United States Prevention, Pesticides EPA712-C-04-371
Environmental Protection and Toxic Substances August 2004
Agency (7101)
&EPA Product Performance
Test Guidelines
OPPTS 810.3800
Methods for Efficacy
Testing of Termite Baits
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INTRODUCTION
This guideline is one of a series of test guidelines that have been
developed by the Office of Prevention, Pesticides and Toxic Substances,
United States Environmental Protection Agency for use in the testing of
pesticides and toxic substances, and the development of test data that must
be submitted to the Agency for review under Federal regulations.
The Office of Prevention, Pesticides and Toxic Substances (OPPTS)
has developed this guideline through a process of harmonization that
blended the testing guidance and requirements that existed in the Office
of Pollution Prevention and Toxics (OPPT) and appeared in Title 40,
Chapter I, Subchapter R of the Code of Federal Regulations (CFR), the
Office of Pesticide Programs (OPP) which appeared in publications of the
National Technical Information Service (NTIS) and the guidelines pub-
lished by the Organization for Economic Cooperation and Development
(OECD).
The purpose of harmonizing these guidelines into a single set of
OPPTS guidelines is to minimize variations among the testing procedures
that must be performed to meet the data requirements of the U. S. Environ-
mental Protection Agency under the Toxic Substances Control Act (15
U.S.C. 2601) and the Federal Insecticide, Fungicide and Rodenticide Act
(7U.S.C. I36,etseq.).
Final Guideline Release: This guideline is available from the U.S.
Government Printing Office, Washington, DC 20402 on disks or paper
copies: call (202) 512-0132. This guideline is also available electronically
in PDF (portable document format) from EPA's World Wide Web site
(http://www.epa.gov/opptsfrs/home/guidelin.htm) under the "Harmonized
Test Guidelines."
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OPPTS 810.3800 Methods for Efficacy Testing of Termite Baits
(a) Scope—(1) Applicability. This guideline describes test protocols
that EPA believes will generally satisfy product performance testing re-
quirements of the Federal Insecticide, Fungicide, and Rodenticide Act
(FIFRA) (7 U.S.C. 136, et seq.) for termite bait products. As explained
in 40 CFR 158.70(a), applicants for pesticide registration may utilize any
appropriate protocol provided that it is of suitable quality and completeness
to provide EPA with sufficient information to adequately assess the prod-
uct. Accordingly, instructions in this protocol directing applicants to con-
duct testing in a specific manner do not and are not intended to establish
a regulatory requirement, but are intended simply to instruct applicants
on what they must do should they choose to follow these particular bait
testing protocols. This guideline is not intended to address labeling for
termite bait products. EPA intends to address labeling guidance in a pes-
ticide registration notice or equivalent document.
(2) Background. The published literature on termiticide bait research
is the source material for issues addressed in this guideline. Paragraph (g)
of this guideline contains a complete list of references cited in this guide-
line.
(b) Definitions. The following definitions are of special importance
in understanding this guideline:
Kills termites. The term kills termites refers to termites dying as result
of feeding or contacting a pesticide.
Post-construction treatment. The term post-construction treatment re-
fers to all pesticide treatments made to kill and/or control termites after
the installation of the final grade.
Preventive or preventative treatment. The term preventive or prevent-
ative treatment refers to all pre-construction or post-construction bait treat-
ments made to provide structural protection before a termite infestation
is present.
Protect(s) a structure, protection, eliminate(s) and control(s). The
terms protect(s) a structure, protection, eliminate(s) and control(s) have
the same meaning as the term structural protection.
Remedial or curative treatment/application. The terms remedial or
curative treatment/application refer to and include all pre-construction or
post-construction pesticide treatments made to kill and control a termite
infestation when present.
Stand alone. The term stand alone refers to a pesticide product that
provides structural protection when applied without other pesticide prod-
ucts for the same purpose.
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Structural protection. The term structural protection refers to the
elimination or prevention of termite activity in a structure as a result of
a preventive or curative application of a pesticide product.
Termite colony. The term termite colony refers to a group of termites
of the same species which constructs a nest (may consist of dispersed gal-
leries and chambers), rears offspring in a cooperative manner, and shares
an interconnected gallery system (Wilson 1971, Su and Scheffrahn 1998b)
(see paragraphs (g)(81) and (g)(71) of this guideline, respectively).
Termite infestation or termite activity. The term(s) termite infestation
or termite activity refer to the presence of live termites in or on a structure.
Termiticide bait or termite bait. The terms termiticide bait or termite
bait refer to any pesticide product that kills or controls termites and has
at least two principal components: A bait matrix, intended to be the equal
of or preferred to other available sources of food and a pesticide incor-
porated into the bait matrix that is intended to kill termites.
(c) Overview. (1) This guideline concerns the product performance
testing for evaluation of pesticides used as baits to kill and control ter-
mites. Good Laboratory Practice Standards (GLP) apply to these laboratory
and field studies as defined in 40 CFR 160.1 to 160.195. Studies which
do not comply with GLP standards may nonetheless be considered if, in
the Agency's judgment, the design and conduct of the study are sufficient
to demonstrate that the results are scientifically reliable. All testing should
be done with the product intended for registration unless EPA approves
testing of the termite bait with bait station prototypes.
(2) This guideline describes specific methods for conducting product
performance testing of termite baits which reflect the Agency's considered
recommendations for minimum steps necessary to develop reliable data
on termite bait product performance. Data should be collected from the
laboratory and both field experiments as described in this guideline to
demonstrate the efficacy of a termite bait product.
(3) A general discussion of criteria for assessing termite bait success
can be found in Thome and Forschler 1999, Thorne and Traniello 1994,
Forschler 1998, Su 1991, Esenther and Beal 1974, Pawson and Gold 1996,
Su 1991 a,b,c, Su et al. 1997, Su and Scheffrahn 1996 a,b,c, Su 1998,
Scheffrahn and Su 1997, Su 1999, Jones 1989, 1991, Lenz 1996, Robson
1996, Rust 1996, Su 1994, Su 1996a,b, Su 1995, Traniello 1994, Grace
1996, Sornnuwat et al 1996a, b; Tsunoda 1998, 1999, and Rust 1996 (see
respective references in paragraph (g) of this guideline). Other references
are cited throughout this guideline.
(d) General considerations for laboratory tests. EPA believes that
laboratory testing for bait products bearing termite control claims should
be designed to evaluate characteristics that are critical to the success of
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the bait. Specifically, such testing should assess whether the bait: Is readily
fed upon by termites, exhibits slow-acting delayed toxicity, is effective
at the proposed product label concentration with a palatable alternative
food source concurrently available, and causes 100 percent mortality in
greater than two weeks but less than ten weeks post-treatment at the tested
dose. Additional or alternative assessments may be considered depending
on the nature of the active ingredient and bait product.
(1) Species. Identify test termites as to genus and species and by sub-
species or strain when possible. Laboratory termite populations should be
colonized from field collected cultures of three subterranean termite spe-
cies from the United States to include Coptotermes formosanus;
Reticulitermes flavipes, and Heterotermes aureus. Collect field cultures ac-
cording to the method of Su and Scheffrahn (1986) (see paragraph (g)(56)
of this guideline) or by an equivalent method. Evaluations against the
dampwood termites, Zootermopis spp., and/or drywood termites,
Cryptotermes and Incistermes spp. should be performed if the bait is in-
tended to control these termite species. Testing with other species may
be considered.
(2) Stage, caste, and age. Test laboratory termite populations cul-
tured from the field no later than 90 days after field collection. Data col-
lected with laboratory populations older than 90 days may be considered
if survivorship in the control treatments is greater than 85 percent. The
selection of the life stage to test will be dependent upon the pesticidal
action of the active ingredient, but should include 100 worker termites
(undifferentiated nymphs of at least the third instar in the genus
Reticulitermes (Thorne 1996) (see paragraph (g)(74) of this guideline) and
fifth or sixth instars of true workers in the genus Coptotermes (Roisin
and Lenz 1999, Forschler and Jenkins 1999) (see paragraphs (g)(49) and
(g)(15) of this guideline, respectively)). This determination should be made
where possible for U.S. species from the same field collection site. In addi-
tion, bioassays may include soldier caste members collected from the same
site as the worker termites. Caste proportion in the test population should
reflect the optimum for the test species (Haverty 1977) (see paragraph
(g)(25) of this guideline).
(3) Test conditions and rearing techniques. Rear termites according
to species specific requirements. A brief description of the test conditions
and rearing methods should accompany laboratory test results.
(4) Bioassay design. All laboratory bioassays should be made using
three colonies of the test species. The selected method(s) should include
no-choice and choice bioassays to assess active ingredient efficacy. Testing
should be done under circumstances in which the termites can forage for
food in a manner similar to their natural behavior. Bioassays with drywood
termites may be done as described by Scheffrahn and Su (1997) (see para-
graph (g)(52) of this guideline).
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(5) Feeding preferences and palatability testing. Testing should be
conducted to show that the bait product is palatable to termites at the active
ingredient concentration(s) on the product label.
(6) Data reporting. See paragraph (c)(l) of this guideline.
(e) General considerations for control of subterranean termites
using termite baits in area-wide field tests not using existing structures
and buildings—(1) Geographic areas. Each termite bait product with its
associated application materials should be, at a minimum, placed in field
sites located in EPA Regions 4, 6 and 9 (EPA Regions are described in
paragraph (f)(2) of this guideline). These sites represent varying climatic
and soil conditions, and are habitats for several subterranean termite spe-
cies (see paragraph (e)(2) of this guideline). Other Regional locations may
be added but should not substitute for testing in Regions 4, 6, and 9.
(2) Termite species. The subterranean termite species that should be
considered as the test subjects for field site testing include species from
the genus Reticulitermes. Data collected with any of the following
Reticulitermes species should be acceptable: Reticulitermes flavipes, R.
virginicus, R. hesperus, R. hageni, and R. tibalis. R. flavipes is preferred
because this species is the principal structural pest in the U.S. Data col-
lected from termites in the genera Heterotermes and Coptotermes should
include Heterotermes aureus and Coptotermes formosanus.
(3) Test design. This test is designed to evaluate baits as termite toxi-
cants at sites of vigorous termite activity in the United States. Impact on
termite populations should be readily demonstrated. See paragraphs
(e)(3)(viii) and (e)(4) of this guideline for types of data to collect and
recommendations for evaluating baiting success. The bait should be evalu-
ated for eliminating subterranean termite populations from termite-infested
monitoring devices/stations. Post-elimination monitoring should continue
at these devices/bait stations for at least one year. For each species tested
in each EPA Region, one colony should be exposed to a location at the
test site with treated baits and one colony should serve as the untreated
control colony and therefore should be in a location at the site without
treated baits. Each test site should include at least one "treated" colony
and one ' 'untreated'' control colony. Treated and untreated colonies should
be differentiated and separation demonstrated by the method specified in
paragraph (e)(3)(i) of this guideline. Testing and monitoring should be
conducted for up to three years.
(i) Establishing the field site. The protocol for establishing each field
site where the tests will be conducted has been derived from a combination
of research studies (Su 1994 (see paragraph (g)(64) of this guideline),
Grace et al 1996 (see paragraph (g)(22) of this guideline), Forschler and
Ryder 1996 (see paragraph (g)(14) of this guideline), Su et al. 1997 (see
paragraph (g)(69) of this guideline), Getty et al. 2000a,b (see paragraphs
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(g)(17) and (g)(18) of this guideline)). Site size and location can vary de-
pending on the termite species present and the size of their respective pop-
ulations. The site selection process should involve the following steps:
Identify the termite species at the site, establish or install monitoring de-
vices (wood stakes, bucket traps, or untreated bait stations), determine
which termites are infesting the monitoring devices, establish colony for-
aging areas (using mark-recapture, mark-release-recapture, DNA analyses
or equivalent methods), assess wood consumption patterns, and assign
colonies to bait treatments or controls (untreated).
(ii) Installation of untreated wood stakes or cellulose material.
Once a site with at least two existing termite colonies has been selected,
wooden survey stakes or other cellulose material should be driven into
the ground to encourage termites to feed at specific sites (Su and
Scheffrahn 1986 (see paragraph (g)(56) of this guideline), Haverty et al.
2000 (see paragraph (g)(31) of this guideline). (Heterotermes aureus may
require a different approach (Jones 1990) (see paragraph (g)(38) of this
guideline)). Wood stakes, preferably southern yellow pine sapwood stakes
or an equally attractive cellulose material, should be arranged in a grid
pattern across the selected site. This should be done for untreated control
and bait treatment sites. It is recommended that wood stakes or other cel-
lulose materials should be spaced 2m (6ft) to 3m (9ft) apart and placed
into the soil to a depth of 15cm (6in) or deeper. Other spacing and soil
depths may be considered by EPA for treated and untreated colonies. The
bait matrix, less the active ingredient, or an untreated wood stake, or other
cellulosic material can be used in the proposed commercial bait station
or a prototype as an additional means of monitoring termite activity. The
wood stakes and other untreated bait matrix can be moistened prior to
installation to make them more attractive to termites. When the wooden
stakes or untreated bait matrix have been fed upon by the termites, an
independent monitoring device (bucket trap) should be placed adjacent to
it or can replace the stake. Two to three foot quartered wood log sections
held together with a strap can also be placed approximately six inches
to one foot deep to recruit termites to these locations and make the obser-
vation of termite activity easier.
(iii) "Bucket Trap" installations. Bucket traps should be used as
independent monitoring tools and should be installed where termites are
actively feeding. These traps can also serve as sites to "mark" termites
during bait evaluation (see paragraph (e)(3)(vi) of this guideline). To con-
struct and establish a bucket trap installation see Pawson and Gold 1996
(see paragraph (g)(46) of this guideline). Into the bucket place a prefab-
ricated pine sandwich prepared according to the method of Su and
Scheffrahn 1986 (see paragraph (g)(56) of this guideline) or equivalent
method. The trap should be covered with a tight fitting plastic or metal
lid and covered with 5cm (2in) of soil. A flag or other means of identifica-
tion should be used to mark the location of the trap. Dyed untreated bait
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matrix can be used in addition to bucket traps to "mark" termites.
Tamashiro et al. 1973 (see paragraph (g)(72) of this guideline), Su and
Scheffrahn 1986 (see paragraph (g)(56) of this guideline), and Grace 1990
(see paragraph (g)(20) of this guideline) provide additional information
and procedures for field detection and monitoring.
(A) Prior to baiting, these monitoring stations should be used to deter-
mine the dispersion and the seasonal foraging and feeding by the termite
colonies (Su and Scheffrahn 1988 (see paragraph (g)(57) of this guideline),
Su et al. 1993b (see paragraph (g)(63) of this guideline), Haverty et al.
1999b, 2000 (see paragraphs (g)(30) and (g)(31) of this guideline)). These
same monitoring stations should be used to measure feeding of the termite
colonies after baiting. For a termite colony to be included in the evaluation,
either as a treated or untreated colony, the foraging area of the target ter-
mite colony should be determined in the study. This is usually done by
documenting that at least three of the monitoring stations must be "con-
nected," i.e., fed upon by termites from the same colony, in order to define
the colony foraging area. However, other methods may be used at the
digression of the study director.
(B) Measure wood consumption and damage at the bucket traps in
the control colonies in order to have a standard of comparison for feeding
cessation at the bucket traps for the baited colonies.
(iv) Bait product installation. Install the bait formulation intended
for registration, sale and distribution to deliver the active ingredient. The
bait station used should be the station to be used for sale and distribution
of the product but a prototype may be considered by the Agency. A min-
imum of five bait stations should be placed within the foraging range of
the colony, i.e., at locations within the perimeter of the "polygon" de-
scribed by the termite occupied independent monitoring stations that are
utilized by or "connected to" that colony or within the area established
by the interconnection of the bait product stations and the bucket traps
(same termite colony foraging at both devices; see also Evans 2001, Evans
et al. 1998, 1999, and Forschler and Jenkins 2000 (see paragraphs (g)(12),
(g)(9), (g)(10) and (g)(16) of this guideline, respectively)). It should be
clearly shown that the termites feeding in the monitoring devices and treat-
ed bait stations are from the same colony.
(v) Termite species identification. Termites should be collected and
identified as to species from at least one bait station or independent moni-
toring device from each test plot that contains termites. Appropriate termite
identification keys such as those published by Weesner 1965 (see para-
graph (g)(80) of this guideline), Nutting 1990 (see paragraph (g)(44) of
this guideline), and Scheffrahn and Su 1994 (see paragraph (g)(51) of this
guideline), should be used to identify termites. Identification using DNA
techniques or cuticular hydrocarbon profiles (Forschler and Jenkins 1999,
(see paragraph (g)(15) of this guideline)), Haverty et al. 1996, 1999) (see
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paragraphs (g)(29), (g)(30), and (g)(27) of this guideline, respectively) can
be used to supplement morphological keys, but should not replace morpho-
logical keys. Termite samples should be collected and identified annually
to note the possible arrival of a different termite species. Voucher speci-
mens (soldiers, workers, and alates (if available) in 80 percent ethanol,
workers and soldiers frozen at -30° C; or dried workers for characterization
of cuticular hydrocarbons) should be kept for each treated and untreated
colony. See also Jenkins et al. 1998, 1999, and 2001 (see paragraphs
(g)(33), (g)(34) and (g)(35) of this guideline) for a discussion of meth-
odologies on use of DNA sequencing and genetic markers for phylogenetic
analyses and genetic structure determination in subterranean termite popu-
lations.
(vi) Colony identification. Identification of termite colonies foraging
in the treated and control plots should be at least once per year. The pur-
pose of marking termites is to verify that the termites feeding in the wood
stakes/bucket traps and in the bait product stations are from the same col-
ony. The preferred method is to dye filter paper with a known concentra-
tion of dye such as Sudan Red 7B (Lai et al. 1983 (see paragraph (g)(41)
of this guideline), Nile Blue A and Neutral Red (Su et al. 1991 a, b, c
(see paragraphs (g)(59), (g)(60) and (g)(61) of this guideline) or other ac-
ceptable dye (see also Delaplane et al. 1989 (see paragraph (g)(5) of this
guideline)), and Evans 1997, 2000 (see paragraphs (g)(8) and (g)(ll) of
this guideline) for a discussion of dyes). The procedure consists of apply-
ing dye dissolved in an appropriate solvent (usually acetone) to filter paper
and then allowing the dyed paper to air dry in a fume hood. Layers of
dyed paper can be compressed between wooden boards or incorporated
into the bait matrix in a bait station. A similar procedure is described by
Atkinson 2000 (see paragraph (g)(2) of this guideline). If this procedure
does not produce enough marked termites to verify identity of the foraging
group, then other techniques may be employed as mentioned in paragraph
(e)(3)(vii) of this guideline.
(vii) Other marking techniques. Other marking techniques such as
using fluorescent spray paint as described by Forschler 1994 (see para-
graph (g)(13) of this guideline), mark-release-recapture techniques as de-
scribed by Jones 1990 (see paragraph (g)(38) of this guideline), or molec-
ular marking techniques (Forschler and Jenkins 1999 (see paragraph
(g)(15) of this guideline); see also Forschler and Jenkins 2000 (see para-
graph (g)(16) of this guideline) for related approaches may also be em-
ployed but there should be laboratory and field data for the termite species
under investigation. If a method using the dyed paper in the monitoring
device does not result in an adequate number of marked foraging termites,
it will be necessary to collect and spray paint, or feed large numbers (thou-
sands) of termites dyed paper in the laboratory, then return these marked
termites to the monitoring device or bait stations from which they were
collected. It should be clearly shown that the termites feeding in the moni-
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toring device and adjacent bait stations are from the same foraging group
or colony. Depending on the species, such data may also include agonistic
behavior, average dry weight of worker termites (minimum of 5 replicates,
10 workers each), and cuticular hydrocarbon profiles (Haverty et al. 1996,
1999a,b (see paragraphs (g)(27), (g)(29) and (g)(30) of this guideline)).
(viii) Data collection and reporting. The following information
should be recorded and reported during the course of these tests in order
to provide the data necessary to evaluate the impact of the bait treatment
on termite populations and their foraging activity:
(A) Purpose of study;
(B) Location and test unit designation;
(C) Date field test unit installed and duration and intervals of moni-
toring and data reporting;
(D) Date of evaluation;
(E) Termite species present;
(F) Termite colony designations, presence or absence of termites in
the wood stakes, bucket traps, split logs (if used) and bait stations;
(G) Presence of dyed termites in the independent monitors and bait
stations;
(H) Amount of wood/bait consumed (estimated percent) in the bucket
traps and bait stations;
(I) Bait and wood weights;
(J) Date termite activity ceased in the bucket traps and bait stations;
(K) Date termite activity resumed in the bucket traps and bait stations;
(L) ASTM damage rating to each pine board/bait matrix following
cessation of termite activity in the bucket traps and bait stations;
(M) Date dyed paper/matrix is first fed upon;
(N) Date of release of laboratory dyed or marked termites into the
bucket trap or bait stations;
(O) Date and location of mud tubing observed on or in a test unit;
(P) Results and discussion;
(Q) Conclusions;
(R) Certification;
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(S) References;
(T) Appendices.
(4) Evaluating baiting success. These recommendations are derived
from recognized pest management practices and bait field test evaluations
found in Jones 1989, 1991, Forschler and Ryder 1996, Grace 1990,
Haverty 1999b, Lenz et al 1996, Pawson and Gold 1996, Su 199la, 1994,
1996a, 1996c, and Thorne and Forschler 2000 (see respective references
in paragraph (g) of this guideline). The study should describe how bait
success will be measured. For a bait to be effective, it should control ter-
mite populations on an area-wide basis. At a minimum, five bait product
stations and their interconnected bucket traps should be active for each
colony tested and evaluated. Generally, treated baits should be evaluated
to determine whether they demonstrate efficacy (i.e., colony extermination;
population suppression) within 12 months after initiation of feeding on
the bait treatment and during the 12-month period following the cessation
of termite activity at the monitoring devices. Su 1996a, Grace et al. 1989
and Forschler and Ryder 1996 (see paragraphs (g)(66), (g)(19) and (g)(14)
of this guideline, respectively) present procedures for making these deter-
minations. The amount of bait eaten/used at each location for each colony
should be documented in the study and effects determined by comparison
with control colonies.
(f) General considerations for field tests using existing structures
and buildings. Field tests at existing structures should use the bait product
and its associated application materials as intended for registration. The
test period under this protocol should be three years from the time that
the bait product stations are installed. See paragraphs (f)(4)(vi) and (f)(5)
of this guideline for types of data to collect and recommendations for eval-
uating baiting success.
(1) Number of homes/structures. Testing should be conducted in
the United States at 100 or more subterranean termite infested wooden
buildings/structures. Testing with drywood termites should be performed
if the product is intended to control these termite species. In this case,
testing should be conducted in 100 drywood termite infested structures.
(2) EPA Regions: The selected structures/buildings for subterranean
termite testing should be distributed (approximately 15 homes per region)
in the following EPA Regions: Region III (DE, DC, MD, PA, VA, WV),
Region IV (AL, FL, GA, KY, MS, NC, SC, TN), Region V( IL, IN, MI,
MN, OH, WI), Region VI ( AR, LA, NM, OK and TX), Region VII (IA,
KS, MO, NE) and Region IX (AZ, CA, HI, NV, AS, GU).
(3) Site selection and inclusion of a building/structure in a field
test. Selected structures should represent the wide range of construction
types existing in the United States and the testing should include post-
application monitoring inside and outside the structure. In-ground and
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above-ground baits should not be tested in the same structure unless they
are to applied as a co-pack.
(i) A wooden frame building must be present. A building can be resi-
dential or commercial and may also include sheds, barns, or garages.
(ii) Based on the results of a thorough inspection, the building should
be infested with one or more of the following subterranean termite species
Reticulitermes spp., Heterotermes aureus, and Coptotermes formosanus
per one or more of the following infestation conditions: Either alates have
swarmed inside the structure or live termites are found to be active within
the building; or there is clear evidence of termite activity or damage on
or in the building (mud tubes, galleries in wood) and live termites or evi-
dence of recent termite activity (recently formed mud tubes and/or galleries
in wood) are found in outside wood attached or immediately adjacent to
the building, i.e., attached fences, wood decks, landscape timbers, etc.
When assessing structures infested with desert termites
(Heterotermesonly), an additional condition should be considered: There
is clear evidence of termite activity or damage on or in the building (mud
tubes, galleries in wood) and the presence of active, live termites found
in bait stations placed in the ground or around the perimeter of the struc-
ture according to label directions.
(iii) The termiticide applications made to the structure should be
known for five years preceding the start of the bait evaluation. Structures
that have been treated within the last five years with a liquid soil
termiticide should not be included in the test because these treatments
could affect bait success.
(iv) Termite species and their identification. See paragraph (e)(3)(v)
of this guideline.
(v) Detection of termites in an existing structure. Acoustical emission
devices, microwave devices, and/or visual inspection should be used to
monitor wood in a structure for termites. Termite "sniffing" dogs may
be considered for detecting termites but should not substitute for the other
methods.
(4) Test design, monitoring termite activity and bait product in-
stallation. A combination of the methods in paragraphs (e)(4)(i), (ii), and
(iii) of this guideline should be used to assess the presence and extent
of the foraging termite population in the vicinity of the foundation for
remedial treatment bait installations. It is recommended that the monitoring
devices/bait stations be placed within one meter of the foundation but their
exact placement should conform with the proposed bait product label's
directions for use. The final experimental design should be determined by
the study director. Additional monitoring stations can be installed farther
away from the structures and used for assessing termite activity provided
that the interconnection of these stations and those within one meter of
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the structure (or as directed by the label) can be established using tech-
niques such as the mark-recapture, mark-release recapture method, and/
or DNA analyses. Generally, monitoring stations should be checked at a
minimum every 30 days during the course of the study.
(i) Installation of untreated wooden stakes or other cellulose mate-
rials. Wood stakes or other cellulose materials should be spaced 2m (6ft)
to 3m (9ft) apart and placed into the soil to a depth of 15cm (6in) or
deeper around the perimeter of the structure. Other spacing and soil depths
may be considered by EPA. The bait matrix, less the active ingredient,
and/or wood stakes cut to the required size, can be used in the proposed
commercial bait station as an additional means of monitoring termite activ-
ity. Wood stakes or cellulose materials, and bait matrix should be used
in combination to monitor activity. They can be moistened prior to installa-
tion to make them more attractive to termites.
(ii) "Bucket Trap" installations. Bucket traps should be used in
addition to the above-monitoring tools and should be installed every 10m
(30ft) or less (these traps can also serve as sites to "mark" termites during
bait evaluation). To establish a bucket trap installation at an existing struc-
ture, see Pawson and Gold 1996 (see paragraph (g)(46) of this guideline).
Into the bucket place a prefabricated pine board sandwich prepared accord-
ing to the method of Su and Scheffrahn 1986 (see paragraph (g)(56) of
this guideline). Do not add the dyed filter paper when monitoring; it should
be used for "marking" termites during bait evaluation as described in
this guideline. The trap should be covered with a tight fitting plastic or
metal lid and covered with 5cm (2in) of soil. A flag or other means of
identification should be used to mark the location of the trap.
(iii) Dyed untreated and treated bait matrix. Dyed untreated and
treated bait matrix can used in addition to bucket traps to "mark" termites.
(iv) Dye and Marking. See (e)(3)(vi) of this guideline.
(v) Termite bait product application and placement. Application
of bait product stations should be done according to proposed label direc-
tions. The number of stations applied will vary with the structure, termite
species to eliminated, extent and location of infestation, size of bait station,
and action of the active ingredient. Either below ground or above-ground
stations can be used. It is recommended that 10 or more bait stations be
applied per structure to ensure baiting success and sufficient information
to adequately assess the efficacy of the bait product.
(vi) Data collection and reporting. Data from all structures included
at any time during the course of the study should be reported. The fol-
lowing information should be recorded and reported during the course of
these tests in order to provide the data necessary to evaluate the impact
of the bait treatment on existing structures:
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(A) Purpose of study;
(B) Location (address);
(C) Date test monitoring units and bait stations installed and duration
and intervals of monitoring and data reporting;
(D) Construction type (split foyer, etc.), square footage, age, founda-
tion type (slab, basement, crawl space, etc.), siding type (brick, block,
foam board/stucco, etc.);
(E) Termiticide treatment history of the structure;
(F) Date of termite inspection;
(G) Termite species present, termite species colony designations, date
and location of placement of monitoring devices and bait stations;
(H) Soil type, soil pH, percent soil moisture and soil surface tempera-
ture at each monthly evaluation, climatic data - temperature and rainfall;
(I) Date of each monthly evaluation;
(J) Termite species present;
(K) Presence or absence of termites in each bucket trap and for other
monitoring stakes/stations;
(L) If applicable, presence of dyed termites in the bucket trap and
bait stations;
(M) Amount of wood/bait matrix consumed (estimated percent) in
bucket trap and each bait station, respectively;
(N) Weight of bait(s) and wood consumed;
(O) Number and percent of bait product stations fed upon;
(P) Date of bait station replacement;
(Q) Date termite activity ceased in the bucket trap and or bait stations
and date termite activity resumed in the bucket trap and or bait stations;
(R) ASTM damage rating to each pine board following cessation of
termite activity in the bucket trap, wood stakes, and bait station(s);
(S) Date dyed paper first fed upon;
(T) Date of release of laboratory dyed or marked termites into the
bucket trap or bait station(s);
(U) Date termites found in the bucket trap, wood stakes, and or bait
station(s);
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(V) Date and location of mud tubing observed in the bait stations,
bucket traps and structure
(W) Conclusions;
(X) Certification;
(Y) References;
(Z) Appendices.
(5) Evaluating bait success, (i) The first evaluation after installation
of the test units should be accomplished by 60 days post-installation. Sub-
sequent evaluations should be conducted at 30-day intervals during the
duration of testing, and should determine presence or absence of termites
(all life stages considered, including alates) in the structure, monitoring
stations, and bait stations. The monitoring devices should be visually ob-
served for mud tubing but should not be disturbed or dismantled. The num-
ber of termites present should not be estimated until they have infested
most of the monitoring device as evidenced by mud tubing in the wood
boards or bait matrix. Bait product station replacement should be done
according to the proposed label directions. Termite presence and activity
must be monitored during the entire field test using methods described
by Su and Scheffrahn 1986 (see paragraph (g)(56) of this guideline),
Pawson and Gold 1996 (see paragraph (g)(46) of this guideline), Haverty
et al. 1975 (see paragraph (g)(24) of this guideline), and Jones 1990 (see
paragraph (g)(38) of this guideline), or equivalent methods. Inspections
of treated infested structures should take place within 12 months of bait
installation and during the times of the year when termites are active, gen-
erally from May to September in much of the United States.
(ii) The bait product treatment should eliminate an existing termite
infestation in 12 months or less following installation of the commercial
product (with or without active ingredient) in the infested structures and
these structures should remain termite free for 12 months following elimi-
nation of the termite infestation. Alate swarms, mud tubing, and presence
of worker or soldier termites in or on the structure are indications of a
termite infestation and may indicate the failure of a bait product to control
termites. Generally, baiting should cease as directed by the product label
but monitoring for termites should continue for an additional 12 months.
If termite activity resumes during the 12-month post-observation period
at the bait product station, bucket trap, or wood stake, additional bait appli-
cation can be made according to the product label directions. Claims of
preventive treatment are generally satisfied when a structure is not infested
with termites for 12 months or more following elimination of a structural
infestation.
(g) References. The following references should be consulted for ad-
ditional background material on this test guideline:
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