May 18, 2026
Physiological evaluation of Quarter Horses undergoing five-month initial training focused on hippotherapy
- Lara Carolynne1,2,
- Emmanuel Arnhold1,2,
- Jorge Passamani3,
- Alexandre Cardoso3,
- Letícia Celeste4,
- Kate Barcelos1,2
- 1School of Veterinary Medicine and Animal Science (EVZ);
- 2Federal University of Goiás (UFG), Goiânia, Goiás, Brazil;
- 3National Association of Equine Therapy (ANDE), Brasília, DF, Brazil;
- 4Universidade de Brasília (UnB), Brasília, DF, Brasil.
Protocol Citation: Lara Carolynne, Emmanuel Arnhold, Jorge Passamani, Alexandre Cardoso, Letícia Celeste, Kate Barcelos 2026. Physiological evaluation of Quarter Horses undergoing five-month initial training focused on hippotherapy. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygxjjxkl8j/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 17, 2026
Last Modified: May 18, 2026
Protocol Integer ID: 317232
Keywords: equine, heart rate, cortisol, thermography, hippotherapy, horsemanship, physiological evaluation of quarter horse, horses for hippotherapy program, preparing horse, horses for hippotherapy, physiological stress, stability of physiological response, horse, physiological response, physiological evaluation, continuous heart rate, heart rate parameter, evidence of acute stress, indicating autonomic adaptation, training stage, cortisol level, autonomic adaptation, hippotherapy program, gradual physiological, quarter horse, plasma cortisol, lacrimal caruncle temperature, maximum heart rate, month initial training
Funders Acknowledgements:
Thermocracy Vet
Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF)
Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG)
Abstract
Preparing horses for hippotherapy programs requires management methodologies that
prioritize well-being and minimize stress in the face of new stimuli. This
study aimed to evaluate the effectiveness of a progressive training protocol
for Quarter Horses intended for hippotherapy, via physiological stress
indicators. Seven animals were monitored for 5 months, including an adaptation
phase and 3 training stages. Continuous heart rate, plasma cortisol, and ocular
infrared thermography were measured before and after exercise. Baseline (p =
0.0466) and minimum (p = 0.0095) heart rate parameters decreased over time,
indicating autonomic adaptation. Maximum heart rate (HRmax) was higher in the
initial phases, associated with the introduction of new stimuli, and decreased
in subsequent months (p = 0.0272). Lacrimal caruncle temperature increased at
the beginning of training (p = 0.002), followed by stabilization, suggesting a
decrease in the organism's reactivity to the new stimuli presented. Plasma
cortisol levels did not vary before and after exercise, nor between months. (p
≥ 0.05). The results indicate that the protocol promoted gradual physiological
adaptation, without evidence of acute stress harming the horse's physical and
emotional health. The progressive approach favored habituation to stimuli and
stability of physiological responses, demonstrating potential applicability in
preparing horses for hippotherapy.
Attachments
Materials
Described in the materials and methods section.
Materials and Methods
Animals:
The project selected seven QHs duly registered with ABQM due to their docile
temperament and three well-marked gaits: walk, trot, and gallop [15]. All animals were
over 3 years old (5.43 ± 0.53 years), had an average withers height ranging
from 1.31 m to 1.42 m (1.38 ± 0.05 m), and body weight between 310 and 415 kg
(354 ± 34 kg). Their age was considered compatible with the young adult phase, when
horses have sufficient physical maturity for mounted work, since horses start
training generally between 2 and 3 years old [6]. The selection also considered morphological
characteristics compatible with use in hippotherapy, prioritizing animals of
medium conformation, with adequate height and weight to facilitate the care of
the practitioner by the future therapist [3].
The QHs were acquired from farms in the region, where they lived on pasture, were
already accustomed to halter handling for general hygiene (grooming) and hoof
trimming and were supplemented with concentrated feed and mineral salt. They
were transported in horse-specific transport to the ANDE-BRASIL facilities at
an equestrian center in Granja do Torto, Federal District, Brasília, Brazil,
where they remained throughout the experiment. The animals were kept in
individual, well-ventilated 4x4 m masonry stalls, where the horses could see
the other animals from the front of the stall and through the door. The stalls
had a cement floor with a drainage grate, wood shavings bedding, a masonry
trough, and an automatic plastic waterer with an individual covered float.
Water was provided ad libitum. The diet corresponded to 2.25% of each animal's
body weight individually in % dry matter (DM), consisting of concentrate
(commercial balanced feed) and roughage (Tifton hay) in a 30:70 ratio. The
concentrate was divided and provided twice a day (7 am and 4 pm). The hay was
divided into portions of similar weight and provided three times a day
(morning, afternoon, and evening). Mineral salt was provided in a separate
trough ad libitum. All animals remained in the stalls until the start of
collection, release into paddocks, and training activities.
Experimental Design: The experiment lasted 5 months, assessing each animal individually
every month, collecting data before (baseline), during, and after training,
according to the type of variable analyzed (HR, plasma cortisol, and ocular IRT).
Thus, the Progressive Training Protocol for Hippotherapy (PTPH) was evaluated
monthly and organized into four main stages and an activity that permeated all
stages, which will be detailed later.
HR: It was assessed continuously, using the Polar H10 device on the horse's chest.
This is a lightweight (21 g), compact (34x65x10 mm) HR monitor with an
adjustable elastic strap made of comfortable textile material, using a CR2025
battery that lasts up to 400 hours. The sensor was connected to the device to
monitor beats per minute (hr) via Bluetooth Low Energy, ANT+ through a coded 5
kHz transmission (Polar Electro OY, 2023). HR was recorded using the Polar
Equine program on a Samsung S7 tablet. The assessment obtained the following
parameters per month of training: baseline HR (HR0), minimum HR (HRMin),
average HR (HRAve), maximum HR (HRMax), and final HR (HRFin).
Thus, it was possible to evaluate variations in physiological stress indicators
throughout the training process, allowing inferences about the animals'
adaptation to the proposed protocol. Data were collected once a month after
selecting the animals randomly, distributed over two consecutive evaluation
days (four animals on the first day and three on the second day). HR0
was measured in the stable, starting at 8 am, before the start of activities.
Subsequent collections were carried out before, during, and immediately after
training. A training session compatible with the training level of each initial
training stage was simulated on the evaluation day. Activities consisted of
leading and groundwork exercises in Phases 1 and 2 and a training circuit in
Phase 3, initially leading the animal on a lead and then riding it.
Ocular IRT: The capture distance was
standardized at 0.5 m from the animal's left eye, directed to the lacrimal
caruncle. The equipment was positioned at 90° in
relation to the sagittal plane, measured with the aid of a digital laser tape
measure [35]. Ocular IRT images were captured with a FLIR E40 camera, with a resolution of 160 x 120
pixels, operating in a temperature range of -20 °C to 650 °C, which offered
accuracy of ±2 °C or ±2% in the readings, ensuring reliable results. Thermal
sensitivity was less than 0.07 °C at 30 °C, detecting minimal temperature
differences. The 3.5-inch color LCD screen provided a clear visualization of
the thermal images. In this stage, the thermograph was calibrated, and the
ambient temperature (21.7 ± 1.3) and relative air humidity (75.2 ± 10.4) were
entered based on information provided by the weather app (Apple Weather‱, iOS)
on all days of the experiment to reduce environmental variables that could
influence the ocular temperature result. Surface emissivity was adjusted to Ɛ = 0.98 in
all collections, following the appropriate value for mammalian skin [34].
The first collection began at 8:30 am inside the stable, sheltered from direct
light, where baseline thermographic images were captured 0.5 m from the left
eye [35]. The images were evaluated immediately after capture to avoid errors and artifacts
(blur, flies, dirt, closed eye), repeating the capture if necessary to ensure
quality during analysis. On training days, the second collection took place
inside the round pen or arena, both covered. The collection and work sequence
was randomized by a draw, evaluating them on 2 consecutive mornings (four
animals on the first day and three on the second day). The images were stored
on a 128 GB SD memory card for later processing on a computer using IRISoft
software (PoliScan‱, FLIR Systems Inc., Wilsonville, OR, USA), selecting only
the images that presented the best technical quality, with sharp focus, absence
of thermal artifacts, and the eye fully open.
Plasmacortisol: Blood samples were obtained via jugular vein venipuncture to measure
plasma cortisol levels [36]. The site was previously cleaned; the samples were collected using disposable needles (25.0 x 0.8 mm) and stored in 10-ml BD Vacutainer‱ tubes
(without anticoagulant) for storage and subsequent processing. They were
collected from 10 am onwards, considering the circadian rhythm of cortisol in
horses [37] and the maximum cortisol peak in the morning (between 6 and 8 am) [38] to standardize the tests and avoid alterations resulting from this factor. The collection and work
sequence was randomized by a draw, evaluating them on two consecutive mornings
(four animals on the first day and three on the second day). Each horse underwent
collections at two different times: first, pre-exercise samples (10 am) were
collected inside the stables (5 to 10 minutes between entering and leaving the
stable), thus obtaining baseline plasma cortisol samples; then, samples were
collected in the round pen or arena (both covered), immediately after
completing the proposed exercises, to compare any changes resulting from the
training. All samples were transported under refrigeration to the Veterinary
Multipurpose Laboratory of the University of Brasília (UnB, Brasília, Brazil),
where they were stored. At this stage, the samples obtained in tubes without
anticoagulant were centrifuged for 10 minutes (Kasvi‱, model
K14-4000, Brazil) at 4000 rpm for plasma separation. Then, the refrigerated plasma
was sent to the specialized Veterinary Endocrinology Laboratory BET Labs
(Niterói, Rio de Janeiro, Brazil), to determine blood cortisol levels. The
technique used was solid-phase radioimmunoassay (RIA) with commercial kits
(Siemens Healthcare Diagnostics Inc., Tarrytown, NY, USA). All tests were
performed in duplicate.
PTPH
Steps: 1. Adaptation – acclimation, familiarization, and coexistence (recommended:
30 to 45 days); 2. Phase 1 – Communication and Integration (recommended: 30 to
45 days); 3. Phase 2 – Desensitization (recommended: 30 to 45 days); 4. Phase 3
– Capacity Building and Skills Development (recommended: 90 to 150 days). A
gymnastics program was also conducted throughout the entire experimental period.
It consisted of physical conditioning exercises, functional gymnastics, and
basic training, carried out continuously (recommended: throughout Phases 1, 2,
and 3 on alternate days to those with the other exercises approached in the
other specific phases).
General
recommendations for the training period: The durations proposed for each
training stage are based on the protocol routinely adopted by the ANDE-BRASIL
team for preparing horses for hippotherapy. Considering that the training is
individualized, the progression time between stages may vary between animals,
being influenced by factors such as previous experience, breed, and
temperament. The stages were standardized to the minimum time necessary to
obtain consistent results and reduce experimental biases, exclusively for the
comparison between individuals. The values adopted per stage in this research
are described in detail in the following topics. The experiment lasted 5
months, including 30 days of adaptation (when no data were collected regarding
the analyzed variables), followed by 4 months of initial training protocol and
experimental evaluations. The PTPH consisted of the following stages:
Adaptation in November (30 days); Phase 1 – Communication and Integration (30
days); Phase 2 – Desensitization (30 days); and Phase 3 – Capacity Building and
Skills Development (60 days), concluding in March. The gymnastics phase was
performed continuously throughout Phases 1, 2, and 3.
Acclimation
Phase – Familiarization and Coexistence (30 days): This period corresponded to
the arrival of the animals at the ANDE-BRASIL facilities, during which clinical
and laboratory examinations, preventive dental treatments, vaccination, and
deworming were carried out to ensure the health of the animals included in the
experiment. Then, the adaptation process to the new environment was conducted,
lasting a total of 30 days. During this period, they underwent activities to
familiarize them with the stalls, daily handling, and the routine of a
hippotherapy center. These activities included individual handling, hygiene,
manipulation, halter training, and release into collective paddocks, aiming at
familiarization with the environment and routine practices. A single person
performed daily handling in all study stages, responsible for leading the
animals in the stalls and general handling, under the supervision of the
trainer. During the training phases, a single trainer was responsible for
leading all animals throughout the entire experimental period to minimize
variations associated with human handling. HR, ocular IRT, and plasma cortisol
levels were collected in the stalls, with a single baseline sample obtained per
animal during this period.
Phase
1 – Communication and Integration (30 days): This stage began with groundwork
in a round pen, characterized by activities without riding to develop
communication between trainer and horse and promote the animal’s body awareness
and physical preparation. This approach aims to facilitate future adaptation to
riding, since the horse already understands the basic commands executed on the
ground [39]. Phases
1 and 2 were conducted progressively, with a structured sequence of ground
exercises. The equipment included a halter, long lead, vocal commands, body
language, stick, and flag stick (training flag). The animals were led to the
round pen by the trainer using a halter and long lead, positioning both
initially in the center of the area. To begin the exercise, the trainer
indicated the direction of movement with one hand and, with the other, moved
the stick posteriorly to the croup area, without direct physical contact,
stimulating the animal's movement based on the principles of pressure and
release [6]. The
exercise consisted of circular movement in the right and left directions, with
five repetitions for each side. At the end of each turn, a vocal command to
stop was given, followed by the animal approaching the center of the round pen
with the aid of the lead. Then, the animals were released from the lead and
kept loose in the round pen, being led to perform circular movements,
maintaining the same principles of direction and control. Later, the trainer
gave a vocal command to approach, initiating movement at a walk to stimulate
voluntary following. This stage allowed the evaluation of the animal's
responsiveness to the trainer's stimuli and its ability to pay attention and
maintain proximity at liberty. During this phase, the animals trained their
walk, trot, and gallop, both with a lead and at liberty. The phase lasted an
average of 30 days, with a frequency of four sessions per week, each session performed
individually, lasting approximately 30 minutes per animal. Continuous HR,
post-exercise ocular IRT, and post-exercise plasma cortisol were collected in a
covered round pen immediately after the sessions. Baseline measurements (HR, IRT,
and cortisol) were taken before exercise, inside each animal's stall.
Phase
2 – Desensitization (30 days): Phase 2 aimed to desensitize the animals to
external stimuli, reducing reactivity to objects, sounds, and different forms
of physical contact, as described in the literature for equine training
protocols [39]. They
were initially handled in the stall, where the halter was placed, and the
animals were led to the round pen at a walk, using a long lead. Upon reaching
the round pen, the halter was removed, and the animals were initially
encouraged to move freely, executing the basic commands previously learned in
Phase 1, for a short period. Then, the stop command was given to replace the
halter and begin the desensitization exercises. The equipment included a
halter, long lead, stick, flag stick, vocal commands, and the trainer's body
language. The desensitization process was conducted progressively. Initially,
tactile stimulation was performed with a stick on different regions of the
animal's body, following a standardized sequence: head, neck, trunk, back,
loin, rump, and limbs. Then, the same procedure was repeated with the trainer's
own hands to promote gradual adaptation to direct physical contact, always
bilaterally. Following this, visual and auditory stimuli were introduced. Using
the stick, movements were made around and above the animal, without direct
contact, as well as movements that generated vibrations in the ground,
promoting additional auditory stimuli. The use of the flag stick allowed the
introduction of more intense visual stimuli through approach and retreat
movements in different body regions, including areas of greater and lesser
sensitivity. These stimuli aimed to increase the animal's tolerance to
situations frequently encountered in hippotherapy contexts. They were initially
presented with the animal in a static position (standing still) and, according
to its acceptance, were applied in motion, during the walk, trot, and gallop
gaits. The progression of stimuli was gradual, respecting the individual
response of each animal and prioritizing the understanding of the trainer's
verbal and body commands. The saddle was introduced to the animals at the end
of the 15th day of the month, progressively, until they allowed it
to be placed on their backs and the girth tightened. As is common in Western
saddles in Brazil, a girth and belly band (two girths) were used during
desensitization. Training continued until the animals could be calm at a walk
and trot in a circle on the lead with the saddle. This phase lasted 30 days,
with sessions held three times a week, lasting approximately 30 minutes per
animal. Continuous HR, post-exercise ocular IRT, and post-exercise plasma
cortisol measurements were taken in a covered round pen immediately after the
sessions. Baseline measurements (HR, IRT, and cortisol) were taken before
exercise, inside each animal's stall.
Phase
3 – Capacity Building and Skills Development (60 days): Phase 3 corresponded to
the initial training and qualification of the animals for use in hippotherapy
sessions. The timeline for this phase is presented in Table 1.
Table 1.
Temporal organization of Phase 3.
| Training period | Duration | Weekly frequency | Location | |
| Beginning | First 30 days | 4 times a week | Covered round pen (twice a week), outdoor track (once a week), and covered arena (once a week) | |
| End | Last 30 days | 4 times a week | Outdoor track or covered arena, alternating as needed for training |
Training period, including
duration, weekly frequency, and location.
In this stage, the animals were introduced to
forward, sideways, circular, stopping, and backing movements, gradually
progressing in the exploration of gaits. Everyday situations, objects, and
experiences that typically occur in hippotherapy sessions were also presented.
The materials were halter, lead rope, complete bridle, blanket, and complete
saddle. Training began with progressive activities, adapting to the riding
equipment, exposure to environmental stimuli, and simulation of routine care
situations to promote behavioral safety, emotional stability, and appropriate
responses to the handler's commands. During the materials presentation phase,
the handler entered the round pen or arena and temporarily assumed the role of
assisting in the training process, sometimes leading the animal and assisting in
initial training throughout the desensitization process with the saddle,
bridle, and other equipment, until the first ride. After this stage, when the
animal was responsive to the commands of the mounted handler at a walk and
trot, specific hippotherapy objects were introduced. Galloping was practiced
during gymnastics training. The materials included a halter, lead rope,
complete bridle, blanket, and complete saddle. In this PTPH stage, the animals
were led through a desensitization activity circuit with the stimuli described
below, where the objects were left randomly or presented by a person known or
unknown to the horse. The adaptation sequence to the objects was performed
continuously, in a sequential circuit format within the location where they
performed their tasks – in this case, a 30 x 40 m covered arena with a nonwoven
fabric floor covered with sand, simulating a real hippotherapy session as it
occurs in Brazil. The materials described below were tested in various
combinations:
Cards and Velcro: A deck of colored cards was
presented and brought close to the animal, followed by tactile (touches on the
neck) and auditory stimuli (opening and closing a bag with Velcro); finally,
the cards and Velcro were moved over the animal's back and loin.
Clipboard with sheets: Moving the clipboard and
sheets near the animal's head and then throwing the clipboard to the ground.
Plastic hoops: Four large, colored plastic hoops
were arranged in a straight line on the ground, so that, when the horse was
led, it should pass through them without showing any refusal or jumping
behavior.
Exercise ball: Positioned on the ground of the
arena, allowing the animal’s spontaneous interaction, such as sniffing, biting,
or avoiding the object, according to its behavioral response.
One or more unknown people interacting from the
ground:
Gentle touch: The person slowly touched the horse on
previously desensitized areas.
Intense touch: The person touched the horse more
firmly on the same areas.
Verbal
interaction: The person established verbal communication with the animal, using
constant intonation and altering vocalization.
Sudden vocal stimulus: After verbal interaction, the
person emitted a high-pitched vocal sound near the animal to evaluate its
reaction to an unexpected sound stimulus.
Soccer ball: Initially presented for the animal to
smell, later used for desensitization through direct contact with the horse's
body, rolling the ball along the body surface. Then, the object was thrown or
rolled over and under the animal, observing its behavioral reactions.
Miniature saddle with figures: Equipment similar to
a racing saddle with colored geometric figures attached by Velcro. The figures
were initially removed and repositioned in front of the animal; then, the
equipment was positioned and slid along its back, repeating the same procedure
(tactile and auditory stimulation).
Mirror: The trainer positioned the horse in front of
a full-length mirror, allowing free observation of its behavioral reactions,
such as vocalizations and behavior, intervening only in case of attack.
Rings: Four colored rings were positioned on the
animal's face and then placed on the ears (two on each ear), while the trainer
led the horse in a circle to the right and left.
Plastic hoop: A plastic hoop was carried along the
animal's body and inserted around the neck, while the horse was led by the
halter, making a complete turn to both sides before the hula hoop was removed.
Auditory
stimuli using a sound device (speaker):
Counting: Playback of a recording containing
numerical counting from one to five.
Days of the week: Playback of a recording with the
names of the days of the week.
Sudden vocal stimulus: Playback of a recording
containing the sound of a child screaming.
The last evaluation simulated a hippotherapy session
under the conditions described above. The animal was initially saddled and
warmed up on the lead, moving to the right and left sides, at a walk and trot,
completing five full turns to each side. Then, each horse, in sequence, was
ridden by the trainer and led on the left side by the assistant guide (who at
the time was the handler), performing a circuit (not timed) and going through
the obstacles arranged in sequence (presented to the horse by an assistant)
inside the arena. The last data collection occurred as soon as the horse
entered the arena, and the final data collection occurred as soon as it
finished the circuit. Continuous HR measurements, post-exercise plasma
cortisol, and post-exercise IRT analysis were all performed in the covered
arena during this phase. The first was performed at the end of the first 30
days, and the second at the end of the experiment (last 30 days) – i.e., in
February and the end of March. Baseline HR, baseline IRT, and baseline cortisol
were measured inside each animal's stall. This phase lasted approximately 60
days, with an average frequency of four sessions per week and an average
duration of 40 minutes per animal, which may vary according to individual
response.
Gymnastics – physical conditioning, functional
gymnastics, and/or basic training: The gymnastics phase consisted of continuous
exercises performed throughout Phases 1, 2, and 3, lasting 30 minutes on
alternate days to the other training activities. Its objective was to promote
and maintain general physical conditioning, encompassing aspects related to
muscle strength, balance, motor coordination, and endurance, while specific
hippotherapy training and initial training were being established. These
activities involved work on a lead or at liberty (in Phases 1 and 2 and part of
the first 30 days of Phase 3) and outdoor or track work with basic training
movements – e.g., 20-15-10-8 m circles, half turns, reverse half turns,
serpentine turns, gallop starts, backing up, shoulder forward, leg yield (the
latter only at the end of the last month), transitions in different gaits
(walk, trot, gallop), small jumps (60-80 cm), poles, and barrels, also only in
Phase 3. The exercises were performed for 30 minutes, on alternate days with
the training days of each specific phase – except Sunday, when no animal was trained;
all had the entire day of rest, loose in paddocks.
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