procedures training large macaws for artificial insemination...artificial insemination procedures...
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Journal of Applied Animal Welfare Science
ISSN: 1088-8705 (Print) 1532-7604 (Online) Journal homepage: http://www.tandfonline.com/loi/haaw20
Training Large Macaws for Artificial InseminationProcedures
Frédérique Leblanc , Géraldine Pothet , Michel Saint Jalme , Mathieu Dorval& Dalila Bovet
To cite this article: Frédérique Leblanc , Géraldine Pothet , Michel Saint Jalme , Mathieu Dorval& Dalila Bovet (2011) Training Large Macaws for Artificial Insemination Procedures, Journal ofApplied Animal Welfare Science, 14:3, 187-210, DOI: 10.1080/10888705.2011.575741
To link to this article: http://dx.doi.org/10.1080/10888705.2011.575741
Published online: 17 Jun 2011.
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JOURNAL OF APPLIED ANIMAL WELFARE SCIENCE, 14:187–210, 2011
Copyright © Taylor & Francis Group, LLC
ISSN: 1088-8705 print/1532-7604 online
DOI: 10.1080/10888705.2011.575741
Training Large Macaws for ArtificialInsemination Procedures
Frédérique Leblanc,1 Géraldine Pothet,1
Michel Saint Jalme,1 Mathieu Dorval,1 and Dalila Bovet2
1Muséum National d’Histoire Naturelle (MNHN), UMR 7204,MNHN-CNRS-Paris VI, Ménagerie du Jardin des Plantes, Paris, France
2Laboratoire d’Éthologie et Cognition Comparées, Université ParisOuest Nanterre La Défense, Nanterre Cedex, France
For some endangered parrot species, captive breeding may be the only insurance for
their survival. However, many individuals in captivity do not reproduce. Artificial
insemination (AI) may help overcome reproductive failures or geographic distance.
For semen collection in birds, massage is the most commonly used method.
However, this process, which usually requires capture and restraint, involves risk of
stress and injuries. The aim of this experiment was to train large macaws to accept
the physical manipulations of their body parts needed for the artificial insemination
process. Within 15 weeks, a male and a female Buffon’s macaw (Ara ambiguus)
learned to accept handling without apparent stress. A pair of green-winged macaws
(Ara chloropterus) progressed more slowly and displayed some signs of stress. This
stress highlights the need to monitor the birds’ possible signs of discomfort during
the training in order to adapt the working protocol. These results demonstrate that
it is possible to train the AI behaviors, thus avoiding the capture, restraint, and
anesthesia. An added benefit to this is the potential for the training to provide a
form of behavioral enrichment.
Parrots are one of the most endangered groups of birds in the world (Juniper
& Parr, 1998; Snyder, McGowan, Gilardi, & Grajal, 2000). The situation in
Latin America and the Caribbean is especially serious: 65 of the 151 species of
parrots are endangered (BirdLife International, 2010). For some species, habitat
Correspondence should be sent to Michel Saint Jalme, Muséum National d’Histoire Naturelle,
57 rue Cuvier, 75005 Paris, France. Email: [email protected]
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188 LEBLANC ET AL.
destruction is the main cause of population decline, and reproduction in captivity
may be the only chance for survival (Snyder et al., 2000). Captive reproduction
requires taking into account the birds’ basic biological and behavioral require-
ments in order for them to successfully adapt to their environment and the need to
manage genetic diversity through international breeding programs (Saint Jalme,
2002). However, optimal genetic management may conflict with free choice of
the partner, which appears to be one of the keys of breeding success (Curio,
1998; Sutherland, 1998). Thus, reproduction in captivity, where mate choice
is limited due to low number of individuals, is particularly challenging for
species of endangered parrots such as Amazona sp. and macaws (Derrickson
& Snyder, 1992). Considering the failures of reproduction-related behavioral
incompatibilities between managed captive pairs, artificial insemination (AI)
may constitute a better alternative. Such an alternative could also allow the
formation of breeding pairs between birds in different breeding facilities. AI has
already been successfully carried out in a variety of bird species:
1. Falconiformes (Boyd, 1978);
2. Galliformes (Caïn, 1978; Saint Jalme, 2002; Saint Jalme, Lecoq, Seigneurin,
Blesbois, & Plouzeau, 2003);
3. Sphenisciformes (O’Brien, Oehler, Malowski, & Roth, 1999);
4. Gruiformes (Jones & Nicolich, 2001; Saint Jalme, Gaucher, & Paillat,
1994); and
5. Psittaciformes (Brock, 1991; Samour, Smith, Moore, & Markham, 1986).
The massage method is the most common procedure used for semen collection
for birds in the wild who are noncooperative and not imprinted on humans (Gee,
Bertschinger, Donoghue, Blanco, & Soley, 2004). Semen collection involves
massage to stimulate the birds with pressure applied on the sides of the cloaca
to extract the sperm.
Female insemination is more challenging. To introduce the sperm, the oviduct
must be localized, either by palpation—everting the cloaca—or by use of a
speculum of appropriate size; use of a speculum may require gaseous anesthesia.
AI is a labor-intensive process that requires caregivers to catch and restrain the
birds. This may pose a problem particularly for large psittacines, who can inflict
serious bites and therefore increase the risk of injuries for both bird and handler.
The only AI attempts documented with macaws describe the harvesting of
semen carried out under sedation with electroejaculation (Harrison & Wasmund,
1983), a technique that is not recommended because of the risks of burns and
convulsions (Gee et al., 2004). Restraint and sedation of nonhuman animals
are sources of stress and can affect their welfare (Morgan & Tromborg, 2007).
Although AI may be an important tool for reproduction in some species, it is
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ARTIFICIAL INSEMINATION PROCEDURES FOR MACAWS 189
not currently used with large parrots because of the difficulty of the procedure,
the stress for the birds, and the risks of injuries for both birds and handlers.
In zoos and aquariums, training has now become a common practice, making
it possible to teach animals to voluntarily accept a variety of potentially stressful
procedures. This avoids the need for using excessive restraint or anesthesia. For
example, macaws have been successfully trained to accept tactile examinations,
blood collection, insertion of a thermometer in the cloaca, and inhalations of
medication (Heidenreich, 2004).
To teach cooperative husbandry behaviors, we applied principles of operant
conditioning with positive reinforcement. We used positive reinforcement, gener-
ally in the form of preferred food items, to shape and increase the probability of
the desired approximations until the target behavior was obtained (Heidenreich,
2007).
In order to train wild birds to accept the procedures necessary for AI, they
must first accept handling by trainers. Birds in general are very sensitive to stress
induced by handling (Straub, Forbes, Pees, & Krautwald-Junghanns, 2003).
Barnett, Hemsworth, Hennessey, McCallum, and Newman (1994) compared a
group of hens subjected to minimal contact with humans to a group subjected
daily to 15 min of additional contact mimicking usual husbandry procedures.
This additional contact caused adverse changes in cell-mediated immunological
responsiveness (thus probably involving a chronic stress response), even though
hens are domestic animals. Consequently, it is important to ensure that the
training carried out to avoid the need for restraint is not in itself a source of
stress that could reduce welfare or reproductive success (Mason, Clubb, Latham,
& Vickery; Swaisgood, 2007). Chronic stress may be revealed by changes in
behavior performed by animals, such as exploratory behavior, feeding, repro-
ductive behaviors, locomotion, apathy, vigilance, aggression, and stereotypic
behaviors (Mason et al., 2007; Morgan & Tromborg, 2007; Swaisgood, 2007).
Stereotypic behaviors in parrots can be locomotive (e.g., pacing, perch circles,
corner flips, and route tracing) or oral (e.g., wire chewing, sham chewing,
food manipulation, and dribbling; Garner, Meehan, Famula, & Mench, 2006).
Stereotypy is important to study because it is associated with frustration and poor
psychological welfare in captive animals (Garner, Meehan, & Mench, 2003).
The experiment was performed on a pair of Buffon’s macaws (Ara ambiguus)
and a pair of green-winged macaws (Ara chloropterus) held at the Ménagerie du
Jardin des Plantes, a zoo belonging to the National Museum of Natural History,
Paris, France. The goal of the experiment was to develop a program to train male,
large macaws so that they could learn to accept the semen collection procedure
without capture and restraint, thereby avoiding acute and chronic stress. For
the females, the objective was to train them to accept the introduction of a
speculum into the cloaca and to introduce their heads into a mask used for
gaseous anesthesia. This latter process was considered as an alternative method
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190 LEBLANC ET AL.
in case the behaviors necessary for insemination could not be trained. In order
to evaluate the subjects’ general welfare, the baseline behavior of birds was
recorded outside of training sessions throughout the program. We wanted to
answer the following questions:
1. Is it possible to train large psittacines to accept AI handling procedure?
2. Would this training be a source of stress?
3. Could the training provide some behavioral enrichment?
MATERIALS AND METHODS
Species, Sample, and Housing
The Buffon’s macaw is classified in the endangered category by the International
Union for Conservation of Nature, and the species is registered in Appendix I
of the Convention on Trade in Endangered Species (CITES). The nominal
subspecies Ara ambiguus ambiguus lives in Central America and ranges from
eastern Honduras to eastern Panama and northern Colombia. The subspecies Araambiguus guayaquilensis, the most threatened, is restricted to Ecuador where
the population is estimated at just 100 individuals (Snyder et al., 2000). The
three major threats to macaws are (a) the loss of their habitat; (b) the poaching
of nests for the pet trade; and (c) to a lesser extent, the poaching of birds
either for food or for the collection of feathers (Grajal, 2000). Unfortunately,
the Buffon’s macaw seldom reproduces in captivity (Juniper & Parr, 1998). The
green-winged macaw (Ara chloropterus) lives in eastern Panama and in South
America from Colombia to northern Argentina. The species is registered in
Appendix II of CITES. Both species are large birds, reaching 95 cm in overall
length and weighing 1,200 g to 1,300 g (Dunning, 2008).
Training was carried out with two pairs of macaws who had never successfully
reproduced. The first one was a pair of Buffon’s macaws (BM) living together
since the beginning of the experiment: a 7-year-old male and a 2-year-old female.
They were held by the Ménagerie for 2 months at the time the experiment began.
The second was a pair of green-winged macaws (GWM) living together at the
Ménagerie for 15 months before the beginning of this experiment: a male almost
3 years old and a female 22 years old. The 4 subjects were all born in captivity.
The two BM and the male GWM had been reared by their parents. The female
GWM was given to a zoo by a private individual when she was 9 years old, and
her rearing condition is unknown.
The aviary of the BM measured 4.6 m � 4.7 m � 5 m. An artificial nest was
fixed in it. Branches were installed at the start of training for both enrichment
and to facilitate the locomotion of the birds. The aviary of the GWM measured
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ARTIFICIAL INSEMINATION PROCEDURES FOR MACAWS 191
6.1 m � 4.7 m � 5 m. An artificial nest was installed 6 weeks after the experiment
began. Both aviaries were visible to the public; the visitors’ path was at a distance
of 1 m. The birds were fed twice a day: at 8:45 a.m. they were given extruded
pellets, and a fresh fruit meal was given to them at 12:00 p.m. Water was
provided ad libitum.
Training Process
Two trainers, a man and a woman, took turns training all 4 subjects. One of the
trainers had no previous contact with the birds prior to the experiment; the other
trainer had minimal contact during transport and medical vaccination.
Training was carried out inside the aviaries, 5 days a week. It was carried out
each day in two sessions of 10 min per bird at midmorning and early afternoon;
the birds continued to receive their customary diet. This training was done from
February 19, 2007, to June 28, 2007, for the GWM and from March 6, 2007,
to June 4, 2007, for the BM.
Verbal cues were used for the first two stages. Then, as a visual signal,
the index finger was shown to the bird during the third stage (as described in
Training Stage 3 following). A food reward preferred by the individual bird
(sunflower seeds, pieces of walnuts, hazelnuts, and extruded pellets) was given
after each correct response or behavior approaching the correct response at the
beginning of a new exercise. A conditioned reinforcer, “very good,” was used
to mark a successful behavioral approximation.
A secondary reinforcer or “bridge” was used between the emission of the
desired behavior and the delivery of rewards. This was a sharp snap emitted
by a clicker held with one hand. This sound constitutes an acoustic secondary
reinforcement, acquiring its reinforcing properties by immediately preceding
each food reward (McCall & Burgin, 2002). Even brief delays are detrimental
to learning; therefore, the function of training practice is to mark the precise
moment of the correct execution of exercises and to indicate that a reward will
follow (Heidenreich, 2007). We used both the verbal reinforcer, “very good,”
and the clicker because the birds were already used to this verbal expression
(sometimes said by the caretakers), but the clicker allowed a more precise
and more standardized signal as a bridge. If the parrot expressed a nondesired
behavior such as attempting to bite trainers, the bird was ignored: the trainer
turned round and ceased interacting with the bird for 10 s.
Training Stages
Stage 1: Approaching and accepting hand-feeding. In this stage, the
birds learned to gradually associate the sound of the clicker to the food reward.
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192 LEBLANC ET AL.
The trainer installed a wooden perch (1.2 m high) beside a large branch. The
trainer called each individual bird by name and presented some food, handheld.
If the bird did not approach, the trainer put the food on a branch situated between
bird and trainer, moved away, and again called the bird by name. As soon as the
bird took the food reward, the trainer activated the clicker. When the subject had
learned to reliably approach the trainer to a specific location, we began Stage 2
of the training process.
Stage 2: Go up the perch. The cue “place” was given. Initial approxima-
tions had the trainer holding the reward toward the bird above the perch. Next,
the reward was placed on the perch. In the final training approximations, the
subject was conditioned to move to the proper location without having to first
see the reward.
Stage 3: Accepting beak touching. A visual signal, the index finger, was
shown to the bird; this indicated to the bird that the trainer was going to attempt
to touch a body part. The bird was rewarded, with acoustic bridging, for staying
on the perch without escaping or displaying defensive or attacking reactions. The
trainer proceeded to approach the beak with the index finger and then calmly
touched the beak.
Stage 4: Accepting touches to the body including the wings, the back,
the tail, and the cloaca. The trainer started by touching the wings, ap-
proached with the index finger, then proceeded to touch gently, and finally
touched with more pressure. The trainer proceeded in the same way for the
touches of the back, the tail, and finally of the cloacal area.
Stage 5a: Accepting massage and for females the introduction of thespeculum. The trainer held the reward in front of the bird’s head and rein-
forced each long caress accepted with no muscle tension or avoidance behaviors
exhibited by the bird. For the females, the speculum was placed in contact with
the cloacal zone initially and then introduced into the cloaca itself. The female
was rewarded on a continuous reinforcement schedule during the introduction
of the speculum, which was lubricated beforehand with Vaseline.
Stage 5b: Accept the mask and a light pressure on the body (females
only). The verbal order, “touch,” was given. At the beginning of this stage, the
reward was put at the entry of a transparent anesthesia mask, and then it was
applied against the mask from the outside. This stage was trained at the same
time as the massage approximations.
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ARTIFICIAL INSEMINATION PROCEDURES FOR MACAWS 193
Evaluation of the Performances
Success of training was evaluated for each stage before progressing to the next
step in the process: a stage was considered mastered if three orders had been
successively executed in the same session over the span of three consecutive
sessions.
During the training process the following parameters were recorded:
1. Number of sessions for engaging or for passing a stage;
2. Number of orders given; and
3. Number of behaviors executed per order, including distress calls, fear
postures, and threatening postures directed toward the trainers.
Participation of the birds in a given training session was measured using the
criteria, “taking food from the hand at least once.”
Behavioral Data Recorded Outside Training Sessions
The behavioral observations collected outside of the training sessions were
carried out from February to the end of June in three daily sessions of 20 min
each for the 2 individuals of each aviary: the first one starting at 9:15 a.m., the
second one after the morning training at 11:15 a.m., and the last one following
the afternoon training at 3:30 p.m. The order of observation of the two aviaries
was alternated each day. Behavioral data were collected with the scan sampling
method (Altmann, 1974) with scans taken every 30 s.
The behavioral categories recorded were resting, vigilance, foraging, loco-
motion, solitary activities others than stereotypic behaviors, possible stereotypic
behaviors, comfort behaviors, and social behaviors. The reproductive behaviors
were also quantified. These behaviors are described in more detail in Table 1.
For each session, the number of visitors passing in front of the aviary was also
noted.
Data Analysis
The behavioral data were used to calculate average frequencies of behaviors per
day for each stage of training. The frequencies of one given behavior for each
day were calculated by dividing the number of observations of this behavior in
that day by 120 (number of scan samples for 1 day). We then averaged these
frequencies per day for each stage of training.
The duration of each stage depended on the progression of the birds and
varied among subjects. Stage 0 corresponded to the period before the start of
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TABLE 1
Ethogram for Ara ambiguus and Ara chloropterus
Resting� Sleep: eyes are closed or beak is under the back or wings feathers
Vigilance� Perched on two feet: the bird is perched high, standing on two feet, head up, eyes open� Leaning on two feet: the bird is standing on two feet, body leaning toward the front, wings a little
spread, looking at a particular point� Perched on one foot: the bird is perched high, standing on one foot, head up, eyes open� Hanging: the bird is hanging from the wire mesh or from a rope by the feet and/or the beak, eyes open
Foraging� Eats food held in foot: the bird holds a piece of food in one foot and eats it� Pecks from feeder: the bird takes food from the feeder with beak� Looks for food: the bird walks looking on the ground and pecking in different directions� Pecks from ground: the bird pecks on food or vegetation that is on the ground� Eats: the bird chews some food or drinks the juice from a fruit� Asks for food: the bird lowers front of body, tilts head toward partner, who holds some food in beak
or foot� Steals food: the bird takes or tries to take food from partner’s foot� Gives food: the bird puts from beak some food in the partner’s beak, after being asked.� Drinks: the bird puts beak in water, and swallows it
Locomotion� Walks: the birds moves around using feet� Climbs: the bird climbs using beak and feet� Flies: the birds moves around using wings
Solitary activities other than stereotypies� Acrobatics: hanging to the ceiling’s mesh by one or two feet, or by one feet and beak, the bird swings
and gesticulates� Tears to shreds: the bird bites, tears to shreds, or tears off some bark, leaves, or twigs� Wings flap: the bird hangs on a branch or the mesh with feet and beak and flaps wings� Nods: the bird repeatedly nods head up and down� Bites metal: the bird bites the wire mesh or some wires� Bites a rope: the bird uses beak to manipulate a rope� Bites rock: the bird uses beak to manipulate a rock� Turns: turns around a branch� Plays alone: gives a peck to a rope or a hosepipe hanging and looks at it while it moves
Stereotypies� Intention of flying: the bird sits on a branch, extends neck, flaps wings slightly, then opens wings
without flapping and vocalizes. Often also shakes head to the right and the left (behavior observed in
the male BM)� Weak intention of flying: same as aforementioned, but the wings are not spread out and the bird does
not vocalize (behavior observed in the male BM and the female GWM)� Shakes head: bird shakes head from right to left many times in a row (behavior observed in the female
GWM)� Swings front of body: the bird sits on a branch, extends neck, and swings the front of body from right
to left in rhythm, many times in a row (behavior observed in the female GWM)
(continued )
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ARTIFICIAL INSEMINATION PROCEDURES FOR MACAWS 195
TABLE 1
(Continued )
Comfort� Preening: the bird cleans and arranges feathers using beak� Beak cleaning: the bird cleans the inside of beak with feet� Beak scrubbing: the bird vigorously scrubs beak on a branch� Feet cleaning: the bird cleans feet with beak� Scratches: the bird scratches himself or herself with foot� Bites himself or herself: the bird bites body and vocalizes� Stretches: the bird stretches down one foot and the wing of the same side� Wings up: the bird raises wings without flapping� Shakes himself or herself: the bird shakes and fluffs feathers� Takes a bath: the bird sits in water and makes head and wing movements to clean himself or herself� Yawns: the bird opens beak wide and inhales deeply� Defecates: the bird discharge feces from the body
Social behavior
Affiliative behaviors� Allopreening: preening of the partner� Asking for preening: the bird leans toward partner and presents nape of neck� Goes toward: the bird goes toward partner, head forward, wings spread� Play together: both birds hold the same rope and pull together in opposite directions� Beak contact: the birds hold each other by their beaks� Slight peck: the bird gives a slight peck on partner’s head or beak� Bows: the birds leans head quickly a few times toward partner with high-pitched vocalization, then
turns a few times looking at partner� Hide together: the birds go together to a small cavity in a trunk, stay there for some time, then go out
of it together
Reproductive behaviors� Regurgitates: the bird regurgitates in partner’s beak, with spasmodic movements� Copulation: the male is on the female’s back and they make cloacal contact� Nest inspection: the bird goes in the nest
Agonistic behaviors� Rises: the bird rises and spreads wings a little, with fan-shaped tail, beak wide open, and sometimes
piercing vocalizations� Feet up: the bird lifts a foot at chest height toward the other, stretches neck, feathers fluffed or not,
often groaning� Wings spread: the bird spreads wings, head feathers fluffed and pupils contracted� Rushes: the bird fluffs feathers, spreads wings slightly, and walks fast toward the other with head
down and open beak� Strong peck: the bird gives strong pecks to the other, often both birds vocalize� Small peck: the bird gives a slighter peck to the other, both birds grunt� Peck exchanges: both birds give pecks to each other, beaks are wide open, both birds grunt or vocalize� Escapes: one of the birds rushes away when the other approaches� Shudders: the bird trembles convulsively� Stiffens: the bird flattens feathers, retracts neck, and stiffens
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the training. Stage 6 (attained only by the Buffon’s macaws) corresponded to
the period after the end of the training program.
The number of visitors passing was calculated as monthly averages based
upon the numbers recorded each day during the three observation sessions. The
differences among the average frequencies of the behaviors across training stages
and also in relation to the monthly averages for visitors were analyzed using the
Kruskall-Wallis test. If significant variations had been noted with the Kruskall-
Wallis test, these were then compared two by two with the test of Permutation
(StatXact software).
RESULTS
Training Stages
We performed 134 training sessions of 10 min each per bird for the BM
(15 weeks) and in 160 sessions (18 weeks) for the GWM. The female GWM
took food from the trainer’s hand from the first session and the male BM from
the second session. The female BM took food reliably from the hand from
Session 19. The male GWM only regularly accepted hand-feeding beginning in,
and continuing from, Session 33. Twenty-one sessions were necessary for the
female GWM to join the trainer near the perch, 24 for the male BM, 28 for the
female BM, and 52 for the male GWM (Table 2).
The male GWM systematically produced distress calls at the entry of the
trainers into the aviary during the first 3 weeks of training. The other birds
emitted distress calls only at the first entry. Only the male GWM and the
female BM expressed fearful body postures. The male BM displayed threatening
postures during the first 10 sessions, tending to precipitate toward the trainer.
We also noted conflicts between the birds: macaws not being rewarded during
training often stole or tried to steal their partner’s food. These conflicts could
impair the training. To limit as much as possible the stress linked to the training,
however, we preferred not to separate the members of a pair during the training
sessions.
The male BM learned to accept the massage with pressure on the sides of
the cloaca in 124 sessions (Table 2). The female BM learned to accept the
massage in 114 sessions and the introduction of a speculum into the cloaca in
134 sessions. She also learned to introduce her head into a mask for anesthesia
and to hold it inside for 1 min, while also accepting the light pressure of the
trainer’s hands on her body, in 122 sessions (Table 2). The male GWM learned to
accept touches of his wings in 143 sessions (Table 2). The female GWM learned
to accept the massage in 147 sessions and to put her head in the anesthesia mask
in 129 sessions (Table 2). For the GWM, the male never accepted massages, and
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ARTIFICIAL INSEMINATION PROCEDURES FOR MACAWS 197
TABLE 2
Number of Sessions for Engaging and for Validating a Stage for
Each Individual of the Two Pairs of Macaws: Buffon’s Macaw (BM)
and Green-Winged Macaws (GWM)
♂ BM ♀ BM ♂ GWM ♀ GWM
Stages of Training IS CS IS CS IS CS IS CS
Stage 1: Accepting food and going
near the perch
1 24 1 28 1 52 1 21
Stage 2: Going up the perch 25 45 29 45 53 87 22 65
Stage 3: Touch of beak 46 108 46 107 88 133 66 129
Stage 4: Touch of wing 69 108 79 107 110 143 104 129
Stage 5(a): Massage 109 115 110 114 129 147
Stage 5(a): Pressure on the cloaca 117 124
Stage 5(a): Speculum in the cloaca 131 134
Stage 5(b): Head inside the mask 85 104 129
Stage 5(b): Remain inside 99 107 129
Stage 5(b): Accept the touch 114 122
Note. IS (Initial Session): session number during which the corresponding order was given
for first time; CS (Criterion Session): session number during which the behavior was regarded as
learned according to the preset criteria (three orders successively obeyed during a session, for three
consecutive sessions).
training did not reach the stage of introducing the speculum into the female’s
cloaca.
The male BM had the highest participation of any subject throughout the
experiment and did not significantly vary (p > .05) according to the training
stages (Figure 1). The participation of the female BM was weak during the first
stages but increased by 150% between Stages 1 and 2 (p < .0001) and did not
significantly vary thereafter (Figure 1).
By comparison, participation of the male GWM was rather poor throughout
the program and did not significantly vary according to the training stages
(Figure 1). For the female GWM, the participation was high at the beginning of
the program and then decreased significantly (p D .048) between Stages 2 and
3, presumably when the physical contact criterion was introduced (Figure 1).
During training, aggressive behaviors toward trainers were sometimes ob-
served. The male BM sometimes rose up and raised his leg toward the trainer
when the visual signal of the touch was given. The reactions to the touch most
frequently performed were quivering and sometimes, in the case of GWM,
leaving the perch. The female GWM showed threats or fear when physical
contact was introduced to training; she often ruffled her head feathers (threat or
fear or both) when the visual signal of the touch was given. Fortunately, serious
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198 LEBLANC ET AL.
*p Kruskall Wallis.
FIGURE 1 Variation of average participation of each bird according to the training stages.
First stage: Approach the trainer, taking food in the hand and approach the perch. Second
stage: Go up the perch. Third stage: Accept to be touched on the beak. Fourth stage: Accept
to be touched on the wings, the back, the tail, and the cloaca. Fifth stage: Accept the massages
and for female the introduction of the speculum. BM D Buffon’s macaws, GWM D green-
winged macaws.
aggression in response to physical contact by the trainer (measured as attempts
to bite) was minimal: male BM (five total), female GWM (one).
Behavioral Data Recorded Outside Training Sessions
For the male BM (Figure 2a), all behavior categories, except locomotion, varied
significantly among training stages. Vigilance decreased between Stage 0 and
Stage 5 (p D .072) as did stereotypic behaviors (p < .0001) even though they
increased again between Stages 5 and 6 (p D .021), and an increase in social
behaviors was observed from Stage 0 to Stage 5 (p D .0002). The female BM
(Figure 2b) never presented any stereotypic behavior. Her vigilance increased
between Stages 0 and 1 (p D .049) and then decreased between Stages 1 and
5 (p D .001). Foraging activity increased between Stages 0 and 3 (p D .02),
then decreased between Stages 3 and 5 (p D .048). We observed an increase of
social activity (similar to the male BM) from Stage 0 to Stage 5 (p D .0001).
No variation of social activity behavior was observed in the GWM during
the training process. For the female, even though vigilance varied significantly
during the program, no general tendency was observed. The vigilance of the
male GWM (Figure 2c) increased between Stages 0 and 1 (p D .011) and then
remained at a relatively high level during all stages of training but decreased
between Stages 1 and 4 (p D .014). The male GWM never expressed stereotypic
behaviors. The stereotypic behaviors of the female GWM (Figure 2d) increased
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*p
<.0
5.
**
p<
.01
.*
**
p<
.00
1.
(a)
FIG
UR
E2
(a,b
);(c
,d).
Co
mp
aris
on
of
the
aver
age
freq
uen
cies
of
the
beh
avio
ral
cate
go
ries
for
each
stag
e,(a
)fo
rth
em
ale
BM
,(b
)fo
rth
e
fem
ale
BM
,(c
)fo
rth
em
ale
GW
M,
(d)
for
the
fem
ale
GW
M.
Fir
stst
age:
Ap
pro
ach
the
trai
ner
,ta
kin
gfo
od
inth
eh
and
and
app
roac
hth
ep
erch
.
Sec
on
dst
age:
Go
up
the
per
ch.
Th
ird
stag
e:A
ccep
tto
be
tou
ched
on
the
bea
k.
Fo
urt
hst
age:
Acc
ept
tob
eto
uch
edo
nth
ew
ing
s,th
eb
ack
,th
eta
il,
and
the
clo
aca.
Fif
thst
age:
Acc
ept
the
mas
sag
esan
dfo
rfe
mal
eth
ein
tro
du
ctio
no
fth
esp
ecu
lum
.B
MD
Bu
ffo
n’s
mac
aws,
GW
MD
gre
en-w
ing
ed
mac
aws.
(co
nti
nu
ed
)
199
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*p
<.0
5.
**
p<
.01
.
(b)
FIG
UR
E2
(Continued
).
200
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*p
<.0
5.
(c)
FIG
UR
E2
(Continued
).
201
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*p
<.0
5.
**
p<
.01
.*
**
p<
.00
1.
(d)
FIG
UR
E2
(Continued
).
202
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ARTIFICIAL INSEMINATION PROCEDURES FOR MACAWS 203
TABLE 3
Variations of the Monthly Average of the Number of Visitors Spending
Each Day in Front of the Macaws’ Birdcage Into 60 Min of Observation
Months Monthly Average of the Daily Number of Visitors ˙ SD
February 22 ˙ 3,21
March 27,05 ˙ 4,08
April 125,76 ˙ 12,77
May 108,42 ˙ 11,69
June 100,94 ˙ 10,26
between Stages 2 and 3 (p D .0005). Behaviors associated with reproduction
were observed in the pair of BM (Figure 3). They increased significantly between
Stages 0 and 5 (p D . 044) for the male and between Stages 0 and 4 (p D .026)
for the female. Very few behaviors that could be associated with reproduction
were observed in the pair of GWM (Figure 3).
The number of visitors (Table 3) passing each day in front of the macaws’
aviaries increased significantly from March to April (p < .0001) and then did
not vary significantly from April to the end of the training program.
DISCUSSION
Our training method using operant conditioning with positive reinforcement
(food) was effective in teaching macaws to accept increasing levels of handling
in a few months, with the aim of training behaviors that allow the collection of
the semen in males and AI of females. The behavioral observations suggest that
the training done here was not a significant source of stress for the 2 birds who
completed the entire training regimen. We found the following in both birds:
1. A decrease in, or no, stereotypic behaviors;
2. No increase or a decrease in vigilance after Stage 1;
3. An increase in social behaviors; and
4. No significant perturbation of other behaviors.
Although the first stage (approach—take from hand) was short for the 2 oldest
subjects (male BM and female GWM), it was longer and was associated with
behaviors related to fear and stress for the 2 youngest macaws (female BM and
male GWM). We observed important individual differences for the behaviors
learned as well as for the behaviors expressed outside the sessions during the
training program. These differences could be related to many factors such as
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*p
<.0
5.
FIG
UR
E3
Co
mp
aris
on
of
the
aver
age
freq
uen
cies
of
the
rep
rod
uct
ive
beh
avio
rsfo
rea
chst
age
for
each
of
the
4su
bje
cts.
BM
DB
uff
on
’s
mac
aws,
GW
MD
gre
en-w
ing
edm
acaw
s.
204
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ARTIFICIAL INSEMINATION PROCEDURES FOR MACAWS 205
species, age, and life experience (rearing conditions—parents-reared vs. hand-
reared). These differences show the necessity of adapting the training to each
individual. We may have moved too fast to the physical-contact stages with the
female GWM; her participation decreased when the physical-contact criterion
was introduced. Because she often left the perch at the sight of the visual signal
preceding the touch, we should probably have slowed down the stage where
touch was introduced and added more reinforcement value for holding still
while food was offered and eaten. In this case, not requiring physical contact
for each training session may have helped to maintain the bird’s participation.
For the male BM, the first stage (approach—take from hand) was very quickly
acquired in spite of his not being familiar with hand-feeding before training
commenced. A long experience of captivity, feeding by caretakers, and human
proximity could explain this fast learning process. The participation of this male
was also very good for all the remaining training stages. The male BM learned
to accept the massage with pressure on the sides of the cloaca; however, no
semen collection was performed because the trainer was not experienced in the
AI process. Semen collection will be the project’s next step.
For the female BM, the first stage was much slower than for the male.
This could be related to the young age of the subject and less habituation to
human proximity. Nevertheless, her participation increased from the beginning
of the second stage and remained excellent without significant variation for the
remainder of the training stages. This female learned to accept the massage and
the introduction of a lubricated speculum into the cloaca. She also learned to
introduce her head into the anesthesia mask, to keep this position for a few
seconds, and to accept a light pressure of the trainer’s hands on her body during
this time.
The female GWM was the quickest of all subjects to go near the trainer and
take food from the hand. This could be explained by the relatively old age of
the subject (22 years) and a long experience with human proximity. However,
we observed a drop of the participation between Stages 2 and 3. That may be
related to the introduction of the touches at Stage 3. Before her acquisition by the
Ménagerie, this bird was kept by a private collector. This could have influenced
her behavior during the training. The influence of negative handling by humans
on later behavioral responses of animals is described by Breuer, Hemsworth,
and Coleman (2003). Hemsworth, Coleman, Cox, and Barnett (1994) discuss
the generalization of responses to other humans for the same stimulus. The
female GWM learned to accept some massage and to introduce her head into an
anesthesia mask; however, because of her behavior during the training (in many
sessions leaving the perch at the sight of the visual signal indicating to the bird
that she would be touched), the following training stages were not reached.
The first stage (approach—take from hand) was very long for the male GWM,
and his participation was weak during all stages of training. This could be
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206 LEBLANC ET AL.
related to his lack of socialization with humans. He seemed to be a fearful
bird who, at the beginning of the program, was startled by the clicker’s sound.
In addition, he was particularly sensitive to the presence of different trainers
(screaming and refusing at every change to participate in the training) and to
many possible sources of disturbances, such as the vocalizations of other macaws
or the presence of noisy visitors. He learned to accept a touch of his wings but
did not proceed beyond this step.
None of the following behavioral variations indicative of chronic stress were
detected in the male BM: alteration of exploratory, feeding, and reproductive
behaviors; locomotion; or an increase in apathy, vigilance, aggression, and
stereotypic behaviors (Morgan & Tromborg, 2007; Swaisgood, 2007). Moreover,
the reduction of stereotypic behaviors shown by the male throughout the program
indicates a possible improvement in the welfare of the bird; this may suggest that
the training represented a behavioral enrichment (Mason et al., 2007). However,
the observations of the BM were conducted just after their installation in the
aviary; this could have generated additional stress in the male; thus, the decrease
of stereotypic behaviors observed during the training could also be explained
by the progressive habituation of the bird to his new environment. The increase
in stereotypic behaviors recorded after Stage 5, when training had stopped,
supports the assumption that training could be considered a form of behavioral
enrichment. Nevertheless, the last increase of stereotypic behaviors could also
be linked to the end of the reproductive season (which was concomitant with
the end of the training program); this could also lead to an increase in inactivity
and, thus, boredom. Because this increase is very small, it also may not be
biologically relevant (even if it is statistically significant).
Concerning the female BM and the male GWM, increased vigilance (as
defined by the behaviors described in Table 1) was recorded for these 2 subjects
between the period preceding the start of training and the first stage and may
constitute a sign of stress (Morgan & Tromborg, 2007; Swaisgood, 2007), which
could be related to the initiation of the training schedule. This assumption
is supported by the weak participation and the fearful behaviors that these
birds expressed in the first stage of training sessions. However, the female
BM’s vigilance decreased from Stage 1 to Stage 5 with a parallel increase in
participation. Reduction in foraging behavior recorded from Stage 4 to the end
of the program may be related to the bird’s increased participation and the fact
that many food rewards were therefore consumed during the training sessions. In
this context, and in the absence of other corollary signs of behavioral changes,
this variation is probably not indicative of a stress response. For this bird, some
initial increase in stress was probably related to the first approaches of the trainer
and the trainer’s presence in the aviary. On the other hand, for the male GWM,
the relatively high level of vigilance and the weak participation during all stages
of training suggest a more chronic stress response.
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ARTIFICIAL INSEMINATION PROCEDURES FOR MACAWS 207
For the female GWM, the increase of stereotypic behaviors observed during
the training process is likely to indicate elevated stress, according to Mason
et al. (2007); the increase seemed to be in direct response to the introduction of
the first touches of the beak. This increase of stereotypies, combined with the
fall of participation in the training observed between these two stages, supports
the assumption of increased stress in relation to the handling carried out during
training. However, Stage 3 began on April 7 and continued until May 7 for the
female GWM, when the monthly average number of visitors passing each day
in front of the macaws’ aviaries increased significantly. It is well known that the
visitors can also constitute a source of stress in captivity (Morgan & Tromborg,
2007).
The behaviors of the BM associated with reproduction increased during the
training program and then decreased after the training was stopped. However,
macaw reproduction is seasonal and the reduction in reproductive behavior
(recorded after the end of the training program for the male and starting from
the first massages for the female) could indicate the end of the season. In
contrast, the reproductive behaviors of the GWM did not develop over the
period. The reproductive behaviors of the male recorded during Stage 2 were
related to inspection of the nest in the week following its installation inside the
aviary. In this context, these inspections could be more related to exploratory
behavior than to reproductive behavior. Although the absence of reproductive
behavior in the GWM could indicate chronic stress, other explanations are
also possible. Large macaws are known to reach their sexual maturity only
around the age of 4 to 5 years (Quemin, 2003): the 3-year-old male was
probably not sexually mature, and the 22-year-old female had never reproduced.
The female’s rearing history was not known; hand-rearing or an absence of
socialization with conspecifics during maturation are known to have detrimental
effects on reproduction (Schmid, Doherr, & Steiger, 2005). Hand-rearing is one
of the possible causes of the difficulties in macaws reproducing in captivity
(Derrickson & Snyder, 1992). Finally, social familiarity and sexual compatibility
are important determinants of the parrots’ pairing process (Derrickson & Snyder,
1992). Thus, the behavioral incompatibility of the partners may also explain the
failure of these subjects’ reproductive behaviors to develop; in this case, the AI
process would probably be necessary.
CONCLUSION
Despite interindividual variation in responses to the training program, we suc-
ceeded in 15 weeks to teach 2 BM to accept various handling procedures
necessary for AI. In less than 4 months, the female accepted the introduction
of the speculum into the cloaca. This suggests that with additional training,
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208 LEBLANC ET AL.
the complete voluntary application of the insemination procedures is feasible
without having to use restraint or gaseous anesthesia. Moreover, the training
did not generate chronic stress, and the birds’ reproductive behaviors developed
over the period, so they were not disturbed by the training. Concerning the
2 GWM, the training produced some positive results; however, in particular
for the male, they were still insufficient with regard to the AI process. For the
female, behavioral indicators revealed signs of stress likely to be linked not
only with the training but also with other sources of stress in captivity such as
the bird’s individual experience or proximity of visitors. Extending the birds’
training periods should be considered.
Therefore, our experiment shows that it is possible to conduct AI procedures
by using training instead of traditional techniques (restraint or anesthesia). As
long as the birds are compatible with human interaction, this procedure offers
the added benefits of being not only a source of behavioral enrichment that may
improve the welfare of some birds but also a key element of assisted reproductive
approaches.
ACKNOWLEDGMENTS
This study was hosted at the Ménagerie du Jardin des Plantes of MNHN
at Paris, France. We thank the director, Jacques Rigoulet, for his hospitality
and zookeepers Betty Knopf, Marie Vergues, and David Buchemeyer for their
assistance in bird managing. We also thank Sarah-Jane Vick and anonymous
reviewers for their suggestions to improve the manuscript. The experiments
comply with the French laws concerning animal care.
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