Zoo Biology 17:25–32 (1998)

TECHNICAL REPORT

Crate Conditioning of Bongo

(Tragelaphus eurycerus) for Veterinary
and Husbandry Procedures at the
Denver Zoological Gardens
Megan Phillips,1 Temple Grandin,2* Wendy Graffam,2 Nancy A. Irlbeck,2
and Richard C. Cambre1
1
Denver Zoological Gardens, Denver, Colorado
2
Department of Animal Sciences, Colorado State University, Fort Collins, Colorado

Crate conditioning of bongo (Tragelaphus eurycerus) at the Denver Zoological

Gardens made it possible to perform veterinary and husbandry procedures without
chemical or manual restraint. Procedures included blood sampling, injections, wound
treatment, milk collection, and caudal tail fold tuberculin testing. Habituation and
positive reinforcement techniques were used to accomplish conditioning. The meth-
ods used were similar to those described by Grandin et al. [1995] for conditioning
of nyala (Tragelaphus angasi). A remote-controlled feed dispenser was mounted
on top of the crate. It allowed the handler to provide food to subordinate animals
selectively, so that dominant animals would not eat all of the rewards. Bongo were
more vigilant and more attentive to activities outside the stall than nyala trained in
previous studies. It took longer to habituate and condition some individual bongo
to particular steps of the process. Bongo were responsive to vocal cues from the
handler. The mean plasma cortisol level in three crate-conditioned, unsedated bongo
was 6.4 ± 3.8 ng/ml. The mean glucose level was 61.25 ± 19.45 mg/dl in crate-
conditioned animals and 166.5 ± 54.59 mg/dl in animals immobilized with either a
dart or a pole syringe. Creatine kinase (CK) levels were 70.75 ± 17.9 IU in crate-
conditioned animals and 288.75 ± 194 IU in immobilized animals. Values for these
blood parameters, commonly associated with stress, appeared to be lower in crate-
conditioned animals. Zoo Biol 17:25–32, 1998. © 1998 Wiley-Liss, Inc.

Key words: handling; habituation; conditioning; bongo; antelope; cortisol

*Correspondence to: Temple Grandin, Department of Animal Sciences, Colorado State University, Fort
Collins, CO 80523.
Received for publication 8 June 1995; revision accepted 24 October 1997.

© 1998 Wiley-Liss, Inc.

26 Phillips et al.

INTRODUCTION
Conditioning techniques using positive reinforcement have been used to facili-
tate veterinary, husbandry, and research procedures in a variety of captive exotic ani-
mals. Such animal taxa include, but are not limited to cetaceans [Laule, 1993],
artiodactyls [Grandin et al., 1995; Kinzley, 1993], perissodactyls [Kinzley, 1993; Michel
and Illig, 1995], primates [Bloomsmith, 1992], felines and pinnipeds [Reichard et al.,
1992]. Conditioning can often replace chemical immobilization or manual restraint
methods, which carry significant safety risks for both handlers and animals. Condition-
ing may also reduce physical and mental stress associated with other restraint methods
[Hastings et al., 1992; Mitchell et al., 1988]. Reduction of stress is important because
handling stress has been shown to impair immune function [Kelley et al., 1981] and
carries safety risks for both animal and handler.
The main objective of this project was to condition bongo (Tragelaphus
eurycerus) to confinement in a handling crate to provide a nonstressful means of
performing veterinary and husbandry procedures without immobilization. Such pro-
cedures included blood sampling, caudal tail fold tuberculin skin testing, injection,
palpation, and milk collection.

MATERIALS AND METHODS

Animals
The group of conditioned animals consisted of two adult females, one adult male,
two juvenile females, and one juvenile male. Two animals were born and conditioned
after conditioning of the adults was in place. An additional animal was acquired from
another institution after conditioning of the group was in place. Before crate condition-
ing, animals varied in their tolerance to being approached and touched by handlers.
The adult male could be petted, but the other animals had large flight zones and could
not be touched. Prior to conditioning, some animals had experience with chemical
immobilization and with being injected via blow dart gun or pole syringe.
Food Rewards
The best food reward for bongo was spinach. It was highly palatable, nutrition-
ally sound, and not part of the regular diet. To accommodate taste preferences of
individual animals or longer procedures, additional items such as apple, yam, and
banana were occasionally offered.
Handling Crate
The handling crate was constructed of stout plywood with vertical slide doors
at each end. The doors of the crate were operated from outside the stall via cables
and pulleys (Fig. 1). Outside dimensions of the crate were 300 cm long, 200 cm
high, and 60 cm wide. A third vertical slide door was installed 190 cm from the rear
door of the crate to accommodate smaller animals. Openings in the crate provided
access to the rear leg for blood sampling and to the rear quarters for injections and
tuberculin skin testing. Two smaller holes were used to provide food rewards. All
openings were equipped with latching covers.
A remote-controlled treat dispenser was installed on top of the crate so that spinach
could be dropped into the crate without the handler entering the stall with the animals
 Bongo Crate Conditioning 27

Fig. 1. Handling crate.

(Fig. 2). The 20×20-cm square dispenser was made of plywood and stood 120 cm high.
It had four flip-down shelves that operated like those of a candy vending machine. Spin-
ach was dispensed by pulling cables that were attached to pins supporting each shelf.
Foam tape on the leading edge of each shelf reduced noise when a shelf was released.
Crate Conditioning
Conditioning was done very carefully and slowly to avoid frightening the ani-
mals and triggering a flight reaction. In general, methods were similar to those used
to condition nyala [Grandin et al., 1995]. First, the animals were enticed into the
crate with food rewards and desensitized to the sounds and movements of the crate
doors and treat dispenser. Next, the animals were captured in the crate and gradually
habituated to longer captures. The animals were then gradually desensitized to being
touched while in the crate. Very light, brief touches were used initially, followed by
progressively firmer touches until the animal did not respond to being touched. Next,
the desired procedure was simulated. Each aspect of the procedure was introduced
slowly until the animal did not respond to the stimulus.
Blood Collection
Habituation for blood collection and actual blood collection were accomplished
using the same methods and equipment described by Grandin et al. [1995] for col-
28 Phillips et al.

Fig. 2. Treat dispenser.

lecting blood from crate-conditioned nyala. Blood samples were taken for plasma
cortisol analysis (RIA, Star Corp., Stillwater, MN) from unsedated animals 10–20
min after capture in the crate. Blood samples were taken for plasma cortisol analysis
from two adult females when they were immobilized for an embryo transfer proce-
dure. They were hand injected in the crate with 1.2 mg of carfentanil citrate (Wild-
life Laboratories, Ft. Collins, CO) and 20 mg of xylazine (Xyla-ject ®, Phoenix
Pharmaceutical, St. Joseph, MO). They were released from the crate immediately
after injection and prior to collapse. The collection of blood occurred 25–30 min
after injection. Time was allowed to lapse before blood was drawn for cortisol analy-
sis because a minimum of 10 min is required for cortisol levels to peak following
stress [Hastings, 1992].
Tuberculin Testing and Injections
The procedure for habituation to caudal tail fold tuberculin tests began with
lifting the animal’s tail and touching the tail fold area. Next, the animal was habitu-
ated to increasingly harder pinches on the tail fold. Once the animal was habituated
to pinches, it was conditioned to keep the tail still using food rewards.
Habituation for injections was similar to that for tuberculin testing. Each ani-
mal was desensitized to increasingly harder pinches on both hips. The animals were
then conditioned to stand still. The food reward was provided when the animal stayed
still and withheld if it moved away from the pinch.
Milk Collection
Experienced dams were desensitized to having their bellies and udders touched
up to 8 weeks prior to parturition. Milk collection began as early as the fifth day of
lactation and continued throughout lactation. Collection of a full milk sample in-
volved administration of a 0.5 ml (20 IU/ml) intramuscular injection of oxytocin
 Bongo Crate Conditioning 29

(Vedco, St. Joseph, MO), and up to 40 min in the handling crate. During such lengthy
procedures, additional food items such as apple and banana were offered to maintain
the dam’s interest throughout the session.

RESULTS
Six animals were conditioned to enter the handling crate and stand still for
blood sampling and injections. The juvenile male and one female were conditioned
to stand still for caudal tail fold tuberculin skin tests. Female animals were habitu-
ated to having the abdomen and udder palpated to assess pregnancy and parturition.
Experienced reproductive females were successfully conditioned to allow manual
milk collection so that milk composition could be analyzed. Milk collection did not
have any negative effect on the successful rearing of calves by their dams. A repro-
ductive female sustained a laceration near the vulva and was successfully treated
without prior desensitization.
Sixteen plasma cortisol tests were performed on three adult, unsedated, crate-
conditioned animals. Cortisol values ranged from a mean of 4.4 ± 3.4 ng/ml in the
male to a mean of 8.5 ± 4.7 ng/ml in the subordinate female. Cortisol values from
the two females that were injected in the crate and immobilized for embryo transfer
were 10 ng/ml and 18 ng/ml. Data were not available in medical records to compare
cortisol levels in animals immobilized with a dart or pole syringe with levels in
crate-conditioned animals. We could not ethically justify using these methods on
crate-conditioned animals to obtain these data.
Glucose and CK (creatine kinase; also CPK, creatinine phosphokinase) lev-
els obtained from four unsedated animals were compared with levels found in hos-
pital records from animals immobilized prior to crate conditioning. The mean
glucose value for crate-conditioned animals was 61.25 ± 19.45 mg/dl versus 166.5
± 54.59 mg/dl for animals immobilized with a dart or pole injection. The mean CK
value was 70.75 ± 17.9 IU in crate-conditioned and 288.75 ± 194 IU in immobi-
lized animals.
Conditioning has been maintained by simulating blood sampling, injections,
and/or tuberculin tests with pinches. Maintenance sessions occur about once a week
for each animal. It is likely that the frequency of these sessions can be reduced in the
future, as nyala conditioning has been maintained with monthly sessions.

DISCUSSION
During early conditioning phases, the use of a treat dispenser was necessary.
Dominant animals prevented subordinates from obtaining food items so that subor-
dinate animals were not able to form a positive association with the crate. It was
unsafe for the handler to enter the room with the bongos, so the dispenser was used
to provide food to subordinate animals when they showed interest and dominant
animals were not present. This feature was not used with the nyala as their social
structure did not interfere with conditioning and because it was safe for the handler
to be in the stall with the animals [Grandin et al., 1995].
On the average, the time required to accomplish each step of conditioning was
the same as the times reported for nyala. Those times were as follows: 21 days to
condition to enter crate; 7–14 days to habituate to presence of crate, to movement of
30 Phillips et al.

crate doors, to being captured up to 10 min, to being touched; and 14 days to condi-
tion to stand still for blood sampling [Grandin et al., 1995]. However, conditioning
of certain individual bongo took much longer. One bongo took up to 7 days longer to
habituate to movement of the crate doors. Another took up to 24 more days to ha-
bituate to captures of 10 min. It was important to proceed slowly to accommodate
these animals and to avoid triggering a flight reaction or negative association with
conditioning.
Additional behavioral differences between nyala and bongo became apparent
during conditioning. Nyala responded less when initial touches to the leg were firm
[Grandin et al., 1995]. The same firm touches elicited a vigorous response from
bongo, so that it was necessary to use gradual desensitization to touches.
Bongo were more vigilant than nyala while captured in the crate. Nyala only
occasionally responded to noises and other disturbances outside the stall. Bongo were
very attentive to all noises and motions occurring outside the crate or stall. They
would turn their heads and ears toward the disturbance, look out through the treat
holes, and would often refuse rewards until the distraction ceased. Keeping the win-
dows and doors to the stall closed helped to ensure that the animals would not be-
come distracted.
Bongo were responsive to vocal cues from the handler. When an animal was
still and a reward was provided, the words “good girl” or “good boy” were spoken
in a soothing voice. If the animal performed an undesirable behavior, such as kick-
ing or moving, a firm, loud no was spoken and the reward was withheld. It is
possible that the high degree of attentiveness of the species is one reason that
vocal cues were successful.
In the wild, the bongo is a forest-dwelling species. It is likely that adaptation
of this species to a forest habitat influenced behavior patterns observed during
crate conditioning. The presence of the solid barrier of the crate had a calming
effect on the animals, but they remained attentive to most external stimuli. These
behaviors are consistent with an adaptive behavior pattern to avoid predation in a
forested environment.
Cortisol levels were determined to provide a physiological indicator of stress
levels of animals handled in the crate. The mean cortisol values obtained in the
unsedated, crate-conditioned animals were within the resting baseline range of do-
mestic cattle [Alam and Dobson, 1986]. Morton et al. [1995] obtained plasma corti-
sol values for both physical restraint and immobilization in six antelope species when
they were translocated in Zimbabwe. Mean cortisol levels ranged from a high of
120.4 ng/ml in physically restrained waterbuck (Kobus eilipsysrymus) to a low mean
of 20.04 ng/ml in sedated eland (Taurotragus oryx). Unfortunately, no comparable
data were collected from wild bongo, but it is possible that cortisol levels from eland
might serve as the best comparison with bongo. The cortisol values from condi-
tioned, unsedated bongo were lower than the range of values reported for many other
restrained and immobilized antelope species [Morton et al., 1995].
Immobilization via hand injection in the crate yielded an elevated cortisol level
in one bongo, but not the other. This may have been due to the order of immobiliza-
tion. The second animal, with the higher cortisol value, would have observed and
heard the disturbances when the first animal was immobilized. Cortisol results from
these two animals were lower than those reported by Morton et al. [1995]. Based on
cortisol values and behavioral observations, injection while in the crate appears to be
 Bongo Crate Conditioning 31

a less stressful procedure than immobilization via dart gun or pole injection. Habitu-
ation and conditioning may have reduced stress in crate-injected immobilized ani-
mals. One study concluded that deer fawns that were hand-reared, and were therefore
habituated to handling, had lower cortisol levels during physical restraint compared
to free-range fawns [Hastings et al., 1992].
Increased glucose levels in immobilized animals may be due, in part, to the
action of cortisol within a stressed animal. Increased CK values in immobilized
animals indicate a greater amount of muscle stress. It is important to note that
glucose and CK values in conditioned, unsedated animals were also lower than
“normal” values reported by the International Species Inventory System (ISIS).
The data used to compile these “normal values” are most likely obtained from
immobilized animals.
The cortisol values from crate-conditioned bongo support behavioral observa-
tions that the animals were not stressed by confinement in the crate. Crate condition-
ing of bongo provided a means to safely perform procedures that would otherwise
have required immobilization. Using behavioral observations and physiological indi-
cators, the method appeared to reduce stress to the animals.

CONCLUSIONS
1. Bongo can be conditioned to confinement in a handling crate and desensi-
tized to a number of husbandry and veterinary procedures.
2. Bongo were very vigilant during conditioning and certain individuals took
longer to condition.
3. Crate conditioning may reduce the stressfulness of veterinary and husbandry
procedures.

ACKNOWLEDGMENTS
This project was funded in part by Grandin Livestock Handling Systems, Inc.,
and by a grant from The Frost Foundation, Santa Fe, NM. The authors gratefully
acknowledge the cooperation of the Denver Zoological Gardens and the keepers and
maintenance personnel who helped with the animals and with design, construction
and maintenance of the handling crate. Particular thanks go to Dallas Meads, Martha
Turnbull, Joan Poston, Dr. David E. Kenny, and Jim Gostele.

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