Alpacas’ behavior during physical examination, and its relationships with health indicators and alpaca caretaker’s questionnaire survey

Abstract

Background

Behavioral responses during examinations can reflect stress and ease of handling. This study aimed to determine alpacas’ behavioral responses to body score and health assessment during physical examination, and to investigate correlations between alpacas’ behavior during physical examination (mean behavior score was computed for each animal based on the assessment of its behavioral responses: ease of introduction to the box, response to fixation, body language, vocalization, kicking, and spitting, on a 3-point scale), body condition score (BCS, on a 5-point scale) and other health indicators (on a 5-point scale), and caretaker questionnaire survey (based on evaluation of the individual’s behavioral characteristics and its ease of handling, using a 5-point Likert scale).

Results

Thirty alpacas (17 females, 13 males) took part in the study. The studied animals did not show many behavioral signs of stress during physical examinations, with a mean behavioral score of 1.33, and most of them were assessed with a BCS of 3.0, indicating their good condition. Spitting was negatively correlated with BCS and eye mucous membrane color, suggesting a potential link between animal stress and health. The duration of the physical examination was most influenced by the ease of introduction to the assessment box. Moreover, shorter examination times were associated with desirable traits such as gentleness and adaptation to new situations, assessed in the caretaker questionnaire survey. Females showed more stress-related behaviors during physical examination, and it took longer to examine them compared to males, although no sex differences were found in BCS and other health indicators or survey traits.

Conclusions

Physical examination is not a significantly stressful procedure for alpacas. However, some difficulties were noted during the introduction of the animal to the examination box, and signs of anxiety were observed in its body language. The results indicate a relationship between behavior assessment during physical examination and desirable traits assessed in the caretaker questionnaire. This study suggests that the future consideration of behavior indicators during human-animal relation could be helpful in evaluating alpacas’ health risks. The proposed assessment protocol combining behavior, the BCS and other health indicators offers a practical tool for breeders to monitor alpaca welfare.

Background

Routine health and body condition monitoring provides the perfect opportunity for alpacas to be evaluated for any signs of disease. Observation of animal behavior is considered a diagnostic tool, providing information about an animal’s physical and mental well-being [1]. Sickness-related behavior can be the first symptom of a health problem observed by an animal caretaker. Sickness reactions include decreased activity, loss of appetite and thirst, reduced grooming and social interaction, decreased exploratory behavior, cognitive dysfunction, as well as reduced sexual and maternal activity [2–5]. However, sickness behavior is less pronounced in alpacas compared to other herbivores, which are more vulnerable to predators and tend to manifest such responses.

The most popular health indicators include body temperature, heart and respiratory rates, the color of the eye mucous membranes, and sclera, as well as the appearance of teeth, toenails, and faeces. A more detailed picture of an alpaca’s health status can be provided by routine blood assessments and faecal monitoring for evidence of parasites [5, 6]. Regular evaluation of the body condition score (BCS) is also important in assessing the health of alpacas [5–8]. The BCS can be assessed via a scale of 1–5 or 1–10. Wagener et al. [8] analyzed many publications on condition assessment in SACs and reported that the 1–5 scale is more often used and 3.0 is considered the correct value. Most alpacas in e.g., Australia have a BCS of 3.0 [9]. Buchallik-Schregel et al. [10] also indicated that using the BCS scale is a reliable method of assessing the condition of alpacas. Regular health assessment aligns with the elements of ensuring welfare from the perspective of the Five Freedoms (freedom from disease, pain, and injury) [11, 12]. Some authors [13, 14] suggest, that behavioral predictors are related with body condition of the animals (e.g., sheep or horses). Thus, behavior indicators can be helpful in predicting some health changes. To our knowledge, no studies have integrated behavioral responses during routine physical examination with both objective health indicators and caretaker-derived temperament assessments in alpacas.

Health assessment can be partly performed by an alpaca’s caretaker, for example, regular BCS assessment, checking eye mucous membranes and sclera, as well as the appearance of teeth and toenails. However, some diagnostic tools can be used only by a veterinarian who is unfamiliar with the animals. Physical veterinary examination and blood sample collection are associated with animal restraint and close contact with an unfamiliar person, which could be potentially stressful for the examined animal [15]. During veterinary and husbandry procedures, alpacas may exhibit stress-related behaviors, such as vocalizations (moaning, screaming, and snorting), kicking, stomping, spitting, abruptly lying down and rising, rearing, and freezing [11, 12, 15–17]. There is also a common recommendation not to develop too intensive animal-human contact, especially in alpacas, before the age of 6–10 months [18], as it could lead to the development of an inappropriate behavior called Berserk Male Syndrome. This abnormal behavior is manifested by human-directed aggression because humans are viewed as conspecifics [15]. However, this behavioral disorder could be observed as a result of excessive socialization in bottle-fed alpacas and frequent youngs’ isolation from the conspecifics [19] and not because of early gentle human contact (e.g. stroking and touching) with young alpacas staying with mothers in their herd. The studies of Windschnurer et al. [15, 20] showed that early gentle human–animal contact (talking to the animals, stroking and touching them, walking through a group of alpacas), as well as tactile contact in later life and using training methods (e.g. Camelidynamics), can have positive effects on the ease of alpacas’ handling.

Human–animal relationship assessment is crucial for the alpacas’ welfare, and it can be measured by direct animal behavior observation, as well as by evaluating an animal handler’s opinion about a particular individual. A caretaker’s evaluation of the individual’s behavioral characteristics and its ease of handling is not an objective method; however, it is based on everyday contact during animal care and handling [21, 22]. The questionnaire completed by the caretaker could be beneficial, as it allows for a comparison of the objective animal behavior assessment with the subjective evaluation provided by a person taking daily animal care [22]. The caretaker’s recognition of easy-to-handle and “difficult” animals could significantly affect human-animal relations, influence animal behavior, and, consequently, have an impact on the physical and mental well-being of the animal.

This study aimed to determine alpacas’ behavioral responses to body score and health assessment during physical examination, and to investigate correlations between alpacas’ behavior during physical examination, body condition score and other health indicators, and caretaker questionnaire survey. We hypothesized that (1) the variables studied in the behavioral responses during physical examination, duration of the physical examination and health assessment score are related, (2) the behavioral traits surveyed in the caretaker questionnaire are correlated to each other and with behavioral responses and duration of the physical examination; and (3) the results of behavioral assessment during physical examination and behavioral traits surveyed in the caretaker questionnaire are different in females and males.

Methods

Animals

This study included 30 alpacas (17 females, 13 unneutered males) aged 1–19 years. The animals were kept on an alpaca farm in Lublin Province (southeastern Poland) and were used for both fibre production and agrotourism. Females and males were kept in separate buildings (the box area per individual was from 3m² for females to 5 m² for males) with hay and water available ad libitum. The animals were fed twice daily a mixed feed composed of dried alfalfa and wheat bran, supplemented with added grain, minerals, and vitamins. In the period from May to October, alpacas spend nine hours a day on pasture (a quarter of the area from 300 to 700 m²). The average temperature during the examination was 18 °C, no rain or strong winds, sunny, average cloud cover. The studied alpacas had daily contact with humans during routine procedures, such as feeding, cleaning, and handling. All animals were trained using the Camelidynamics method at least 6 months before the start of the experiment. The alpacas were under constant veterinary care and, were clinically healthy as well as they showed no behavioral symptoms of Berserk Male Syndrome.

Assessment of alpacas’ behavior, body condition score and health during the physical examination

The assessment was conducted during the day between 10am and 2pm. The assessment of behavior, BCS, and other health indicators during the physical examination was carried out in a small box (3 × 2 m) directly adjacent to the large box in which the given group of animals was housed. The procedure was performed by three (always the same) people (the caretaker – a person known to the animals, the assessor – a person unknown to the animals, and the person recording the results – a person unknown to the animals). The assessor was unfamiliar to the animals intentionally, to simulate the unknown veterinarian’s or professional breeder’s visit, for the assessment of animals’ response to an unfamiliar person performing physical examination activities. The order of the assessment was randomized and each animal was examined once. Before introducing the animal to the box, the caretaker put a halter on it, led it on a leash to the assessment box from the directly adjacent large box, and then fixed the alpaca using a “midline catch” (a handle with the hands from behind on the occiput and under the jaw) or by grabbing the neck and holding it in the withers area if necessary. None of the animals was sedated. The assessor (with animal behavior education and a few years’ experience in alpacas’ husbandry practices) had been trained previously, and all examination activities were performed in the same order and the same way for each animal. The assessor palpated the animal to assess its body condition (BCS) by checking the state of musculature and fat on the side behind the last lumbar vertebra, and other health indicators by checking the color of the eye mucous membranes, as well as the appearance of the toenails and the incisors. All elements were assessed on a scale of 1–5 points, where 3 points indicated the normal body condition/normal eye mucous membranes’ color/normal appearance of toenails and incisors, scores below 3 points represented the under conditioned animal/pale membranes/too short or broken toenails and incisors and scores above 3 points indicated the over conditioned animal/congested membranes/overgrown toenails and incisors. During the physical examination, the animals were also assessed for their behavioral responses (ease of introduction to the box, response to fixation, body language, vocalization, kicking, and spitting) on a scale of 1–3 points, where 1 indicated the most desirable behavior in terms of ease of handling. Additionally, the duration (s) of physical examinations were measured from the moment the animal’s led to the box till the end of the assessment. The mean health score was extracted based on body condition and other indicators of health assessment, whereas the mean behavior score was computed for each animal based on the behavioral assessment.

Detailed descriptions of body condition and health, as well as the behavioral assessment, are presented in Tables 1 and 2. The experimental procedures followed the body condition and health assessment methods described by Storey et al. [23], Van Saun and Herdt [24], Van Saun [25] as well as the authors of this study adapted the description of some health indicators: eye mucous membranes’ color, toenails and incisors appearance (based on Storey et al. [23], Van Saun and Herdt [24]) (Table 1). The behavior assessment (Table 2) followed the method described by Windschnurer et al. [15] and the authors of the study adapted the description of body language as a part of behavior assessment (based on Kapustka and Budzyńska [16, 26]).

Table 1.

Assessment of BCS and other health indicators during the physical examination in alpacas (based on storey et al. [23], Van Saun and Herdt [24], Van Saun [25])

Variable	Description
BCS (body condition score)	1 – emaciated 2 - thin 3 – moderate 4 - excess 5 - obese
Eye mucous membranes’ color	1 – mucous membranes very pale 2 – mucous membranes slightly pale 3 – mucous membranes pink 4 – mucous membranes reddened 5 – mucous membranes bloodshot and/or with petechiae
Toenails’ appearance	1 – more than one nail is too short, broken 2 – one nail is too short/broken 3 – nails trimmed evenly, of the correct length 4 – one nail slightly overgrown 5 – more than one nail is overgrown, curving sideways
Incisors’ appearance	1 – incisors very short, worn or broken 2 – incisors slightly too short 3 – incisors of correct length, straight 4 – incisors slightly overgrown 5 – incisors overgrown, crooked, protruding beyond the jaw, visible when the mouth is closed

Table 2.

Behavioural assessment during physical examination of body score and health in alpacas, with regard to particular behavioral responses (based on windschnurer et al. [15], Kapustka and Budzyńska [16, 26])

Variable	Description
Ease of introduction to the box	1 - Alpaca is easy to introduce to the box; it can be led without resistance 2 – Alpaca allows being led into the box, but they resist 3 – Alpaca puts up much resistance, does not want to enter the box, and requires the use of considerable force from the handler so that the animal does not escape
Fixation	1 – Alpaca stands still; the owner holds it only using the ‘midline catch’ 2 – Alpaca requires more substantial fixation around the neck and at the height of the withers 3 – Alpaca is very difficult to keep; it rears up and tries to escape
Body language	1 – Ears loose, body posture not too tense 2 – Ears flattened, tense body posture, tail slightly raised 3 – Ears flattened, tense body posture, tail held high, head raised (warning against spitting)
Vocalization	1 – No vocalization 2 – Vocalize 1–3 times during the examination 3 – Vocalize more than 3 times or continuously during the examination
Kicking	1 – No kicking 2 – Kick 1–3 times during the examination 3 – Kick more than 3 times during the examination
Spitting	1 – No spitting 2 – Spit 1–3 times during the examination 3 – Spit more than 3 times or continuously during the examination

Alpaca caretaker questionnaire survey

The alpaca caretaker agreed to complete a short survey questionnaire for each animal examined in this study. The survey questionnaire for the alpaca caretaker included 9 statements regarding the behavioral characteristics of the alpacas in terms of the ease of animal handling. Each point of the questionnaire was presented in the form of a statement: “Alpaca is.“, e.g. “Alpaca is curious”. The statements were divided into two groups describing: (1) positive (facilitating handling, desirable) traits of the alpaca, such as curious, gentle, calm, friendly towards people, quickly adapting to new situations and (2) negative (troublesome, undesirable) traits, such as dangerous, unpredictable, difficult to handle, and aggressive. The caretaker could indicate answers using a 5-point Likert scale, where: 5 meant ‘I strongly agree’, 4 meant ‘I rather agree’, 3 meant ‘I have no opinion’, 2 meant ‘I rather disagree’, and 1 meant ‘I strongly disagree’. Based on the results, the medians of positive and negative behavioral traits were computed for each animal assessed, as well as overall positive (all positive traits combined) and overall negative (all negative traits combined). The questionnaire for the alpaca caretaker was based on the survey developed by Windschnurer et al. [20].

Statistical analysis

Statistical analysis was performed using Statistica 13.3 software. The results were presented as descriptive statistics with the median (med.) and upper and lower quartiles (Q1, Q3) provided for the variables without a normal distribution and mean (x) and standard deviation (SD) or the variables with a normal distribution. The Shapiro-Wilk test was performed to assess the normality of the dependent variable distribution. The results showed that one trait (duration of physical examination (s)) had a normal distribution (p > 0.05), while others did not (p < 0.05). The Student’s t-test was then conducted for the variable with a normal distribution, and the Mann-Whitney U test and Wilcoxon paired test were used for those without a normal distribution, with t and p values as well as Z and p values provided from the respective tests. The Spearman R rank correlation analysis was performed to check for correlations between the studied variables. Differences were considered statistically significant at p < 0.05.

Results

The median of the behavior score of all alpacas during the physical examination did not exceed 2.0 points in any of the assessed behavioral responses (med.=1.33, Q1 = 1.17, Q3 = 1.50). The differences of results between particular forms of observed behaviors are presented in Table 3. The median BCS condition score of the entire herd was 3.0 points (Q1 = 3.0, Q3 = 3.0), and the median of the other health indicators was 3.0 points (Q1 = 2.88, Q3 = 3.13). The mean duration of the physical examination was 75.00 s (1 min 15 s; SD = 18.00 s), whereas the median of positive traits assessed in the survey questionnaire was 4.0 points (Q1 = 3.6, Q3 = 4.4), and the median of negative traits was 1.25 (Q1 = 1.0, Q3 = 2.0).

Table 3.

Results of the behavior assessment during the physical examination of alpacas

Variable	Med	Q1-Q3
Ease of introduction to the box	2.0A, a,b	1.0–2.0
Fixation	1.0a, c	1.0–1.0
Body language	2.0B, c,d	1.0–2.0
Vocalization	1.0e	1.0–2.0
Kicking	1.0A, B,e	1.0–1.0
Spitting	1.0b, d	1.0–1.0

Statistical significances: A,B - values marked with the same uppercase letter differ significantly at p < 0.001, a-e- values marked with the same lowercase letter differ significantly at p < 0.05

Relationships between behavioral responses during physical examination, duration of the physical examination and health assessment score

The color of eye mucous membranes was positively correlated with BCS condition (r = 0.433, p = 0.017) and with the appearance of incisors (r = 0.394, p = 0.031). The health assessment score was correlated with only a few behavioral traits, both during the behavioral assessment and in the questionnaire survey. Significant, negative correlations were found between BCS condition (r = −0.579, p < 0.001), color of eye mucous membranes (r = −0.455, p = 0.012), and health assessment score (r = −0.487, p = 0.006) and spitting frequency. A negative correlation between the nail appearance and the body language scores (r = −0.363, p = 0.049) was also found.

The duration of the physical examination was the most significantly correlated with the ease of introduction to the box (r = 0.522, p = 0.003).

Relationships between behavioral responses during physical examination, duration of the physical examination and behavioral traits from the alpaca caretaker’s questionnaire survey

The overall score of positive traits was negatively correlated with the overall score of negative traits (r = 0.420, p = 0.017). The correlations of positive and negative behavioral traits are provided in Table 4.

Table 4.

Correlations between positive and negative traits of alpacas assessed in the survey

Variables	Curiosity	Gentleness	Calmness	Friendly attitude towards people	Adaptati-on to new situations	Dangerous	Unpredictable	Difficult to use	Aggressive
Curiosity		0.243	0.022	0.540*	0.639**	0.084	−0.073	−0.114	0.169
Gentleness	0.243		0.806**	0.613**	0.469*	−0.532*	−0.411*	−0.417*	−0.363*
Calmness	0.022	0.806**		0.346	0.286	−0.715**	−0.539*	−0.592**	−0.553*
Friendly attitude towards people	0.540*	0.613**	0.346		0.539*	−0.204	−0.137	−0.194	−0.039
Adaptation to new situations	0.639**	0.469*	0.286	0.539*		0.026	−0.148	−0.167	0.291
Dangerous	0.084	−0.532*	−0.715**	−0.204	0.026		0.782**	0.793**	0.801**
Unpredictable	−0.073	−0.411*	−0.539*	−0.137	−0.148	0.782**		0.844**	0.696**
Difficult to use	−0.114	−0.417*	−0.592**	−0.194	−0.167	0.793**	0.843**		0.667**
Aggressive	0.169	−0.363*	−0.553*	−0.039	0.291	0.801**	0.696**	0.667**

* p-values with an asterisk are considered significant at p < 0.05

** p-values with two asterisks are considered significant at p < 0.001

Gentleness was a behavioral trait positively correlated with overall positive traits and negatively correlated with overall negative traits, with the exception of curiosity. In turn, the curiosity trait was positively correlated with a friendly attitude towards people and the adaptation to new situations. All negative traits were positively correlated with each other, and negatively with gentleness and calmness (Table 4). There was a statistically significant correlation between alpaca traits and alpaca behavior during physical examination, when the aggression trait was positively correlated with kicking (r = 0.421, p = 0.020).

The duration of the physical examination was negatively correlated with gentleness (r=−0.378, p = 0.040), adaptation to new situations (r=−0.414, p = 0.023) and the overall score of positive traits (r=−0.417, p = 0.022) (Fig. 1), but was not correlated with any negative trait (0.009 < r < 0.220, 0.243 < p < 0.963).

Fig. 1.

Scatterplot between the duration of physical examination and selected positive traits of alpacas assessed in the survey

Sex-specific behavior and health assessment score

Higher median behavioral scores during the physical examination were given to females (med.=1.5, Q1 = 1.33, Q3 = 1.67) compared to males (med.=1.17, Q1 = 1.00, Q3 = 1.33, Z=−2.846, p = 0.004) (Fig. 2). The mean of the examination duration was also higher in females than in males (F: x = 83.00 s, SD = 17.00; M: x = 64.00 s, SD = 12.00, t=−3.403, p = 0.002) (Fig. 3). The results of individual behavioral responses in female and male alpacas are presented in Table 5. No differences were found in the health assessment scores as affected by sex, (for example, body condition scores (BCS) F: med.=3.125, Q1 = 2.75, Q3 = 3.25, M: med.=3.0, Q1 = 3.0, Q3 = 3.125, Z=−0.021, p = 0.983).

Fig. 2.

Behavioral assessment of females and males during a physical examination

Fig. 3.

Mean duration of behavioral assessment of females and males during physical examination

Table 5.

Results of behavior assessment during physical examination in alpacas with regard to their sex

Variable		Females N = 17	Males N = 13	Z	p-value
Ease of introduction to the box	Med	2.0	1.0	−1.988	0.035*
	Q1-Q3	1.5–2.5	1.0–2.0
Fixation	Med	1.0	1.0	0.711	0.258
	Q1-Q3	1.0–1.0	1.0–1.5.0.5
Body language	Med	1.5	1.5	−1,569	0.082
	Q1-Q3	1.5–2.0.5.0	1.0–1.5.0.5
Vocalization	Med	1.0	1.0	−1.025	0.616
	Q1-Q3	1.0–2.0	align="left"1.0–1.5.0.5
Kicking	Med	1.0	1.0	−0.251	0.802
	Q1-Q3	1.0–1.0	1.0–1.0
Spitting	Med	1.0	1.0	−0.523	0.427
	Q1-Q3	1.0–1.0	1.0–1.0

* p-values with an asterisk are considered significant at p < 0.05

a - markings with the same letters differ significantly at p < 0.05.

a - markings with the same letters differ significantly at p < 0.05.

There were no statistically significant differences between females and males in the assessment of positive behavioral traits (F: med.=3.8, Q1 = 3.4, Q3 = 4.4, M: med.=4.5, Q1 = 3.8, Q3 = 4.6; Z = 0.867, p = 0.393) and negative behavioral traits (F: med.=1.25, Q1 = 1.0, Q3 = 2.0, M: med.=1.0, Q1 = 1.0, Q3 = 1.75; Z=−0.419, p = 0.676) in the questionnaire survey. Although not statistically significant, the median of positive traits was higher in males, and that of negative traits was higher in females.

Discussion

Most of the studied alpacas were assessed as calm and not causing any difficulties during the physical examination. The highest median of behavioral score occurred for two observed variables: ease of introduction to the box and body language. Introduction to the box requires separation from the herd, which causes stress for the alpacas [27]. Body language scored as two meant that the alpaca was stressed, but did not warn against spitting. Spitting is a kind of defensive aggression in alpacas, before it, the animals warn raising their head [26]. In this study, most alpacas did not warn before spitting, nor did they spit. It can be concluded that the animals were stressed due to separation from the herd, but they did not feel threatened seriously enough to defend themselves. Similar reactions were observed by Windschnurer et al. [15].

The body condition of the studied alpacas was proper. The body condition of alpacas can be rated on a 10-point scale [6] or a 5-point scale [28] using the BCS scale, but the 5-point scale is more common [8]. In this study, a score on a 1–5 scale was used. It defines the level of musculature and fatness, which was originally defined for cattle or sheep and also adapted for South American camelids [8, 28]. It is determined by palpation at the lumbar level, complemented by the area above the rib cage, between the hind and front legs, and around the vulva [6, 8, 28]. Additionally, the BCS score was supplemented with other health scores, which included the color of eye mucous membranes and the condition of the nails and incisors. On a 1–5 scale, a score of 3 was considered correct [8, 9]. Buchallik-Schregel et al. [10] emphasize that the BCS scale can also be used to create a model for body weight calculation in alpacas. In our study, we used the health assessment protocol extended by the color of eye mucous membranes, toenails and incisor appearances. This protocol can contribute to developing a practical tool for alpaca breeders, which could result in a more detailed yet easy-to-perform basic health assessment.

Relationships between behavioral responses during physical examination, duration of the physical examination and health assessment score

The first hypothesis was partially confirmed. In our research, we observed that two of six studied behavioral traits were correlated with indicators of alpaca health assessment. The frequency of spitting during physical examination was negatively correlated with BCS, the color of the eye mucous membranes, and the overall health assessment score. The higher the scores in these elements, the less the alpacas spit, i.e. they show stress-related behavior with less intensity. However, it is important to mention that our findings deal with health and temperament assessment on one alpaca farm and the studied animals were characterized by good overall health status and trained using the Camelidynamics method. To our knowledge, this is the first approach to highlight the combining of behavioral and health measures in alpacas in the aspect of human-animal interactions and animal welfare. Therefore, higher variability of studied traits and more potential relationships between studied indicators could be revealed in the future research including higher number of animals in different alpaca farms. The research by Stockman et al. [13] emphasizes that behavior traits may influence the level of food motivation and, thus, the animal’s body condition. On the other hand, the study on sheep indicated that the traits such as feed utilization, growth rate, and body condition score were similar in animals with low and high behavioral reactivity [29].

We found that the physical examination duration was significantly correlated with the ease of alpaca introduction to the box, and this relationship suggests that the ease of alpaca handling during this introduction had a significant impact on the duration of the physical examination. In the study on vicuñas it was also observed that the longer the animals were fixed, the more agonistic behaviors they showed [30]. However, in the study on shearing stress in alpacas, no such relationship was observed [31].

Relationships between behavioral responses during physical examination, duration of the physical examination and behavioral traits from the alpaca caretaker’s questionnaire survey

The second hypothesis was also partially confirmed. The behavioral assessment during the physical examination and the assessment using the caretaker questionnaire were not correlated, except for a negative correlation between the duration of the physical examination and gentleness, as well as adaptation to new situations. An assessment of an alpaca’s health by a stranger may be perceived by the animal as a stressor, triggering defensive behavior [15]. This type of examination did not cause significant difficulties in handling and assessing their body condition and other health indicators. Most of the studied alpacas were assessed as calm animals. Windschnurer et al. [15] noted that alpacas which had contact with humans from the beginning of their lives were much easier to handle and showed fewer symptoms of stress when performing such activities.

The owners emphasize the importance of tactile contact, staying with the animals, and talking to them during the later management of the herd. For these domesticated animals, the presence of humans and tactile contact with them is natural, and they should not be afraid of it. A study by Kapustka and Budzyńska [32] suggests that young animals exhibit higher curiosity than fear of novelty, making it easier to start working with them when they are young. In this study, all alpacas were trained using the Camelidynamics method. This method utilizes the habituation mechanism for touch and subsequently for equipment [19]. Therefore, the studied herd can be considered accustomed to human touch and presence earlier development of positive interactions based on animal learning may influence their responses during the physical examination procedure. That’s why we can state that working with alpacas in one area can facilitate handling in another. A similar relationship can also be observed in other species, such as cattle [33] and horses [34].

In this study, the authors demonstrated that positive features, especially gentleness and adaptability to new situations, were negatively correlated with the duration of the physical examination, which may confirm the accuracy of the owner’s assessment of the animals. Windschnurer et al. [20] also point out that individual features of alpacas influence their subjective perception by their owners (being with the animal is more/less comfortable).

An analysis of temperament traits reveals a correlation between positive and negative traits. In the study by Windschnurer et al. [20], respondents indicated that the most crucial animal features in the context of human-animal relations are curiosity and owner recognition. In this study, gentleness was the most correlated trait with the other ones, so it can be assumed to be the most universal factor. Most of the alpacas were rated as very gentle, calm, and friendly towards people, and they easily adapted to new situations, while their negative traits were rated as low. Gentleness and curiosity are significantly related to other traits perceived as positive, particularly in terms of ease of handling animals. The correlation between all negative traits suggests that such animals may be difficult to handle. Kelly et al. [35] indicate that in horses, human-animal relationships and the assessment of the horse’s reaction to the rider depend on many factors (e.g. horse emotional state and training experience, handling management practices). In the case of alpacas, no such studies have been conducted in this field, unlike those on horses. Therefore, each subsequent research of this type contributes to the development of the issue related to the impact of human-animal interactions on the behavior and welfare of alpacas.

Sex-specific behavior during physical examination

The third hypothesis was partially confirmed. The sex of the studied alpacas influenced the results of the behavioral assessment during the physical examination. Females had higher scores, indicating more frequent stress-related behaviors and higher reactivity levels. These findings suggest that females were more challenging to handle than males. The duration of the physical examination was also longer for females. Although these differences were no longer as significant in the survey assessment, stallions were still described as having a milder temperament and being easier to handle. In the study on shearing, alpaca females showed more intensive stress-related behaviors (abrupt movement, vocalizing, spitting) and physiological reactivity, having higher salivary cortisol levels [31]. Studies in other SAC species, such as vicuñas [30] and guanacos [36], have also confirmed higher behavioral reactivity in females compared to males. In contrast, the results of Pragai and Kovacs [37] on shearing stress demonstrated that alpaca males showed higher behavioral and physiological reactivity compared to females. Alpaca owners suggest that differences in behavioral reactivity could be due to the stronger bond that females have with the herd, and therefore, they can show greater stress during isolation (vocalization). Pollard and Littlejohn [27] confirmed that social isolation in alpacas was an intense stressor for them, but they did not determine whether there were any sex differences. Therefore, the issue related to sex differences requires further investigation.

Limitations of the study

The small sample size of animals and the fact that all the studied alpacas were from the same farm are the main limitations of the present study. The results of sex differences need to be interpreted with caution due to the limited number of animals studied. In the present study, the body condition and overall health of the herd were good, and few associations between behavioral indicators and poor condition or health could be found. In our study we used the questionnaire completed by the caretaker that was based on the subjective evaluation provided by a person directly involved in daily animal care. On the one side there was a risk of a potential caretaker bias in questionnaire responses but on the other side using this type of method is valuable, as it allows to compare the subjective temperament measures to the objective animal behavior assessment. In the further research, it would be interesting to study behavioral and health parameters in alpacas from different farms and to look if there are differences, when examining animals trained, as well as, not trained by the method of Camelidynamics. Using training methods (e.g. Camelidynamics), could increase the ease of alpacas’ handling and decrease the signs of stress-related behavior. Moreover, the limitations include also a lack of physiological stress measures (e.g. cortisol or the other hormones). The monitoring of these indicators could be an interesting topic in future research as combining physiological and behavioral measures can be more effective in detecting stress-related symptoms.

Conclusions

Physical examination is not a significantly stressful procedure for alpacas. While testing behavioral responses to handling during physical examination, the animals did not exhibit many stress-related behavior (vocalization, kicking, spitting). However, some difficulties were noticed during the introduction of the animal to the examination box, and signs of anxiety were observed in its body language. The results suggest a possible relationship between behavior assessment (e.g. easier handling and shorter physical examination time) and desirable traits (e.g. gentleness and adaptability to new situations) assessed in the caretaker questionnaire survey. The results also confirm that spitting was negatively associated with the body condition score and the color of the eye mucous membranes. Moreover, females were more reactive and more complex to handle than males. This study suggests that the future consideration of behavior indicators during human-animal relation could be helpful in evaluating alpacas’ health risks. However, as a small number of animals was included into this study, and they all originated from one farm, in the future, studying a larger number of alpaca farms could be involved to indicate a more general trend of the studied relationships. The proposed assessment protocol, which combines behavior, BCS, and health indicators, offers a practical tool for breeders to monitor alpaca welfare, facilitating the rapid detection of changes in the health status of animals, especially when assessments are conducted regularly.

Authors’ contributions

MB designed the work, analyzed the data, supervised and revised the work, and aquisited the funding. JK designed and conducted the study, analyzed the data, and drafted the work. MP conducted the study and analyzed the data, drafted the work. All authors have approved the submitted version and have agreed both to be personally accountable for the author’s own contributions and to ensure that questions related to the accuracy or integrity of any part of the work, even ones in which the author was not personally involved, are appropriately investigated, resolved, and the resolution documented in the literature.

Funding

This research received no external funding.

Data availability

The datasets analyzed during the current study are available from the corresponding author upon reasonable request.

Declarations

Ethics approval and consent to participate

The animal study protocol was approved by the Ethics Committee of the University of Life Sciences in Lublin, Poland, protocol code 78/2020, 11 December 2020.

The owners gave informed consent to participate and informed consent to the use of their animals in research.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.