Developing Emergency Management Plans for University Laboratory Animal
Programs and Facilities. Contemporary Topics 37 (5): 052.
1996 Guide for the Care and Use of Lab Animals specifically recommends
development of a disaster plan.
Four steps involved:
1. Establish a planning team and get authorization
A. Define an incident commander - policeman or fireman in charge
B. Establish a command center
C. Team should include: personnel from facilities and environmental
health and safety, and animal facility managers, caretakers
and veterinarians
2. Analyze response capabilities and identify specific hazards that
may occur
3. Devolop the plan
(4 components should be included)
A. executive summary / mission statement
B. authorizing signatures
C. elements of the plan
D. appendices
4. Implement the plan
involves training and orientation sessions and drills
Resources available for assistance:
Federal Emergency Management Agency (FEMA)
State Emergency Management Agency (SEMA)
AVMA Emergency Preparedness and Response Guide
Questions:
1. List 4 steps that are involved in developing an emergency management
plan.
2. List 3 resources that can be helpful in developing the plan.
Answers:
1. Establish a planning team and get authorization
Analyze response capabilities and identify specific hazards that may
occur
Devolop the plan
Implement the plan
2. Federal Emergency Management Agency (FEMA)
State Emergency Management Agency (SEMA)
AVMA Emergency Preparedness and Response Guide
Survery of Current Techniques in the Care and Maintenance of the
African Clawed Frog (Xenopus laevis). Contemporary Topics 37 (5):
057.
There is a relative lack of consensus in the literature on optimal
husbandry procedures for Xenopus spp. An electronic survey was conducted
to investigate current standards for maintaining these aquatic anurans.
The survey specifically queried: tank types and transparency, water quality,
temperature, flow and volume/frog, use of tank cover, light cycle, food
type and feeding schedule. A wide range of practices were apparent in the
responses. In general, the majority of respondents housed Xenopus in opaque,
plastic containers of static water, without cover, using a 12h/12h light/dark
schedule at 19-23C. Animals are fed on a range of schedules, depending
upon research use, with most facilities feeding a commercial fish/frog
chow. The results of the survey support the need for more research into
optimizing Xenopus husbandry practices and the development of Standard
Operating Procedures.
Questions:
1. The National Academy of Science recommends what volume of water/frog?
a) 1L
b) 2L
c) 4L
d) 4.45 L
e) 5L
2) Chlorine is toxic to aquatic amphibians at concentrations (mg/L)
exceeding:
a) 0.9
b) 1.7
c) 2.4
d) 3.8
e) 4.1
Answers:
1 - b
2 - d
Comparison of in vitro monoclonal antibody production methods with
an in vivo ascites production technique. Contemporary Topics 37 (5): 061.
The purpose of this study was to determine whether in vitro techniques
are an efficient and practical means of generating monoclonal antibodies
(mAb). For protocols requiring a gram or more of mAb, in vitro system alternatives
to in vivo ascites production were considered. Three in vitro methods were
compared with an in vivo ascites production method with respect to cost,
time, quality, and technical skill. In vitro systems included hybridoma
growth in tissue culture flasks, hybridoma growth in gas permeable bags,
and a hollow fiber cartridge bioreactor (HFC). The In vivo ascites method
produced the highest amount of mAb of all methods evaluated. The mean yield
per mouse differed for each mAb produced. Although rodent health was monitored
daily, morbidity and mortality was not reported. mAb concentration of the
supernatant from the HFC system was ~31% of the concentration produced
from the in vivo system. Concentrations of mAb were lowest for the gas
permeable bag and tissue culture flask systems, ~ 1% and .65% of the in
vivo system, respectively. Intuitively, mAb production in vitro was directly
related to hybridoma cell growth. There were no differences detected in
the quality of mAb produced by each method. Labor costs were included within
the total cost analysis of the different methods evaluated. Depending on
the hybridoma used, in vitro methods were similar to, or up to five times
more expensive than, the ascites method. Cost estimates predict that the
HFC method would be equal to or double the cost of the ascites method.
The authors point out that a disadvantage of the HFC bioreactor is the
initial investment of the apparatus. However, they recommend it as a viable
alternative to the in vivo ascites method. In summary, the authors conclude
their results document the use of in vitro technology for the production
of monoclonal antibodies can be adapted by most laboratories to provide
sufficient resources for the most commonly performed experimental protocols.
They also note that many institutional core facilities and some commercial
vendors offer in vitro mAb production services. The National Center for
Research Resources of the National Institutes of Health also subsidizes
a national cell culture center that provides these services at minimal
costs (http://www.nccc.com).
QUESTIONS:
1. What is a disadvantage of the in vitro HFC bioreactor system?
2. Is their a cost effective alternative to the use of the ascites
method for monoclonal antibody production?
3. What were the three in vitro methods compared in this study?
ANSWERS:
1. The initial investment of the apparatus.
2. Yes, in vitro systems are a cost effective alternative.
3. Hybridoma growth in tissue culture flasks, hybridoma growth in gas
permeable bags, and a hollow fiber cartridge bioreactor (HFC).
A Chronic Arterial Cannula for Blood Sampling in Conscious, Unrestrained
Rats. Contemporary Topics 37 (5): 067.
The authors describe a "simple, practical, and reliable" technique
to facilitate the withdrawal of blood for at least 10 days in conscious,
unrestrained rats. The technique had little or no effect on some of the
hematological and immune parameters commonly measured to assess the immune
status of experimental animals.
No questions
A Simple Exsanguination Method That Minimizes Acute Pre-Anesthesia
Stress in the Mouse: Evidence Based on Serum Corticosterone Concentrations.
Contemporary Topics 37 (5): 073.
Reported serum corticosterone concentrations vary widely in the literature.
Many are > 100ng/ml, suggesting substantial stress. Factors which have
been implicated in this elevation of corticosterone include: environment
(caging, bedding, food, companions, etc), photoperiod, duration of disturbance,
new surroundings, and inhalant anesthestics (pungent odor and irritant
action on the eyes and respiratory tract).
This study compared CO2 and methoxyflurane as anesthetic agents and
the effect of inducing anesthesia in the animal room versus transport to
an adjacent room (travel time of less than 30 seconds). Mice in the treatment
group "holding room/CO2" had significantly lower (32 +/- 17 versus 103
+/- 32, etc) corticosterone levels than the other groups. This finding
was confirmed in subsequent trials. They also determined that distress
communicated by the mice (when anesthetized in the holding room) did not
affect adjacent mice (used subsequently).
They recommend the use of CO2 with familiar routine and in a familiar
environment to refine procedural stress preceeding anesthesia in the mouse.
QUESTIONS:
1) T F Mice housed in ventilated rack systems have been reported to
have lower mean serum corticosterone concentration levels than mice housed
conventionally.
2) Part A: To decrease un-necessary stress prior to exsanguination,
what has been the reported recommendation for a "safe" duration of disturbance
prior to the procedure (choose from choices below)?
Part B: What is the recommendation based on the findings of this paper?
(choose from choices below)
a. < 30 seconds
b. 1-2 minutes
c. 2-4 minutes
d. 4-6 minutes
e. 30 minutes
3) T F Moving mice to new surroundings immediately prior to inducing
anesthesia can increase serum corticosterone concentrations levels several
fold.
4) T F Inhalant anesthetics, like methoxyflurane, do not cause increased
levels of serum corticosterone.
5) T F CO2 produces minimal irritant action on the respiratory tract
and
has rapid induction time for anesthesia (<15 sec).
ANSWERS:
1) True
2) Part A: c
Part B: a
3) True
4) False (irritant and take 3-4 minutes to induce anesthesia)
5) True
Type B Ventricular Preexcitation With Abnormal Contraction of the
Ventricular Septum in a Dog. Contemporary Topics 37 (5): 078.
Ventricular preexcitation (VP) develops when impulses originating in
the sinoatrial node or atrium prematurely activate a portion of the ventricles
through an accessory pathway that bypasses the normal conduction system.
This is a report of VP in a beagle and echocardiographic and atropine induced
tachycardia effects were described. The heart rate of this dog with VP
increased after atropine administration. The onset of the change in heart
rate happened faster than the change in heart rate exhibited by normal
dogs when given atropine. Echocardiography revealed abnormal septal contration
without histologic correlates.
No questions
Development of a System for Improved Transportation and Handling
of Full-Sized Pregnant Sows at a Metropolitan Research Facility. Contemporary
Topics 37 (5): 082.
Researchers who normally used pregnant minipigs ($$$) or went to farms
to harvest domestic crossbred neonatal pig due to the physical difficulties
of transporting 200-250 kg sows to the research facility. Ergonomist developed
walk ramps, rolling carts and winch systems to reduce man power requirements
from 6 to 2. Solid sides on single file chutes decreased stress. Loading
is the most stressful action on the sow.
No questions
Changes in Hematologic Values During Infection of New World Monkeys
with Plasmodium falciparum and Plasmodium vivax. Contemporary Topics 37
(5): 086.
Anti-malarial vaccines are tested in Aotus nancymai (owl monkeys) experimentally
induced with Plasmodium falciparum (Vietnam Oak Knoll (FVO) strain or Uganda
Palo Alo (FUP) strain) in the blood stage and in Saimiri sciureus boliviensis
(squirrel monkeys) experimentally induced with P. vivax in the mosquito
stage, respectively. P. falciparum was harvested from infected erythrocytes.
P. vivax was harvested from mosquito salivary glands. Both have a 48-hour
life cycle, after which the red cell containing the parasite is destroyed.
P. falciparum infection often results in high-density parasitemia which
is responsible for massive erythrocyte destruction in both splenectomized
and intact hosts. Because Plasmodium vivax only grows in very young erythrocytes
(reticulocytes), changes in erythrocyte counts in animals infected with
the organisms were expected to be much lower than those infected with P.
falciparum infection.; however, the reduction of erythrocyte count was
greater than expected. A lancet stick to the thigh technique was used to
collect blood samples for malaria count. A "thin and thick" film was prepared.
The malaria count was determined from the thick film by using the Earle
and Perez method for parasite quantitation. Multiple linear regression
was used to examine the hematologic changes observed over a 14-day period
in spleen-intact and splenectomized malaria-infected owl monkeys and splenectomized
malaria-infected squirrel monkeys while controlling for the effect of parasitemia
present in each animal. During 14-day periods: the numbers of erythrocytes,
platelets, and granulocytes decreased; the values of hemoglobin and hematocrit
decreased; the numbers of lymphocytes and monocytes increased. However,
squirrel monkeys infected with P. vivax had less degrees of changes in
white blood cells mentioned above. Decreased platelet count was also seen
in malaria infected humans and rhesus monkeys. For humane treatment, animals
treated with FVO strain were given 20 mg mefloquine and 50 mg quinine in
a single dose. Animals treated with FUP strain were given 30 mg chloroquine
over 2 or 3 days. Anemia may result from increased destruction or decreased
production of red cells. Increased destruction may be due to the removal
of damaged red cells by spleen or liver or due to autoimmune condition
from malaria infection to primary destruction of red cells or to renal
disease with an associated anemia. Decreased production of red cells can
be from kidney disease or inherited kidney disease which may have normal
BUN.
Questions:
1. T or F. Because Plasmodium vivax only grows in very young erythrocytes,
changes in erythrocyte counts in animals infected with the organisms were
expected to be much lower than those infected with P. falciparum infection.;
however, the reduction of erythrocyte count was greater than expected.
2. T or F. The malaria count is performed on the thick film of a "thin
and thick" film.
3. Plasmodium falciparum and P. vivax have a ____ life cycle.
a.) 24 hour (quotidian) b.) 48 hour (tertian) c.) 72 hour (quartan)
Answers: T; T; b.
A New Method to Collect Bile and Access the Duodenum in Conscious
Dog. Contemporary Topics 37 (5): 089.
The authors describe a novel, implantable catheter system inserted
into the canine bile duct which allows both bile collection and duodenal
access in conscious dogs. This represents a refinement from previously
used systems in that it has no exteriorized components, allows quantitative
bile collection, is associated with normal liver function, accommodates
long-term use of animals, provides selective infusion access to the biliary
system, duodenum, and the catheter occluding mechanism, and allows unrestricted
movement of the animal. A T-tube catheter is used with consists of a short
T piece attached to 3 long catheters for (1) bile collection, (2) opening
and closing of the diaphragm which serves to occlude the duct in the middle
of the T piece, and (3) the distal flush catheter on the duodenal aspect.
The diaphragm is a flexible occluding diaphragm manipulated by fluid pressure
introduced into the middle catheter.. When open, bile flows freely from
the gall bladder into the duodenum. When occluded by infusion of saline
into the catheter, the diaphragm is pushed outward into the "closed" position,
allowing collection of bile via the bile collection catheter. The free
ends of the catheters are channeled out of the abdominal cavity using a
trocar and attached to subdermally positioned access ports.
The surgical procedure involves the following: location for the T-piece
is identified by positioning the collection arm tip below the most caudal
junction of a hepatic duct with the common duct. A 3 mm length of common
duct is next isolated at the collection arm's insertion point into the
common duct. The duct is incised and the collection arm of the T-piece
is advanced into the duct and secured in place with a preplaced circumferential
ligature. The duct isolation and catheterization is repeated with the distal
T-piece arm. Oversewing the bile duct area with omentum and performing
a duodenopexy have been used to try to limit post-operative adhesions.
Two methods were used to collect bile: a collection bag protected by an
outer case and jacket, and a modified jacket and tether system which allows
bile to be collected outside the cage. Animals were checked daily for clinical
signs of biliary obstruction and serum chemistries were collected weekly
for the first 12 months after surgery and every other week thereafter.
The collection catheter is accessed every other week to confirm bile flow
and the distal flush catheter was accessed and flushed once monthly.
A validation study involved intravenous infusion of 14C-glycocholic
acid which is excreted 100% in the bile. Bile was collected for 12 hr and
urine and feces were collected for 24 hr. Contrast radiography was also
used to evaluate placement of the catheter system and patency of the catheters.
Diatrizoate meglumine and diatrizoate sodium solution were infused into
the collection and/or distal flush catheters and serial radiographs taken
to track movement of the contrast solution.
A transient increase in ALT and AP was seen after surgery. Functional
bile collection was available from all 21 dogs for an average of a 212
days (0-1010 days). Complications resulting in removal from study included
collection catheters torn or detached from the ports, adhesions blocking
bile flow through the common duct, persistently elevated liver enzymes,
pancreatitis, solids plugging a T-piece arm, surgery failures, and aspirated
dose solution. Bacteria were cultured from bile samples from 11 dogs. Three
of these persisted with follow-up cultures and were suggestive of an intestinal
origin (Lactobacillus fermentum, E. coli). Transcutaneous access of the
ports represents another potential route for bacterial infection.
Questions:
1) Increased liver enzyme levels were caused by
a) bacterobilia
b) blockage of bile flow into the duodenum
c) both of the above
2) The type of needle used to collect fluid from vascular access ports
is called a ____________needle.
3) List 3 clinical signs of biliary obstruction.
Answers:
1) c
2) Huber
3) Jaundice, discolored urine and feces, decreased food consumption
A Method for Immobilizing the Forelimbs of Rabbits. Contemporary
Topics 37 (5): 094.
This article describes the problem of immobilizing the forelimbs of
rabbits after tendon repair surgery. It offers a technique which has been
successful in immobilizing the limb with 97% of the rabbits leaving the
cast on. The technique involves casting the leg with the radiohumeral and
radiocarpal joints placed in a flexed position. With the animal sedated
3 stages are performed: cotton dressing, plaster casting, and casting tape.
2-3 layes of the 1.5" wide cotton dressing, 2-4 layers of the 2"wide quick-setting
plaster, and at least 2 layers of the 2" wide casting tape was applied.
The joints were flexed at 70-100 degrees flexion.
QUESTIONS:
1. The NZW rabbits' forelimb was immobilized by placing the:
a. radiohumeral joint in extension and the radiocarpal joint in flexion
b. radiohumeral joint in flexion and the radiocarpal joint in extension
c. radiohumeral joint in extension and the radiocarpal joint in extension
d. radiohumeral joint in flexion and the radiocarpal joint in flexion
e. scapulohumeral joint in flexion and the radiohumeral joint in flexion
2. The forelimb should be cast with joints placed at an angle of how
many degrees?
a. 45-90
b. 60-120
c. 70-100
d. 90-135
e. 120-150
3. The genus and species of the rabbit is:
4. Clinically, what instument is used to measure joint angles of flexion
and extension?
ANSWERS:
1. The NZW rabbits' forelimb was immobilized by placing the:
d. radiohumeral joint in flexion and the radiocarpal joint in flexion
2. The forelimb should be cast with joints placed at an angle of how
many degrees?
c. 70-100
3. The genus and species of the rabbit is: Oryctolagus cuniculus
4. Clinically, what instument is used to measure joint angles of flexion
and extension? Goniometer
Short-term Care of White-Tailed Deer Fawns (Odocoileus virginianus)
in a Conventional Laboratory Animal Facility. Contemporary Topics 37 (5):
096.
Four orphaned fawns were obtained and kept in an indoor facility for
3-4 months to study the role of white-tailed deer as a reservoir for a
strain of lungworm (Dictyocaulus viviparus) that effects cattle. An animal
care program was created and implemented for the fawns by extrapolating
information available for housing deer in an outdoor environment.
The staff veterinarian examined fawns, shortly after their arrival
to the facility. Dehydration and hypothermia are common conditions of orphaned
neonates and therefore ensuring adequate hydration and body temperature
were of immediate concern. Parasitological examination (fecal floatation
and direct examination of fecal smears) indicated two species of coccidia
and an unidentified Stongyloides, however fawns were not treated for parasites
because of concerns of residual drug affecting experimental parasitic infections.
To minimize autoinfection holding areas were sanitized twice each week
to remove ova and oocysts. Pyrethrin soaked gauze sponges and manual extraction
removed ticks attached to the fawns' ears. Tattoos were placed in left
ear with a clamp-like tattoo instrument. Fawns were housed in pairs and
the pens were constructed of portable chain-link fencing. Ambient temperature
was maintained between 68-72 F, relative humidity between 65-70%, and the
light dark cycle was 12:12. To minimize injury from poor footing, pen floors
were lined with rubber-slatted mats and covered with aspen shavings. Portable
kennels were lined with shavings and placed in a corner of the pen to provide
a place of refuge. The fawns were fed a milk replacer formulated for goats
and small amounts of vegetation. Lactobacillus sp. was been added to milk
for the first few feedings to promote colonization of intestinal flora.
Milk replacers formulated for cattle should be avoided, because fawns are
unable to adequately digest them and diarrhea often results. When fawns
were 7-10 days old they were supplemented with clover hay and alfalfa pellets.
Manual restraint was possible when the fawns were small, by placing a towel
over the fawn's head, which produced a calming effect. Chemical restraint
with a combination of ketamine (6.2 mg/kg) and xylazine (1.2 mg/kg) was
also effective.
One fawn was infected with Theileria cervi, a protozoan that is transmitted
by the tick, Amblyomma americanum. The disease is usually subclinical,
however stress can exacerbate the disease. Low PCV, icteric serum, increased
bilirubin concentrations and numerous intra-erythrocytic protozoan parasites
can be identified.
Questions:
1. What is the genus and species of the white-tailed deer?
Answer:
1. Odocoileus virginianus
Bulbourethral Gland Cysts in Three Mice. Contemporary Topics 37 (5):
101.
This report describes bulbourethral gland cysts in 3 male mice--a DBA/2J,
a nude mouse and a chimera.
Bulbourethral glands are also known as Cowper's glands. They secrete
mucin for semen coagulation and copulatory plug formation. Disorders are
rarely reported in mice. In this report, at least one mouse did not produce
any progeny. The mice presented with slowly growing subcutaneous nodules
in the dorsal perineal region. The masses were fluctuant and moveable with
the base attached to the urethra. On cut surface, the nodules were polycystic
and filled with either blood or clear viscous fluid.
The etiology of bulbourethral gland cysts is unknown and may be due
to blockage of the duct. Cysts of bulbourethral glands or their ducts can
occur at all ages and are often asymptomatic and likely underdiagnosed
in humans and animals. In some cases they may become infected and rupture.
The cysts described in this report were asymptomatic except for the perineal
swelling.
Bulbourethral gland cysts should be included in the ddx for perineal
nodules or swellings in mice and other animal species.
Questions:
1. What is the scientific term for Cowper's glands?
2. Cysts of Cowper's glands only occur in aged male animals. T or F
3. What is the function of the Cowper's glands?
Answers
1. Bulbourethral glands
2. False--any age
3. In rodents--secretion of mucin for semen coagulation and copulatory
plug formation
Periorbital Mass in a Squirrel Monkey (Saimiri sciureus).
Contemporary Topics 37 (5): 103.
This short note consists of a case report describing the findings associated
with a mass on the left cheek below the lateral canthus of the eye of an
adult male squirrel monkey. The monkey was fed standard primate diet and
was housed single within the sight and sound of other monkeys. No physical
contact was allowed between monkeys. The monkey was used in behavioral
studies and had been conditioned to use of pole and collar capture and
a restraint apparatus.
No clinical signs were associated with the mass. CBC and chemistry
profile were unremarkable. Radiographs demonstrated a round pedunculated
nodule, 0.5 cm in diameter, and attached to the frontal process of the
zygomatic bone. An excisional biopsy was performed and histopathology described
lamellar bone with normal adipocytes and hematopoietic elements in a cystic
space. The site of attachment consisted of dense fibrous tissue.
Differential diagnoses for the mass included osteoma, fibrous dysplasia,
ossifying fibroma, heterotopic ossification, and exostosis. The final diagnosis
based on histopathology findings was osteoma. Prognosis after removal of
mass was good.
Questions:
1. Trabeculae of woven bone and lack of osteoblastic rimming of trabeculae
distinguish fibrous dysplasia from other fibro-osseous lesions. T or F
2. The finding of a benign tumor composed of well-differentiated bone,
most commonly found on the mandible, and craniofacial bones of domestic
animals, and consisting of wide trabeculae of lamellar bone is consistent
with the diagnosis of:
a) exostoses
b) ossifying fibroma
c) osteoma
d) fibrous dysplasia
Answers:
1. T
2. c)
Fasting and Feeding Effects on Gastric Emptying Time in Juvenile
Swine. Contemporary Topics 37 (5): 106.
Swine are commonly used for bioavailability studies of toxic compounds
due to their many biological similarities to humans. The effect of differing
intervals between feeding and dosing on gastric retention was the purpose
of this pilot study. To determine the effect of this interval, the authors
used a technique involving administering barium-containing doughballs at
pre-and post-feeding time periods and tracking the movement of the barium
with serial radiographs.
Three different protocols were followed using 2 male juvenile swine.
For Study A, there was an overnight fast of 16 hrs followed by a barium
doughball. In Study B, animals received their regular amount of feed 4
hrs prior to barium dosing. For Study C, barium dosing followed an overnight
fast. This was followed by feeding their regular ration of feed 2 hrs post-dosage.
Results: Studies A and C showed 95% clearance of the barium by 2 hrs
post-treatment. In Study B, gastric emptying was delayed, with the majority
of the barium still in the stomach at 2 hrs post-dosage. In Study C, the
added feeding of the normal ration after treatment excelerated the clearance
of the rest of the barium from the stomach (when compared to study A).
In summary, prior feeding and fasting and their relationship in time
to administration of a dosed material affect gastric emptying in juvenile
pigs, as measured by barium residence and transit time through the stomach.
Questions:
1. Fasting a juvenile pig prior to dosing with a toxic substance will
most likely:
a. decrease the gastric retention time of the toxic substance
b. increase the gastric retention time of the toxic substance
c. have no affect on transit time
2. Feeding a juvenile pig after dosing with a toxic substance will
most likely:
a. decrease the gastric retention time of the toxic substance
b. increase the gastric retention time of the toxic substance
c. have no affect on transit time
Answers:
1. a.
2. a.