Pain behaviors before and after treatment of oral disease in cats using video assessment: a prospective, blinded, randomized clinical trial.

BACKGROUND: Specific behaviors associated with pain in cats with oral disease have not been consistently studied. The aim of this exploratory study was to identify pain-induced behaviors in cats before and after treatment of oral disease using video assessment. Twenty-four cats (6 ± 3.3 years old; 4.9 ± 1.7 kg) were included in a prospective, blinded, randomized clinical trial. Cats were equally divided into minimal (G1: minimal dental treatment) or severe (G2: multiple dental extractions) oral disease groups. After acclimation at day 0, they underwent oral examination, radiographs, scaling, and dental extractions under general anesthesia (anesthetic protocol: acepromazine, hydromorphone, propofol, isoflurane, meloxicam, and local anesthetic blocks; day 1), and were discharged at day 6. Cats were filmed remotely for 10 min using a wide-angle glass lens camera before surgery (baseline) and throughout the study at different time points (36 h of video recording). The videos consisted of four parts namely general, playing, feeding and post-feeding behaviors. A board-certified behaviorist evaluated the duration/frequency of different behaviors based on an ethogram, which were analyzed using linear mixed models and a generalized linear model, respectively (p < 0.05). RESULTS: In comparison with baseline, duration of "not pawing the face" was significantly shorter at day 3 in G2. These cats spent significantly longer time "standing" and "laying" at days 3 and 6, respectively; G1 spent significantly less time "walking" and "standing" at days 3 and 4, respectively and significantly longer time "immobile" at day 3. Duration of "no/slow tail movement" was significantly longer in G2 than G1 at day 5. Duration of "pawing the ribbon" (playing) was significantly shorter in G2 than G1 at day 1. Feeding and post-feeding behaviors with soft food were not significantly different between groups or over time. Frequency of "difficulty grasping dry food" was significantly higher in G2 than G1 up to day 6. Frequency of post-feeding "head shaking" was significantly higher in both groups at day 6 when compared with baseline. CONCLUSIONS: This study identified pain-induced behaviors in cats undergoing treatment of oral disease. These behaviors may be used to differentiate painful versus pain-free cats in clinical practice.

An automated approach to the quantitation of vocalizations and vocal learning in the songbird.

Studies of learning mechanisms critically depend on the ability to accurately assess learning outcomes. This assessment can be impeded by the often complex, multidimensional nature of behavior. We present a novel, automated approach to evaluating imitative learning. Conceptually, our approach estimates how much of the content present in a reference behavior is absent from the learned behavior. We validate our approach through examination of songbird vocalizations, complex learned behaviors the study of which has provided many insights into sensory-motor learning in general and vocal learning in particular. Historically, learning has been holistically assessed by human inspection or through comparison of specific song features selected by experimenters (e.g. fundamental frequency, spectral entropy). In contrast, our approach uses statistical models to broadly capture the structure of each song, and then estimates the divergence between the two models. We show that our measure of song learning (the Kullback-Leibler divergence between two distributions corresponding to specific song data, or, Song DKL) is well correlated with human evaluation of song learning. We then expand the analysis beyond learning and show that Song DKL also detects the typical song deterioration that occurs following deafening. Finally, we illustrate how this measure can be extended to quantify differences in other complex behaviors such as human speech and handwriting. This approach potentially provides a framework for assessing learning across a broad range of behaviors like song that can be described as a set of discrete and repeated motor actions.

When and Where Learning is Taking Place: Multisynaptic Changes in Strength During Different Behaviors Related to the Acquisition of an Operant Conditioning Task by Behaving Rats.

Although it is generally assumed that brain circuits are modified by new experiences, the question of which changes in synaptic efficacy take place in cortical and subcortical circuits across the learning process remains unanswered. Rats were trained in the acquisition of an operant conditioning in a Skinner box provided with light beams to detect animals' approaches to lever and feeder. Behaviors such as pressing the lever, eating, exploring, and grooming were also recorded. Animals were chronically implanted with stimulating and recording electrodes in hippocampal, prefrontal, and subcortical sites relevant to the task. Field synaptic potentials were evoked during the performance of the above-mentioned behaviors and before, during, and after the acquisition process. Afferent pathways to the hippocampus and the intrinsic hippocampal circuit were slightly modified in synaptic strength during the performance of those behaviors. In contrast, afferent and efferent circuits of the medial prefrontal cortex were significantly modified in synaptic strength across training sessions, mostly at the moment of the largest change in the learning curve. Performance of behaviors nondirectly related to the acquisition process (exploring, grooming) also evoked changes in synaptic strength across training. This study helps to understand when and where learning is being engraved in the brain.

Reevaluating cases of trait-dependent diversification in primates.

OBJECTIVES: Previous investigations have identified several ecological traits that may have shaped differences in species richness among primate clades by influencing speciation and extinction probabilities. In this study, I reevaluate these cases in light of concerns about high false-positive rates of the methods used to detect trait-dependent diversification. MATERIALS AND METHODS: Data for six traits were taken from the literature: activity pattern, sociality, mating system, terrestriality, frugivory, and mutualistic interactions with angiosperms (i.e., seed dispersal and pollination). These traits were treated as binary variables and analyzed in a phylogenetic framework using likelihood-based and nonparametric methods for estimating trait-specific diversification rates. RESULTS: Of the traits analyzed here, only activity pattern was found by all methods to be correlated with diversification: diurnal primates diversify at higher rates than nocturnal primates. Solitary foragers have a lower rate of diversification than group-living primates, but this link is driven by the association between solitariness and nocturnality. DISCUSSION: These results indicate that ecological traits have had a more limited influence on primate diversification than suggested by previous studies. The false positives detected here can be linked with heterogeneity in diversification caused by sources other than the focal traits. One of these sources is the very high rate of diversification in Cercopithecoidea found in previous investigations. Another is the correlation between activity patterns and diversification. The underlying causal mechanism for the contrast in diversification dynamics between diurnal and nocturnal primates is unclear, but this correlation highlights the important role of activity pattern in shaping primate evolutionary history.

Resource-Efficient Pet Dog Sound Events Classification Using LSTM-FCN Based on Time-Series Data.

The use of IoT (Internet of Things) technology for the management of pet dogs left alone at home is increasing. This includes tasks such as automatic feeding, operation of play equipment, and location detection. Classification of the vocalizations of pet dogs using information from a sound sensor is an important method to analyze the behavior or emotions of dogs that are left alone. These sounds should be acquired by attaching the IoT sound sensor to the dog, and then classifying the sound events (e.g., barking, growling, howling, and whining). However, sound sensors tend to transmit large amounts of data and consume considerable amounts of power, which presents issues in the case of resource-constrained IoT sensor devices. In this paper, we propose a way to classify pet dog sound events and improve resource efficiency without significant degradation of accuracy. To achieve this, we only acquire the intensity data of sounds by using a relatively resource-efficient noise sensor. This presents issues as well, since it is difficult to achieve sufficient classification accuracy using only intensity data due to the loss of information from the sound events. To address this problem and avoid significant degradation of classification accuracy, we apply long short-term memory-fully convolutional network (LSTM-FCN), which is a deep learning method, to analyze time-series data, and exploit bicubic interpolation. Based on experimental results, the proposed method based on noise sensors (i.e., Shapelet and LSTM-FCN for time-series) was found to improve energy efficiency by 10 times without significant degradation of accuracy compared to typical methods based on sound sensors (i.e., mel-frequency cepstrum coefficient (MFCC), spectrogram, and mel-spectrum for feature extraction, and support vector machine (SVM) and k-nearest neighbor (K-NN) for classification).

What does variation in primate behavior mean?

Interest in intraspecific behavioral variation has grown with concerns about the ability of primates to adapt to the rapidly changing ecological and demographic conditions that threaten their survival. Now, in addition to identifying the causes and phylogenetic distribution of normative, species-specific behavior patterns for interspecific comparisons, there is widespread recognition of the need to incorporate intraspecific variation. This variation is evident across groups and populations of the same species as well over the long histories of single groups of long-lived, socially complex animals with overlapping generations. Yet, analyses of both cross-sectional and longitudinal data require explicit criteria about how to classify and interpret behavioral variation, and must be sensitive to the limitations of space-for-time substitutions in these comparisons. Current approaches have made great advances, but there remains an urgent challenge of understanding intraspecific variation in a way that will facilitate the development of new predictive models to assess population resilience and extinction risks in the face of climate change and other anthropogenic influences.

A dictionary of behavioral motifs reveals clusters of genes affecting Caenorhabditis elegans locomotion.

Visible phenotypes based on locomotion and posture have played a critical role in understanding the molecular basis of behavior and development in Caenorhabditis elegans and other model organisms. However, it is not known whether these human-defined features capture the most important aspects of behavior for phenotypic comparison or whether they are sufficient to discover new behaviors. Here we show that four basic shapes, or eigenworms, previously described for wild-type worms, also capture mutant shapes, and that this representation can be used to build a dictionary of repetitive behavioral motifs in an unbiased way. By measuring the distance between each individual's behavior and the elements in the motif dictionary, we create a fingerprint that can be used to compare mutants to wild type and to each other. This analysis has revealed phenotypes not previously detected by real-time observation and has allowed clustering of mutants into related groups. Behavioral motifs provide a compact and intuitive representation of behavioral phenotypes.

Altered hippocampal function before emotional trauma in rats susceptible to PTSD-like behaviors.

Posttraumatic stress disorder (PTSD) is an anxiety disorder that occurs after experiencing a traumatic event. Susceptibility to PTSD exists, as only some trauma-exposed individuals develop this condition. Investigating susceptibilities in animal models can contribute to understanding the etiology of the disorder. We previously reported an animal model which allows reliable pre-classification of rats as susceptible (Sus) or resistant (Res) to developing a PTSD-like phenotype after a later trauma. Here we report that Sus, compared to Res, rats have altered hippocampal function, along the septo-temporal axis, prior to experiencing a traumatic event. In Experiment I, Res and Sus rats explored a novel box twice. Using a cellular imaging method for assessing plasticity-related immediate-early gene expression in large neuronal ensembles, Arc/Homer1a catFISH, we show that Sus rats have smaller vCA3 ensembles during the second exploration. This suppressed vCA3 activation in Sus rats was not due to a difference in exploratory behavior, or to a difference in Arc/Homer1a expression in the basolateral amygdala (BLA). BLA is a main source of inputs to vCA3, but both the ensemble size and overlap of BLA ensembles activated during the two explorations was similar to that of Res rats. Additionally, Sus rats had significant 'infidelity' in their dorsal hippocampal representations of the second event: a lower overlap, compared to Res rats, of Arc/Homer1a-expressing ensembles activated during the two explorations (the size of the ensembles were similar to those of Res rats). These differences were revealed only in conditions of relatively low stress, because they were not observed when Sus and Res rats experienced fear conditioning (Experiment II). Combined, the findings show that altered hippocampal function exists before experiencing emotional trauma in susceptible rats and suggest that this is a risk factor for PTSD.

Identification of blood meal sources of Lutzomyia longipalpis using polymerase chain reaction-restriction fragment length polymorphism analysis of the cytochrome B gene.

An analysis of the dietary content of haematophagous insects can provide important information about the transmission networks of certain zoonoses. The present study evaluated the potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial cytochrome B (cytb) gene to differentiate between vertebrate species that were identified as possible sources of sandfly meals. The complete cytb gene sequences of 11 vertebrate species available in the National Center for Biotechnology Information database were digested with Aci I, Alu I, Hae III and Rsa I restriction enzymes in silico using Restriction Mapper software. The cytb gene fragment (358 bp) was amplified from tissue samples of vertebrate species and the dietary contents of sandflies and digested with restriction enzymes. Vertebrate species presented a restriction fragment profile that differed from that of other species, with the exception of Canis familiaris and Cerdocyon thous. The 358 bp fragment was identified in 76 sandflies. Of these, 10 were evaluated using the restriction enzymes and the food sources were predicted for four: Homo sapiens (1), Bos taurus (1) and Equus caballus (2). Thus, the PCR-RFLP technique could be a potential method for identifying the food sources of arthropods. However, some points must be clarified regarding the applicability of the method, such as the extent of DNA degradation through intestinal digestion, the potential for multiple sources of blood meals and the need for greater knowledge regarding intraspecific variations in mtDNA.

A quick assessment tool for human-directed aggression in pet dogs.

Many test series have been developed to assess dog temperament and aggressive behavior, but most of them have been criticized for their relatively low predictive validity or being too long, stressful, and/or problematic to carry out. We aimed to develop a short and effective series of tests that corresponds with (a) the dog's bite history, and (b) owner evaluation of the dog's aggressive tendencies. Seventy-three pet dogs were divided into three groups by their biting history; non-biter, bit once, and multiple biter. All dogs were exposed to a short test series modeling five real-life situations: friendly greeting, take away bone, threatening approach, tug-of-war, and roll over. We found strong correlations between the in-test behavior and owner reports of dogs' aggressive tendencies towards strangers; however, the test results did not mirror the reported owner-directed aggressive tendencies. Three test situations (friendly greeting, take-away bone, threatening approach) proved to be effective in evoking specific behavioral differences according to dog biting history. Non-biters differed from biters, and there were also specific differences related to aggression and fear between the two biter groups. When a subsample of dogs was retested, the test revealed consistent results over time. We suggest that our test is adequate for a quick, general assessment of human-directed aggression in dogs, particularly to evaluate their tendency for aggressive behaviors towards strangers. Identifying important behavioral indicators of aggressive tendencies, this test can serve as a useful tool to study the genetic or neural correlates of human-directed aggression in dogs.

Social tolerance in wild female crested macaques (Macaca nigra) in Tangkoko-Batuangus Nature Reserve, Sulawesi, Indonesia.

In primates, females typically drive the evolution of the social system and present a wide diversity of social structures. To understand this diversity, it is necessary to document the consistency and/or flexibility of female social structures across and within species, contexts, and environments. Macaques (Macaca sp.) are an ideal taxon for such comparative study, showing both consistency and variation in their social relations. Their social styles, constituting robust sets of social traits, can be classified in four grades, from despotic to tolerant. However, tolerant species are still understudied, especially in the wild. To foster our understanding of tolerant societies and to assess the validity of the concept of social style, we studied female crested macaques, Macaca nigra, under entirely natural conditions. We assessed their degree of social tolerance by analyzing the frequency, intensity, and distribution of agonistic and affiliative behaviors, their dominance gradient, their bared-teeth display, and their level of conciliatory tendency. We also analyzed previously undocumented behavioral patterns in grade 4 macaques: reaction upon approach and distribution of affiliative behavior across partners. We compared the observed patterns to data from other populations of grade 4 macaques and from species of other grades. Overall, female crested macaques expressed a tolerant social style, with low intensity, frequently bidirectional, and reconciled conflicts. Dominance asymmetry was moderate, associated with an affiliative bared-teeth display. Females greatly tolerated one another in close proximity. The observed patterns matched the profile of other tolerant macaques and were outside the range of patterns of more despotic species. This study is the first comprehensive analysis of females' social behavior in a tolerant macaque species under natural conditions and as such, contributes to a better understanding of macaque societies. It also highlights the relevance of the social style concept in the assessment of the degree of tolerance/despotism in social systems.

Are personality differences in a small iteroparous mammal maintained by a life-history trade-off?

Despite increasing interest, animal personality is still a puzzling phenomenon. Several theoretical models have been proposed to explain intraindividual consistency and interindividual variation in behaviour, which have been primarily supported by qualitative data and simulations. Using an empirical approach, I tested predictions of one main life-history hypothesis, which posits that consistent individual differences in behaviour are favoured by a trade-off between current and future reproduction. Data on life-history were collected for individuals of a natural population of grey mouse lemurs (Microcebus murinus). Using open-field and novel-object tests, I quantified variation in activity, exploration and boldness for 117 individuals over 3 years. I found systematic variation in boldness between individuals of different residual reproductive value. Young males with low current but high expected future fitness were less bold than older males with high current fecundity, and males might increase in boldness with age. Females have low variation in assets and in boldness with age. Body condition was not related to boldness and only explained marginal variation in exploration. Overall, these data indicate that a trade-off between current and future reproduction might maintain personality variation in mouse lemurs, and thus provide empirical support of this life-history trade-off hypothesis.

A meta-analysis of sex differences in animal personality: no evidence for the greater male variability hypothesis.

The notion that men are more variable than women has become embedded into scientific thinking. For mental traits like personality, greater male variability has been partly attributed to biology, underpinned by claims that there is generally greater variation among males than females in non-human animals due to stronger sexual selection on males. However, evidence for greater male variability is limited to morphological traits, and there is little information regarding sex differences in personality-like behaviours for non-human animals. Here, we meta-analysed sex differences in means and variances for over 2100 effects (204 studies) from 220 species (covering five broad taxonomic groups) across five personality traits: boldness, aggression, activity, sociality and exploration. We also tested if sexual size dimorphism, a proxy for sex-specific sexual selection, explains variation in the magnitude of sex differences in personality. We found no significant differences in personality between the sexes. In addition, sexual size dimorphism did not explain variation in the magnitude of the observed sex differences in the mean or variance in personality for any taxonomic group. In sum, we find no evidence for widespread sex differences in variability in non-human animal personality.

Alcohol-induced behavior change in zebrafish models.

Zebrafish are at the forefront of neurobiological research and have been gaining popularity as a viable and valid behavioral model in a variety of research applications (e.g., assessing drug induced behavioral changes). This model becomes even more attractive when considering the behavioral changes that follow exposure to compounds that are water-soluble. As such, several studies have implicated both acute and chronic ethanol exposure in the modulation of zebrafish behavior. Within this arena there appears to be a common trend across multiple studies. As with many drugs ethanol appears to influence behavior in a dose-dependent manner. In this review, we compare and contrast several studies that measure behavior as a result of alcohol exposure. Appended to this review are pilot data that report zebrafish blood alcohol concentrations as a function of acute exposure.

Effects of stress and motivation on performing a spatial task.

Learning is ubiquitous in the animal kingdom but has been studied extensively in only a handful of species. Moreover, learning studied under laboratory conditions is typically unrelated to the animal's natural environment or life history. Here, we designed a task relevant to the natural behavior of male African cichlid fish (Astatotilapia burtoni), to determine if they could be trained on a spatial task to gain access to females and shelter. We measured both how successfully animals completed this task over time and whether and how immediate early gene and hormone expression profiles were related to success. While training fish in a maze, we measured time to task completion, circulating levels of three key hormones (cortisol, 11-ketotestosterone, and testosterone) and mRNA abundance of seven target genes including three immediate early genes (that served proxies for brain activity) in nine brain regions. Data from our subjects fell naturally into three phenotypes: fish that could be trained (learners), fish that could not be trained (non-learners) and fish that never attempted the task (non-attempters). Learners and non-learners had lower levels of circulating cortisol compared to fish that never attempted the task. Learners had the highest immediate early gene mRNA levels in the homologue of the hippocampus (dorsolateral telencephalon; Dl), lower cortisol (stress) levels and were more motivated to accomplish the task as measured by behavioral observations. Fish that never attempted the task showed the lowest activity within the Dl, high stress levels and little to no apparent motivation. Data from non-learners fell between these two extremes in behavior, stress, and motivation.

Locomotor behavior of wild orangutans (Pongo pygmaeus wurmbii) in disturbed peat swamp forest, Sabangau, Central Kalimantan, Indonesia.

This study examined the locomotor behavior of wild Bornean orangutans (P. p. wurmbii) in an area of disturbed peat swamp forest (Sabangau Catchment, Indonesia) in relation to the height in the canopy, age-sex class, behavior (feeding or traveling), and the number of supports used to bear body mass. Backward elimination log-linear modeling was employed to expose the main influences on orangutan locomotion. Our results showed that the most important distinctions with regard to locomotion were between suspensory and compressive, or, orthograde (vertical trunk) and pronograde (horizontal trunk) behavior. Whether orangutans were traveling or feeding had the most important influence on locomotion whereby compressive locomotion had a strong association with feeding, suspensory locomotion had a strong association with travel in the peripheral strata using multiple supports, whereas vertical climb/descent and oscillation showed a strong association with travel on single supports in the core stratum. In contrast to theoretical predictions on positional behavior and body size, age-sex category had a limited influence on locomotion. The study revealed that torso orthograde suspension dominates orangutan locomotion, concurring with previous studies in dipterocarp forest. But, orangutans in the Sabangau exhibited substantially higher frequencies of oscillatory locomotion than observed at other sites, suggesting this behavior confers particular benefits for traversing the highly compliant arboreal environment typical of disturbed peat swamp forest. In addition, torso pronograde suspensory locomotion was observed at much lower levels than in the Sumatran species. Together these results highlight the necessity for further examination of differences between species, which control for habitat.

Multi-View Mouse Social Behaviour Recognition With Deep Graphic Model.

Home-cage social behaviour analysis of mice is an invaluable tool to assess therapeutic efficacy of neurodegenerative diseases. Despite tremendous efforts made within the research community, single-camera video recordings are mainly used for such analysis. Because of the potential to create rich descriptions for mouse social behaviors, the use of multi-view video recordings for rodent observations is increasingly receiving much attention. However, identifying social behaviours from various views is still challenging due to the lack of correspondence across data sources. To address this problem, we here propose a novel multi-view latent-attention and dynamic discriminative model that jointly learns view-specific and view-shared sub-structures, where the former captures unique dynamics of each view whilst the latter encodes the interaction between the views. Furthermore, a novel multi-view latent-attention variational autoencoder model is introduced in learning the acquired features, enabling us to learn discriminative features in each view. Experimental results on the standard CRMI13 and our multi-view Parkinson's Disease Mouse Behaviour (PDMB) datasets demonstrate that our proposed model outperforms the other state of the arts technologies, has lower computational cost than the other graphical models and effectively deals with the imbalanced data problem.

A framework for studying behavioral evolution by reconstructing ancestral repertoires.

Although different animal species often exhibit extensive variation in many behaviors, typically scientists examine one or a small number of behaviors in any single study. Here, we propose a new framework to simultaneously study the evolution of many behaviors. We measured the behavioral repertoire of individuals from six species of fruit flies using unsupervised techniques and identified all stereotyped movements exhibited by each species. We then fit a Generalized Linear Mixed Model to estimate the intra- and inter-species behavioral covariances, and, by using the known phylogenetic relationships among species, we estimated the (unobserved) behaviors exhibited by ancestral species. We found that much of intra-specific behavioral variation has a similar covariance structure to previously described long-time scale variation in an individual's behavior, suggesting that much of the measured variation between individuals of a single species in our assay reflects differences in the status of neural networks, rather than genetic or developmental differences between individuals. We then propose a method to identify groups of behaviors that appear to have evolved in a correlated manner, illustrating how sets of behaviors, rather than individual behaviors, likely evolved. Our approach provides a new framework for identifying co-evolving behaviors and may provide new opportunities to study the mechanistic basis of behavioral evolution.

Protocol for behavioral tests using chemogenetically manipulated mice.

Behavioral analyses using mice chemogenetically manipulated by designer receptors exclusively activated by designer drugs (DREADDs) are powerful tools to elucidate neural functions. Here, we describe the detailed protocols for stereotaxic surgery, adeno-associated virus (AAV)-mediated introduction to Gq-DREADDs in mice, and for behavioral testing and analyses related to anxiety, risk assessment, and burying behaviors. A series of these tests are useful in evaluating animal anxiety and their defensive response patterns to potential threats. For complete details on the use and execution of this protocol, please refer to Horii-Hayashi et al. (2021).

Impacts of early life exposure to estrogen on subsequent breeding behavior and reproductive success in zebrafish.

Impacts of exposure to environmental estrogens on reproductive development are well documented, but recently wider concern has been raised due to evidence that such exposures can disrupt normal patterns of reproductive behavior, dominance, and parentage, with potential population level implications. It is fundamental therefore to understand any such effects for effective risk assessment. This study investigated the impact of a transient exposure to ethinylestradiol (EE(2)) during early life (from 20-60 days post fertilization), including at a dosing level within the environmental range, on the subsequent reproductive behavior and success in both male and female zebrafish (Danio rerio) in competitive breeding scenarios. There were no obvious effects of the early life EE(2) exposures on the subsequent gonadal phenotypes in either mature males or females. In fact, reproductive success in males exposed to 2.76 ng EE(2)/L was increased in competitive spawning scenarios. In contrast, exposure of females to EE(2) (9.86 ng/L) during early life reduced their subsequent reproductive success in competitive spawning scenarios. Mate choice experiments suggested this was a consequence of the females' diminished courting behavior toward males, rather than any male preference for unexposed females. Reproductive capability of females is generally considered a key determinant in population demographics and dynamics, and therefore the effect of exposure to EE(2) on female reproductive success may have significant implications for exposed fish populations.