Dynamics of changes in broiler spatial distribution induced by a robot with autonomous navigation along the growing cycle.

Welfare problems in broiler chickens are associated with accelerated growth in high density and barren environments. Encouraging broiler movement yields benefits by increasing locomotion, foraging, and environmental exploration. Robot sensors with autonomous navigation capabilities developed to collect husbandry information could collaterally induce movement of birds while traversing the chicken houses. This study examines the short-time dynamic of changes in broiler spatial distribution within the robot's zone of influence throughout the growing cycle. Two batches of mixed-sex Cobb-500 were raised in a commercial broiler farm until 42 d of age, in 2 houses divided into 4 equally sized sectors. In half of the sectors an AviSense robot sustained 2-h per day of autonomous navigation. The minute prior and the 4 min following the robot entering the zone of influence were video recorded weekly. Control sectors without a robot were analyzed equivalently. Number of individuals within the zone of influence of the robot were obtained at 1-s intervals and relative density (%) was estimated. Physical interactions between broilers and the robot, as well as interactions with the environment were also recorded. The entrance of the robot triggers within seconds a strong depopulation of the zone with birds walking to neighboring areas (P < 0.03, in all ages). The decreases in relative density induced by the robot appears more pronounced, and repopulation of the zone was slower, in younger than in older birds (P < 0.05). Broilers´ showed physical interactions towards the robot and were also touched and/or slightly pushed by the robots (19 and 84% of videos recorded, respectively). They were also found scratching and/or pecking the ground after the robot passed (64% of videos). Findings strongly suggest that robots, beyond their specific capabilities as environmental sensors, were effective in promoting increased movement in broilers along the growing cycle and could also favor additional exploratory behaviors. Thus, these robots could be considered as environmental enrichment elements that contribute to welfare improvements during intensive rearing.

Dietary restriction modulates ultradian rhythms and autocorrelation properties in mice behavior.

Animal behavior emerges from integration of many processes with different spatial and temporal scales. Dynamical behavioral patterns, including daily and ultradian rhythms and the dynamical microstructure of behavior (i.e., autocorrelations properties), can be differentially affected by external cues. Identifying these patterns is important for understanding how organisms adapt to their environment, yet unbiased methods to quantify dynamical changes over multiple temporal scales are lacking. Herein, we combine a wavelet approach with Detrended Fluctuation Analysis to identify behavioral patterns and evaluate changes over 42-days in mice subjected to different dietary restriction paradigms. We show that feeding restriction alters dynamical patterns: not only are daily rhythms modulated but also the presence, phase and/or strength of ~12h-rhythms, as well as the nature of autocorrelation properties of feed-intake and wheel running behaviors. These results highlight the underlying complexity of behavioral architecture and offer insights into the multi-scale impact of feeding habits on physiology.

High-resolution ethograms, accelerometer recordings, and behavioral time series of Japanese quail.

Although many small vertebrates are capable of performing high-speed behaviors, most studies continue to focus on low-resolution temporal scales (>>1 s). Herein, we present video-recordings, behavior time series, and the computer software for video-analysis of Japanese quail within social groups. Home-boxes were monitored using both top and side video-cameras. High-resolution ethograms were developed for analyses. Pairs of females were assigned as either controls or using one of two methods for attachment of an accelerometer (patch or backpack). Behavior was recorded during 1 h on the first 2-days, sampled at 1 s intervals (days 1 and 2). On day 8, an unfamiliar male was placed in the home-box and its behavior was recorded during the first 10 min, sampled every 1/15 s. Male accelerometer recordings were also obtained. Video-recordings and resulting detailed high-resolution behavioral time series are valuable for reuse in comparative studies regarding the temporal dynamics of behavior within social environments. In addition, they are necessary for the assessment of novel machine learning algorithms that could be used for deciphering the output of accelerometer recordings.

Dietary supplementation with chia polyphenols alleviates oxidative stress and improves egg nutritional quality in Japanese quails under heat stress.

Heat stress in poultry is a major concern, especially in regions with hot summers and scarce cooling infrastructure. Dietary supplementation with antioxidants, such as polyphenols, has risen as a strategy to mitigate the physiological consequences of heat stress. A by-product of the extraction of oil from chia seeds, which is discarded if not used, could be a possible source of polyphenols. The aim of the present study was to evaluate the effects of dietary supplementation with polyphenols from defatted chia seed cake on the general performance, and oxidative status of Japanese quail exposed to heat stress. Furthermore, productive performance, egg quality and yolk fatty acid composition were also assessed. A total of 36 females (96 days of age) were randomly assigned to different diets: BASAL (control), LDCP (low dose of chia polyphenols), or HDCP (high dose of chia polyphenols). Half the animals in each diet group were exposed to 34 °C for 9 h a day (Heat Stress; HS), while the other half remained at the standard 24 °C (No Heat Stress; NHS). After 23 days of experimental conditions, animals under HS showed higher body temperatures and time spent panting, but lower egg laying rate. Moreover, HS modulated the activity of catalase and glutathione peroxidase enzymes, increasing lipid peroxidation in serum and liver; and increased saturated fatty acids in egg yolk. Supplementation with chia polyphenols helped to mitigate the HS effects, especially on glutathione peroxidase activity, decreasing lipid peroxidation. In addition, supplementation with HDCP showed the highest proportion of polyunsaturated fatty acids in liver and egg yolk. In conclusion, the use of defatted chia seed cake could represent a sustainable strategy to mitigate heat stress effects on Japanese quail, due to its capacity to decrease oxidative stress and improve the nutritional quality of egg, while decreasing the amount of waste generated by the food industry.

Computational Approaches and Tools as Applied to the Study of Rhythms and Chaos in Biology.

The temporal dynamics in biological systems displays a wide range of behaviors, from periodic oscillations, as in rhythms, bursts, long-range (fractal) correlations, chaotic dynamics up to brown and white noise. Herein, we propose a comprehensive analytical strategy for identifying, representing, and analyzing biological time series, focusing on two strongly linked dynamics: periodic (oscillatory) rhythms and chaos. Understanding the underlying temporal dynamics of a system is of fundamental importance; however, it presents methodological challenges due to intrinsic characteristics, among them the presence of noise or trends, and distinct dynamics at different time scales given by molecular, dcellular, organ, and organism levels of organization. For example, in locomotion circadian and ultradian rhythms coexist with fractal dynamics at faster time scales. We propose and describe the use of a combined approach employing different analytical methodologies to synergize their strengths and mitigate their weaknesses. Specifically, we describe advantages and caveats to consider for applying probability distribution, autocorrelation analysis, phase space reconstruction, Lyapunov exponent estimation as well as different analyses such as harmonic, namely, power spectrum; continuous wavelet transforms; synchrosqueezing transform; and wavelet coherence. Computational harmonic analysis is proposed as an analytical framework for using different types of wavelet analyses. We show that when the correct wavelet analysis is applied, the complexity in the statistical properties, including temporal scales, present in time series of signals, can be unveiled and modeled. Our chapter showcase two specific examples where an in-depth analysis of rhythms and chaos is performed: (1) locomotor and food intake rhythms over a 42-day period of mice subjected to different feeding regimes; and (2) chaotic calcium dynamics in a computational model of mitochondrial function.

Short- and long-term dynamics of the physiological and behavioral response to heat stress and thymol supplementation in Japanese quail.

Organisms have evolved endogenous timing systems that enable them to predict temporal changes and to coordinate complex internal processes. However, temporal dynamics of biological responses are most often ignored in fields such as dietary supplementation of farm animals exposed to artificial environmental challenges. Herein, we hypothesized that the potential for thymol (2-isopropyl-5-methylphenol) to alleviate physiological and behavioral consequences of heat stress is time-dependent on both long-term (i.e. weeks) and short-term (i.e. within day) time scales. First, during 3-weeks adult female Japanese quail (Coturnix japonica) were exposed daily to 9h of increased environmental temperature (34.2 ± 0.1 °C). Controls remained at standard temperatures (23.6 ± 0.1 °C). Simultaneously, half received thymol dietary supplementation and the other half a control basal diet. On day 4, both thymol and heat stress decreased body weight and feed intake respect to controls (basal, standard temperature). After three weeks, feed intake recovered for thymol groups. Therefore, we performed a second experiment focused on the critical first week of treatment, sampling variables three times a day. The beneficial effects of thymol supplementation were mainly observed during the morning, including prevention of high respiratory rates and reduction in the weight of droppings induced by heat stress, and increased walking under both temperatures. In summary, thymol's potential for alleviating heat stress consequences is time-dependent, and can be conceived as an emergent property resulting from the complex interplay between the dynamics of the biological response to thymol and heat stress. Findings highlight the importance of considering time-related factors when developing supplementation protocols to mitigate environmental challenges.

Aggressive dominance can decrease behavioral complexity on subordinates through synchronization of locomotor activities.

Social environments are known to influence behavior. Moreover, within small social groups, dominant/subordinate relationships frequently emerge. Dominants can display aggressive behaviors towards subordinates and sustain priority access to resources. Herein, Japanese quail (Coturnix japonica) were used, given that they establish hierarchies through frequent aggressive interactions. We apply a combination of different mathematical tools to provide a precise quantification of the effect of social environments and the consequence of dominance at an individual level on the temporal dynamics of behavior. Main results show that subordinates performed locomotion dynamics with stronger long-range positive correlations in comparison to birds that receive few or no aggressions from conspecifics (more random dynamics). Dominant birds and their subordinates also showed a high level of synchronization in the locomotor pattern, likely emerging from the lack of environmental opportunities to engage in independent behavior. Findings suggest that dominance can potentially modulate behavioral dynamics through synchronization of locomotor activities.

High-resolution behavioral time series of Japanese quail within their social environment.

The behavioral dynamics within a social group not only could depend on individual traits and social-experience of each member, but more importantly, emerges from inter-individual interactions over time. Herein, we first present a dataset, as well as the corresponding original video recordings, of the results of 4 behavioral tests associated with fear and aggressive response performed on 106 Japanese quail. In a second stage, birds were housed with conspecifics that performed similarly in the behavioral tests in groups of 2 females and 1 male. By continuously monitoring each bird in these small social groups, we obtained time series of social and reproductive behavior, and high-resolution locomotor time series. This approach provides the opportunity to perform precise quantification of the temporal dynamics of behavior at an individual level within different social scenarios including when an individual showing continued aggressive behaviors is present. These unique datasets and videos are publicly available in Figshare and can be used in further analysis, or for comparison with existing or future data sets or mathematical models across different taxa.

Expression of aggressiveness modulates mesencephalic c-fos activation during a social interaction test in Japanese quail (Coturnix japonica).

It is well known that during a social conflict, interactions are dependent on the animal's propensity to behave aggressively as well as the behavior of the opponent. However, discriminating between these two confounding factors was difficult. Recently, a Social Interaction (SI) test using photocastrated males as non-aggressive stimuli was proposed as a useful tool to evaluate aggressiveness. The avian Intercollicular- Griseum centralis complex (comparable to mammalian periaqueductal gray) has been reported as a crucial node in the descending pathways that organize behavioral and autonomic aspects of defensive responses and aggressiveness. Herein, using the SI test, we evaluated whether mesencephalic areas are activated (expressed c-fos) when photostimulated adult males are confronted with non-responsive (non-aggressive) opponents. Furthermore, we also examined whether mesencephalic activation is related to male performance during the SI test (i.e., aggressive vs. non-aggressive males) in birds reared in enriched or in standard environments. Five mesencephalic areas at two anatomic levels (intermediate and rostral) and locomotion during SI testing were studied. Aggressive males showed increased c-fos expression in all areas studied, and moved at faster speeds in comparison to their non-aggressive and control counterparts. Non-aggressive males and the test controls showed similar c-fos labeling. In general, rearing condition did not appear to influence c-fos expression nor behavior during the SI test. Findings suggest that mesencephalic activation is involved when males are actively expressing aggressive behaviors. This overall phenomenon is shown regardless of both the environmental stimuli provided during the birds´ rearing and the potentially stressful stimuli during the SI trial.

Chronic stress in Lizards: Studies on the Behavior and Benzodiazepine Receptors in Liolaemus koslowskyi and Cnemidophorus tergolaevigatus.

Behavioral and physiological adaptive responses of animals facing chronic exposure to a single stressor may allow them to overcome its negative effects for future exposures to similar stressful situations. At chemical level, the GABAA /benzodiazepine complex is considered one of the main receptor systems involved in the modulation of stress-induced responses. Here, we describe the behavioral responses of two different lizard species, Liolaemus koslowskyi and Cnemidophorus tergolaevigatus exposed to three potential chronic stressful treatments: (a) high temperature, (b) forced swimming, and (c) simulated predator. Additionally, we aimed to determine in those lizards whether the central-type benzodiazepine receptor (CBR; an allosteric modulator site of the GABAA receptor) is related to adaptive responses to those stressful stimulations. Our results revealed that the simulated predator was the stress condition that showed the largest difference in behavioral responses between the two species, resembling previously described strategies in nature. The basal affinity of CBRs (obtained from undisturbed animals) showed differences between both species, and the simulated predator was the only stressor that altered the affinity of CBRs. L. koslowskyi CBRs showed a decreased receptor affinity, whereas C. tergolaevigatus showed an increased receptor affinity in comparison to their respective control groups. We show for the first time the effects of different types of stressors upon behavioral responses and CBR biochemical parameters in two lizard species. Our findings suggest a potential GABA/benzodiazepine role in the ability of lizards to cope with a repeated exposure to a stressful (e.g., predator) condition.

Glutathione/thioredoxin systems modulate mitochondrial H2O2 emission: an experimental-computational study.

The net emission of hydrogen peroxide (H(2)O(2)) from mitochondria results from the balance between reactive oxygen species (ROS) continuously generated in the respiratory chain and ROS scavenging. The relative contribution of the two major antioxidant systems in the mitochondrial matrix, glutathione (GSH) and thioredoxin (Trx), has not been assessed. In this paper, we examine this key question via combined experimental and theoretical approaches, using isolated heart mitochondria from mouse, rat, and guinea pig. As compared with untreated control mitochondria, selective inhibition of Trx reductase with auranofin along with depletion of GSH with 2,4-dinitrochlorobenzene led to a species-dependent increase in H(2)O(2) emission flux of 17, 11, and 6 fold in state 4 and 15, 7, and 8 fold in state 3 for mouse, rat, and guinea pig mitochondria, respectively. The maximal H(2)O(2) emission as a percentage of the total O(2) consumption flux was 11%/2.3% for mouse in states 4 and 3 followed by 2%/0.25% and 0.74%/0.29% in the rat and guinea pig, respectively. A minimal computational model accounting for the kinetics of GSH/Trx systems was developed and was able to simulate increase in H(2)O(2) emission fluxes when both scavenging systems were inhibited separately or together. Model simulations suggest that GSH/Trx systems act in concert. When the scavenging capacity of either one of them saturates during H(2)O(2) overload, they relieve each other until complete saturation, when maximal ROS emission occurs. Quantitatively, these results converge on the idea that GSH/Trx scavenging systems in mitochondria are both essential for keeping minimal levels of H(2)O(2) emission, especially during state 3 respiration, when the energetic output is maximal. This suggests that the very low levels of H(2)O(2) emission observed during forward electron transport in the respiratory chain are a result of the well-orchestrated actions of the two antioxidant systems working continuously to offset ROS production.