Archivos Latinoamericanos de Producción Animal. 2022. 30 (4)
Welfare assessment of horses ridden in the
Costa Rica National Horse Parade
Recibido: 20211203. Aceptado: 20220822
1 Corresponding author: ana.ariasesquivel@ucr.ac.cr
2 Department of Animal Sciences, University of Florida, Gainesville, FL
3 Instituto Clodomiro Picado, Universidad de Costa Rica, San José 11501, Costa Rica
311
Luis A. VillalobosVillalobos
Ana Margarita AriasEsquivel1
Abstract. The National Horse Parade of Costa Rica (NHPCR) is an equestrian event held annually to celebrate the
national day of the horse rider. Public concern regarding the welfare of horses and riders during the event has
increased due to the removal of horses in poor body condition, spine injuries, and hoof problems. An Assessment
of horse behavior and stress response during these events has never been conducted. The objective of this study
was to evaluate the horse behavior, fecal glucocorticoid metabolites and body weight as indicators of welfare
during the NHPCR. In the Experiment 1, body weight and fecal glucocorticoid metabolites were measured in 13
Costarricense de Paso horses participating in the NHPCR. All measures were recorded at the horses' home stable 48
hours before and 24 hours after the event. In Experiment 2, three stations (equally spaced along the 3kilometer
parade route) were used to randomly conduct a scan sampling of the behavior of 513 horses. Eyewrinkle and eye
white presence were recorded on 48 horses using photographs taken during the event. Fecal glucocorticoid
metabolites showed no significant differences (p > 0.05) at the two time points in the horses evaluated, and there
was a significant weight loss (7.08 kg) 24 hours after the event (p = 0.00034). The scan sampling provided evidence
that traits such as sweating (60.74 %), bit chewing (54.70 %), and active gait (65 %) were more common than head
tossing (28.88 %), neck above the withers (6.83 %), hyperflexion (28.71 %), and ears pinned backwards (31.19 %). A
strong eye wrinkle was detected on 70 % of the horses, while only 16 % had the presence of eyewhite. Although
our results for fecal glucocorticoid metabolites were not conclusive, other behavioral traits indicated that over fifty
per cent of the horses attending the NHPCR experience some level of stress.
Keywords: body weight, behavior, stress response, glucocorticoid metabolite
Evaluación del bienestar de caballos montados en el desfile nacional
de caballos de Costa Rica
Resumen. El Tope Nacional de Costa Rica (NHPCR) es un evento ecuestre realizado anualmente para celebrar el
día nacional del caballista. La preocupación pública sobre el bienestar de los caballos y jinetes ha aumentado debido
a la cantidad de caballos sacados del evento debido a mala condición corporal, lesiones en columna, problemas de
cascos y dificultades en carga y descarga. Aunque se han implementado acciones para salvaguardar el bienestar de
los caballos en años recientes, no se ha evaluado su comportamiento y los niveles de estrés durante este evento. El
objetivo del estudio fue evaluar el comportamiento, los metabolitos de glucocorticoides en heces y el peso corporal
como indicadores de bienestar de caballos durante el NHPCR. En el Experimento 1 se determinó el peso corporal y
los niveles de cortisol fecal en 13 caballos Costarricense de Paso 48 horas antes y 24 horas después del evento. En el
Experimento 2, tres estaciones (equidistantes a lo largo de la ruta de 3 kilómetros del recorrido) se usaron para
evaluar el comportamiento de 513 caballos. Se registró la presencia de arrugas y conjuntiva del ojo en 48 caballos
con fotografías tomadas en el evento. No hubo diferencias significativas (p > 0.05) en el análisis de metabolitos de
glucocorticoides fecales en los dos momentos de muestreo y hubo una pérdida significativa (p = 0.00034) de peso 24
horas después del evento (7.08 kg). Se halló evidencia de que características como la sudoración (60.74 %),
mordisqueo del bocado (54.70 %) y paso activo (65 %) fueron más comunes que la sacudida de cabeza (28.88 %),
cuello levantado (6.83 %), hiperflexión del cuello (28.71 %) y orejas posicionadas hacia atrás (31.19 %) durante el
evento. Se detectó una arruga fuerte en los ojos (70 %) y presencia de la conjuntiva (16 %) en los caballos. Aunque
los resultados de glucocorticoides fecales no fueron concluyentes, otras características de comportamiento indicaron
que más de la mitad de los caballos asistentes al NHPCR experimentan algún nivel de estrés.
Palabras clave: peso corporal, comportamiento, respuesta de estrés, metabolitos de glucocorticoides
www.doi.org/10.53588/alpa.300404
Escuela de Zootecnia. Universidad de Costa Rica. San José, San Pedro Montes de Oca, Costa Rica
Carissa L. Wickens2 Erika Camacho3
312
The National Horse Parade of Costa Rica (NHPCR)
is a cultural event celebrated on December 26 to
celebrate the National Horseman's Day. Around 2000
horses participate in this event every year, being
ridden in a threekilometre route in the streets of San
José, Costa Rica. In recent years, incidents have raised
welfare concerns for equines participating in the event.
According to Camacho (2017), the National Service for
Animal Health (SENASA) had to remove sixteen
horses from the event in 2017 due to various traumas.
Likewise, five sets of nonpermitted spurs carried by
the riders were seized. Even though SENASA and the
Municipality of San José enforce local regulations to
safeguard the welfare of participating animals, the
question of whether horses undergo stress during the
event has not been addressed empirically (Pena, 2015).
Physiological measures provide insight into how
animals perceive environmental challenges (Wikelski
and Cooke 2006). The two main stress axes involved in
triggering the release of stress hormones are the
autonomic nervous system and the hypothalamic
pituitaryadrenocortical (HPA) axes (Flauger 2010).
The neuroendocrine stress response is a cascade of
events mediated by the HPA axis. When an organism
encounters a stressor, the HPAaxis is activated to
initiate an appropriate centralized stress response
(Möstl et al. 2002). The end products of the HPA axis
are glucocorticoids (cortisol and corticosterone),
steroid hormones, that play a key role in mobilizing
glucose, mediating energy balance and regulating
animal metabolism, nutrition, reproduction, and
immunity (Cox et al. 2010). Thus, glucocorticoid
concentrations are not equivalent to measuring a stress
response directly, rather, they represent an end
product from a signaling cascade (MacDougall
Shackleton et al. 2019).
Glucocorticoid metabolites can be used as indicators
of stress in horses. Kruguer (2010) found a positive
correlation between fecal glucocorticoid metabolites
and serum cortisol in horses. In addition, fecal
glucocorticoid metabolites have been used in other
equine species, such as zebras. Franceschini et. al (2008)
reported that fecal glucocorticoid metabolites have
been helpful in measuring stress caused by
transportation, management, and acclimation to
captivity in zebras. The same authors reported that
fecal glucocorticoid metabolites were higher during the
first days of captivity and then decreased when the
animals became accustomed to the unfamiliar
environment.
The behavior assessment is also a reliable indicator
of stress in horses. The position of the neck, ears and
nostrils have been used as indicators of welfare (Hintze
et al. 2016). According to Hintze et al. (2016), eye
wrinkles and eye expression could be useful visual
indicators that may allow the identification of stress in
Introduction
Avaliação do bemestar de cavalos montados no desfile nacional de cavalos da Costa Rica
Resumo. O Tope Nacional de Costa Rica (NHPCR) é um evento equestre realizado anualmente para celebrar o Dia
Nacional do Cavaleiro. A preocupação do público com o bemestar dos cavalos e cavaleiros aumentou devido ao
número de cavalos retirados do evento com inadequadas condições corporais, lesões na coluna vertebral, problemas
nos cascos e dificuldades de carga e descarga. Embora ações tenham sido implementadas para salvaguardar o bem
estar dos cavalos nos últimos anos, seu comportamento e os níveis de estresse durante este evento não foram
avaliados. O objetivo do estudo foi avaliar ocomportamento, os metabolitos de glicocorticoides em fezes e o peso
corporal como indicadores de bemestar dos equinos durante o NHPCR. No Experimento 1, o peso corporal e os
níveis de cortisol fecal foram medidos em 13 cavalos da Costa Rica Paso 48 horas antes e 24 horas após o evento. No
Experimento 2, três estações (igualmente espaçadas ao longo do percurso de 3 quilômetros) foram usadas para
avaliar o comportamento de 513 cavalos. A presença de rugas e conjuntiva ocular foi registrada em 48 cavalos com
fotografias tiradas no evento. Não houve diferenças significativas (p > 0.05) na análise dos metabólitos fecais de
glicocorticoides nos dois momentos de amostragem e houve perda de peso significativa (p = 0.00034) 24 horas após
o evento (7.08 kg). Evidenciouse que características como sudorese (60.74 %), mordeduras (54.70 %) e marcha ativa
(65 %) foram mais comuns do que sacudir a cabeça (28.88 %), pescoço levantado (6.83 %), hiperflexão do pescoço
(28.71). %) e orelhas posicionadas para trás (31.19 %) durante o evento. Forte enrugamento dos olhos (70 %) e
presença da conjuntiva (16 %) foram detectados nos cavalos. Embora os resultados de glicocorticoides fecais tenham
sido inconclusivos, outras características comportamentais indicaram que mais da metade dos cavalos atendidos no
NHPCR experimentam algum nível de estresse.
Palavraschave: peso corporal, comportamento, resposta ao estresse, metabolitos de glicocorticoides
Arias et al.
ISSNL 10221301. Archivos Latinoamericanos de Producción Animal. 2022. 30 (4): 311319
Materials and Methods
Ethics Approval, local and animals
This study was performed in San José, Costa Rica,
from November 2019 through August 2020. Thirteen
Costarricense de Paso breed horses (312 years) were
randomly selected from a single farm where horses
were kept under similar conditions of housing (3.5 x
3.5 m stalls), feeding (forage and concentrate twice per
day), and exercise (once per week). All the horses had
previously attended the NHPCR.
Experiment 1: Body weight and fecal glucocorticoid
metabolites measurements
Both horses' body weight and fecal samples were
taken at twotime points at their individual stables (48
h before and 24 h following the event) as
recommended by Palme et al. (1996) and Möstl et al.
(1999). Horses were weighed by using a heavyduty
scale that was placed on a pliable wooden platform.
Fresh fecal samples were handcollected from the
stalls and pieces of grass, hay, feed, or litter were
carefully removed from the sample using a brush to
avoid contamination from other materials in the
analyses. Feces were mixed and homogenized by
reducing their particle size by hand in individual
aluminum containers and 100 grams of each sample
were weighed and placed in individual plastic bags
for later storage at 20 °C in a freezer in the Anatomy
and Physiology Laboratory of the Department of
Animal Sciences of the University of Costa Rica.
Glucocorticoid metabolite analysis was performed
using a commercially available 11
oxoetiocholanonolone ELISA kit (Cayman chemical
®). The analytical sensitivity of the assay was 0.2 ng/
ml and the intraessay coefficient was 11.4 %,
following the methodology suggested by Flauger et al.
(2010).
Experiment 2: Observational study
An observational study was performed during the
NHPCR of 2019 across three stations equally spaced
(approximately 500 m) along the 3kilometer parade
route (start, the halfway point, and end of the event).
Three trained observers per station performed scan
sampling using an ethogram (Table 1 and Table 2) on
both horses and their respective riders for a total of
513 observations. Observers were positioned
approximately 2 meters away from the analyzed
horses and observations were recorded every 3
minutes (and lasted 3 minutes) on a random horse.
Observations were taken from 12p.m. to 5p.m., which
corresponds to the duration of the event.
313
Welfare Assessment of Horses Participating in a National Horse Parade
horses. Moderate to high stress levels have been
reported to increase oral behaviors, the presence of
flared nostrils, and flattened or pinned ears, which has
been correlated with an increase in heart rate and
salivary cortisol (Young et al. 2012). Therefore, learning
how to recognize eye wrinkles and behavioral
indicators of stress during horse events could be a
valuable tool for horse owners and entities responsible
for regulating animal welfare.
Given the incidents presented in previous years
related to horses' welfare during the NHPCR,
additional research can provide insight into the level of
stress the animals experience as well as identify
strategies to amend such problems. Therefore, the
objective of this study was to evaluate the horse
behavior, fecal glucocorticoid metabolites and body
weight as indicators of welfare during the NHPCR.
Table 1. Ethogram utilized for conducting horse behavior measures during the National Horse Parade of Costa Rica.
Variable Status Ethogram interpretation
Sweating Present Visible and evident humidity (wet coat) was present under the saddle and /or the neck and flanks.
Absent Nonvisible or evident humidity was present under the saddle and/or the neck and flanks.
Bit chewing Present Actively chewing the bit and moving the lips side to side at least once during the 3 minutes of observation.
Absent Nonvisible active chewing of the bit or moving the lips side to side during the 3 minutes of observation.
Head tossing Present Head tossed up and down or sided to side at least once during the 3 minutes of observation.
Absent Nonvisible head tossing during the 3 minutes of observation.
Active gait Present The horse maintains an active gait (prancing) in the same spot or while in movement during the 3 minutes of
observation.
Absent No presence of active gait.
Ear position Normal Ears at any other position except pinned backwards during the 3 minutes of observation.
Pinned Ears pinned very close to its head at least once during the 3 minutes of observation.
backwards
Neck position Normal The horse maintains a balanced position on the neck compared to the withers.
Upwards The horse keeps the neck at an angle greater than 45° from the withers.
Hyperflexion The horse's mouth is as close to the chest during the 3 minutes of observation.
ISSNL 10221301. Archivos Latinoamericanos de Producción Animal. 2022. 30 (4): 311319
314
Experiment 1: Body weight and fecal glucocorticoid
metabolites
The feces collected in this experiment had similar (p
> 0.05) concentrations of glucocorticoid metabolites at
the two time points for each horse. Body weight of
horses was significantly (p < 0.05) lower after the event
(Table 3).
Results and Discussion
Table 2. Ethogram utilized for conducting rider behavior measures during the 2019 NHPCR.
Variable Status Ethogram interpretation
Excessive kicking Present The rider is excessively kicking on the flanks of the horse during the 3 minutes of observation
Absent No evident excessive kicking on the flanks of the horse during the 3 minutes of observation
Excessive use
of the whip Present The rider is actively hitting the horse with the whip during the 3 minutes of observation
Absent The rider may have a whip, but it is not actively hitting the horse with it during the 3 minutes of observation.
Aggressive
behavior Present The rider is noisy (yelling) or bothering other horses and/or riders during the 3 minutes of observation.
Absent Calm rider
Presence of
alcohol Present Alcoholic beverages were visible in the rider's hands or in the horse's saddle. Rider was drinking during the 3
minutes of observation.
Absent Alcoholic beverages were not visible in the rider's hands or in the horse's saddle. Rider iwa not drinking
during the 3 minutes of observation.
Eye wrinkles and eyewhite assessment
The visual assessments for eye wrinkles and eye
white were taken at the second station through a
photographic record by using a semiprofessional
digital camera (Panasonic Lumix DMCFZ70). Pictures
of 50 random horses were taken during the event
every 10 minutes to collect data from different sections
and time points throughout the day. After the event,
the photographic record went through a screening
process (pictures with low resolution) to choose the
best pictures to be analyzed, following the
methodology of Hintze et al (2016). The eye wrinkles
were assessed and classified into three categories (no
wrinkle, weak or strong), while eyewhite was
assessed by presence or absence.
Statistical analysis
Experiment 1
Data were analyzed with R Studio software R v3.6.1
(R Core Team, Vienna, Austria). The experimental
design consisted of a onefactor analysis
corresponding to the time points when data were
collected (pre and postevent). A paired ttest was
conducted to evaluate changes in both fecal
glucocorticoid metabolites and body weight of the
horses pre and post event.
Experiment 2
Data for the observational variables were analyzed
by a Pearson Chi Squared test in R Studio software.
The variables were treated differently for each
observation station, allowing them to estimate the
probabilities of the behavior occurring at each station.
Descriptive statistics were used to describe the eye
wrinkles and eyewhite present in the horses
participating in the NHPCR.
Table 3. Fecal glucocorticoid metabolites (FGM) and body weight of thirteen horses attending the National Horse Parade of
Costa Rica.
Variable (units) Timepoint Mean Standard deviation pvalue
FGM Before 0.80 ng.ml1 0.29 0.1889
(8.0 ng.g1)
After 1.02 ng.ml1 0.29
(10.2 ng.g1)
Body weight Before 451 Kg 53.2 0.0013
After 446 Kg 49.8
Arias et al.
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315
Previous studies have found that animals learn to
cope with situations expected to generate stress by
manifesting different behaviors (Fureix et al. 2013).
Stressful situations can trigger the release of
glucocorticoid metabolites, remaining at levels higher
than normal during events such as the NHPCR. Merl
et al. (2000) measured fecal glucocorticoid metabolites
in stallions following castration, known to be a factor
that causes stress in equines. Before castration, these
authors report median concentrations of 10.5 nmol/kg
feces (3.19 ng/g1) and after castration values increased
to 26.2 (7.97 ng/g1) and 50 nmol/kg feces (15.22 ng/g
1). In addition, Hinchcliffe et al. 2021 found that fecal
glucocorticoid metabolites were elevated from the
second day following a stressor (a roundup) and then
slowly returned to basal levels over the next 2 weeks.
Franceschini et al. (2007) monitored fecal
glucocorticoid metabolites in zebras from the time of
capture, during captivity and postrelease as an
indicator of the stress of translocation and acclimation
to a new environment. These authors found that when
held in pens at Meru Park 3–4 and 5–6 weeks after
capture, the zebra had higher fecal glucocorticoid
metabolites than either at capture or compared to non
translocated controls.
Despite the similar concentrations of fecal
glucocorticoid metabolites found in this study pre and
post event, horses showed behavioral indicators of
stress, which are also considered stressors that could
activate the HPA axis during the NHPCR. Pawluski et
al. (2017) compared horses' baseline levels of cortisol
and several welfare indicators and revealed that horses
whose welfare was clearly compromised (as indicated
by unusual ears backward position, presence of
vertebral problems or hematological anomalies, e.g.,
anaemia) also had lower levels of cortisol (and
metabolites). The results of our study expand earlier
findings evidencing low or no change in cortisol levels
after intensive training or racing of Thoroughbred
fillies (Nogueira et al. 1997) or chronic suboptimal
living conditions of horses (Visser et al. 2008). This
could be evidence that the HPA axis could have an
attenuated activity when welfare is compromised.
Low levels of glucocorticoids and/or decreased
HPA reactivity after different types of chronic
stressors have been reported in several species,
including farm animals and humans (Herman et al.
2016). The results of our study can be explained by the
"general adaptation syndrome", a threestage process
that describes the physiological changes the body goes
through when under stress (Selye 1956). The third
phase of the "general adaptation syndrome" is
exhaustion, where the organism is no longer able to
cope with stress, thus making stress hormones not to
be released as normal (Selye 1956).
Another factor that could explain our results is the
"allostatic overload" mechanism in the case of chronic
stress and the development of pathologies in humans
(McEwen et al. 2003), which may be extended to other
species, such as horses. According to this mechanism,
stress hormones play a key role in maintaining
homeostasis through change (allostasis). When
allostasis mediators, such as cortisol, are released in
response to stressors or lifestyle factors (diet, sleep
wake cycles and exercise), they facilitate adaptation
and are generally beneficial. However, when the
stressor is sufficiently intense or prolonged (chronic),
then the allostatic load can increase dramatically until
it reaches "allostatic overload". Allostatic overload can
then lead to wear and tear of the system on the body
and brain such that it no longer responds
appropriately to the change in allostasis (McEwen et al.
2003). It might be possible that chronic confrontations
with repeated welfarerelated stressors in horses
involving an allostatic overload, may be reflected by
maintaining the levels of fecal glucocorticoid
metabolites when exposed to an event such as the
NHPCR. The horses evaluated had restricted access to
social contact and roughage (except hay and alfalfa) as
well as intensive riding techniques (rein tension, high
hands or amplified aids, strong control attempts on
horses' movements) were used in the stables, which
can alter welfare and animal's perception of its
environment, creating an attenuated HPA axis as
explained previously.
The lack of difference in fecal glucocorticoid
metabolites could also be attributed to an adequate
management during the event (i.e., careful
transportation and acclimation before and during the
event), which might have reduced stress and
maintained the comfort of the horses without altering
the concentration of fecal glucocorticoid metabolites.
Also, the horses that participated in this study have
been previously exposed to or participated in events
like the NHPCR, which can help to decrease the
likelihood of having stressed animals. Stull and Rodiek
(2000) indicate that a decrease in body weight could
occur due to dehydration during transport or exercise.
The horses used in this study spent at least 12 hours
since they left the stables for the NHPCR and until
they returned later that same day, limiting the forage
and concentrate intake for half a day.
Harris (2015) reports that the energetic response to
acute stress is determined by the nature and severity
of the stressor. Typical responses to acute stressors
Welfare Assessment of Horses Participating in a National Horse Parade
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316
include inhibition of food intake, increased heat
production, and increased activity with sustained
changes in body weight, behavior, and HPA reactivity.
These responses were common in horses attending to
the NHPCR, which can explain body weight loss
found after the event.
Experiment 2: Observational study
The behavior indicators evaluated were significantly
different (p < 0.05) between the stations (Table 4).
Except for head tossing and active gait, all the
behaviors consistently showed higher probabilities of
occurring in the second and third stations. The high
probabilities (> 0.50) found for sweating, bit chewing
and ear position in the second and third stations,
indicate that horses may be experiencing stress and
exhaustion as they progress in the NHPCR.
Table 4. Behavioral recordings on horses participating in the National Horse Parade of Costa Rica.
Condition Absence Presence Chi Square Station 1 Station 2 Station 3
( %) ( %) pvalue probability probability probability
Sweating 39.3 60.7 < 0.01 0.345 0.712 0.655
Bit chewing 45.3 54.7 0.044 0.477 0.592 0.587
Head tossing 71.1 28.9 0.032 0.347 0.288 0.221
Active Gait 65.1 34.9 0.013 0.376 0.256 0.399
Ear position 31.2 68.8 < 0.01 0.637 0.655 0.784
* Probabilities are shown only for variables with a significant pvalue.
* Probabilities correspond to the chances of a horse showing a specific behavior at each of the stations.
Almost one third (p < 0.05) of the horses visually
assessed showed a hyperflexion of the neck during the
NHPCR (Table 5). Several studies have suggested that
hyperflexion may negatively affect the horse and
compromise welfare, causing impaired vision, airway
obstruction, overstretched muscles and ligaments and
delayed neuromuscular transmission (Von Borstel et
al. 2009, Cehak et al. 2010, Elgersma et al. 2010).
Christensen et al. (2014) mentioned that not only
hyperflexion of the neck is uncomfortable and painful
for the horse while riding, but it is also a concern in
terms of animal welfare.
Table 5. Neck position on horses attending the National Horse Parade of Costa Rica.
Condition Normal ( %) Upwards ( %) Hyperflexion ( %) Chi Square
p value
Neck Position 64.5 6.8 28.7 < 0.01
Alcohol consumption by the riders participating in
the NHPCR represented a small proportion in this
study (Table 6), indicating that strict enforcement was
applied during the event. The rider's behavior may
result in stress, exhaustion, or pain in the horses.
Overall, rider behavior was mostly nonaggressive,
with low use of the whip or kicking, which could
indicate that this factor did not trigger stress in the
horses attending NHPCR.
Table 6. Rider behavior evaluated during the National Horse Parade of Costa Rica.
Condition Absence Presence Chi Square Station 1 Station 2 Station 3
( %) ( %) pvalue probability probability probability
Alcohol consumption 91.4 8.6 < 0.01 0.1077 0.0196 0.1212
Excessive kicking 97.3 2.7 0.6243
Excessive whip use 96.3 3.7 0.8946
Aggressive rider behavior 97.1 2.9 0.2926
* Probabilities are only shown for variables with a significant pvalue.
* Probabilities correspond to the chances of a horse showing a specific behavior at each of the stations.
Visual assessment of eyewhite and eyewrinkles
showed that roughly twothirds of the horses
evaluated had strong eye wrinkles (Table 7). In
contrast, a small percentage (16.7 %) had a visible eye
white presence.
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Conclusions
This project was feasible thanks to the collaboration
of the students from the University of Costa Rica and
the San Francisco de Asís University, as well as the
invaluable help of PhD. José Ramón Molina and DVM.
Paula Capella. Any opinions, findings, conclusions, or
recommendations expressed in this publication are
those of the author(s) and do not necessarily reflect the
view of the National Institute of Food and Agriculture
(NIFA) or the United States Department of Agriculture
(USDA).
Despite the lack of difference in fecal glucocorticoid
metabolites found in this study, indicators such as the
presence of eye wrinkles and eyewhite indicate that a
moderate proportion of the horses attending the
NHPCR experience some level of stress. In addition,
the loss in body weight of the horses from experiment
1 can be mainly attributed to dehydration due to the
number of hours spent in the NHPCR, which creates a
different environment for the horse, affecting their
feeding and drinking schedule and intake.
Implementing "rehydration stations" (utilizing
disposable recipients for each horse to avoid sanitary
issues) during the event can attenuate the effects of
this issue, allowing horses to drink small doses of
water throughout the event. Horse owners must be
aware that previous conditioning (by progressive and
consistent training) and offering hay during
transportation may be factors that could positively
impact animal welfare during the NHPCR. The
presence of active sweating, bit chewing, and ears
pinned backwards found in the observational study
revealed that a higher percentage of the horses might
be presenting stress responses during the NHPCR. The
higher probabilities found for sweating, bit chewing,
active gait and, ears pinned backwards at stations 2
and 3 can be attributed to exhaustion. A closer follow
up of each horse at different moments through the
NHPCR could provide further information on the
potential of fecal glucocorticoid metabolites to assess
stress in horses. Other variables, such as heart rate
variability or eye temperature, could support our
findings of whether the NHPCR is causing stress to the
horses participating. With a larger scale welfare
assessment, the organizing authorities could identify
stress in the overall event. Eyewhite and eyewrinkles
are rapid and lowcost indicators that may be
evaluated during the NHPCR, allowing the organizing
authorities to assess stress in horses during the event.
Specific training about animal welfare before the event
would allow the organizing authorities of the NHPCR
to reduce the percentage of horses presenting welfare
issues or discomfort and promptly address the issue or
remove the animal from the event.
Acknowledgements
Conflicts of interest: The authors of this article disclose no conflicts of interest
Table 7. Eye wrinkle condition in horses attending the
National Horse Parade of Costa Rica.
Wrinkle condition Percentage ( %)
n = 48
No wrinkle 12.5
Weak 16.7
Strong 70.8
Eye wrinkles (also known as worry wrinkles) have
been empirically associated with stress and pain
among equine owners, professionals, and
veterinarians (Merkies et al. 2019). A negative
emotional state tends to be associated with wrinkles
formed above the eyeball (Hintze et al. 2016). The
Equine Facial Action Coding system describes this
expression as the inner brow raiser (levator angulu
oculi medialis muscle and corrugator supercili
muscle), which increases the perceived size of the eye,
but not the aperture of the eye (Wathan et al. 2015).
Hintze et. al (2016) conducted a study using diffe
rent tests to provoke a positive or negative stimulus
on the horse and found that eye wrinkles and eye
white can be useful indicators of fear or pain. They
evaluated qualitative impression (first subjective
impression of the expression of eyewrinkles), eyelid
shape, markedness of the wrinkles, presence of eye
white, number of wrinkles, and the angle between the
line through the eyeball and the highest wrinkle. In
our study, we followed Hintze et al. (2016)
methodology but only the presence of eyewrinkles
and eyewhite were evaluated. Within the
randomized pictures taken during the event, we
found that most horses had a strong eye wrinkle (70
%), and a smaller percentage had visible eyewhite
(16.7 %), indicating that the NHPCR could be
generating stress in the animals (Table 7).
Ethics statement: This study was approved by the University of Costa Rica Animal Care and Use Committee
(CICUA).
Welfare Assessment of Horses Participating in a National Horse Parade
ISSNL 10221301. Archivos Latinoamericanos de Producción Animal. 2022. 30 (4): 311319
318
Author contributions: All the authors contributed equally to the writing of this paper. All the authors revised the
manuscript critically and approved its final version.
Funding: This work was supported by The Hatch Act of 1887 (Multistate Research Fund), project accession no.
1018072 from the USDA National Institute of Food and Agriculture.
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