Behavioral Observations Research Articles

Machine Learning Approaches for ADHD Detection

This report explores the application of deep learning convolutional neural networks (CNNs) for analyzing brain MRI scans to assist in the diagnosis of Attention Deficit Hyperactivity Disorder (ADHD) syndrome. ADHD is a neuro developmental disorder that affects millions of individuals worldwide, characterized by difficulties in attention, hyperactivity, and impulsivity. Traditional methods of diagnosis rely on subjective assessments and behavioral observations, leading to inaccuracies and delays in treatment. Leveraging the power of deep learning CNNs for MRI analysis offers the potential for more objective and efficient diagnosis, facilitating timely intervention and personalized treatment strategies

Diurnal predators in dim light: the ability of mantids to prey for supper.

Many insects rely heavily on visual cues in foraging and other life activities. Mantids are insect predators that usually ambush prey. The sophisticated visual system of mantids allows them to spot, track, and strike at prey with high accuracy. Mantids are categorized as diurnal animals in most cases, while our field observations suggested that they were active in foraging both day and night. Therefore, we hypothesize that predation in dim light is possible for mantids, while mantids are unable to capture prey in complete darkness. In this study, we experimentally examined whether different light conditions could affect the predation success and efficiency of mantid nymphs and adults, Hierodula chinensis Werner (Mantodea: Mantidae), through behavioral observations. Individual mantids were placed in individual chambers in complete darkness, simulated moonlight (0.1 lux), or simulated dusk (50 lux) conditions and were allowed to forage for prey items for 10min. Our observations showed no evidence that H. chinensis could capture any prey in complete deprivation of light. The proportion of nymphs with successful predation in simulated moonlight was 50% higher than that in complete darkness and 45.83% lower than that in simulated dusk. The proportion of adults with successful predation in simulated moonlight was 42.11% higher than that in complete darkness and 57.89% lower than that in simulated dusk. Overall, the results provide new insights into the behavioral ecology of diurnal predators at night, with potential association with moonlight, starlight, and light pollution.

Ontogenetic changes in jaw leverage and skull shape in tufted and untufted capuchins.

The ontogeny of feeding is characterized by shifting functional demands concurrent with changes in craniofacial anatomy; relationships between these factors will look different in primates with disparate feeding behaviors during development. This study examines the ontogeny of skull morphology and jaw leverage in tufted (Sapajus) and untufted (Cebus) capuchin monkeys. Unlike Cebus, Sapajus have a mechanically challenging diet and behavioral observations of juvenile Sapajus suggest these foods are exploited early in development. Landmarks were placed on three-dimensional surface models of an ontogenetic series of Sapajus and Cebus skulls (n = 53) and used to generate shape data and jaw-leverage estimates across the tooth row for three jaw-closing muscles (temporalis, masseter, medial pterygoid) as well as a weighted combined estimate. Using geometric morphometric methods, we found that skull shape diverges early and shape is significantly different between Sapajus and Cebus throughout ontogeny. Additionally, jaw leverage varies with age and position on the tooth rowand is greater in Sapajus compared to Cebus when calculated at the permanent dentition. We used two-block partial least squares analyses to identify covariance between skull shape and each of our jaw muscle leverage estimates. Sapajus, but not Cebus, has significant covariance between all leverage estimates at the anterior dentition. Our findings show that Sapajus and Cebus exhibit distinct craniofacial morphologies early in ontogeny and strong covariance between leverage estimates and craniofacial shape in Sapajus. These results are consistent with prior behavioral and comparative work suggesting these differences are a function of selection for exploiting mechanically challenging foods in Sapajus, and further emphasize that these differences appear quite early in ontogeny. This research builds on prior work that has highlighted the importance of understanding ontogeny for interpreting adult morphology.

Parenting styles in caregivers of children with Autism Spectrum Disorder and effects of service dogs

ObjectiveParents of children with Autism Spectrum Disorder (ASD) have parenting styles that differ from parents of typically developing children. Integration of a service dog (SD) at home has been demonstrated as having multiple effects within families of children with ASD. Our aims were to investigate if (a) specific parenting styles can be identified during parents’ interactions with their child with ASD using ethological methods, and (b) integration of a SD have an effect on these styles.MethodsBehavioural coding was performed on videos recorded at home by 20 parents of 6-12-years old children with ASD before SD integration. Parents were asked to record themselves and their child while making a puzzle. 14 parents performed a second similar recording 3-6 months after SD integration. Data were analysed using Principal Component Analysis, Hierarchical Cluster Analysis and non-parametric tests.ResultsThree parenting styles emerged: Parents Involved in the Task (PIT), Parents Relaxed in the Interaction (PRI), and Parents Disengaged from the Interaction (PDI). PIT were characterised as more controlling and verbally focused on the activity. PRI were less controlling and talk about things other than the activity. The same applied to PDI, except that they were less warm in their interactions. Analysis performed after SD integration revealed that these groups also diverged in the evolution of certain behaviours.ConclusionThis study is the first to demonstrate that behavioural observations can highlight different parenting styles in caregivers of children with ASD, and that the integration of a SD has effects on these styles, with variation according to parents’ style prior to SD integration. Indeed, a decrease in activity control behaviours was observed in parents with an initial profile characterise by higher expression of such behaviours (i.e., PIT), while an increase of those behaviours was observed in parents initially with an initial profile characterise by a weaker expression of such behaviours (i.e., PRI). Interestingly, the last profile characterized by less engagement in the interaction and activity (i.e., PDI) did not seem to show significant changes.

Impact of feeding Artemia franciscana enriched with various oil resources on growth, blood biochemical and behavioral indices, and survival of Oreochromis niloticus

Abstract The main objective of the present trial was to examine the efficacy of feeding tilapia fry fish on Artemia franciscana diets supplemented with various oil emulsion resources in terms of performance, behavior indices, survival rate, blood biochemical parameters, and immunological response. Four hundred Nile tilapia fry (weighing 0.15±0.05 g and measuring 2.17±0.08 cm) were randomly allocated into four equal groups (each with five repetitions) and acclimatized for fifteen days. The first group served as the control and received unenriched Artemia franciscana (G0), while the remaining three groups were fed Artemia franciscana diets enriched with different oil resources (0.5 mL oil per Liter for 6 hours): soybean oil (G1), sesame oil (G2), and rice bran oil (G3). Behavioral observations were recorded during the 45-day experimental period. At the end of the feeding trial, the chemical composition and fatty acid content of both Artemia and fingerlings were analyzed. Furthermore, the growth performance, survival, and immune response of the fingerlings were evaluated. The results indicated noticeable improvements in behavioral measurements (feeding, foraging and schooling), performance (final length, final weight, net weight gain, feed conversion ratio and specific growth rate), survival, and immune response among fry fish supplemented with enriched Artemia, particularly those enriched with soybean oil. Additionally, the chemical composition and fatty acid content of both Artemia and fish fry were significantly enhanced when oil emulsions are applied, with soybean oil demonstrating the most prominent improvements. Whereas, supplementing fry fish Artemia diets with oil resulted in lower liver enzyme activity and higher protein component levels in plasma in comparison to the control group. In brief, feeding Nile tilapia fry fish Artemia diets enriched with a soybean oil emulsion (0.5 mL/L) is recommended for promoting high performance, immunological activity, and survival throughout the early stage till fingerlings phase.

Behavioural faecal exposure pathways in a nomadic lifestyle :

Background: Water, sanitation and hygiene (WASH) is the cornerstone for health and growth at all stages of life in maintain health and increase in life span. Poor sanitation has led to disease increase in microorganisms such as Escherichia coli. Sanitation is influenced in a community by factors such as perception, behavioral practices such as open defecation and disposal of these waste. This study aimed to identify behavioral faecal exposure pathways in the nomadic community of Turbi ward. Methods: Mixed method approach using qualitative and quantitative research methods was applied in this study to assess exposure pathways and related behavior among the nomadic community of Turbi ward. Thematic data analysis was used to analyze data generated from behavioural observations. Data was presented using graphs and pie charts. Results: A total of 100 households were sampled and about 68% (n=100) of population use surface water for domestic. Turbi manyatta had the highest with  30% (n=100), followed by Kambi Nyoka at 16%(n=100) and the least was Shurr manyatta at 2% (n=100). Surface water was the dominant source of water for these community. About 76% (n=100) of the households did not treat their water before use for domestic purposes and due to scarcity of water the community store water in jerricans for more than one week. About 87% of the residents reported not washing hands before milking animals. Open defecation was the predominant method of waste disposal among the five selected Manyattas, accounting for 95% (n= 100). There was significant relationship between practicing open defecation and using surface water without treatment for domestic purpose at p-value 0.032 (95% CI), using Wilcoxon Signed-Rank test. Conclusion: The study revealed that the study community’s behavioral practices exposes them to faecal exposure due to habitual practice of open defecation, throwing waste to the bush, open burning, drinking untreated surface water and not washing hands before milking animals. There is need for advocacy on social behavior change, and strategies on health promotion targeting Water, Sanitation and Hygiene (WASH).

Effects of artificial rearing on behaviour, welfare, and immune function in piglets

With the steady increase in litter sizes, artificial rearing is becoming widely used to raise surplus piglets. The study aimed to evaluate the potential impacts of artificial rearing on behaviour and immune function in piglets. A total of 240 piglets were divided into three groups: piglets reared by the sow until weaning on day 35 (CON group), piglets artificially reared from day 7 (AR7 group), and piglets artificially reared from day 21 (AR21 group). Behavioural observation was conducted for 8 hours weekly, and the elimination position was recorded on days 7, 21, and 35. Serum and hair samples were collected to measure immunoglobulins and cortisol concentration, respectively. The results revealed that piglets in the AR7 group exhibited less frequent milk-drinking, exploring, and playing, but more frequent belly nosing and oral manipulation of conspecifics. Moreover, artificial rearing affected the elimination sites of AR7 piglets on day 21, but this phenomenon was not observed on day 35. In addition, the AR7 piglets had higher levels of cortisol and CRP, as well as lower levels of IgG and IgA compared to the CON piglets. The AR21 piglets also had a higher CRP level compared to the CON piglets. In conclusion, artificial rearing, initiated either from day 21 or day 7, negatively affected behavioural expression and immune function. Artificial rearing from day 7 was found to be the most physiologically detrimental.

Open-cup nesters in the Kalahari: Incubation and egg-shading behaviour in passerines cannot be detected with temperature dataloggers during hot periods

Birds that build open-cup nests in semi-arid or arid habitats have difficulty in maintaining clutch temperatures during incubation in a suitable incubation range due to high air temperatures and solar radiation, resulting in either embryo death or behavioural responses to preserve clutch viability. We investigated how Southern Fiscals (Lanius collaris), open-cup nesting passerines with female-only incubation in the Kalahari basin, allocated time for self-care (off-bouts) and incubation (on-bouts) and whether egg shading, a previously described behaviour in this species, is a widespread response during hot periods. We used a dual approach combining temperature dataloggers in the nest and behavioural observations during the hottest hours of the day. We found that in the early morning and late evening, Southern Fiscals behaved similarly to species incubating in temperate habitats by alternating off- and on-bouts, but during the hottest hours of the day, shading became the main activity. Behavioural observations were key to describing this behaviour indicating that it is necessary to combine different data collection strategies to successfully assess the behaviour of open-cup nesters in hot environments.

Quantification of play behaviour in calves using automated ultra-wideband location data and its association with age, weaning and health status

Play behaviour can act as an indicator of positive animal welfare. Previous attempts to predict play behaviour in farmed calves are limited because of the classification methods used, which lead to overestimation, and the short time periods that calves are observed. The study aimed to automatically classify and quantify play behaviour in farmed calves using location data from ultra-wide band sensors and to investigate factors associated with play behaviour. Location data were collected from 46 calves in three cohorts for a period of 18 weeks. Behavioural observations from video footage were merged with location data to obtain a total of 101.36 h of labelled data. An AdaBoost ensemble learning algorithm was implemented to classify play behaviour. To account for overestimation, generally seen in low-prevalence behaviours, an adjusted count technique was applied to the outputs of the classifier. Two generalized linear mixed models were fitted to investigate factors (e.g. age, health) associated with duration of play and number of play instances per day. Our algorithm identified play behaviour with > 94% accuracy when evaluated on the test set with no animals used for training, and 16% overestimation, which was computed based on the predicted number of samples of play versus the number of samples labelled as play on the test set. The instances and duration of play behaviour per day significantly decreased with age and sickness, whilst play behaviour significantly increased during and after weaning. The instances of play also significantly decreased as mean temperature increased. We suggest that the quantification method that we used could be used to detect and monitor other low prevalence behaviours (e.g. social grooming) from location data, including indicators of positive welfare.

Attachment-like behavioral expressions to humans in puppies are related to oxytocin and cortisol: A comparative study of Akitas and Labrador Retrievers

This study investigated the relationship between urinary hormone concentrations and attachment-related behaviors in two dog breeds, the Akitas and Labrador Retrievers, to elucidate the hormonal and behavioral mechanisms underlying domestication and interspecies attachment to humans. By measuring cortisol and oxytocin concentrations, and conducting the Strange Situation Test (SST), we aimed to investigate breed differences in endocrine secretions associated with domestication and how these differences influence dog behavior toward humans. Our results showed significant breed differences in urinary cortisol concentrations, with Akitas exhibiting higher levels than Labrador Retrievers. This suggests a breed-specific stress response related to genetic proximity to wolves. However, oxytocin concentrations did not differ significantly, which suggests a complex interplay between factors influencing the domestication process and the formation of attachment behaviors. Behavioral observations during the SST revealed breed-specific patterns, with Labrador Retrievers showing more playful and attachment-like behaviors and Akitas showing more exploratory and passive behaviors. The study found correlations between hormones and behaviors within breeds, particularly in Labrador Retrievers, where oxytocin concentrations were associated with attachment-like behaviors, and cortisol concentrations reflected individual differences in physical activity rather than stress responses to social situations. These findings contribute to the understanding of the evolutionary and adaptive processes underlying the ability of domestic dogs to form close relationships with humans while highlighting the role of hormonal mechanisms in mediating attachment behaviors and the influence of breed-specific genetic backgrounds on these processes.

An LSTM-based adversarial variational autoencoder framework for self-supervised neural decoding of behavioral choices.

Objective. This paper presents data-driven solutions to address two challenges in the problem of linking neural data and behavior: 1) unsupervised analysis of behavioral data and automatic label generation from behavioral observations, and 2) extraction of subject-invariant features for the development of generalized neural decoding models.

Approach. For behavioral analysis and label generation, an unsupervised method, which employs an autoencoder to transform behavior data into a cluster-friendly feature space is presented. The model iteratively refines the assigned clusters with soft clustering assignment loss, and gradually improves the learned feature representations. To address subject variability in decoding neural activity, adversarial learning in combination with a long short-term memory-based adversarial variational autoencoder (LSTM-AVAE) model is employed. By using an adversary network to constrain the latent representations, the model captures shared information among subjects' neural activity, making it proper for cross-subject transfer learning.

Main results. The proposed approach is evaluated using cortical recordings of Thy1-GCaMP6s transgenic mice obtained via widefield calcium imaging during a motivational licking behavioral experiment. The results show that the proposed model achieves an accuracy of 89.7% in cross-subject neural decoding, outperforming other well-known autoencoder-based feature learning models. These findings suggest that incorporating an adversary network eliminates subject dependency in representations, leading to improved cross-subject transfer learning performance, while also demonstrating the effectiveness of LSTM-based models in capturing the temporal dependencies within neural data.

Significance. Results demonstrate the feasibility of the proposed framework in unsupervised clustering and label generation of behavioral data, as well as achieving high accuracy in cross-subject neural decoding, indicating its potentials for relating neural activity to behavior.

Novel behavioral observations and body scarring for the bluntnose sixgill shark (Hexanchus griseus) offer clues to reproductive patterns and potential mating events

Novel behavioral observations and body scarring for the bluntnose sixgill shark (Hexanchus griseus) offer clues to reproductive patterns and potential mating events

The Use of Guanfacine to Mediate Anxiety-related Reactivity and Reduce Associated Agonistic Behavior in Two Pigtail Macaques (Macaca nemestrina).

Guanfacine, an α₂ adrenoceptor agonist, has been used to successfully treat self-injurious behavior in nonhuman primates, including macaques (Macaca mulatta) and baboons (Papio anubis). It does so by facilitating a correction to the dopaminergic system that mediates a reduction in impulsivity and reactivity. Given this, we assessed the potential efficacy of guanfacine to treat socially directed agonistic behavior in primates with an apparent reactive behavioral phenotype. We present data from 2 pigtail macaques (Macaca nemestrina): an intact adult male housed in a breeding group, and an experimentally naive adult female living in a research setting with her social partner. Baseline behavioral assessments suggested that both macaques showed extreme responses to external stressors that triggered them to aggress social partners often leading to wounding that required veterinary intervention. Both animals were tracked during the course of 1 y. Once treated regularly with guanfacine, both animals showed significant reduction in their agonistic behavior and the rate at which they wounded other animals. Indeed, in the year since the female has been treated with guanfacine she has never wounded her cagemate. By collecting regular and detailed behavioral observations on the male in the breeding colony, we were able to identify triggers for his aggression and to track the behavioral changes evidenced after guanfacine treatment. These data supported our hypothesis that his aggression reflected extreme reactivity to external stressors, rather than general anxiety. Importantly, we saw only a limited and short-lived reduction in the male's affiliative behavioral rates, and thus guanfacine had no sedative effect, but did successfully reduce his reactivity and resultant agonism and wounding.

Yingxiang Acupoint Embedding Improves Mucosal Barrier Function in Rats with Local Allergic Rhinitis

Introduction: Epithelial barrier disruption is the initial cause of various diseases. We previously reported that acupoint catgut embedding (AE) improves tight junction proteins (TJs) in rats with allergic rhinitis. However, whether AE improves the epithelial barrier in local allergic rhinitis (LAR) remains unknown. Methods: A total of 36 Sprague Dawley (SD) male rats aged 5–7 weeks were divided into 6 groups with 6 rats each: control group, LAR model group, false acupoint embedding + LAR group, acupoint embedding + LAR group, capsaicin + LAR group, and tunicamycin + acupoint embedding + LAR group. Behavioral observation, ELISA to detect inflammatory factors in nasal lavage fluid and serum IgE, nasal mucosal permeability test, hematoxylin-eosin staining, PCR to detect Substance P (SP), Western blot, and immunofluorescence to detect endoplasmic reticulum stress (ERS) index and TJs were used to investigate the mechanism of AE in LAR. Results: AE improved the symptoms and pathological features of nasal mucosa of LAR rats, reduced the inflammatory factors (IL4, IL5, IL13) of nasal lavage fluid, and showed no significant change in serum IgE levels in all groups. In addition, AE decreased the expression of SP in nasal mucosa of LAR rats, inhibited ERS, increased the expression of tight junction protein, reduced the permeability of nasal mucosa, and improved the function of nasal mucosal barrier. Conclusion: This study confirms that AE can improve the nasal mucosal barrier function of LAR by reducing the expression of SP, inhibiting ERS and increasing the expression of TJs, thus enhancing the nasal mucosal barrier function.

Individual differences in short-term social dynamics: Theoretical perspective and empirical development of the social dynamics scale

AbstractPeople have a need to form and maintain fulfilling social contact, yet they differ with respect to with whom they satisfy the need and how quickly this need is deprived or overly satiated. These social dynamics across relationships and across time are theoretically delineated in the current article. Furthermore, we developed a questionnaire to measure individual differences in three aspects of such social dynamics: (a) Family-friends interdependence, (b) Social deprivation, and (c) Social oversatiation. In a longitudinal study spanning 9 weeks in spring 2020, in total 471 participants (18–75 years, 47% women) answered the newly developed items on social dynamics, questionnaires on social dispositions (e.g., affiliation motive, need to be alone, social anxiety), and questions on personal and indirect contact with family and friends during nationwide contact restrictions related to COVID-19. The results showed that individual differences in Family-friends interdependence, Social deprivation, and Social oversatiation can be measured reliably, validly, and with predictive value for changes in daily contact as contact restrictions were loosened. We discuss potential applications of the Social Dynamics Scale (SDS) for studying social relationships in healthy and clinical populations, and conclude that the brief self-report questionnaire of social dynamics can be useful for situations and samples where direct behavioral observations are not feasible.

Automated Observations of Dogs' Resting Behaviour Patterns Using Artificial Intelligence and Their Similarity to Behavioural Observations.

Although direct behavioural observations are widely used, they are time-consuming, prone to error, require knowledge of the observed species, and depend on intra/inter-observer consistency. As a result, they pose challenges to the reliability and repeatability of studies. Automated video analysis is becoming popular for behavioural observations. Sleep is a biological metric that has the potential to become a reliable broad-spectrum metric that can indicate the quality of life and understanding sleep patterns can contribute to identifying and addressing potential welfare concerns, such as stress, discomfort, or health issues, thus promoting the overall welfare of animals; however, due to the laborious process of quantifying sleep patterns, it has been overlooked in animal welfare research. This study presents a system comparing convolutional neural networks (CNNs) with direct behavioural observation methods for the same data to detect and quantify dogs' sleeping patterns. A total of 13,688 videos were used to develop and train the model to quantify sleep duration and sleep fragmentation in dogs. To evaluate its similarity to the direct behavioural observations made by a single human observer, 6000 previously unseen frames were used. The system successfully classified 5430 frames, scoring a similarity rate of 89% when compared to the manually recorded observations. There was no significant difference in the percentage of time observed between the system and the human observer (p > 0.05). However, a significant difference was found in total sleep time recorded, where the automated system captured more hours than the observer (p < 0.05). This highlights the potential of using a CNN-based system to study animal welfare and behaviour research.

Relationship between Ear Temperature, Behaviour and Stress Hormones in Guinea Pigs (Cavia porcellus) during Different Interactive Activities in Zoos.

Guinea pigs (Cavia porcellus) are used for interactive activities in zoos; therefore, it is important to investigate their welfare. This study aimed to evaluate the validity of measuring the guinea pigs' body temperature of guinea pigs through the ear canal and investigate the relationship among changes in the expression of negative behavior, changes in body temperature, and changes in salivary cortisol concentration, and examine the effects of different interactive activities. In the normal interactive activities performed at the site, the decreased body temperature of pigs was observed over time. In contrast, increased body temperature was observed in excessive interactive activities, which are not recommended. Among the negative behaviors, "Head turning" and "Locomotion" increased significantly in excessive interactions compared to normal interactions, but "Head tossing" decreased significantly over time in both types of interactions. "Freezing" was observed only in excessive interactions. Salivary cortisol concentration increased significantly for all activities. Investigating the relationship between the individual expression of negative behavior and changes in body temperature and changes cortisol level made it possible to uncover the potential for inferring an animal's physiological state. Combining ear temperature monitoring and behavioral observation during zoo interaction activities is recommended as an ethical and scientifically supported practice.

The infant–doctor relationship: an examination of infants’ distress reactions in the presence of a doctor

Fear of doctors is a common source of distress among infants; however, the underlying sources of this distress are unknown. To investigate the doctor-infant relationship, the behaviors of 61 healthy infants (176–617 days old) were observed in a simulated examination room. Their behaviors and electrocardiograms were recorded. Two groups of infants were analyzed: those who cried and those who did not. When an experimenter dressed in the doctor’s attire entered the room, all 9 infants who were crying (14.8% of all infants) stopped crying, all infants gazed at the experimenter, and their mean heart rate (HR) decreased. After the auscultation started, 29.5% of all infants cried, and the HRs of infants who cried were higher than those of infants who did not cry. During the auscultation, 80.0% of infants who cried averted from the experimenter, while 34.4% of infants who did not cry. Within 5 s of gazing at the stethoscope, the number of infants who cried increased from 3 to 12, and their mean HR also increased. Our findings suggest that the fear of doctors is not due to the appearance of doctors but rather to specific actions performed by doctors, such as auscultation. Infants may regard a doctor’s appearance as a source of interest. Furthermore, a stethoscope is a possible trigger for infants’ crying. These behavioral observations suggest the potential for patient-centered care for infants.

Evaluation of Anti-epileptic Activity of Leaves of Leucas Cephalotes

Epilepsy is a neurological disorder characterized by recurrent seizures, affecting millions of people worldwide. Despite advancements in therapeutic interventions, there remains a need for novel and effective treatments with fewer side effects. This study aimed to evaluate the anti-epileptic potential of Leucas cephalotes leaves through experimental models. The leaves of Leucas cephalotes were collected, dried, and subjected to extraction using suitable solvents. The obtained extract was then evaluated for acute toxicity using standard guidelines. Subsequently, anti-epileptic activity was assessed using chemically induced seizure models in rodents. Behavioral observations, seizure latency, duration, and severity were recorded to determine the extract's efficacy. Preliminary phytochemical analysis of the extract revealed the presence of alkaloids, flavonoids, terpenoids, and phenolic compounds, which are known for their neurological effects. Acute toxicity testing indicated the safety of the extract at tested doses. In seizure models, the extract demonstrated a significant increase in seizure latency, reduction in seizure duration, and alleviation of seizure severity compared to control groups. These findings suggest that the leaves of Leucas cephalotes possess anti-epileptic properties, possibly attributed to the presence of bioactive compounds. Further studies are warranted to elucidate the underlying mechanisms and optimize dosage regimens for clinical application. Leucas cephalotes could emerge as a promising natural source for developing anti-epileptic agents with improved safety profiles.

Highly diverse cuticular hydrocarbon profiles but no evidence for aggression towards non-kin in the ambrosia beetle Xyleborinus saxesenii.

Animal societies use nestmate recognition to protect against social cheaters and parasites. In most social insect societies, individuals recognize and exclude any non-nestmates and the roles of cuticular hydrocarbons as recognition cues are well documented. Some ambrosia beetles live in cooperatively breeding societies with farmed fungus cultures that are challenging to establish, but of very high value once established. Hence, social cheaters that sneak into a nest without paying the costs of nest foundation may be selected. Therefore, nestmate recognition is also expected to exist in ambrosia beetles, but so far nobody has investigated this behavior and its underlying mechanisms. Here we studied the ability for nestmate recognition in the cooperatively breeding ambrosia beetle Xyleborinus saxesenii, combining behavioural observations and cuticular hydrocarbon analyses. Laboratory nests of X. saxesenii were exposed to foreign adult females from the same population, another population and another species. Survival as well as the behaviours of the foreign female were observed. The behaviours of the receiving individuals were also observed. We expected that increasing genetic distance would cause increasing distance in chemical profiles and increasing levels of behavioural exclusion and possibly mortality. Chemical profiles differed between populations and appeared as variable as in other highly social insects. However, we found only very little evidence for the behavioural exclusion of foreign individuals. Interpopulation donors left nests at a higher rate than control donors, but neither their behaviours nor the behaviours of receiver individuals within the nest showed any response to the foreign individual in either of the treatments. These results suggest that cuticular hydrocarbon profiles might be used for communication and nestmate recognition, but that behavioural exclusion of non-nestmates is either absent in X. saxesenii or that agonistic encounters are so rare or subtle that they could not be detected by our method. Additional studies are needed to investigate this further.