Safety Testing - An Essential Use of Laboratory Animals. Contemporary
Topics 36 (5): 40.
This article reviews the basic rationale for safety testing of consumer
products and the importance of animal studies in this process. It places
laboratory animal-based safety testing into historic and present day contexts
of protecting public health and safety.
Safety testing permits manufacturers to label their products with appropriate
instructions on the safe use of that product, precautions for preventing
injury, and measures to take in case of accidental misuse.
Safety testing identifies products and substances that may be safely
introduced into the global environment and those that require special handling
and disposal.
Product safety requires biologic testing methods such as:
1. clinical, human volunteers
2. in vivo, using nonhuman living organisms
3. in vitro, tests that do not use whole animal systems
Safety testing keeps toxic products off the market and also determines
safe levels of exposure and methods of use.
Toxicity studies obtain information about adverse effects and possible
treatments for problems caused by acute exposure to a product as well as
exposure over a period of time.
1938 Federal Food, Drug, and Cosmetic Act: mandates that before marketing,
the safety of prescription drugs must be tested with adequate methods,
including the use of experimental animals. Enforced by Food and Drug Administration.
1960 Federal Hazardous Substances Labeling Act: requires that a manufacturer
determine if a product for household use should bear appropriate conspicuous
labeling such as "Poison", "Danger", or "Caution" on the basis of testing
in "normal living tissue" or "laboratory white rats". Animal testing is
not required if determination of safety and labeling can be made on the
basis of previous human experience, existing animals study data, or expert
opinion.
The Environmental Protection Agency can ban or restrict the use of
any substance that represents an "unreasonable risk of injury to health
or the environment." To make these judgments, the EPA can require that
specific animal safety testing be conducted by the manufacturer.
Questions:
1. Three biological testing methods that may used to test product safety
are:
2. Who enforces the Federal Food, Drug, and Cosmetics Act?
Answers:
1. in vitro, in vivo (nonhumans), clinical (human volunteers).
2. FDA (Food and Drug Administration)
Institutional Animal Care and Use Committees: A Survey Covering 10
Years of Experience. Contemporary Topics 36 (5): 42.
A survey was performed of IACUCs at U.S. institutions that maintain
an animal welfare assurance with OPRR. Questions were asked about the IACUC
composition, function and procedures; training and education programs;
controversial issues; protocol review and semi-annual program review and
inspection; surgical or potentially painful procedure; characteristic of
participating institutions.
>80% of institutions did not reimburse their non affiliasted members.
Questions:
1. Do the OPRR and the USDA allow institutions to reimburse the IACUC
non-affiliated community members?
2. What three criteria must be fulfilled if IACUCs use electronic communication
(fax, audio or videoconferencing, email) for the review
process?
Answers:
1. Yes but the amount of compensation, if any, should not be so substantial
as to be considered an important source of income or to influence voting
on the IACUC.
2. (i) must be fully described in the institution's PHS assurance statement
(ii) must allow a high degree of interaction and deliberation
(iii) a quorum of IACUC members must be in direct communication with
each other and be given full opportunity to participate for the duration
of the meeting
Barrier Film Protects Skin of Incontinent Rats. Contemporary Topics
36 (5): 46.
Urinary incontinence is often seen in spinal cord injury animal models.
The sequelae of urinary incontinence can lead to removal of significant
numbers of animals from a study. The paper reports the effectiveness of
the use of a skin protectant barrier film product in preventing urine scalding.
The barrier film product was a spray on product, containing polydimethylsiloxane
in hexamethyldisiloxane carrier. It is non-alcohol based and non-irritating.
It allows gas and mositure permeability, decreasing skin irritation.
Questions:
1) List three complications associated with urine scalding.
2) List examples of neurogenic urinary incontinence.
3) List examples of non-neurogenic urinary incontinence.
Answers:
1) Intractable skin ulceration; secondary bacterial dermatitis; self-trauma;
urosepsis; urinary tract infections
2) Neoplasia or trauma to the sacral spinal cord; cauda equina syndrome;
intervertebral disk protrusions; neoplasia or trauma to the upper regions
of the spinal cord; meningitis; cerebral or cerebellar disease; over-distension
of the bladder and associated depolarization of the detrusor muscle.
3) Hormonally induced; congenital genitourinary abnormalities; urethral
strictures or calculi; senility
A Practical In-house Method for Bile Duct Cannulation of Rats. Contemporary
Topics 36 (5): 49.
The author developed a technique to implant cannula in the bile duct.
The goals were to improve failure rate of commercially available vendor-cannulated
rats, shipping stress, weight loss and general health, and increase the
longevity of patent canulas.
Catheters are constructed of PE-50 tubing and silicone tubing is used
as beads. Both ends of catheters, bile duct and duodenum canulas, are linked
by a customized U-Shaped piece off a 23-gauge needle and sutured dorsal
to the scapulas.
All goals were attained without contamination of pancreatic secretions.
Bile is used to evaluate drug absorption, bioavailability, pharmacokinetics,
and metabilism of experimental compounds. With the improvement of general
health as well as substantial improvement in reliability, the number of
surgically instrumented rats required for each study protocol will be reduced.
No questions
Use of Computed Tomography to Aid in Diagnosing Transient Blindness
in a Rhesus Macaque (Macaca mulatta). Contemporary Topics 36 (5):
51.
Computed Tomography (CT) uses coordinate movement between the x-ray
source and detector keeping the orientation of the x-ray source and detector
combination perpendicular to the long axis of the patient. The thickness
of the each plane examined is governed by the x-ray beam width. Usually
this width is 1 to 10 mm. These planes are further divided into volume
subunits called pixels. By using complex mathematics and geometric processes,
the relative density as compared to water of each pixel is determined.
The calculated densities are assigned a shade of gray. The available shades
of gray discernable by the human eye can be redistributed among these various
relative densities to permit visual density discrimination far greater
than that of general radiographic films. Sagittal, dorsal, and oblique
orientations can be generated by computer manipulation of the raw data
since algorithms originally generated it. Another name for CT is CAT (Computed
Axial Tomography) scan. CT scans are used extensively to diagnose intracranial
diseases in humans. In nonhuman primates (NHP), diagnosis of intracranial
disease is difficult due to the inability to perform a neurologic exam
on a conscious animal. Observations are limited usually to passive observations.
CT has been used in NHP to diagnose cerebral aneurysms and cerebral infarcts.
This paper examines and follows the progression of intracranial abnormalities
potentially related to neurologic abnormalities seen in a female Macaca
mulatta.
A 3-year female M. mulatta had a cephalic recording stainless-steel
cylinder implanted surgically. The monkey showed no abnormalities as result
of the surgery or after the stereotatic placement of electrodes in the
cylinder was performed. The monkey initially developed lethargy, anorexia,
weakness, and vomiting. The monkey then developed dysmetria and the inability
to grasp foods that were offered. Clinical chemistries and CBC were normal.
Observing the monkeys response to new people entering the room, to tracking
visual objects, and to placement in a new cage suggested to the authors
that the monkey was blind. Twenty-four days after the onset of clinical
signs the monkey was anesthetized. A complete ophthalmic examination and
CSF analysis were performed and found to be normal. At this time, a CT
scan was performed without contrast enhancement. Enlarged lateral ventricles,
cerebral aqueduct, third ventricle, and fourth ventricle were the only
abnormalities found. A tentative diagnosis of hydrocephalus was made. Without
treatment, the monkeys condition did not progress. Fifty-one days after
the first examination the monkeys eye sight returned and the she became
able to resume her experimental activity. Seven months after the initial
CT scan a second CT scan was performed. There was a marked decrease in
the size of all the ventricles especially in the fourth ventricle.
Hydrocephalus has been reported in Papio cynocephalus, Saimiri sp,
macaques (M. mulatta, M. fascicularis, M. artoides), Aptus nancymai. In
this monkey magnetic resonance imaging was not chosen since the indwelling
metal implants precluded its use. The implants did limit analysis to the
level of the implants. Apparently, the algorithms used in CT scan can be
set to accommidate metal objects to avoid streaking and other artifacts
as a result.
Questions:
1. Only one orientation can be generated with CT scans. T or F
2. What does CT and CAT stand for?
3. Magnetic resonance imaging can easily replace CT scan analysis in
all instances. T or F
Answers:
1. F
2. Computed tomography and computed axial tomography
3. F
Urinary Excretion of Cortisol from Rhesus Monkeys (Macaca mulatta)
Habituated to Restraint. Contemporary Topics 36 (5): 55.
The introduction of this study specifically addressed the issues surrounding
long term (12- to 14- day) restraint of monkeys for research conducted
during space flights... ie, muscle atrophy, neuroendocrine stress. The
authors premise was that prior habituation to restraint would attenuate
cortisol excretion in urine (an index of stress) and decrease muscle atrophy.
Urinary excretion of cortisol, creatinine, electrolytes, and food and water
intake were compared between nonrestrained monkeys (ie, samples taken before
and after a restraint phase) and restrained monkeys. Restraint devices
used were Primate Products restraint (PPR) system and a flight chair restraint
(FCR)system. (Please refer to the article for a description of this restraint
system.) Because the flight chair system was in development during the
study, monkeys were initially trained to the PPR system, then switched
to the FCR system for more advanced training. Training was conducted over
a 4-year period typically for 4-8 hours/day initially and progressing up
to 20 days. Thereafter, they were maintained on 10-day cycles of alternating
restraint and nonrestraint. When not restrained, monkeys were maintained
in metabolism cages. Five weeks prior to study, monkeys were instrumented
for EMG and had muscle, lymph node, and bone biopsies performed as part
of another study being conducted.
Study design: 4 male 9 to 10 year-old monkeys were used;
Prestudy - nonrestraint; phase I - monkeys in metabolism cages for
7 days; urine collected via metab cage ;
Day 8 - placed in FCR system for restraint phase II for 18 days; urine
collect via sheath attached to penis.
Day 26 - returned to metabolism cage for 5 days of phase III-postrestraint
phase.
Urine samples were analyzed for cortisol, sodium, potassium, creatinine.
Excretion rates of each were calculated. Statistical comparisons were made
between the 3 phases. Values for the nonrestraint phases (I and III) were
used as baseline control values to which restraint values were compared.
Results: One monkey was removed from restraint prematurely ( d 16 of
restraint) because of an ulcer on its callosity. Body weights did not change
between start of phases I and III. Mean excretion rate of creatinine, sodium,
and potassium were also not altered.
Conclusions: Prior habituation to restraint attenuated neuroendocrine
responses and the chair restraint system used (FCR) allowed adequate activity
to prevent muscle wasting.
No questions
A Rapid and Simple Technique for Serial or Continous Collection of
Blood Samples and Intravenous Administration of Drugs to Consious Swine.
Contemporary Topics 36 (5): 58.
This paper describes a procedure to construct a catheter using a butterfly
catheter, non-thrombogenic polyethylene tubing and a syringe with a 23
ga. needle. The catheter was placed in the auricular vein along the caudal
ear margin of a conscious pig immobilized in a sling.
The pig was acclimated to handling. They were placed in an up-right
sling and the ear was clipped and cleaned. Lidocaine ointment was applied
to the ear margin. A rubber band was placed around the base of the ear.
The catheter was placed and then taped to the ear. Heparin was infused
in order to maintain catheter patency. These catheters were used for infusion,
and periodic blood sample collection. 74 catheters were placed. Average
time required for a successful catheterization was 30 minutes. The maximum
volume of blood that could be collected was approx. 1 ml/min. Some irritation
around the catheter site was noted 4-6 hours after cannulation. Patency
was maintained up to 28 hrs. No major complications were noted.
Questions:
1) What is the mechanism of heparin as an anticoagulant?
a) It combines with antithrombin III to inactivate factor Xa,
preventing the conversion of prothrombin to thrombin.
b) it inactivates thrombin
c) it blocks the conversion of fibrinogen to fibrin
d) it combines with anatithrombin III and inactivates factors IX,
X, XI, XII
e) all of the above
2) What is the origin of heparin?
a) pig intestine
b) horse hooves
c) cow lungs
d) recombinant E. coli
e) a and c
f) all of the above
Answers
1) e
2) e. Heparin is a sulfated glycosaminoglycan found naturally in mast
cells. It has a MW of 12,000.
The Influence of Position and Orientation of Racks on Airflow Dynamics
in a Small Animal Room. Contemporary Topics 36 (5): 62.
A series of 7 studies were conducted to assess the effects of rack
placement on animal room ventilation. Heating, ventilation, and air conditioning
(HVAC) room configurations were studied. Computational fluid dynamics (CFD)
was used to evaluate the data. 3 animal rooms were used to conduct the
studies. One was equipped with the newly developed air capture and containment
(ACC) system. The other 2 rooms were conventionally configured. The ACC
room had 2 radial diffusers and 6 extract vents as a single overhead unit.
One conventional room had 2 radial diffusers placed in the center of the
long axis of the room with 4 low-level extract vents. The other room had
a linear-slot diffuser with 4 low level extract vents. An additonal set
of studies were conducted to compare the effects of rack orientation with
a heavy animal load (6 racks @200 watts/rack; 3 racks on each wall). Parallel
and perpendicular rack placements were compared. The low to moderate load
was 2-3 racks of rats; 100 rats/rack x 200g/rat= 200watts/rack.
Study 1 - 2 racks (1 on each side of wall) were placed on opposite sides
of the room adjacent to a low-level extract vent in a corner.
Study 2 - 2 racks on opposite walls located away from the extract vent
in a position such that they were centered on a radial diffuser.
Study 3 - 2 racks adjacent to each other on same side of wall and located
under an extract vent.
Study 4 - 2 racks on same side of the wall centered under the radial-type
inlet diffusers.
Study 5 - 2 racks on same wall under an extract vent and one rack on
the opposite wall also under an extract vent.
Study 6 - racks positioned similar to 5 but centered under the radial-type
inlet diffusers.
Study 7 - 3 racks on same wall and each positioned under an ACC system
extract vent.
Each study was done with the racks placed perpendicular to or parallel
with the side walls for each placement/number combination and HVAC configuration.
42 separate studies were actually conducted.
CONCLUSIONS
Position and geometric orientation had substantial effects on airflow
and balance. Conventional room-radial type diffusers and 4 corner exhaust
vents with racks in close proximity and centering them under the diffusers
resulted in best air circulation with lowest amount of re-entrainment and
dead space. Placing the racks parallel or perpendicular had no impact.
The ACC system had the best ventilation for animals and staff regardless
of geometric positioning.
STUDY QUESTIONS
1. When did orientation have little or no effect on ventialtion?
2. What can cause a deterioration in airflow around a rack?
3. What is the rat breeze effect?
4. How can you minimize this effect?
ANSWERS
1. when there were 2 opposing racks in a room
2. moving it further away from a corner extract vent or a perpendicular
arrangement along the same wall
3. areas of low velocity flow and dead space developed immediately
in front of the racks. Cool, fresh air flowed down from the inlet to the
floor while the air warmed by the animals rose above the rack and proceeded
to the ceiling, where in a conventional setting, is mixed with fresh air,
re-entrained, and re-circulated thoughout the room.
4. balancing opposite sides of the room
Spontaneous Hypertrophic Osteopathy in a Wistar Rat. Contemporary
Topics 36 (5): 68.
Report is first case of hypertrophic osteopathy reported in a rodent.
A colony of SPF WI/HicksCar rats were being monitored during a study of
aging. No experimental procedures had been performed on the rats. A 34month
old male rat was presented for lethargy, pale mucous membrans, bilateral
swelling of rear metatarsal regions, bilateral rear limb and tail paresis.
Necropsy revealed bilateral swollen, firm, fused hock joints, enlarged
distal tail, nodule in right caudal lung, distended urinary bladder, enlarged
pituitary gland, hepatomegaly, red mesenteric mass. Radiology = periosteal
new bone growth in hocks and tail. Serology and bacteriology were negative.
Histology = periosteal new bone formation, pulmonary adenoma, multifocal
granulomatous pneumonia, cardiac fibrosis and heart failure, bilateral
interstitial cell tumors, pituitary adenoma, mesenteric hemangioma, bilateral
nephrosis, chronic periodontitis, squamous metaplasia of nasal respiratory
epithelium, axonal degeneration and polyradiculoneuropathy of cauda equina.
Hypertrophic osteopathy (HO),[ aka Marie's disease, acropachia, and
hypertrophic pulmonary osteoarthropathy], is characterized by increased
blood flow to limbs, thickened parosteal connective tissue, and periosteal
new bone formation. Condition is usually associated with intrathoracic
neoplasia, but also reported with bladder, liver, and ovarian tumors in
man and other animals.
HO had been induced in rats by chronic administration of an isoquinoline,
but had not been seen as a spontaneous condition. In dogs and man, HO has
been associated with heart disease, pneumonia, intrathoracic and extrathoracic
neoplasms. This rat had congestive heart failure, multifocal granulomatous
pneumonia, and multiple neoplasms; contribution of these lesions to development
of HO is suspected but unproven; developing an animal model to study HO
may be possible.
Questions:
1) What is hypertrophic osteopathy?
2) What infectious agent(s) might be considered in a rat with arthropathy?
Answers:
1) HO is a disease of unknown etiology, reported in many different
animals and man, consisting of increased blood flow to limbs, thickened
parosteal connective tissue, and periosteal new bone formation associated
with degenerative and neoplastic conditions.
2) Mycoplasma arthritidis
Streptobacillus moniliformis
Corynebacterium kutscheri
Hemangiopericytoma in a Rat. Contemporary Topics 36 (5): 71.
This is the first report of a spontaneous hemangiopericytoma in a rat.
A 13month old male SpragueDawley rat developed a mass on the right pinna.
The rat had not been used experimentally. The mass was excised; histo =
fusiform cells arranged in a whirling fingerprint pattern which converged
on a central vascular structure; this pattern is characteristic of hemangiopericytomas.
Hemangiopericytomas are uncommon in all species. They are thought to
arise from the pericyte of Zimmerman cells found adjacent to capillaries.
They have been experimentally induced in the medaka (Oryzias latipes) and
mice, and been reported spontaneously in man and dogs. They are slowgrowing
tumors, but have incidence of metastasis of 50%. Not all hemangiopericytomas
display the characteristic fingerprint pattern; immunohistochemistry and
ultrastructure can be used to achieve a diagnosis. Pericytes are pluripotential,
and may express a variety of antigens depending on the location of the
cells in a given species, so diagnosis can be challenging.
Questions:
1. T or F All hemangiopericytomas display a characteristic fingerprint
pattern, so diagnosis is easy.
2. T or F All hemangiopericytomas express the same antigens.
Answers:
1. F; the pattern is characteristic, but some hemangiopericytomas do
NOT display this patterns; conversely, some other neoplasms may display
this pattern, so diagnosis cannot be based on this alone.
2. F; pericytes are pluripotential, and show different patterns of
differentiation and express different antigens depending on location and
species.
Spontaneous Carotid-Cavernous Sinus Fistula in an Aged Rhesus Monkey
(Macaca mulatta). Contemporary Topics 36 (5): 73.
A Carotid Cavernous Sinus Fistula (CCSF) results from an abnormal shunting
of blood between an intracranial branch of the carotid artery and the venous
cavernous sinus located just caudal to the eye.
Direct fistulas--arterial blood passes through a defect in the wall
of the intracavernous segment of the internal carotid artery. Usually due
to trauma involving fractures or to the rupture of an aneurysm or as a
result of atherosclerosis in the internal carotid (humans). The classical
clinical picture is exophthalmos, severe orbital vascular congestion, limited
ocular motility and bruit (abnormal sound or murmur heard on auscultation).
Spontaneous CCSF--usually dural shunts between branches of the carotid
and dural veins in the region of the cavernous sinus. Clinical symptoms
are: lesser degree of congestion, conjunctival arterialization, ocular
hypertension, abduction deficit; less likely to detect bruit.
This case involved a 35 year old rhesus monkey with clinical signs
of a direct fistula, but diagnosis of a spontaneous CCSF. The clinical
history is similar to an older female human suggesting that macaques may
be a good model for this disease.
History: Swollen right eye (no previous history of surgery or therapy
other than routine testing and amputation of a digit). Initially, mild
periorbital edema and erythema with slight scleral and conjunctival injection.
Rapidly progressed to chemosis (conjunctival edema), and hyphema (blood
in the anterior chamber), exophthalmos, engorgement of vessels, decreased
ocular motility and increased intraocular pressure. Culture, radiography,
conjunctival scrapings and blood values led to no significant findings.
A bruit was discovered on auscultation of the eyebrow. A CCSF was diagnosed
by orbital ultrasound ( dilated superior ophthalmic vein, low reflective
widening of Tenon's capsule, thickening of extraocular muscles, enlargement
of the retrobulbar optic nerve positions) and indirect fundic exam (massively
dilated retinal veins, disc edema, retinal hemorrhage, venous stasis retinopathy,
and choroidal effusions).
Treatment: Analgesics, ophthalmic beta blocker to relieve glaucoma.
Clincal signs worsened the first week, then regressed with disappearance
of the bruit during the second week. US findings revealed normal orbital
structures, but follow-up exam revealed blindness, optic atrophy, afferent
pupillary defect, and anisocoria.
Histological exam: Anterior synechiae, loss of some retinal layers,
marked cupping of the optic nerve head, and atrophy of the optic nerve.
Discussion: Once a fistula has formed between the cavernous sinus and
the carotid arterial supply, the AV shunting of blood results in increased
venous pressure and decreased arterial perfusion. The orbital veins (outflow
from the sinuses) becomes engorged and there is secondary congestion of
the orbital soft tissues. Decreases arterial perfusion causes hypoxic changes
that contribute to soft tissue swelling, muscle paralysis, and ischemia
of the anterior chamber (can result in protein flare, cells in aqueous
humor, corneal edema, glaucoma, iris rubeosis, cataracts).
In humans, spontaneous CCSF have developed most commonly in postmenopausal
women. Predisposing factors include aneurysm or AV malformation, atherosclerotic
changes, collagen vascular disease, hypertension, and childbirth. The carotid
vasculature of the macaque in this case was not analyzed, but there was
atherosclerosis of the abdominal aorta and the uterine vessels. The ovarian
and uterine changes were consistent with postmenopausal changes in rhesus
monkeys and human beings.
Up to 60% of spontaneous CCSF in human beings will resolve naturally,
are generally not considered life threatening, and the clinical course
is usually benign. Treatments are aimed at relieving pain and ocular signs,
preventing glaucoma and ischemic retinopathy. Currently intravascular closure
using detachable balloon catheterization is performed if there is a threat
of visual loss.
Questions:
1. Spontaneous carotid cavernous sinus fistula occur most frequently
in:
a. Aged sprague dawley rats
b. Post-menopausal primates
c. Guinea pigs with vitamin C deficiency
d. Rabbits with cholesterolemia
2. A common clinical sign detected in an animal with a direct carotid-cavernous
sinus fistula is:
a. seizures
b. cerebral edema
c. bruit
d. epistaxis
QUESTIONS:
1) Define bruit:_________________________________________
2) Genus, species of a Rhesus Monkey: ________ __________
3) Which of the following diagnostic procedures can be used to diagnose
a CCSF
a. ultrasound
b. angiography
c. fine-needle aspiration
d. auscultation
e. a & b
f. a,c, & d
g. a,b, & d
Answers:
1. b (post-menopausal primates)
2. c (bruit)
ANSWERS:
1) A sound or murmur heard during auscultation, especially an abnormal
one; aneurysmal bruit: a blowing sound heard over an aneurysm.
2) Macaca mulatta
3) G
In-House Development of an Integrated Management Information System
For a Laboratory Animal Facility. Contemporary Topics 36 (5): 77.
Establishing a computerized system to manage a laboratory animal facility
can be accomplished through the use of commercial software packages, or
software developed in-house.
To identify a suitable software package from the commercially available
products, compare the programs based on the following questions:
1.Does the software provide the basic functionality required to manage
the animal facility (example: does it manage research protocols, animal
procurement, census, billing, and cost accounting )?
2. Will the software adapt to the animal facility requirements, or
will facility procedures have to be changed in order for the software to
work ?
3.How expandable and flexible will the software be to future facility
changes ?
4.How much would the system cost for installation and customization,
and would upgrades, support and future maintenance incur additional charges?
If the expense or extra work required to install and customize commercially
available software is too great, the development of software in - house
may be more time and cost effective. The following is an outline for the
development of in-house software:
1. Identify the major objectives the new system has to address
2. Select an application development tool for software development
3. Identify major operational and functional areas; an operational
area performs a specific duty, a functional area provides support for one
or more operational areas
4. Develop a software program to include the operational and functional
areas
QUESTIONS
1. Which of the following is not an operational area?
a. Animal Procurement Office
b. Census
c. Animal Facility
d. Business Office
2. When identifying the system objectives, which staff members should
be consulted (list 3 major groups)?
1.______________________
2.______________________
3.______________________
3. True or False: Future flexibility and adaptability of a software
package is not a major concern during system design, or selection.
ANSWERS: 1. B | 2. husbandry, clinical, and administrative |
3. False