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The aim of the present study was to test the cognitive ethology approach, which seeks to link cognitions and behaviours as they operate in everyday life with those studied in controlled lab-based investigations. Our test bed was the understanding of first-person and third-person perspectives, which in lab-based investigations have been defined in a diverse and multi-faceted manner. We hypothesized that because these lab-based investigations seek to connect with how first- and third-person perspective operates in everyday life, then either some of the divergent lab-based definitions are missing their mark or the everyday conceptualization of first- and third-person perspective is multi-faceted. Our investigation revealed the latter. By applying a cognitive ethology approach we were able to determine that a) peoples' everyday understanding of perspective is diverse yet reliable, and b) a lab-based investigation that applies these diverse understandings in a controlled setting can accurately predict how people will perform. These findings provide a 'proof of concept' for the cognitive ethology approach. Moreover, the present data demonstrate that previous lab-based studies, that often had very different understandings of first- and third-person perspective, were each in and of themselves valid. That is, each is capturing part of a broader understanding of perspective in everyday life. Our results also revealed a novel social factor not included in traditional conceptualizations of first-person third-perspective, that of eye gaze, i.e., eye contact is equated strongly with first-person perspective and the lack of eye-contact with third-person perspective.
To investigate the mechanism of Huannao Yicong Decoction (HYD) extract on improving of learning memory of transgenic amyloid precursor protein (APP)/presenilin 1 (PS1) mice, we randomly divided 60 transgenic APP/PS1 mice of 3 months old into 4 groups: the model group, the Donepezil group, the HYD-L group, and the HYD-H group, with 15 C57BL/6J mice of the same genetic background as the control group. These mice were gavaged for 6 months in a row. The results showed that the latency was significantly shortened and the number of passing through the original platform was increased. HYD extract can increase the amount of neurons and improve the morphological structure of Nissl body obviously. The γ-secretase activity and the expression of phosphorylated APP, Aβ1-40, and Aβ1-42 in hippocampal CA1 were significantly decreased. The expressions of protein and mRNA of PEN-2 and CREB in hippocampal were significantly downregulated. These results demonstrated that HYD extract can improve the memory ability of transgenic APP/PS1 mice, which was related to the protection of neurons and structure of Nissl body, reducing cleavage of APP and production of Aβ and inhibiting the activity of γ-secretase by decreasing CREB activity because of downregulated expression of PEN-2.
Traditional Chinese herbs (TCH) are currently gaining attention in disease prevention and health care plans. However, their general bitter taste hinders their use. Despite the development of a variety of taste evaluation methods, it is still a major challenge to establish a quantitative detection technique that is objective, authentic and sensitive. Based on the two-bottle preference test (TBP), we proposed a novel quantitative strategy using a standardized animal test and a unified quantitative benchmark. To reduce the difference of results, the methodology of TBP was optimized. The relationship between the concentration of quinine and animal preference index (PI) was obtained. Then the PI of TCH was measured through TBP, and bitterness results were converted into a unified numerical system using the relationship of concentration and PI. To verify the authenticity and sensitivity of quantified results, human sensory testing and electronic tongue testing were applied. The quantified results showed a good discrimination ability. For example, the bitterness of Coptidis Rhizoma was equal to 0.0579 mg/mL quinine, and Nelumbinis Folium was equal to 0.0001 mg/mL. The validation results proved that the new assessment method for TCH was objective and reliable. In conclusion, this study provides an option for the quantification of bitterness and the evaluation of taste masking effects.
Fast object tracking in real time allows convenient tracking of very large numbers of animals and closed-loop experiments that control stimuli for many animals in parallel. We developed MARGO, a MATLAB-based, real-time animal tracking suite for custom behavioral experiments. We demonstrated that MARGO can rapidly and accurately track large numbers of animals in parallel over very long timescales, typically when spatially separated such as in multiwell plates. We incorporated control of peripheral hardware, and implemented a flexible software architecture for defining new experimental routines. These features enable closed-loop delivery of stimuli to many individuals simultaneously. We highlight MARGO's ability to coordinate tracking and hardware control with two custom behavioral assays (measuring phototaxis and optomotor response) and one optogenetic operant conditioning assay. There are currently several open source animal trackers. MARGO's strengths are 1) fast and accurate tracking, 2) high throughput, 3) an accessible interface and data output and 4) real-time closed-loop hardware control for for sensory and optogenetic stimuli, all of which are optimized for large-scale experiments.
Ciwujia Tablets (CWT) are produced by concentrating and drying the extract solution of the dried rhizome of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim [Araliaceae; E. senticosus radix et rhizoma]. Besides, CWT is included in the 2020 edition of Chinese Pharmacopoeia and is widely used in the treatment of insomnia. It mainly contains eleutheroside B, eleutheroside E, isofraxidin, eleutheroside C, ciwujiatone, and chlorogenic acid, as well as other chemical components. Although the clinical efficacy of CWT in treating insomnia has been confirmed, its functions and pharmacological effects have not been systematically evaluated and its mechanism of action in the treatment of insomnia remains unclear. Therefore, in this study, behavioral, energy metabolism, and metabonomics methods were applied to systematically evaluate the effect of CWT on insomnia. Additionally, urine metabonomics based on UPLC-Q-TOF-MS/MS were utilized to identify potential endogenous biomarkers of insomnia, detect the various changes before and after CWT treatment, explore the metabolic pathway and potential target of CWT, and reveal its pharmacological mechanism. Results revealed that CWT increased inhibitory neurotransmitter (5-HT and GABA) content and reduced the content of excitatory neurotransmitters (DA and NE). Moreover, CWT enhanced autonomous behavioral activity, stabilized emotions, and promoted the return of daily basic metabolic indexes of insomniac rats to normal levels. The urine metabolomics experiment identified 28 potential endogenous biomarkers, such as allysine, 7,8-dihydroneopterin, 5-phosphonooxy-L-lysine, and N-acetylserotonin. After CWT treatment, the content of 22 biomarkers returned to normal levels. The representative markers included N-acetylserotonin, serotonin, N-methyltryptamine, and 6-hydroxymelatonin. Additionally, the metabolic pathways in rats were significantly reversed, such as tryptophan metabolism, folate biosynthesis, phenylalanine metabolism, and tyrosine metabolism. Ultimately, it is concluded that CWT regulated tryptophan metabolism, folate biosynthesis, phenylalanine metabolism, and other metabolic levels in the body. This drug has been confirmed to be effective in the treatment of insomnia by regulating the content of serotonin, 6-hydroxymelatonin, N-acetylserotonin, and N-methyltryptamine to a stable and normal level in tryptophan metabolism.
Despite being closely related, dogs perform worse than wolves in independent problem-solving tasks. These differences in problem-solving performance have been attributed to dogs' greater reliance on humans, who are usually present when problem-solving tasks are presented. However, more fundamental motivational factors or behavioural traits such as persistence, motor diversity and neophobia may also be responsible for differences in task performance. Hence, to better understand what drives the differences between dogs' and wolves' problem-solving performance, it is essential to test them in the absence of humans. Here, we tested equally raised and kept dogs and wolves with two unsolvable tasks, a commonly used paradigm to study problem-solving behaviour in these species. Differently from previous studies, we ensured no humans were present in the testing situation. We also ensured that the task was unsolvable from the start, which eliminated the possibility that specific manipulative behaviours were reinforced. This allowed us to measure both persistence and motor diversity more accurately. In line with previous studies, we found wolves to be more persistent than dogs. We also found motor diversity to be linked to persistence and persistence to be linked to contact latency. Finally, subjects were consistent in their performance between the two tasks. These results suggest that fundamental differences in motivation to interact with objects drive the differences in the performance of dogs and wolves in problem-solving tasks. Since correlates of problem-solving success, that is persistence, neophobia, and motor diversity are influenced by a species' ecology, our results support the socioecological hypothesis, which postulates that the different ecological niches of the two species (dogs have evolved to primarily be scavengers and thrive on and around human refuse, while wolves have evolved to primarily be group hunters and have a low hunting success rate) have, at least partly, shaped their behaviours.
Persistence in object manipulation has been consistently associated with problem-solving success and it is known to be affected, at the individual level, by life experience. Differences in life experiences are particularly poorly studied in the problem-solving context and mainly refer to the comparison between wild and captive animals. Dogs represent interesting study subjects, since dog populations differ widely in their life experiences. In this comparative study we investigated subjects' persistence when presenting a novel object containing food that could not be accessed (impossible task) to three dog populations with very diverse life experiences: free-ranging village dogs (in Morocco), pet dogs (in Vienna) and captive pack living dogs (Wolf Science Center-WSC). We found that pet dogs and captive dogs (WSC) were more manipulative and persistent than free-ranging dogs. The low persistence of free ranging-dogs is unlikely the effect of a lack of exposure to objects, since they are confronted with many human' artefacts in their environment daily. Instead, we suggest that the higher persistence of captive dogs and pet dogs in comparison to free-ranging dogs might be due to their increased experience of human-mediated object interaction. This provides subjects with a socially guided experience in manipulating and interacting with objects increasing their motivation to engage in such tasks.
Automated monitoring of the movements and behaviour of animals is a valuable research tool. Recently, machine learning tools were applied to many species to classify units of behaviour. For the monitoring of wild species, collecting enough data for training models might be problematic, thus we examine how machine learning models trained on one species can be applied to another closely related species with similar behavioural conformation. We contrast two ways to calculate accuracies, termed here as overall and threshold accuracy, because the field has yet to define solid standards for reporting and measuring classification performances. We measure 21 dogs and 7 wolves, and find that overall accuracies are between 51 and 60% for classifying 8 behaviours (lay, sit, stand, walk, trot, run, eat, drink) when training and testing data are from the same species and between 41 and 51% when training and testing is cross-species. We show that using data from dogs to predict the behaviour of wolves is feasible. We also show that optimising the model for overall accuracy leads to similar overall and threshold accuracies, while optimizing for threshold accuracy leads to threshold accuracies well above 80%, but yielding very low overall accuracies, often below the chance level. Moreover, we show that the most common method for dividing the data between training and testing data (random selection of test data) overestimates the accuracy of models when applied to data of new specimens. Consequently, we argue that for the most common goals of animal behaviour recognition overall accuracy should be the preferred metric. Considering, that often the goal is to collect movement data without other methods of observation, we argue that training data and testing data should be divided by individual and not randomly.
Dogs are looking at and gaining information from human faces in a variety of contexts. Next to behavioral studies investigating the topic, recent fMRI studies reported face sensitive brain areas in dogs' temporal cortex. However, these studies used whole heads as stimuli which contain both internal (eyes, nose, mouth) and external facial features (hair, chin, face-outline). Behavioral studies reported that (1) recognition of human faces by dogs requires visibility of head contour and that (2) dogs are less successful in recognizing their owners from 2D pictures than from real human heads. In contrast, face perception in humans heavily depends on internal features and generalizes to 2D images. Whether putative face sensitive regions in dogs have comparable properties to those of humans has not been tested so far. In two fMRI experiments, we investigated (1) the location of putative face sensitive areas presenting only internal features of a real human face vs. a mono-colored control surface and (2) whether these regions show higher activity toward live human faces and/or static images of those faces compared to scrambled face images, all with the same outline. In Study 1 (n = 13) we found strong activity for faces in multiple regions, including the previously described temporo-parietal and occipital regions when the control was a mono-colored, homogeneous surface. These differences disappeared in Study 2 (n = 11) when we compared faces to scrambled faces, controlling for low-level visual cues. Our results do not support the assumption that dogs rely on a specialized brain region for processing internal facial characteristics, which is in line with the behavioral findings regarding dogs inability to recognize human faces based on these features.
The results of current wolf-dog studies on human-directed behaviors seem to suggest that domestication has acted on dogs' general attitudes and not on specific socio-cognitive skills. A recent hypothesis suggests that domestication may have increased dogs' overall sociability (hypersociability hypothesis). The aim of the present study was to test one aspect of the hypersociability hypothesis, whereby dogs should be more interested in social human contact compared to wolves, and to investigate the relative roles of both domestication and experience on the value that dogs attribute to human social contact. We compared equally raised wolves and dogs kept at the Wolf Science Center (WSCw, WSCd) but also dogs with different human socialization experiences i.e., pet dogs and free-ranging dogs. We presented subjects with a simple test, divided in two phases: in the Pre-test phase animals were exposed to two people in succession. One person invited the animal for a social/cuddle session (contact provider) and the other fed the animal (food provider). In the Test phase, animals could choose which of the two persons to approach, when both stood in a neutral posture. We directly compared WSCd with WSCw and free-ranging dogs with pet dogs. We found that in the Pre-test, WSCd and free-ranging dogs spent more time with the contact provider than WSCw and pet dogs, respectively. The results regarding the free-ranging dog and pet dog comparison were surprising, hence we conducted a follow-up testing pet dogs in a familiar, distraction-free area. Free-ranging dogs and this group of pet dogs did not differ in the time spent cuddling. In the test phase, WSCd were more likely than WSCw to approach the two experimenters. However, neither for the WSCd-WSCw comparison nor for the free-ranging dogs-pet dogs comparison, we could find a clear preference for one person over the other. Our findings support the idea that domestication has affected dogs' behavior in terms of their overall interest in being in proximity with a human partner also in case of dogs with a relatively sparse socialization experience (free-ranging dogs). However, it remains unclear what the driving motivation to interact with the human may be.
Assistance dogs must manage stress efficiently because they are involved in challenging tasks. Their welfare is currently a fundamental issue. This preliminary study aimed to compare assistance dogs (AD; n = 22) with pet dogs (PD; n = 24), using blood neuromodulator indicators to help find biomarkers that can improve the AD breeding, selection, training, and welfare monitoring. Both populations originated from different breeds, are of different ages, and had different lifestyles. Basal peripheral concentrations of prolactin (PRL), serotonin (5-HT), free (fOT) and total (tOT) oxytocin were measured by immunoassays. Multiple linear regressions were performed to assess the effect of activity, age, sex, and their interactions on these parameters. Correlations between neurohormonal levels were analyzed. No interactions were significant. fOT and tOT concentrations were significantly influenced by age (p < 0.0001 and p = 0.0002, respectively) and dogs' activity (p = 0.0006 and p = 0.0277, respectively). A tendency was observed for age effect on PRL (p = 0.0625) and 5-HT (p = 0.0548), as well as for sex effect on tOT (p = 0.0588). PRL concentrations were heterogenous among AD. fOT and tOT were significantly but weakly correlated (Pearson's r = 0.34; p = 0.04). Blood prolactin, serotonin, and oxytocin may represent biomarkers to assess workload and chronic stress-related responses in ADs and eventually improve their selection and training.
The impossible task paradigm has been extensively used to study the looking back behaviour in dogs. This behaviour is commonly considered a social problem-solving strategy: dogs facing an unsolvable task, soon give up and look back at the experimenter to ask for help. We aimed to test if the looking back in an impossible task does indeed represent a social problem-solving strategy. We used a modified version of the classic impossible task, in which the subjects simultaneously faced three possible and one impossible trials. Additionally, subjects were tested in four different conditions: social condition (with an unknown experimenter); asocial condition (subject alone); 'dummy' human condition (with a 'dummy' human); object condition (with a big sheet of cardboard). Finally, we compared two populations of dogs differing in their experience of receiving help from humans: 20 pet dogs tested in their houses and 31 free-ranging dogs tested in Morocco. We found that the pet dogs and free-ranging dogs had similar persistence in interacting with the impossible task in all conditions. Moreover, subjects looked back with similar latencies at the human, at the dummy human and at the object. Overall, pet dogs looked back longer at the human than free-ranging dogs. This could be interpreted as pet dogs being more attracted to humans and/or having a stronger association between humans and food than free-ranging dogs. Concluding, the looking back in an impossible task does not represent a problem-solving strategy. This behaviour seems rather linked to the subject's persistence, to the salience of the stimuli presented, and potentially to the past reinforcement history.
While personality and cognition are distinct domains, some personality traits may affect the capacity for problem-solving. It was suggested that there is a positive association between the Playfulness trait and problem-solving performance in humans. Studies on giftedness (extremely good capacity in the case of a specific skill), typically aimed to reveal the genetic, experiential, and mental origins of such extreme inter-individual variation. We exploited recent findings on giftedness in a specific cognitive skill, object label learning, in dogs to explore the potential association between this exceptional skill and personality traits. We administered the Dog Personality Questionnaire to 21 gifted dog owners and compared the personality traits of their dogs to those of matched samples of 43 Hungarian and 101 Austrian typical dogs, i.e., dogs lacking this exceptional capacity. Since most Gifted Word Learner dogs are Border collies, we restricted our analysis to dogs of this breed. We hypothesized that the Gifted Word Learner dogs may show higher levels of Playfulness. As expected, we found that the gifted Border collies were rated as more playful than both the Hungarian and Austrian typical ones. Our results suggest that an extremely high level of Playfulness is associated with giftedness in a specific cognitive trait in dogs: the capacity to learn object verbal labels, thus opening new possibilities for comparative research on the relationship between giftedness and personality.
Exceptional performance is present in various human activities but its origins are debated and challenging to study. We report evidence of exceptional performance and qualitative variation in learning object-names in dogs. 34 naïve family dogs and 6 knowledgeable individuals that knew multiple toy names, found in 2 years of search around the Globe, were exposed to 3 months of training to learn two novel toy-names and were tested in two-way choice tests. Only 1 naïve and all 6 knowledgeable dogs passed the tests. Additionally, only these dogs learned at least 10 new toy names over the 3 months, showing qualitative variation in this capacity. Although previous object-name knowledge could provide an explanation for the superior performance of the knowledgeable dogs, their rarity and the absence of previous training of this skill point to exceptional giftedness in these individuals, providing the basis to establish dogs as a model-species for studying talent.
When addressing preverbal infants and family dogs, people tend to use specific speech styles. While recent studies suggest acoustic parallels between infant- and dog-directed speech, it is unclear whether dogs, like infants, show enhanced neural sensitivity to prosodic aspects of speech directed to them. Using functional magnetic resonance imaging on awake unrestrained dogs we identify two non-primary auditory regions, one that involve the ventralmost part of the left caudal Sylvian gyrus and the temporal pole and the other at the transition of the left caudal and rostral Sylvian gyrus, which respond more to naturalistic dog- and/or infant-directed speech than to adult-directed speech, especially when speak by female speakers. This activity increase is driven by sensitivity to fundamental frequency mean and variance resulting in positive modulatory effects of these acoustic parameters in both aforementioned non-primary auditory regions. These findings show that the dog auditory cortex, similarly to that of human infants, is sensitive to the acoustic properties of speech directed to non-speaking partners. This increased neuronal responsiveness to exaggerated prosody may be one reason why dogs outperform other animals when processing speech.
During social interactions, acoustic parameters of tetrapods' vocalisations reflect the emotional state of the caller. Higher levels of spectral noise and the occurrence of irregularities (non-linear phenomena NLP) might be negative arousal indicators in alarm calls, although less is known about other distress vocalisations. Family dogs experience different levels of stress during separation from their owner and may vocalise extensively. Analysing their whines can provide evidence for the relationship between arousal and NLP. We recorded 167 family dogs' separation behaviour including vocalisations, assessed their stress level based on behaviour and tested how these, their individual features, and owner reported separation-related problems (SRP) relate to their whines' (N = 4086) spectral noise and NLP. Dogs with SRP produced NLP whines more likely. More active dogs and dogs that tried to escape produced noisier whines. Older dogs' whines were more harmonic than younger ones', but they also showed a higher NLP ratio. Our results show that vocal harshness and NLP are associated with arousal in contact calls, and thus might function as stress indicators. The higher occurrence of NLP in older dogs irrespective to separation stress suggests loss in precise neural control of the larynx, and hence can be a potential ageing indicator.
The neuropeptide oxytocin (OT) has been shown to enhance dogs' ability to perform an object choice task (OCT) involving the use of human pointing cues, when delivered intranasally. This study aimed at further investigating whether OT enhances task performance by increasing choices made, or by increasing correctness of choices made, and to compare these treatment effects to dog appeasing pheromone (DAP), known to balance emotional activation in dogs. Hence, we compared OCT performance between three groups of dogs: (i) dogs administered OT and a sham collar, (ii) dogs administered a saline placebo and a DAP collar, and (iii) control dogs administered a saline placebo and a sham collar. All three groups consisted of a combination of male and female pet dogs and assistance-dogs-in-training currently living with a volunteer carer. The study also evaluated the effect of intranasal OT and/or DAP on plasma levels of OT, and prolactin; which has previously been linked with anxiety in dogs. The dogs' emotional state was measured using the Emotional Disorders Evaluation in Dogs (EDED) scale. The owners'/carers' degree of anxious- and avoidant-style attachment to their dogs was accessed using the Pet Attachment Questionnaire (PAQ). Interesting descriptive data appeared for both treatment groups. Particularly, in OT group, we obtained significant results demonstrating that intranasal OT enhances OCT performance in dogs compared to control, by increasing the percentage of correct choices, but not the number of choices, made. Results also support that the mode of action of intranasal OT is via direct access to the brain and not via the blood, since no elevation of plasma OT (or prolactin) levels were observed after intranasal administration in this study. Similarly, DAP application did not significantly alter OT or prolactin peripheral concentrations. Several differences were observed between fostered and pet dogs, namely: fostered dogs demonstrated higher levels of serum prolactin, made more choices on the OCT compared to pet dogs but were not more likely to be correct, and were fostered by carers with higher avoidant attachment scores than pet dog owners. These findings implicate consideration of potential carer and training consequences for assistance dogs.
Resting-state networks are spatially distributed, functionally connected brain regions. Studying these networks gives us information about the large-scale functional organization of the brain and alternations in these networks are considered to play a role in a wide range of neurological conditions and aging. To describe resting-state networks in dogs, we measured 22 awake, unrestrained individuals of both sexes and carried out group-level spatial independent component analysis to explore whole-brain connectivity patterns. In this exploratory study, using resting-state functional magnetic resonance imaging (rs-fMRI), we found several such networks: a network involving prefrontal, anterior cingulate, posterior cingulate and hippocampal regions; sensorimotor (SMN), auditory (AUD), frontal (FRO), cerebellar (CER) and striatal networks. The network containing posterior cingulate regions, similarly to Primates, but unlike previous studies in dogs, showed antero-posterior connectedness with involvement of hippocampal and lateral temporal regions. The results give insight into the resting-state networks of awake animals from a taxon beyond rodents through a non-invasive method.
Chasing motion is often used to study the perception of inanimate objects as animate. When chasing interaction and independent motions between two agents are displayed simultaneously on a screen, we expect observers to quickly perceive and recognise the chasing pattern (because of its familiarity) and turn their attention to the independent motion (novelty effect). In case of isosceles triangles as moving figures, dogs and humans both display this behaviour, but dogs initially preferred to look at the chasing pattern whereas humans started to increase their gaze towards the independent motion earlier. Here, we compared whether family cats perceive moving inanimate objects as animate and whether their looking behaviour is similar to that of small family dogs. We displayed a chasing and independent motion side by side on a screen in two consecutive trials and assessed subjects' looking behaviour towards the motions. Similarly to previous studies, we found that dogs eventually looked longer at the independent motion, but cats preferred to look at the independent motion at the beginning of the video display and only later shifted their attention to the chasing motion. No difference was found in the frequency of gaze alternation of the two species. Thus, although cats discriminate between the chasing and independent motions, it is not clear whether this discrimination is controlled by animate motion cues. The difference may originate from their ecological situation and/or may be explained by specific perceptual mechanisms.
The olfactory acuity of domestic dogs has been well established through numerous studies on trained canines, however whether untrained dogs spontaneously utilize this ability for problem solving is less clear. In the present paper we report two studies that examine what strategies family dogs use in two types of olfaction-based problems as well as their success at various distances. In Study 1, thirty dogs were tasked with distinguishing a target, either their covered owner (Exp 1) or baited food (Exp 2), from three visually identical choices at distances of 0m (touching distance), 1m, and 3m. There were nine consecutive trials for each target. We found that in Exp 1 the dogs successfully chose their owners over strangers at 0m and 1m, but not at 3m, where they used a win-stay strategy instead. In Exp 2 the dogs were only successful in choosing the baited pot at 0m. They used the win-stay strategy at 1m, but chose randomly at 3m. In Study 2, a different group of dogs was tested with their owners (Exp 1) and baited food (Exp 2) at just the 3m distance with two possible targets in 10-10 trials. In Exp 1 the dogs' overall performance was at chance level; however, when analyzed by trial, we noticed that despite tending to find their owners on the first trial, they generally switched to a win-stay strategy in subsequent trials, only to return to correctly choosing their owners based on olfaction in the later trials. In Exp 2, the dogs chose randomly throughout. We also found that dogs who relied on visual information in the warm-up trials were less successful in the olfaction-based test. Our results suggest that despite their ability to successfully collect information through olfaction, family dogs often prioritize other strategies to solve basic choice tasks.
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