1. Vol. 14/ No. 6 December 2009
Husbandry of theVeiled
Chameleon (Chamaeleo
calyptratus)
By Chrystal L Redding, MS, RLATG Northern Arizona University,
Flagstaff, AZ
Veiled chameleons (Chamaeleo calyptratus),
indigenous to Yemen and Saudi Arabia, are
hardy; they can withstand temperatures from
20 °Cto43.3 °Candelevationsto9,500 feet.The
veiled chameleon iswidelyavailable in the pet
trade due to ease of breeding, egg incubation,
and hatching. These same qualities make the
veiled chameleon a great choice in studies of
environmental impact on sex determination during their long incubation.1,2
Emis-
sionof bodyvibration has led tostudiesonpossiblecommunication throughsubstrate bythesechameleons.3
Chameleons, likemanyamphibiansand reptiles,growquicklyduring thefirstfewyears. Afterhatching, males
are easily identified by a tarsal spur on the posterior of each rear foot. These tarsal spurs, and head crest
dimorphism between males and females, make them easy models for mate and sexual selection studies.4
The total body length of adult maleveiled chameleons ranges from 30.5–48.3 cm, whileadult females range
from 20.3–30.5 cm. Both males and females have a wide variety of color patterns; however, males tend to
have brighter colors.
Within a year of hatching, chameleons can increase their body mass by two orders of magnitude. Rapid
growth requires a diet rich in calcium and vitamins A and D to prevent weak, brittle bones and to support
muscle movement, including the ballistic tongue projection forwhich chameleonsare best known. While
fresh-caught wild insects from pesticide-free areas provide variety and high nutrition, most laboratory
animal professionals lack the time for collection. Therefore, it is best to feed adult chameleons a diet of
crickets sprinkled with calcium/vitamin powder 3–4 times per wk. Adults will eat most insects, small
mammals, lizards, or amphibians and plants, including leaves, greens, fruits, vegetables and flower
blossoms.
A well-ventilated terrarium with a water bowl provides primary housing for these arboreal chame-
leons. Newly hatched young may be housed together for the first year, but adults, especially males,
are very territorial and should not be housed together. Veiled chameleons are intense baskers
that prefer temperatures from 23.8 °C to 32.2 °C; a 50- to 100-watt bulb set on a timer should be
included in each cage as an additional light source. Real and
plastic plants inside the terrarium provide color and cover, and
help to stabilize temperature and humidity. Do not use sand,
dirt, or wood chips as terrarium substrate. These materials
stick to the chameleon's tongue during feeding and can cause
intestinal blockage. Cloth, newspaper, or brown paper towels
provide adequate coverage and make the terrarium easy to
clean. Veiled chameleons prefer 50% to 60% percent humid-
ity, which can be maintained by daily misting or with a drip
system that allows water to drip from the top of the cage into
the water bowl.
Rabbit Intubation. . . . . . . 2
Gavage Alternative. . . . . . 2
NHP Conditioning. . . . . . . 4
Dear Labby. . . . . . . . . . . . . 5
Crossword Puzzle . . . . . . . 6
PRO-Files in LAS. . . . . . . . 7
The Newsletter for Laboratory Animal Science Technicians
American Association for
Laboratory Animal Science
9190 Crestwyn Hills Drive
Memphis, TN 38125-8538
www.aalas.org Chameleon continued on page 8

2. 2
Food forThought: AViable Alternative to the Gavage Method
By H. B. Moak, M. Robinson, R. J. Kastenmayer, andW. R. Elkins, DHHS/NIH/NIAID/DIR Comparative Medicine Branch, Bethesda, MD
While the esophageal gavage method is a commonly accepted procedure to orally administer medication, it has
several disadvantages: it requires a specialized needle, must be performed by a trained professional, risks perfo-
ration of the esophagus, and potentially induces stress. In seeking to identify an alternative delivery method in
accordance with the 3 Rs, we tried administering medication using a highly palatable food.
Toestablishareadilyconsumed substance, weconducted an initial studyevaluating thepalatabilityof fruit juice,
pureed meat, milk products, peanut butter, and cornstarch. Of the preliminary offerings, cream, margarine,
and sweetened condensed milk (SCM) proved highly palatable. To identify the most palatable substance, we
offered cream, margarine, and SCM in randomized trials to 4- to 8-wk old females from three commonly used
laboratory strains of mice (BALB/C, C57BL/6, and Swiss Webster). Nine mice per strain were weighed and
placed in individual cages with no bedding orwater. Following an acclimation period of 15 min, a weigh boat
containing 0.5 ml of cream, margarine, or SCM was placed in the cage, and the mice were observed for 30
min. Once the feeding period was terminated, any remaining
food was weighed and consumption data recorded.
Using ANOVA analysis, cream was shown to be prefer-
entially consumed (P < 0.05) over SCM and margarine.
When we presented the mice with all three substances
simultaneously, cream proved to betheunconditional
favorite. However,with increased familiaritywiththe
weigh boats and food options, the mice displayed
progressive willingness to consume all three
substances. Weight remained a significant
confounding factor in our consumption
data. Onaverage, a mouseconsumed 0.12
g/g body weight (P = 0.0009).
A Step-by-Step Approach for
Blind Intubation of the Rabbit
By William Dyckman, AAS, LAT, SRS, Hartford Hospital, Hartford, CT
The anatomy of the oropharynx in rabbits, which includes a right angle bend
at the caudal aspect, a narrow mouth opening, a large tongue, large incisors,
and limitedrangeof motioninthejaw, makesintubationdifficult.Additionally,
reflex laryngospasm is not uncommon when attempting to intubate rabbits.
Our blind intubation technique has worked well for us, and it is easily accom-
plished with training. We use it routinely for our rabbit surgery and have had
good results. With practice, it is not a difficult technique to learn, and it does
not require an assistant.
We used New Zealand White rabbits weighing 2–3 kg. Rabbits were housed
individuallyincageswitha12-h light/darkcycle. Rabbitswereoffered restricted
access to pelleted feed and ad libitum access to water. The study was approved
by our institutional IACUC Committee.
Step 1
Therabbit isweighed, and buprenorphine 0.01–0.05 mg/kg IM isgiven 1 h prior
to anesthetic administration. We then administered xylazine 5–10 mg/kg IM.
Ten minutesafterxylazineadministration, wedosed therabbitswith ketamine
33–35 mg/kg IM and waited for it to takeeffect. This dose regime induces total
relaxation in the rabbits, which is crucial for this type of intubation.
Step 2
The rabbit is brought into the prep room, weighed again, and shaved for the
procedure.
Step 3
Fora 1- to 3-kg rabbit, weusuallyusea 2.0–3.0mm OD endotracheal tube. For
a 3- to 7-kg rabbit, we use a 3.0–6.0, OD tube. The end of the endotracheal
tube cuff is lubricated with 5% lidocaine ointment. The valve of the cuff is
checked by inflating and deflating the cuff using a 3-cc syringe attached to
the valve.
You'll need:
• Cuffed endotracheal
tube
• 5% lidocaine ointment
• 3 cc syringe for
inflating cuff
• Stethoscope
• Adhesive tape to secure
tube

3. 3
Ultimately, thisexperimentsuggests thata feasible
alternativetooralgavageexists. Inconjunctionwith
the3 Rs,ourproposed method terminatestheneed
for the gavage needle and eliminates the resulting
stress. However, it is unclear if oral medications
can be successfully administered in a highly pal-
atable substance; it is also unknown whether
combining a medication with food impacts the
drug’s metabolism or stability. Future studies
are needed toevaluatetasteresponsivenessof
an ordinarily palatable substrate combined
with medication and the compatibility of
the drug with the substrate of choice.
Also, theaddition of water may increase
the consumption of offered food.
In contrast with the direct deposit mechanism of the gavage needle, this process does not
guarantee absolute consumption. This study was completed in the early morning; we sus-
pect that if medication is presented before the ordinary feed or if the mice are placed on
a timed eating schedule, they would more readily consume the offered substances. It is
essential that futurestudiesscrutinizethepracticalityof delivering medication insuch
a method; however, the implications of a dosing alternative remain vast.
Acknowledgements
This research was supported by the Intramural Research Program of the NIH,
NIAID.
Step 4
The anesthetized rabbit is placed in sternal recumbency on a
warming blanket on the operating room table. The eyes are lu-
bricated withophthalmicointment, and apulseoximeterprobe
is attached to the shaven front paw. The rabbit's head is held
by grasping around the base of skull with the non-dominant
hand. The thumb and index finger are used to grasp the upper
jaw and extend it straight up, hyperextending the oropharynx
to straighten it for intubation.
Step 5
Withtheneckextendedverticallyandthehead hyperex-
tended, theendotracheal tubeisslowlyinsertedoverthe
tongue and into the pharynx. The technician perform-
ing the procedure listens forair movement at the end of
the tube; the tube is directed toward the breath sounds
coming through the glottis. As the tube passes into the
larynx and trachea, the sounds usually become louder,
andcondensationmaybeseeninsidethetube;sometimes
acoughisheard throughthetube. If anyresistanceisfelt,
thetubeiswithdrawnslightlyand reinsertedagainusing
gentle manipulation; pushing against the resistance can
injure the rabbit. Sometimes it helps to gently turn the
tube clockwise slightly, then counterclockwise during
insertion. If the first attempt at insertion is unsuccessful,
a respiration mask is used to assist the rabbit's breathing
before reattempting insertion.
Step 6
With the endotracheal tube inserted, the technician
checks for bilateral breath sounds, pulling back the tube
if necessary. A 3-cc syringe is used to inflate the cuff. The
tube is secured with 0.25-in. wide adhesive tape in front
of the upper incisors and around the upper jaw; it can
also be tied in place with a gauze sponge.
Hold the rabbit's head
as shown to hyperextend
and straighten the
oropharynx.

4. 4
OperantTraining Differences between Male and Female Group-Housed Nonhuman Primates
By Jayson Egeler, BS, LAT, MPI Research, Mattawan, Michigan
Operanttrainingof animals isawidelyaccepted practicetotrainorconditionanimalstocooperate
with technical procedures or perform certain tasks in a laboratory setting. This type of training
has been proven to reduce stress on the animal and reduces potential injuries to the animal or
its handlers. However, it has been suggested that female cynomolgus monkeys are more difficult
to train than males.
In conjunction with our goal of offering group housing that meets European standards for non-
human primates at our facility, we implemented an operant training program to train animals
to be removed from the new, larger-style caging. Our data support the theory that females are
more difficult to train than males. This article outlines the necessary steps in developing and
implementing a successful operant training program in untrained monkeys, and describes how
we assessed program progress and adapted the program to meet various challenges.
Training Program
Ourstudygroupconsistedof 16 male(2.24–2.80 kg)and 16 female(1.96–2.91 kg) Chinesecynomolgus macaques (Macaca fascicularis), 2.5–3.5
years of age, that were housed in groups of 4. Our goal was to train the animals to come to the front of the cage, station, and stay at a target
thatwas placed onto the frontof thecage. Training sessionswereconducted onceaday, 3–5 days perwk, forapproximately 10 min pergroup.
Clicker training was used to connect a food reward with the desired behavior. We did this approximately twice a day during our normal
cageside observations. Once animals showed no fear of the clicker (approximately 1 wk), we introduced the PVC pipe. Once or twice a day,
we stuck a small length of 0.5-in. PVC pipe into the cage; when the animal touched the pipe, we pressed the clicker button and offered a
reward. As animals became conditioned to touch the PVC pipe to receive a treat, we taught them to station. We attached PVC elbows to the
front of the cage (one per animal). When animals came forward and touched the elbow, we pressed the clicker button and offered a reward.
If animals wouldn’t touch the elbow, we placed the food reward on the elbow and clicked when they came forward to retrieve it. To keep
the training new and fresh, we varied the time of day training was conducted, and moved the targets to various spots of the cage to train
the monkeys to follow the elbow instead of learning to stay at one particular place.
Issues Encountered
Shortly into the training program, it became evident that the decision to house the monkeys in groups of four was having an effect on
the training of the females. The females seemed to be much more cautious and reserved than the males. Although both sexes developed
hierarchies within their groups, the difference was much more noticeable with the females. The gregarious nature of the males negated
any effect dominance had on training; the drive for food seemed to outweigh any pressure the subordinate animal would receive from the
dominant animal. The females' hierarchy, on the other hand, was so rigid that it prevented progress with the more subordinate animals.
Approximately 8 wk into the training program, we moved the animals into slightly larger banks and separated males and females into their
own rooms. The males adjusted quickly, and resumed their normal training schedules. The females, however, became more difficult to
work with and even regressed in terms of the progress they had made prior to the move.
Program Adjustments and Results
To improve the success rate of our training program, we housed the females in pairs. We also provided positive human interaction to both
males and females during handling by petting them, speaking to them in a soothing voice, then offering a treat before returning them to
their home cage. Many of the females responded favorably to these changes. For some of our more difficult female pairs, separating the
pair for the training session seemed to help. This separation allowed the submissive female to “come out of her shell” and participate in
the training without the disapproval of her dominant companion. For other females, improvement was achieved by trying different types
of enrichment until we found one that they would respond to.
Recommendations
There are several factors to consider when starting an operant conditioning program: the time needed to train animals, the necessity
of training in groups (smaller groups are easier to train), and the animal's sex (males are easier to train). As we learned, some types of
enrichment worked better for certain animals. You have to experiment with a variety of enrichment types to find out which kinds work
best for your animals. Using a wide range of food items prevents the animals from losing interest in the training. It’s also important
to train in slowly, in small segments that maximize the time you can train without filling up the animals and losing their attention.
Some animals responded better to the PVC elbow than to the pipe. If you have a difficult animal that won’t
touch the pipe, try skipping ahead to the elbow and see if that works.
It is important to be patientwhen training monkeys. Thework described above took placeovera 6-mo period.
Although many of our males were doing well by the 3-mo mark, training was a continual process. We had
days where all of our work seemed to be heading nowhere, and other days where everything seemed to come
together. Give your animals some time—they will surprise you!
Acknowledgements
This projectwould not have been possiblewithout mygroup housing teammates Harriet Hoffman and Sarah
Hoekwater, along with the thoughtful guidance of our manager, Crystal Diabo. Many thanks to Iris Bolton
and Kelsey Neeb, who started us off in the right direction and were great resources. Also thanks to the many
individuals who lent their expertise along the way at MPI Research.
Editor’s Note
Please remember to get approval
from the principal investigator,
your IACUC, and your facility
veterinary staff before imple-
menting any new program.

5. 5
Now that some of the governmental restrictions have been lifted on stem cell research, where are we on
that? Has it resulted in all the great things it was supposed to prior to the funding ban?
Looking for Cures
Dear Looking,
Stem cell research, despite the lack of federal funding, continued to make headway from 2000 till now,
although at a slower pace. Many dedicated researchers continued to forge ahead during the ban and
stood poised and ready to move forward when the time came for renewed opportunities.
Stem cells can be obtained from other sources, such as bone marrow, but the most valuable of all
sources is the unformed, undifferentiated embryonic stem cell. These cells are capable of giving rise
to all organs and tissue types. In the early stages of stem cell research, the only way to harvest these
blank-slate cells was to sacrifice the embryo itself, resulting in the political and social stigma attached
to the research. Even during the near halt in stem cell research, scientists found ways to create and
harvest embryonic stem cells without sacrificing the embryo. Although not a “cure” per se, these
breakthroughs will help pave the way for even greater discoveries in the future.
As far as cures go, stem cell treatments are currently a reality for some diseases of the blood, such
as leukemia and sickle cell anemia. There are some very impressive success stories of patients being
cured of the above diseases, as well as at least one case where HIV was banished during the course
of treating a leukemia patient. Currently, there are some very ambitious projects, led by the Army, to
use stem cells to develop replacement organs. The ability to regenerate tissue and organs may result
in restoring lost limbs one day. A recently published study has demonstrated the advances that have
been made along these lines by enlisting stem cells to regenerate teeth in mice.With all the positive news concerning stem cells, there is still much to learn before treatments will
become everyday options for fighting disease. There are three essential elements needed for stem cell
therapies to be successful: the stem cells themselves, a protein “scaffolding” for the stem cells to build
upon, and the complex signaling molecules that regulate their growth. Without effective control
signals, stem cells may grow out of control, causing extremely serious complications. This third and
crucial element is the current focus of most research today.There is still tons work to be done before stem cell therapies become mainstream, everyday cures.
Despite this fact, the stem cell remains one of the most promising weapons we have in our arsenal to
fight some of the most debilitating health issues plaguing humankind today.

6. 66
Crossword Puzzle

7. 7
PRO-filesinLAS
Name:
Catherine J. Bernstein, MS, RLATG, CMAR
Facility:
Rockefeller University, NYC
JobTitle:
Assistant Manager
Years in animal science:
20 years
How long have you been a national AALAS member?
15+ years
What branch (es) have you belonged to?
Metro New York and New England Branch
What is your level of AALAS certification?
RLATG, CMAR
What awards have you received?
I won the essay contest for an ILAM scholarship in 2002. I
alsowon “bestactress”atILAM formyincredibleportrayal
of Ann Murray.
How did you get in this field?
As an undergraduate student at NYU, I had a part-time
job at a used bookstore. I had my own apartment in
midtown Manhattan at the time, and one part-time
job wasn't enough. I answered an on-campus ad for
a job in the lab animal facility, and the rest is history!
What did you want to be when you were growing up?
At first I wanted to be a nun (I admired my teachers). But later I wanted to be a veterinarian.
Who were your mentors?
Shawn Rousseauwas the firstsupervisorI had whoencouraged metoget into management. Jack Hesslerand Fred Quimby
made me happy and grateful to be involved in lab animal science.
What are your current interests in animal science?
My current interests are team-building and positive management techniques. I enjoy taking a group of husbandry or
cagewash technicians and guiding them into becoming a unified team where the results are far greater than the sum of
the individual parts.
What is one thing no one would guess about you?
As a kid, I convinced my sister to join me in a bird rescueoperation in mygrandmother's backyard. We rescued baby birds
that fell out of the trees and put them in an old bird cage we salvaged from the trash.
What companion animals do you have?
I have two cats—one is a shelter rescue mutt and the other is a goofy Siamese.
What are your career goals?
My goal is to keep working at enjoying what I do because if you have fun at work, every day is like a vacation!
What advice do you have for others entering this field?
Try different areas of the field—it's more diverse than people think!

8. EDITORIAL
COMMITTEE
EDITOR
Patricia Denison, AS, LVT, LATG
Wayne State University,
Div. of Laboratory Animal Resources
CHAIR
Sylvia Banks, BS, RLATG
University of Oregon
VICE CHAIR
Melissa Dyson, DVM
University of Michigan
COMMITTEE MEMBERS
Verda Davis, RLAT
University of Kentucky
Kenneth Pyle, RLATG
State University of NewYork Delhi
Jennifer C. Smith, DVM
Taconic Farms
Julie Watson, DVM
Johns Hopkins University
Sonia Doss, BS, MEd, RLATG, BOT Liaison
John Farrar, BA, AALAS Staff Liaison
Nicole Brown, MA, Graphic Design
Article Recruitment
Sub-Committee
Grace Aranda, LAT
Dawn Conover, AS, LATG, LVT
Leta Eng, BS, RLATG
Amy Ingraham, BS, RLATG
Gene Rukavina, AAS, RVT, RLATG
GailThompson, RLATG
Crossword Solution
Chameleon continued from page 1
While each individual has different personali-
ties, common behaviorsprovide important keys
to health:
• Pale color with gaping mouth indicates that
the chameleon is overheated.
• Closed eyes and a refusal to eat, possibly ac-
companied by brightcoloration (often shades
of yellow) indicate that the chameleon is sick.
• Vivid coloration and sunken eyes are an in-
dication that the chameleon is thirsty. Pinch
the belly skin; if the skin remains pinched,
the chameleon is dehydrated. Mist until the
chameleon stops drinking.
• Swelling (edema) is generally a sign of Vitamin
A or D deficiency. Weakness, shaking, and in-
ability to project the tongue during feeding is
often a calcium deficiency.
References
1. Andrews, RM. 2007. Effects of temperature on
embryonic development of the veiled chameleon,
Chamaeleocalyptratus. Comparative Biochemistry
and Physiology, Part A, Molecular and Integrative
Physiology 148(3):698–706.
2. Andrews, RM, and S Donoghue. 2004. Effects of
temperature and moisture on embryonic diapause
of the veiled chameleon (Chamaeleo calyptratus).
Journal of Experimental Zoology 301A(8):629–635.
3. Barnett, KE, RB Cocroft, and LJ Fleishman. 1999. Possible communication by substrate vibration in a
chameleon. Copeia 1:225–228.
4. Kelso, EC, and PA Verrell. 2001. Do male veiled chameleons, Chamaeleo calyptratus, adjust their courtship
displays in response to female reproductive status? Ethology 108(6):495–512.