PI) agreed to the changes requested by ORD. The 

final versions of the consent forms are not included in the file. Both the PI and EPA agree that 

the changes to the consent forms must have been made because receipt of grant funding was 

contingent on addressing EPA comments. Given that the grants were awarded, EPA reasonably 

assumes that comments were addressed. This ethics review takes into account, when 

appropriate, the 16-page description of the research project, the draft consent forms and the IRB 

correspondence included in the ORD file.

D. Contact with IRB and Primary Investigator

To supplement the ethics information provided in the journal article, I contacted the 

relevant institutional review board, the Institutional Review Board for Research on Human 

Subjects at Mississippi State University, and requested copies of the available records about the 

two studies profiled in the Davis et al article. Because study 2 was conducted in 2002, the IRB 

did not maintain documentation past the required retention time. As a result, the IRB did not 

have copies of their reviews. However, the ORD file included IRB-related documents listed on

page 4 of this memo. 

After speaking with the IRB, I contacted the Primary Investigator for the Research, Dr. 

Janice E. Chambers, Professor and Director of the College of Veterinary Medicine Faculty at 

MSU. Dr. Chambers confirmed relevant background information from the ORD file on how the 

study was conducted; that background information is incorporated into this ethics review. My 

interview with Dr. Chambers is documented in Attachment 2. Dr. Chambers was a member of 

the Human Studies Review Board (HSRB) for 8 years, from March 27, 2006 until August 31, 

2014.

E. Role of Veterinary Students in Rubbing Dogs and EPA’s Position

As discussed on page 566 of the Davis et al article, “Samplers utilized during both studies 

were students enrolled in the veterinary medicine curriculum at the College of Veterinary 

Medicine, Mississippi State University.” Primary investigator Dr. Chambers confirmed that both 

the researchers and IRB viewed the samplers as technicians in the study and not as human 
Page 6 of 18

subjects. EPA considers the view of the researchers’ and IRB on this pre-rule research to be a 

reasonable one.

Looking at the information in the research article and ORD file, the primary investigator 

did not obtain data about the technicians, nor did they ever intend to do so. Human subject 

means a living individual about whom an investigator conducting research obtains data. The 

researchers were not focusing on the technicians in this study. Instead, the researchers were 

collecting data only about the residues on the gloves.

The researchers did not question the technicians about how they rubbed the animals. If it 

were a situation where the dog owners were petting their own animals throughout the day and the 

researchers had to interview and interact with the dog owners to collect data about how long they 

interacted with their pets, where they petted their dogs, and in what manner, the researchers 

would be collecting data about the human subjects, not just about the residues. This was not the 

case in this situation. As stated in the research article, the samplers/technicians rubbed the dogs 

“in a marked 10 x 4 inch area with clean, white, cotton gloves for a continuous 5-min period.” 

The dogs were rubbed in three specific locations (near the base of the tail, at the neck with the 

collar removed, and at the neck with the collar in place). In the ORD file, page 14 of the 

research abstract further states that “the samplers will be trained so that consistency in the 

sample collection is maintained among dogs and among samplers.” The samplers/technicians 

were wearing gloves and stroking the animals in a standardized and prescribed way. The 

researchers did not collect information or data about the samplers. EPA agrees with the primary 

investigator and IRB that the samplers were not human subjects in these studies.

F. Ethical Considerations

1. Value of the Research to Society: 

The objective of this research was to assess the amount of exposure to TCVP that could 

occur in children and adults from the use of a TCVP-containing collar on a pet dog. 

According to the authors, one research area that had not been adequately explored is the 

possibility that residues of insecticides remaining on pet fur from flea control collars could 

be a significant source of pesticide exposure in children who pet or hug treated animals. 

EPA is considering some of the data from this research in its risk assessment for 

tetrachlorvinphos and to potentially take a more stringent regulatory action than could be 

taken without the data.

2. Subject Selection: 

a. Demographics. The studies were conducted in Oktibbeha County, Mississippi, with 

volunteer households whose pet dogs routinely wore flea collars. For study 1, the dogs 

from 23 families were used. For study 2, one child and one adult participated from 

each of the 22 volunteer households, which equates to 44 human subjects for study 2. 

The participating children ranged in age from 3 to 13. Approximately half of the 

participating children and adults were male and half were female. 

b. Pregnancy and Nursing Status and Participation of Children. There is no indication 

in the research article or ORD file that pregnant or nursing women participated in the 
Page 7 of 18

studies. However, 22 children, ranging in age from 3 to 13, participated in study 2. 

EPA can only rely on research involving children if the criteria and procedures in 40 

CFR §26.1706 are met.

c. Recruitment. The article states that, “the studies were conducted in Oktibbeha County, 

Mississippi (USA), with volunteer households having pet dogs” and that “participating 

families were volunteers who routinely used flea control products on their pet dogs.” 

The article states that, “One child and one adult were selected from each participating 

family (study 2 only) as previously described by Chambers et al (2007).” However, the 

2007 article to which this citation refers does not include additional information 

regarding recruitment or subject selection. 

The ORD file includes the following pertinent information. As stated on page 13 of the 

research abstract, “Dogs selected for this study will be owned by professional (DVM) 

or graduate students enrolled in the College of Veterinary Medicine, or staff/faculty 

members of Mississippi State University with a child aged 4-10 years in the household 

who routinely plays with this dog. Students or staff should be the most reliable group of 

owners (in contrast to the general public) in that they are accessible daily, their dogs 

can readily be treated and sampled when the students are in class or the staff members 

are at work, and as members of the academic community, the compliance and 

appreciation of the value of research should be high. Dogs participating in this study

must be enrolled in the Small Animal Community Practice Health Maintenance 

Program, so that their health status and vaccination history are known..”. Primary 

Investigator Janice Chambers confirmed that MSU’s College of Veterinary Medicine is 

located in Oktibbeha County, Mississippi. 

Page 14 of the research abstract includes a section entitled, “Selection of human test 

subjects” which reads as follows: “The human subjects will be residents in the same 

household as the canine test subject. The household must have a child (either sex) in 

the age range of 4-l 0 years who regularly plays with the dog, and an adult (probably a 

parent; either sex); both subjects must be willing to provide the samples required by the 

study protocol. The samples will be tee shirts from the child and first morning urine 

samples from the child and the adult. The age, sex, height and weight of both child and 

adult test subjects will be recorded, and the adult will be asked to give an estimate of 

the amount of time and degree of contact that the child and the adult had with the dog 

on the day of the tee shirt sample; these data will be available for later correlations with 

residue/metabolite data. A description of the protocol and an informed consent form 

will be developed, and the subjects will be assured of anonymity. These protocols and 

approval forms will be approved by the Institutional Review Board for Research on 

Human Subjects.”

3. Risks and Benefits: 

a. Risks. The Davis et al article is silent about risks to subjects from their participation 

in the study. The article states that, “Because TCVP is an organophosphate 

insecticide, its primary mechanism of toxicity is the inhibition of the nervous system 

enzyme, acetylcholinesterase (Ecobichon, 1996).” However, only families who 
Page 8 of 18

routinely used flea control products on their pet dogs participated in the studies. The 

researchers purchased, for use in the studies, over-the-counter flea collars that were 

already registered with the U.S. EPA.

b. Benefits. There were no benefits to the subjects and none were noted in the research 

article.

c. Risk-Benefit Balance. The research article does not explicitly discuss the risk-benefit 

balance. However, the stated objective of this research was to assess the amount of

exposure to TCVP that could occur in children and adults from the use of a TCVPcontaining collar on a pet dog. According to the authors, one research area that does 

not seem to have been explored is the possibility that residues of insecticides remaining 

on pet fur from flea control collars could be a significant source of pesticide exposure 

in children who pet or hug treated animals. The article provides study results in this 

area of research.

4. Independent Ethics Review: The Institutional Review Board for Research on Human 

Subjects at Mississippi State University reviewed and approved the sampling protocols and 

informed consent forms. Furthermore, EPA’s Office of Research and Development 

(ORD), the National Center for Environmental Research and Quality Assurance 

(NCERQA) reviewed the STAR grant proposal focusing on this research because it 

involved the use of human subjects and requested funding by EPA. ORD supported the 

research dependent on incorporation of NCERQA comments on the consent forms. 

5. Informed Consent: The article states that “A copy of the protocol was distributed to each 

participating household, and informed consent was obtained from the adults. Children 

were informed verbally of the procedures and oral or written assent was obtained from 

them. The Institutional Review Board for Research on Human Subjects at Mississippi 

State University approved all sampling protocols and informed consent forms.” 

The ORD file contains a draft consent form for adult human subjects entitled, 

“Authorization for Participation in Research Project (collar)” and a draft “Minor’s Assent 

Form,” both provided in Attachment 2. The “authorization for participation” form states 

that the study involves research and identifies its purpose, the expected duration, number of 

urine and tee shirt samples to be provided, that research results will be coded, participants 

are free to withdraw, provides a contact for information and identifies compensation of 

$150 for each participating household. The consent form also states that, “no risks are 

anticipated to the participants” as a result of participating in the study. The implication is 

that since the families already used flea collars on their dogs, there was no added risk from 

participating in the study. However, the researchers proposed the following hypothesis on 

the bottom of page 10 and top of page 11 of the research abstract submitted to ORD: “The 

residues of insecticides available for intermittent transfer to children from the fur of dogs 

treated by either a spot treatment or a collar for flea control will be appreciable and of a 

magnitude necessitating inclusion in cumulative risk assessments of pesticides to children; 

secondly, that the fur rubbing procedure developed to quantify dislodgeable residues 

provides a useful estimate of insecticide residues which could be transferred from the fur of 

dogs to children.” Although the families involved already used flea collars which were 
Page 9 of 18

registered by EPA, in the interest of transparency, the researchers should have shared their 

hypothesis with the parents of the participating children and included it in the consent form. 

This information may have been stated in the protocol provided to the families but we do 

not know.

The minors’ assent form states that researchers “will specifically obtain assent from the 

children recruited to our project ‘Assessing levels of intermittent exposures of children to 

flea control insecticides from fur of dogs’ for their participation in the project...We will 

explain that the child's parent or guardian has given us permission to request his/her help 

participation in the research project. We will then explain the urine collection protocol and 

the tee shirt protocol to the children in language appropriate to the age of the child and 

obtain his/her assent to participate. We will not explain the connection to the pesticide 

residues on the dog so as not to alter the behavior of the child with the dog. We will obtain 

the children's assent orally because of the age range of the children involved.”

6. Respect for Subjects: The article does not reveal the subjects’ identifies. The article 

states that, “A copy of the protocol was distributed to each participating household, and 

informed consent was obtained from the adults. Children were informed verbally of the 

procedures and oral or written assent was obtained from them. The Institutional Review 

Board for Research on Human Subjects at Mississippi State University approved all 

sampling protocols and informed consent forms.” The tee shirts worn by the children as 

part of study 2 were provided by the researchers. The ORD file includes the following 

additional information relevant to respect for subjects.

Researchers demonstrated respect for subjects by providing light-weight, short-sleeve (as 

opposed to long-sleeve) tee shirts to the children participating in study 2. Page 15 of the 

research abstract in the ORD file states that, “The child will wear the tee shirt during the 

afternoon and evening of the sampling day for a 5 hour period. (While long sleeve shirts 

might give a better estimate of exposure, long sleeves would not be tolerable and could be a 

health risk because of the heat during 6-8 months of the year in this region.) The child will 

not be instructed to alter his/her normal behavior with respect to the dog.” Page 12 of the 

abstract states that the tee shirt “is a piece of clothing which the child would readily wear 

without embarrassment in front of his/her peers and siblings.” These statements illustrate 

that the researchers took into account the feelings and comfort of the children participating 

in study 2. 

Furthermore, page 20 of the abstract in the ORD file states that, “The protocols do not 

require any invasive procedures and are not expected to cause any physical or mental 

distress to the individuals involved. The test subjects will be asked to continue their routine 

activities and to not modify their activities in any way. They will be asked to provide urine 

samples on a set schedule; we will give them written assurance that the urine sample will 

be used only for quantitation of insecticide urinary metabolites and will not be used for any 

other purposes, and that all data will remain anonymous.”

With regard to compensation, the consent form states that each participating household will 

receive $150. The ORD file, page 20 of the abstract, states that, “Dogs participating in this 

study will be required to be enrolled in MSU's Small Animal Community Practice Health 
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Maintenance Program whereby the dog's health will be known by physical examination and 

vaccination history. If the dog was not previously enrolled in the program, the cost of the 

initial physical will be borne within the $100 incentive offered to each participant.” Page 

19 of the abstract has a section on incentives for dog owners which states, “In order to 

provide incentives for the cooperative participation of the dog owners in this study, we

propose to offer $100 equivalent of veterinary care provided by the Animal Health Center

of the College of Veterinary Medicine for each dog participating in the study. In addition, 

the spot treatments, shampoos, and collars constituting the study will be provided to the 

owners without additional charge. An additional $150 cash incentive will also be given to 

each participating household because of the shirt and urine samples. We predict that these 

incentives will lead to more than an adequate number of households to participate in this 

study. We currently estimate that there will be at least 500 dogs owned by veterinary and 

graduate students and faculty and staff at the College of Veterinary Medicine at any given 

time, and many of these households would have a child of the appropriate age. In the 

unlikely event that insufficient numbers of households can be enrolled for the study, we 

will open the participation to students, faculty, and staff members of Mississippi State 

University outside the College of Veterinary Medicine; these individuals would be offered 

the same incentives.”

II. Applicable Standards

A. Definition of Research

The relevant definitions are discussed below: 

• In order to meet the regulatory definition of research involving human subjects, the 

regulatory definition of both research and human subject must be met. 

• Studies 1 and 2 described in the Davis et al article are systematic investigations designed 

to develop or contribute to generalizable knowledge and so meet the regulatory definition 

of research.

• Through collection of urine and tee shirt samples, study 2 obtained data about individuals 

and so meets the regulatory definition of human subject research. 

• Although the families involved already used flea collars, the researchers bought and 

provided specific flea collars to the participating families and asked that their dogs wear 

the flea collars during the studies. As a result, study 2 constitutes intentional exposure. 

• These studies determined, in part, the amount of transferable TCVP residues and the 

urinary TCMA concentration. The purpose of these studies was not to identify or 

measure a toxic effect in humans. The research was conducted prior to the effective date 

of the human research rule. 

As a result, the only reason that EPA is bringing this pre-rule research to the HSRB for review is 

because study 2 involved children as participants in providing tee shirt and urine samples.

B. Standards Applicable to the Conduct of the Research

The research article was published in 2008 and focuses on two studies. Study 1 was 

conducted in 1998 and study 2 was conducted in 2002, both before EPA’s Rule for Protection of 

Human Subjects of Research became effective in 2006. Thus, 40 CFR part 26 did not apply 
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when this research was conducted. However, the Common Rule was in place and applies to the 

underlying research which received EPA STAR grant funding. 

Key elements of the Common Rule are IRB oversight and prior approval, an acceptable 

informed consent process and consent form, risk minimization, a favorable risk-benefit balance, 

equitable subject selection, and fully informed, fully voluntary participation by subjects.

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

(FIFRA) existed at the time of these studies and reads: 

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

human beings unless such human beings (i) are fully informed of the nature and 

purposes of the test and of any physical and mental health consequences which 

are reasonably foreseeable therefrom, and (ii) freely volunteer to participate in the 

test. 

C. Standards Applicable to EPA’s Reliance on the Research

The Agency’s rule (40 CFR part 26 subpart Q) defines standards for EPA to apply in 

deciding whether to rely on research—like study 1 and 2—involving intentional exposure of 

human subjects. The applicable acceptance standards from 40 CFR part 26 subpart Q are these:

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

subject to this subpart involving intentional exposure of any human subject who is a 

pregnant woman (and therefore her fetus), a nursing woman, or a child. 

§26.1704: Except as provided in §26.1706, in actions within the scope of 

§26.1701, EPA shall not rely on data from any research initiated before April 7, 

2006, if there is clear and convincing evidence that: (1) the conduct of the 

research was fundamentally unethical (e.g., the research was intended to seriously 

harm participants or failed to obtain informed consent), or (2) the conduct of the 

research was deficient relative to the ethical standards prevailing at the time the 

research was conducted in a way that placed participants at increased risk of harm 

(based on knowledge available at the time the study was conducted) or impaired 

their informed consent.

§26.1706: This section establishes the exclusive criteria and procedure by which 

EPA may decide to rely on data from research that is not acceptable under the 

standards in §§26.1703 through 26.1705. EPA may rely on such data only if all 

the conditions in paragraphs (a) through (d) of this section are satisfied:

(a) EPA has obtained the views of the Human Studies Review Board concerning the 

proposal to rely on the otherwise unacceptable data,

(b) EPA has provided an opportunity for public comment on the proposal to rely on the 

otherwise unacceptable data,
Page 12 of 18

(c) EPA has determined that relying on the data is crucial to a decision that would impose 

a more stringent regulatory restriction that would improve protection of public health, 

such as a limitation on the use of a pesticide, than could be justified without relying on 

the data, and

(d) EPA has published a full explanation of its decision to rely on the otherwise 

unacceptable data, including a thorough discussion of the ethical deficiencies of 

the underlying research and the full rationale for finding that the standard in 

paragraph (c) of this section was met.

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

Rodenticide Act (FIFRA) applies. This passage reads: 

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

human beings unless such human beings (i) are fully informed of the nature and 

purposes of the test and of any physical and mental health consequences which 

are reasonably foreseeable therefrom, and (ii) freely volunteer to participate in the 

test. 

D. Completeness of Ethics Information

EPA ensured compliance with §26.1303 as demonstrated by the checklist in Attachment 

3; the checklist summarizes the completeness of available information pertaining to the ethical 

conduct of the completed research. 

E. Compliance with Applicable Standards

§26.1703. Except as provided in §26.1706, prohibition of reliance on research involving 

intentional exposure of human subjects who are pregnant women (and therefore their 

fetuses), nursing women, or children.

Study 1 did not involve children or pregnant or nursing women. As part of study 2, 

conducted in 2002, researchers collected three types of data: TCVP glove residue data, tee shirt 

samples, and urine samples. Children participated in study 2 by wearing the tee shirts and 

providing urine samples. As such, EPA is prohibited from relying on study 2 unless the 

conditions of §26.1706 are met.

§26.1704. Prohibition of reliance on unethical human research with nonpregnant adults 

conducted before April 7, 2006.

There is no clear and convincing evidence that this pre-rule research was 

fundamentally unethical; that is, the research was not intended to harm the participants 

and did not fail to obtain informed consent. Similarly, the conduct of the research was 

not deficient relative to the ethical standards prevailing at the time the research was 

conducted; the studies did not place participants at increased risk of harm (based on 

knowledge available at the time the study was conducted) or impair their informed 

consent. As a result, EPA is not prohibited from relying on this research under §26.1704.
Page 13 of 18

§26.1706: Criteria and procedures for decisions to protect public health by relying on 

otherwise unacceptable data.

As of the date of this memo, OPP wishes to rely on the TCVP glove residue data

generated in study 1 and 2. The data may be crucial to a potential EPA decision to improve 

public health protection by imposing a more stringent regulatory restriction than could be 

justified without the data. If EPA proceeds under §26.1706, EPA needs to obtain the views of 

the Human Studies Review Board, provide an opportunity for public comment, and publish a full 

explanation of its decision to rely on the data, including a thorough discussion of the ethical 

deficiencies of the underlying research and the full rationale for finding that EPA met the 

standard in 40 CFR §26.1706 (c) (i.e., that the research is essential to a more stringent regulatory 

action to improve protection of public health). 

FIFRA §12(a)(2)(P): 

The researchers complied with FIFRA §12(a)(2)(P). The human subjects were 

informed of the nature and purposes of the research and volunteered to participate in the 

studies.

III. Conclusion

I defer to others for a full review of the scientific validity of the TCVP glove residue data 

on which EPA wishes to rely. If the data were determined not to have scientific validity, it 

would also not be ethically acceptable and EPA could not use it.

The TCVP glove residue data was one component of broader research that also included 

tee shirt and urine samples. Children and adults were participants in study 2 and provided tee 

shirt and urine samples. Because of this, EPA can only rely on this research if EPA determines 

that reliance on the data may be crucial to EPA’s decision to improve public health protection by 

imposing a more stringent regulatory restriction than could be justified without the data. If OPP

determines this, under 40 CFR §26.1706, EPA must first complete three required steps. EPA 

must obtain the views of the Human Studies Review Board, provide an opportunity for public 

comment, and publish a full explanation of its decision to rely on the data, including a thorough 

discussion of the ethical deficiencies of the underlying research and the full rationale for finding 

that EPA met the standard in 40 CFR §26.1706 (c) (i.e., that the research is essential to a more 

stringent regulatory action to improve protection of public health). 

The conduct of studies 1 and 2 was not deficient relative to the ethical standards 

prevailing at the time the research was conducted. If EPA’s Office of Pesticide Programs makes 

the aforementioned determination and implements the required steps identified above, there is no 

regulatory barrier to EPA relying on this research in actions taken under FIFRA or §408 of 

FFDCA. 

Attachment 1: ORD file with relevant information (Note: Please see separate ORD file.)

Attachment 2: OPP Interview with Primary Investigator 

Attachment 3: Completeness Checklist for Ethics Documentation
 Attachment 2 

Questions for Principal Investigator Dr. Chambers – November, 2015

This attachment provides responses to questions posed to Dr. Janet Chambers regarding Study1 

and 2 referenced in the Davis et al research article.

Background: 

The EPA supported the research described in the published article through the STAR grant 

program and issuance of two grants (R8251770 and R828017). The Office of Research and 

Development (ORD) had a copy of ORD’s review of the proposed research entitled, “Assessing 

Levels of Intermittent Exposures to Children to Flea Control Insecticides from the Fur of Dogs,” 

funded under R8251770. This ORD file of research is referenced in some of the questions.

1) Were you the primary investigator for study 1 and 2 referenced in the Davis article?

Response: I was the principal investigator.

2) The research related to TCVP described in the ORD file appears to relate to the two 

studies discussed in the Davis article. Do you agree that this is the case?

Response: Yes, I agree.

3) A) Were the consent forms included in the ORD file the same as the consent forms used 

in Study 2 referenced in the Davis et al article? These consent forms include the minor’s assent 

form and the adult’s authorization for participation in the research involving flea collars. 

Response: The consent forms would have been updated based on the comments from ORD on 

the draft.

B) Do you recall if the families received signed copies of the consent forms? 

Response: I do not recall.

C) Do you recall if you addressed ORD’s comments on the consent forms before using them? 

Response: We would have had to address ORD comments in order to receive the STAR grant 

funding. Given that we received the STAR grant funding, I’m assuming we addressed their 

comments before finalizing the consent forms.

D) Do you recall if you resubmitted the revised forms to the IRB for their approval? 

Response: I don’t recall. But it would have been standard procedure to resubmit the revised 

consent form(s) to the IRB so I’m assuming that we did that.

4) A) Regarding the Davis et al article and studies 1and 2, were the students enrolled in the 

veterinary medicine curriculum at the College of Veterinary Medicine, Mississippi State 

University the only “samplers” who petted the dogs? 

Response: As far as I know, yes, they would have been the only samplers.
Page 15 of 18

B) Did you consider these students to also be human subjects? 

Response: No, we did not. We considered them to be technicians involved with the study.

C) Do you know whether the MSU IRB considered the petting element of the study to be 

“human research”? 

Response: No, they did not.

D) Do you recall if the students were compensated for their participation?

Response: It’s likely that the students would have been conducting clinical veterinary rotations. 

We probably compensated them in some way for their time.

5) Regarding the compensation discussed on page 19 in the ORD file, do you recall if this 

same compensation was used for study 2 discussed in the Davis article?

Response: Yes, this would have been the compensation. (Dr. Chambers noted that part of the 

compensation discussed in the ORD file only related to Study 2.)

6) Were the names of participating subjects kept confidential? 

Response: Yes.

5) Here is some background information included in the ORD file: For the study 

“Assessing Levels of Intermittent Exposures to Children to Flea Control Insecticides from the 

Fur of Dogs” submitted to ORD, page 13 of the ORD file indicates that dogs involved in the 

study must be enrolled in the Small Animal Community Practice Health Maintenance program 

and would belong to professional (DVM) or graduate students enrolled in the College of 

Veterinary Medicine or staff/faculty members of MSU with children in the household who 

routinely play with their pet dog. Page 19 of the ORD file states that, “in the unlikely event that 

insufficient numbers of households can be enrolled for the study, we will open the participation 

to students, faculty, and staff members of Mississippi State University outside of the College of 

Veterinary Medicine; these individuals would be offered the same incentives.”

The Davis article talks about 2 studies that involved families in Oktibbeha County, Mississippi 

and the families’ pet dogs.

a) Do you recall how the families in Study 2 were recruited? 

Response: I do not. I’m assuming that we probably circulated a flyer.

Do you recall how the adult and child from each household was selected? 

Response: I do not.
Page 16 of 18

If not, the ORD file indicates that Dr. John Tyler was responsible for recruitment. Do 

you know how we can reach Dr. Tyler?

Response: You could try the address included in the footnote of the article.

b) Do you recall if the participating families included adults who were members of the 

University? 

Response: Yes, they probably would have been. Oktibbeha County is where the 

University is located. 

7) The paragraph on page14 of the ORD file that’s entitled “selection of human test subjects,” 

states in part that: The age, sex, height and weight of both child and adult test subjects will 

be recorded, and the adult will be asked to give an estimate of the amount of time and degree 

of contact that the child and adult had with the dog on the day of the tee shirt sample. Do 

you recall if this information recorded was also recorded during study 2? 

Response: Yes, this information would have been recorded. We gave a sheet to the 

participating family and asked the adult to record the estimated amount of time and contact 

the child and adult had with the dog on the day of the tee shirt sample. 

8) Are the raw data which support the exposure outcomes of the studies available for further 

evaluation by EPA?

Response: I don’t know. I will look to see if I can access it and will get back to EPA and let 

you know.
Attachment 3

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

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

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

EPA, such information should include:

Requirement Y/N Comments/Page References 

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

IRB

§1115(a)(1): Copies of 

• all research proposals reviewed, 

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

• approved sample consent documents, 

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

subjects.

Y

n/a

Y

n/a

The Davis et al article describes 

the research. The ORD file 

provided to the HSRB contains a 

research proposal and draft 

consent forms that were reviewed 

by EPA.

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

• attendance at the meetings; 

• actions taken by the IRB; 

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

for, against, and abstaining; 

• the basis for requiring changes in or disapproving research; 

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

resolution.

N

Mississippi State University’s 

(MSU’s) Assistant Director for

Human Research Protection 

Program / Office of Research 

Compliance confirmed that the 

MSU IRB did not retain records 

beyond the required retention 

time. She noted that:

“Per 45 CFR 46.115(b) ‘the 

records required by this policy 

shall be retained for at least 3 

years, and records relating to 

research which is conducted shall 

be retained for at least 3 years 

after completion of the research’. 

This requirement is consistent with 

our Standard Operating Procedure 

01-27 Records of the HRPP which 

states that records will be 

maintained for at least 3 years 

after the study has closed.”

§1115(a)(3): Records of continuing review (CR) activities. N See comment above.

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

investigators. Y

The ORD file provided to the 

HSRB with the ethics memo 

provides “certification of IRB 

approval and institutional 

endorsement.”

§1115(a)(5): 

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

representative capacity; indications of experience such as board 

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

anticipated contributions to IRB deliberations; 

• any employment or other relationship between each member and the 

institution

Y

Y

The ORD file includes a list of IRB 

members at the time the research 

proposal was submitted to EPA. It 

references affiliations with MSU.

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

26.1108(a) and § 26.1108(b). Y

The written procedures for the IRB 

are posted on the MSU IRB 

website.

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

required by § 26.1116(b)(5).

n/a

(b) 

Copies of 

all of the 

§1125(a)

A 

discussio (1) The potential risks to human subjects; Y

Statement regarding primary 

mechanism of toxicity of TCVP is 

included in research article. 

Toxicity of TCVP discussed in 

research proposal in ORD file
Page 18 of 18

Requirement Y/N Comments/Page References 

The consent form in ORD file 

discusses researchers’ 

perspective on risk to participants.

(2) The measures proposed to minimize risks to the human subjects; N

The consent form in ORD file 

discusses researchers’ 

perspective on risk to participants.

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

and to whom they would accrue;

Y

Benefits are discussed in Davis et 

al research article and in ORD file 

in research proposal (e.g. 

approach section and section on 

improvements in risk 

assessment/management).

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

be collected through the proposed research; and

N

The research proposal in ORD file 

explains why study as proposed is 

needed (in section on 

improvements in risk assessment /

Management.)

(5) The balance of risks and benefits of the proposed research. N

The benefits of the proposed 

research is discussed. 

Researchers did not see risk to 

participants in study given that the 

families involved already used flea 

collars on their dogs.

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

as originally provided to the IRB, and as approved by the IRB. Y

There is evidence of IRB approval 

and draft consent forms in ORD 

file provided to HSRB.

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

advertisements proposed to be used. Y

There is information in the Davis 

et al research article and in the 

research proposal in the ORD file.

§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

The draft consent forms are 

included in the ORD file provided 

to the HSRB.

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

sponsors.

 Y

But 

see 

note.

Note: The ORD file includes 

certification of IRB approval and 

institutional endorsement.

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

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

been reviewed and approved by an IRB.

Y

The ORD file includes certification 

of IRB approval and institutional 

endorsement.

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

§26.1117, but not identifying any subjects of the research Y The ORD file includes draft consent forms.

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

provided, the person shall describe the efforts made to obtain the information.

Y

EPA reached out to the MSU IRB 

to attempt to obtain 

documentation. MSU IRB 

confirmed that they did not 

maintain records beyond required 

retention time.