ASTM E2045-99(2009)

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
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Designation: E2045 − 99(Reapproved 2009)
Standard Practice for
Detailed Clinical Observations of Test Animals
This standard is issued under the fixed designation E2045; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3. Significance and Use
1.1 This practice describes the terms used in observing and 3.1 This practice pertains to all forms of toxicological
testing (acute, subchronic, or chronic) performed by any route
recordingcutaneous,gastrointestinal,respiratory,reproductive,
neuromuscular, ocular, and general clinical signs of animals of administration (inhalation, oral, dermal, ocular, or other).
undergoing toxicological testing. This practice also assists in
3.2 The U.S. Environmental Protection Agency, Good
properly observing and assessing laboratory animals for signs
Laboratory Practices for Nonclinical Laboratory Studies, as
of disease or adverse effects of compound administration.
listed in 40 CFR, requires that a testing facility maintain
specific standard operating procedures (SOPs) including an
1.2 This practice includes codes and descriptions for a wide
SOP covering clinical observations in test animals.
variety of clinical signs, anatomical locations, and other
descriptive qualifiers, and a technique for scoring the extent or
3.3 This practice serves as a basis for consistency in clinical
severity of clinical signs.
observations. Actual procedures and forms to be used in
recording observations must be described in individual study
1.3 This practice assumes that the reader is knowledgeable
protocols.
in animal toxicology and related pertinent areas and is trained
in making clinical observations.
4. Procedure
1.4 This standard does not purport to address all of the
4.1 Observe the health of an animal at a distance and of its
safety concerns, if any, associated with its use. It is the
housing environment to gain a general impression of its health.
responsibility of the user of this standard to establish appro-
Alsonoteenvironmentalfactorssuchastemperature,humidity,
priate safety and health practices and determine the applica-
ventilation, air quality and hygienic conditions.
bility of regulatory limitations prior to use.
4.2 Observe each animal and note any subtle changes in
animal behavior, physical appearance, posture, gait,
2. Referenced Documents
vocalization, food and water consumption, and waste produc-
2.1 Federal Standards:
tion. See Section 5 for details.
Title 40, Code of Federal Regulations (CFR), Environmental
4.3 Observe control animals first, followed by test groups in
Protection Agency, Subchapter E, Pesticide Programs,
order of increasing level of treatment. Observe positive control
Part 160, Good Laboratory Practice Standards
group, if any, last.
Title 40, Code of Federal Regulations (CFR), Toxic Sub-
stances Control Act, Part 792, Good Laboratory Practice 4.4 Note any dead animals and collect necessary tissues and
Standards data before decomposition occurs.
Title 40, Code of Federal Regulations (CFR), Environmental
4.5 Report animals that show signs of sickness so that
Protection Agency, Part 798, Health Effects Testing
appropriate diagnosis, treatment, or euthanasia, if appropriate,
Guidelines
can be performed.
5. General Clinical Signs
This practice is under the jurisdiction of ASTM Committee E50 on Environ-
5.1 Note the overall activity, behavior, and condition of the
mental Assessment, Risk Management and Corrective Action and is the direct
animal. Determine the hydration status by examining skin
responsibility of E50.47 on Biological Effects and Environmental Fate.
Current edition approved Nov. 1, 2009. Published March 2010. Originally
turgor, position of the eyes such as normal or sunken, mucous
approved in 1999. Last previous edition approved in 2003 as E2045 – 99(2003).
membrane color, and capillary refill time. Look for asymmetry
DOI: 10.1520/E2045-99R09.
or the presence of abnormal swellings, hemorrhage or signs of
Available from U.S. Government Printing Office, Superintendent of
Documents, Washington, DC 20402. pain.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E2045 − 99 (2009)
5.2 The following are some general conditions along with 6.1.3 Swelling (SW) is an increase in tissue size or in-
suggested codes for record keeping that do not fall into any creased abnormal shape of the skin or other organs from
specific organ system. Refer to Annex A1-Annex A3 for a abnormal presence of: air, that is, emphysema (SWA); fluid or
detailed listing of the codes and their descriptions. Other water, that is, edema (SWE); solid tissue or tumor (SWT);
3,4,5
general reference material will also be helpful. blood, that is, hematoma (SWH); pus, that is, abscess (SWB).
5.2.1 Activity may be described as: decreased (ACD);
6.1.4 Skin condition (SK) may be described as: thickened
increased (ACI); hyperexcitable (HX); hyperactive (HYP);
(SKT); thinned (SKH); scaly (SKS); dry (SKD); or red (SKY)
lethargic (LE); irritable (IRR); moribund (MB), that is near
(see 6.1.5).
death; prostate (PRO), that is, exhibiting inability or unwill-
6.1.5 Erythema (ERY) is an increased pink or red color on
ingness to maintain upright posture.
smooth skin.
5.2.2 Body condition may be described as: obese (OBS);
6.1.6 Rash (RAS) is small red, pink or white dots or
thin (THN); decreased rectal temperature (BTD); increased
pustules on the skin; petechiae (PET) are red dots formed from
rectal temperature (BTI); hypothermia that is cold to touch
blood.
(HPO); hyperthermia that is warm to touch (HPR).
6.1.7 Blisters (BLS) are fluid-filled vesicles. The fluid is
5.2.3 Death may be described as: accidental death (AD);
usually clear, but can be pink (blood-tinged) or red/brown
euthanized (ETH); found dead (FD).
(filled with blood). White-filled vesicles are either pustules
5.2.4 Examineskinfordehydration(DHY).Theskinshould
(RAS) (< 5 mm) or abscesses (SW) (> 5 mm).
fall back into place immediately after it is pulled out of
6.1.8 Color change (CC) may be other than pink or red, for
position; if the skin is pulled out of position and tends to stick
example, bluish-black as in brushing, green as in bruising or
together or slowly fall back into place, the animal may be
severe infection, brown as in increased pigmentation, or white
dehydrated. Other signs of dehydration include sunken eyeball
as in blanching.
(SUN), pale or dry mucous membranes (MM), and a capillary
6.1.9 Abrasions (ABR) are denuded skin or mucous mem-
refill time of >3 s (CR4). Distinguish between dehydration and
brane.
various types of shock.
6.1.10 Lacerations(LCN)arecutsintheskinfrommechani-
5.2.5 Generalized edema (EDE) may appear as swelling of
cal injury, that is, bite, scratch, foreign object, and so forth.
the limbs, lower abdomen, head or under the mandible. When
6.1.11 Ulceration (ULC) is an open sore accompanied by
the apparently fluid-filled tissue is pressed, an indentation may
the disintegration of tissue, usually with necrosis, that is, death
persist for a short time.
of tissue.
5.2.6 Evidence of hemorrhage (HE) may appear on the
6.1.12 Scab (SCB) is an eschar formed from sloughed skin.
haircoat (HEH) or underlying skin or nails (HES), in urine
6.1.13 Pruritis (PRU) is itching evidenced by scratching,
(HEU) or feces (HEF), from the mouth (HEM), nose or
with or without a rash or abrasion.
epistaxis (EPI), eyes (HEO), ears (HEE), genitalia (HEG), or
6.1.14 Urticaria (URT) is a transient appearance of smooth,
anus (HEA).
slightly elevated bumps which are redder or paler than the
5.2.7 Jaundice (JAU) is an overall slight yellow to pale
surrounding skin and often accompanied by severe itching.
orange tinge to the skin and mucous membranes.
Uricaria often appears as localized, discrete or confluent areas
5.2.8 Mucous membrane condition (MM) is noted by the
of edema.
color and condition of the mucous membranes of the eye, nose,
6.1.15 Purpura (PUR) is confluent petechiae, that is, pin-
mouth, or external genitalia.
5.2.9 Swelling (SW) is noted by the size, location and point hemorrhages, which form ecchymoses, that is, blotchy
hemorrhages over any part of the body.
probable cause, such as edema (SWE) from: a solid tissue or
tumor (SWT); blood (SWH); air (SWA); or pus (SWB). 6.1.16 Common manifestations of dermal sensitivity reac-
tions are:
6. Specific Clinical Signs
6.1.16.1 Contact dermatitis (COD) is pruritis, erythema and
6.1 Inspect the entire haircoat and underlying skin for
vesiculation that may be followed by pustulation and necrosis
integumentary signs. Some common clinical signs and their
and that has a pattern consistent with the touch of a foreign
suggested codes are as follows:
object or substance.
6.1.1 Alopecia, that is, hair loss (ALO), includes hair
6.1.16.2 Exanthema (EXA) is macular or papular redness in
thinning, patchy/focal hair loss or balding.
discrete areas.
6.1.2 Haircoat condition (HC) may be described as: oily
6.1.16.3 Exfoliation (EXF) is loss of superficial skin layers
(HCO); rough (HCR); wet (HCW); soiled (HCS); dry (D);
with redness, swelling, and presence of free blood.
bloody (HEH); or piloerection (HCP), that is, distinctly raised
6.1.16.4 Bullous eruption (BUL) is the presence of discrete
fur, excluding the vibrissae, giving a bristled or porcupine-like
serous or seropustular areas.
appearance.
6.1.16.5 Erythema multiform (EMF) is the presence of
multiple types of macules, papules and nodules.
Taylor, E.J., ed., Dorland’s Illustrated Medical Dictionary, W.B. Saunders,
Philadelphia, PA, 27th edition, 1988.
6.2 Gastrointestinal signs are observed during external
Stedman’s Medical Dictionary, Williams and Wilkins, Baltimore, MD, 25th
evaluations of the gastrointestinal system conducted from the
edition, 1990 .
oral cavity to the anal area. Visually inspect the teeth and
Thomas, C.L., ed., Tabor’s Cyclopedic Medical Dictionary, F. A. Davis Co.,
Philadelphia, PA, 17th edition, 1993. mucousmembranesoftheoralcavity,palpatetheabdomenand
E2045 − 99 (2009)
inspect the perianal area. An inspection of the animal’s cage 6.4.1.2 Penile discharge is either none (PDA), normal
will allow evaluation of the volume, color, and consistency of (PDN), increased (PDI), bloody (PDB), serous (PDS), or
the stool. mucous (PDM).
6.2.1 Oral cavity signs and codes are: 6.4.2 Female examinations include:
6.2.1.1 Salivation may be described as: increased salivation 6.4.2.1 In external genitalia look for mucous membrane
(SAL); or lack of salvia, that is, xerostomia (XER). abnormalities (MM) or swellings (SW) of the vagina (VA) or
vulva (VA).
6.2.1.2 Dentition (TE) includes: missing teeth (TEM); loose
teeth (TEL); discolored teeth (TEC); damaged teeth (TED); or 6.4.2.2 In vaginal discharge types or consistency of dis-
malocculation (TEO). charge are: none (VDN), normal (VDN), increased (VDI),
decreased (VDD), bloody (VDB), serus (VDS), or mucous
6.2.1.3 For mucous membranes, note capillary refill time
(CR), color, and condition, such as erosions or vesicles. (VDM).
6.4.3 For urine or urination note the following conditions:
6.2.1.4 Gums may be described as: healthy, intact (GUH);
or gingivitis (GUI), that is, inflamed or bleeding gums. 6.4.3.1 Hematuria (HEU) is bloody urine. Use caution with
rabbitsastheirurinemayrangefromambertolightredwithout
6.2.2 For the abdomen, look at the overall symmetry and
the presence of blood.
size. The abdomen may be described as: smaller or more
“tucked-in” (STA) if the animal is dehydrated; larger as 6.4.3.2 Anuria (ANU) is the absence of urine for a pro-
distended or pendulous abdomen (OPA); or asymmetrical as if longed period.
an enlargement or swelling in a focal area (SW).
6.4.3.3 Dysuria (DYU) is difficulty urinating; be sure to
6.2.3 In the perianal area, look for: abnormal anal sphincter differentiate with tenesmus.
(OEA);fecal(FEF)orurinestaining(PEU);matterhair(PEH); 6.4.3.4 Polyuria (PLY) is excessive volume of urine.
mucous (PEM); or rectal prolapse (RPR), that is, a red to dark
6.4.4 Fertility signs include the following:
red tubular protrusion from the anus.
6.4.4.1 Breeding efficiency may be failure to breed (FTB),
6.2.4 For feces, note: consistency, such as normal (FEN),
conceive (FTC), or low litter size or weight (LLS).
hard or dry (FEH), soft or watery (FED), oily (FEO); amount,
6.4.4.2 Abortion (ABO) is premature delivery of dead
such as none (FEA), small (FES), normal (FEN) or large
offspring.
(FEL); content, such as normal (FEN), blood (FEB), mucous
6.4.4.3 For pregnancy status, animal appears pregnant
(FEM) or foreign material (FEF); or evidence of straining to
(APR).
defecate, that is, tenesmus (TEN). Hard feces may indicate
6.4.4.4 Mothering may be described as: poor care of young
constipation. Soft or watery feces may indicate diarrhea.
(PCY); evidence of cannibalism (CAN); or poor milk produc-
6.2.5 Function may be anorexia or loss of appetite (ANO),
tion (PMP).
emesis or vomiting (EM).
6.5 Neuromuscular signs include disease, trauma, and com-
6.3 Respiratory signs are found by evaluating the following:
poundswhichadverselyaffectthecentralorperipheralnervous
6.3.1 Rate of breathing (RR) may be slow (RRS), normal
system resulting in structural or functional changes being seen
(RRN), or fast (RRF).
in the neuromuscular system, or degree of alertness or activity
6.3.2 Depth of breathing (RD) may be shallow (RDS),
in animals. Evaluation of locomotion and coordinated move-
normal (RDN), or deep (RDD).
ments may reflect the status of the animal’s neuromuscular
6.3.3 Difficulty in breathing is dyspnea (DYS). system.
6.3.4 Periodic cessation of breathing is apnea (APN).
6.5.1 Musculoskeletal signs may be lameness or weight
6.3.5 Nasal discharge (ND) is either none (NDN), clear bearing (LMW) or non-weight bearing (LMN), limb paralysis
(NDC), yellow (NDY), green (NDG), or white (NDW). (LP), or enlarged appendage (ENA).
6.3.6 Respiratory sounds include rales (RAL), coughing 6.5.2 Posture or head carriage may be normal (HDN), tilted
(COU), gasping for air (G), and sneezing (SNE). (HDT), raised (HDR), lowered (HDL), or hunched posture
6.3.7 Epistaxis(EPI)isthefreeflowofbloodorhemorrhage (HP); gait may be normal (GAN), exaggerated (GAE), or slow
(GAS).
from the nose. This is sometimes indicated by the presence of
dry blood or dark material around the nose. 6.5.3 Central nervous systems signs include comatose
(COM), tremors (TR), convulsions (CON), ataxia (ATX),
6.4 For reproductive or urogenital signs, carefu
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