United States       Prevention, Pesticides     EPA 712-C-98-414
         Environmental Protection    and Toxic Substances     March 1998
         Agency        (7101)
4»EPA    Product Performance
          Test Guidelines
          OPPTS 810.3200
          Livestock, Poultry, Fur-
          and Wool-Bearing
          Animal Treatments

     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

     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 The Federal Bul-
letin  Board.   By  modem  dial   202-512-1387,  telnet   and   ftp:
fedbbs.access.gpo.gov  (IP, or  call 202-512-0132 for disks
or paper copies.  This  guideline is also available electronically in ASCII
and PDF (portable document format) from EPA's World Wide Web  site
(http://www.epa.gov/epahome/research.htm) under the heading "Research-
ers and  Scientists/Test Methods and Guidelines/OPPTS  Harmonized Test

OPPTS 810.3200  Livestock,  poultry, fur- and  wool-bearing animal
     (a) Scope—(1) Applicability. This guideline is intended to meet test-
ing requirements of the Federal Insecticide, Fungicide, and  Rodenticide
Act (FIFRA) (7 U.S.C. 136, et seq.)

     (2) Background. The  source materials  used in  developing  this har-
monized OPPTS test  guideline are OPP guidelines  95-8 Livestock, poul-
try, fur and wool bearing animal treatments and 95-30 Acceptable methods
(Pesticide Assessment Guidelines, Subdivision G:  Product Performance,
EPA report 540/9-82-026, October 1982).

     (b) Overview. This guideline concerns efficacy testing of invertebrate
control pesticides used on cattle, horses, sheep, goats, swine, chicken, tur-
keys, other  domestic  fowls, and fur-bearing animals, such as mink and
rabbits,  for control of the major arthropod pests that parasitize these ani-
mals, and are additionally considered to be  public health pests. See OPPTS
810.7000 for a definition of public health invertebrates.

     (c) General considerations—(1)  Site selection. Pests  of livestock,
poultry, fur- and wool-bearing animals include,  but are not limited to, nose
bot, sheep ked, various biting flies (horn fly and stable fly), ticks, housefly,
face fly, and mosquitoes. Evaluating an insecticide that is to be applied
directly to such  animals to control these pests should generally be  based
on adequate tests on representative animals from herds or flocks in at least
5 widely-separated regions where the insecticide is  to be marketed, unless
applicability so limits use of the pesticide in special representative regions.
Tests on dairy cattle may not be necessary if comparable to efficacy data
are reported from tests using beef cattle.

     (2) Sample size. Sample size should be representative of the number
of animals in a particular treatment  group, which can range from one to
a herd or flock of thousands.

     (3) Number  of trials. A minimum of 5  large-scale geographically-
separated  trials are generally necessary, but the number of trials  can vary
somewhat due to the accessibility of infestations, fluctuations in pest popu-
lation pressures, behavior,  and other important considerations in  the biol-
ogy of the target pest.

     (4) Dosage selection. Dosage  levels and concentrations should be
identified in  the laboratory or small-scale tests before field testing. The
data should include  support of the  schedule of repeated applications  as
indicated in the directions for use of the product.

     (5) Application  techniques and equipment—(i)  Single oral dose.
For insecticides  administered orally  as drenches, boluses,  or  in capsules,
care should be taken that animals receive the entire dose. Record the for-
mulation,  final concentration of active ingredient,  total amount  adminis-

tered, and  dosage in terms of mg of active ingredient per  kg of body
weight of  animal. The regulation of many  internal  applications  comes
under the authority of the Food and Drug Administration, Bureau of Vet-
erinary Medicine.  Some of these pesticides may fall into this category.

     (ii) Feed treatment. Insecticides administered to animals  as part of
a feeding regimen should be mixed into the entire feed ration or fed in
a small amount of feed which, when consumed, is followed by untreated
feed.  Record the formulation, final percent content of active  ingredient
(ppm in  feed), or total mg of active ingredient per kg body  weight of
animal, amount of feed or treatment consumed, and length of treatment

     (iii)  Water treatment. Some insecticides may be administered to ani-
mals through drinking water. Animals may be treated individually or in
groups, which, when having consumed the treated water, are  given  un-
treated water. Regardless  of the size of the group, the  following  should
be recorded:  The formulation, final concentration of active ingredient in
terms of ppm in water or mg of active  ingredient per kg of body weight
of animal, average consumption of water per animal, and duration of treat-

     (iv)  Mineral, salt, or protein  supplement. Insecticides  of this type
are generally formulated at low concentrations in mineral, salt,  or protein
supplements and are  offered free choice to animals.  Because consumption
of salt, mineral,  and protein supplement varies considerably from animal
to animal, it is important to determine whether or not all animals  consumed
treated materials. Record the formulation, final concentration of active in-
gredient  in terms  of percentage or ppm  of treated supplement, average
consumption  per animal per day, dosage in terms of mg active  ingredient
per kg body weight per day, and length of treatment period.

     (v)  Injections.  For insecticides  given to  animals in the form  of
intramuscular, intraperitoneal, or subcutaneous injections, record the for-
mulation, amount  of material  injected per animal,  location of injection,
and dosage in terms  of mg of active ingredient per kg of body weight.

     (vi)  Whole-body  sprays. For insecticides applied to  animals  as
whole-body sprays,  care should be taken that animals  are treated thor-
oughly and that enough pressure is used to penetrate hair coat and assure
wetting  of the skin.  A variation  of the whole-body spray is the  use of
spray-dip machine to apply spray to animals. With  either method,  record
the formulation,  final concentration of active ingredient, equipment used
and application techniques, and average volume  of spray applied per ani-

     (vii) Dip. For insecticides used to charge dipping vats, animals  should
be immersed thoroughly in the dip fluids. Record the  formulation, final
concentration  of active ingredient,  volume  of liquid in the  vat, age of

charge at time of dipping, number of animals dipped, and data on recharg-
ing (if necessary). Chemical analyses of active ingredient in vat fluids be-
fore and after  dipping  are  necessary  to  determine the  actual amount of
active ingredient in the vat fluid.

     (viii) Pour-on treatment. Insecticides applied to animals by the pour-
on technique, ready-to-use  formulations, or emulsifiable concentrates  di-
luted with water or oil,  are poured down the backline  of animals in ounce
(milliliter) rates. In an  extension of this technique, ready-to-use formula-
tions are applied to a spot on the backline at milliliter rates.  Record formu-
lation,  diluent, final concentration of active ingredient, amount applied per
animal, area treated,  and dosage based on mg  active  ingredient  per  kg
body weight of animal.

     (ix) Dust treatment (livestock).  Insecticides are applied as dusts by
power  duster or by hand or contained in dust bags and  placed in the pas-
ture for free-choice use or placed  in openings  to feed, mineral, and/or
water sources so that animals are forced to  use them  on a daily basis.
With cattle  grub control, it is important that dust bags  be located  so that
animals are  forced to  treat themselves  on a daily basis to insure that suffi-
cient insecticide is applied  for cattle  control. In tests with dust bags  for
control of ectoparasites, such daily treatment is not essential. Record for-
mulation, final concentration of active ingredient, average amount  of dust
per animal, location of dust bags, and length of treatment period.

     (x) Back spray. In tests to control cattle grubs, the animal's backs
are sprayed  thoroughly  with the contact insecticide. Care is taken to force
insecticide into the warble openings in the animal's backs.  Record formu-
lation, final  concentration of active ingredient, equipment used, application
techniques, and average volume of spray applied per back.

     (xi) Floor or  litter treatment. Insecticides are  applied as mists or
fogs, dust, or granules directly to floor areas, litter, or nest areas.  Record
formulation, final  concentration  of active ingredient,  equipment used,
amount of insecticide per square meter of surface treated, and total  surface
area treated.

     (xii) Dust box treatment (poultry). Insecticides formulated as dusts
or granules  are placed into  dust box containers and fowl allowed  to treat
themselves.  Record formulation,  diluent, final concentration of active  in-
gredient, amount of material per dust box, length of treatment period, num-
ber of birds, and average amount of material used per bird during treatment

     (xiii) Vapor treatment. Strands,  cords, or other devices impregnated
with insecticides are  attached underneath or around cages containing  in-
fested birds. Insecticides volatilize from the impregnated surfaces and kill
ectoparasites on birds. Record the formulation, impregnated material, final
concentration of active  ingredient, length or weight of impregnated mate-

rial per bird or per cage containing a specific number of birds, and length
of treatment  period. Low volume or ultra low volume application  must
be evaluated if these methods are to be specified on the label.

     (6) Record of toxicity and other adverse effects. Adjusted average
daily gains (ADG) on the test and control groups should be recorded and
reported. The pesticide product's possible toxic effects on the animal must
be evaluated. For example, small amounts of certain solvents cause toler-
able minor itching and burning for short periods after application. Higher
concentrations may cause death if not diluted before application. The rec-
ommended method of treatment for pour-on and other ready-to-use prod-
ucts containing oils should be tested to ensure that the recommended meth-
od of treatment will not result in excessive dosages of oil which may evoke
adverse reactions in treated animals. The final use dilution of emulsifiable
concentrations should be tested to ensure that it does not contain dangerous
amounts  of oil. Insecticides which are tested for use on livestock, poultry,
of fur- or wool-bearing animals should not injure the animal even when
application is repeated over a long  period of time; the margin of safety
to the  treated animal is  a vital  consideration  in determining usefulness.
Neither should the  insecticides appear illegally in or on meat,  meat by-
products, milk, or eggs. If the treated animal is less than 3 months of
age (excluding poultry), the effects of stress  caused by a particular oper-
ation such as castration,  dehorning, or other  similar procedures, should
be evaluated in conjunction with toxicity data. This is  especially important
for feed-through insecticides. For  other requirements  on toxicity, refer to
OPPTS guideline series 870, Health Effects Test Guidelines.

     (7)  Evaluation and reporting procedures.  The  evaluation proce-
dures used should be  specified  in the presentation  of the  data. Reports
should include formulations, final  concentrations of  active ingredient in
terms of ppm in water or percent active ingredient  in dusts,  dosage in
terms of mg  of active ingredient per kg of body weight of animal, length
of treatment period, equipment used, and other similar factors. The average
daily gain in  weight should be reported for any tests of 7 to  14 days dura-

     (8) Untreated controls and comparative treatments. When it is im-
practical  to maintain untreated control animals, the  effectiveness of the
test product may be measured by comparative treatments with products
of well-known efficacy used as reference standards. To evaluate the effect
of treatments, records should be kept on the comparative changes in meat,
milk, or  egg  production or some similar measurement; and insect counts
following each treatment.  The numbers of animals in the control or com-
parative treatment groups must be equal to the numbers of animals  in a
treatment group in small scale tests; in large-scale tests, only a  small  por-
tion of the animals need to be treated or given a standard treatment.  Con-
trol groups should contain animals of the same general size, age, condition,

and origin as those in the treatment groups  and have similar infestations
of parasites or  be exposed to similar populations of arthropod parasites.

    (d) Suggested performance standards. Unless otherwise specified,
these  standards are presented on the basis of pest population counts from
treated compared to untreated animals. All percentages  of control refer
to the performance of the  test product (as determined by  insect counts,
yield  of meat and milk, and any  other measures correlated to insect popu-
lation pressures) against the vulnerable stage(s) of the target pest, when
evaluated  according to a recognized treatment program under actual field

    (1) Cattle  (beef and dairy)—(i) Horn fly. The percentages of control
for the horn fly are based upon pre- and post-treatment counts coupled
with a comparison to separate untreated control groups. Such percentages
are derived from actual counts of the number of adult flies per side per
animal,  and may additionally be correlated with average daily weight or
milk production.

    (A) Feed  treatment, drinking-water treatment, and mineral salt
or protein supplements. A minimum of 90% control of emerging adults
along with a 70% reduction of  adults  on the  cattle as measured by side
or whole-body counts.

    (B) Whole-body sprays, dips,  pour-ons.  A minimum of 90% reduc-
tion in infestation.

    (C) Dusts. A minimum of 95%  reduction in infestation.

    (D) Backrubbers. A minimum  of 90% reduction in infestation, under
continued use for one month.

    (E) Low volume (LV), ultralow volume (ULV) and waxed-bar ap-
plications. A minimum of 90% reduction in infestation under continued

    (ii) Other biting flies: (Stable fly, deer flies, and horse flies). The
percentages of control given for these pests are based upon the feeding
rates  of flies before treatment compared to  the  feeding rates  of flies on
the same  cattle after treatment as correlated to  the feeding rates on un-
treated animals taken at the same time. Such feeding rates  should be ex-
pressed as populations of each separate pest  species  observed consistently
at a particular time of day associated with fly activity.

    (A) Whole-body sprays, dips, pour-on,  and dusts. A minimum of
90% reduction  in infestation one day after application and 75% reduction
in infestation one week after application.

    (B) Backrubber, LV  and  ULV applications. A  minimum of 90%
reduction  in infestation under continued use.

     (iii) Face  fly: Whole-body sprays, dips,  pour-ons, dusts,  back-
rubbers, LV and ULV applications, smears, baits, ointments, and face
wipes. Reduction in infestation should range from 20-60% for 2-3 days
from a  single application, based  upon a comparison  of the numbers of
flies per animal pre- and post-treatment correlated with populations  of the
pest occurring on adjacent untreated herds observed at the same time.

     (iv) Ticks. The percentage of control listed as suggested performance
standards for ticks are based upon pre- and post-treatment counts on the
same animals or as the average number counted on untreated controls dur-
ing the  same post-treatment  intervals. If pre- and post-treatment counts
are utilized as a basis for computing the percentage, then untreated control
counts should still be reported to permit correlation with natural tick popu-
lation dynamics for the site/pest complex.

     (A) Whole-body sprays, dips, pour-ons, and dusts. A minimum of
90% reduction in infestation one day after application  and  75% reduction
in infestation for one week after application.

     (B) Backrubber,  LV and ULV applications. A minimum of 90%
reduction in infestation under continued use.

     (2) Horses—(i) Ticks. A minimum  of 90% reduction in  infestation
for one  week and 75% reduction in infestation for one  month, based upon
pre- and post-treatment counts in comparison with untreated controls.

     (ii) Flies: Dips, whole-body sprays, fine-mist sprays,  toxicants and/
or  repellents in aerosols,  sponge-ons,  wipe-ons, smears,  baits, and
treated  halters. The percentage given as a suggested performance  stand-
ard for  fly  control are  based  upon a reduction in infestation numbers for
a minimum period of 3 hours or that period of protection to  be claimed
on the label, whichever is shorter. A minimum of 90% reduction in infesta-
tion under continued use.

     (3) Chickens,  turkeys,  and other domestic fowl—(i) Manure-in-
habiting fly larvae—(A) Feed treatment. A minimum of 90% reduction
in infestation under continued use, based upon both adult fly  reductions
determined either by manure emergence or manure bioassay testing, com-
paring both treated and control groups.

     (B) Fly larvicides to manure. A minimum of 90% reductions in in-
festation for period of 2 weeks after treatment, based  on the criteria pre-
sented in paragraph (d)(3)(i)(A) of this guideline.

     (ii) [Reserved]