Jackelyn Melissa Kembro

Open-Field Temporal Pattern of Ambulation in Japanese Quail Genetically Selected for Contrasting Adrenocortical Responsiveness to Brief Manual Restraint

Japanese quail selected for a low-stress (LS), rather than a high-stress (HS), plasma corticosterone response to brief restraint have been shown to possess lower fearfulness and a nonspecific reduction in stress responsiveness. Detrended fluctuation analysis provides information on the organization and complexity of temporal patterns of behavior. The present study evaluated the temporal pattern of ambulation of LS and HS quail in an open field that represented a novel environment. Time series of 4,200 data points were collected for each bird by registering the distance ambulated every 0.5 s during a 35-min test period. Consistent with their known reduced fearfulness, the LS quail initiated ambulation significantly sooner (P < 0.02) and tended to ambulate more (P < 0.09) than did their HS counterparts. Detrended fluctuation analyses showed a monofractal series (i.e., a series with similar complexity at different temporal scales) in 72% of the birds. These birds initiated their ambulatory activity in less than 600 s. Among these birds, a lower (P < 0.03) autosimilarity coefficient (alpha) was found in the LS quail than in their HS counterparts (alpha = 0.76 +/- 0.03 and 0.87 +/- 0.03, respectively), suggesting a more complex (less regular) ambulatory pattern in the LS quail. However, when the patterns of ambulation were reexamined by considering only the active period of the time series (i.e., after the birds had initiated their ambulatory activity), monofractal patterns were observed in 97% of the birds, and no differences were found between the lines. Collectively, the results suggest that during the active period of open-field testing, during which fear responses are likely less strong and other motivations are the driving forces of ambulation, the LS and HS lines have similar ambulatory organization.

Network dynamics: quantitative analysis of complex behavior in metabolism, organelles, and cells, from experiments to models and back

Advancing from two core traits of biological systems: multilevel network organization and nonlinearity, we review a host of novel and readily available techniques to explore and analyze their complex dynamic behavior within the framework of experimental–computational synergy. In the context of concrete biological examples, analytical methods such as wavelet, power spectra, and metabolomics–fluxomics analyses, are presented, discussed, and their strengths and limitations highlighted. Further shown is how time series from stationary and nonstationary biological variables and signals, such as membrane potential, high‐throughput metabolomics, O2 and CO2 levels, bird locomotion, at the molecular, (sub)cellular, tissue, and whole organ and animal levels, can reveal important information on the properties of the underlying biological networks. Systems biology‐inspired computational methods start to pave the way for addressing the integrated functional dynamics of metabolic, organelle and organ networks. As our capacity to unravel the control and regulatory properties of these networks and their dynamics under normal or pathological conditions broadens, so is our ability to address endogenous rhythms and clocks to improve health‐span in human aging, and to manage complex metabolic disorders, neurodegeneration, and cancer. WIREs Syst Biol Med 2017, 9:e1352. doi: 10.1002/wsbm.1352

Complex oscillatory redox dynamics with signaling potential at the edge between normal and pathological mitochondrial function

The time-keeping properties bestowed by oscillatory behavior on functional rhythms represent an evolutionarily conserved trait in living systems. Mitochondrial networks function as timekeepers maximizing energetic output while tuning reactive oxygen species (ROS) within physiological levels compatible with signaling. In this work, we explore the potential for timekeeping functions dependent on mitochondrial dynamics with the validated two-compartment mitochondrial energetic-redox (ME-R) computational model, that takes into account (a) four main redox couples [NADH, NADPH, GSH, Trx(SH)2], (b) scavenging systems (glutathione, thioredoxin, SOD, catalase) distributed in matrix and extra-matrix compartments, and (c) transport of ROS species between them. Herein, we describe that the ME-R model can exhibit highly complex oscillatory dynamics in energetic/redox variables and ROS species, consisting of at least five frequencies with modulated amplitudes and period according to power spectral analysis. By stability analysis we describe that the extent of steady state—as against complex oscillatory behavior—was dependent upon the abundance of Mn and Cu, Zn SODs, and their interplay with ROS production in the respiratory chain. Large parametric regions corresponding to oscillatory dynamics of increasingly complex waveforms were obtained at low Cu, Zn SOD concentration as a function of Mn SOD. This oscillatory domain was greatly reduced at higher levels of Cu, Zn SOD. Interestingly, the realm of complex oscillations was located at the edge between normal and pathological mitochondrial energetic behavior, and was characterized by oxidative stress. We conclude that complex oscillatory dynamics could represent a frequency- and amplitude-modulated H2O2 signaling mechanism that arises under intense oxidative stress. By modulating SOD, cells could have evolved an adaptive compromise between relative constancy and the flexibility required under stressful redox/energetic conditions.

Social interaction of juvenile Japanese quail classified by their permanence in proximity to a high or low density of conspecifics

One challenge faced by ethologists in poultry welfare is helping birds to form functionally successful groups over time through the expression of appropriate behaviors. Searching for phenotypic variability, we developed in Japanese quail a density-related permanence (DRP) test that enables us to classify young birds (while in groups) according to their individual permanence in boxes containing a high or low density of confined conspecifics (HD or LD, respectively). This study addressed the question of whether contrasting DRP quail behavior may reflect underlying differences in social responses. Birds were classified at 11 d of age in an apparatus consisting of 2 boxes interconnected by a central region delimited by sliding doors. Each box contained at its distal end either 12 or 3 conspecifics confined behind a glass (high or low density, respectively). The doors were closed 9 times every 1 h, and positioning of 36 experimental birds was registered. If birds were found in the box containing high density, low density, or in the central region, they received a 1, -1, or a 0 score, respectively. Birds with final summed scores of ≥3 or ≤-3 were categorized as HD or LD, respectively. Same category groups (HD or LD) were evaluated in their home box (undisturbed) and in a resident/intruder test when 38 d old. A higher proportion of LD than HD groups (5/6 vs. 1/6, respectively) showed at least one aggressive pecking event during a 1-h trial. The LD groups also showed a higher number of aggressive pecking events than HP groups. When an unfamiliar intruder (either HD or LD) was incorporated during 5 min in the HD or LD box, LD resident quail showed shorter latencies and a higher number of aggressive pecking events toward the intruder bird than their HD counterparts. The early individual permanence in the DRP test could be considered a consequence of a different adaptability strategy for group living. This novel test could be relevant for selection programs aiming to obtain birds better suited for rearing in high-density conditions.

Assessment of long-range correlation in animal behavior time series: The temporal pattern of locomotor activity of Japanese quail (Coturnix coturnix) and mosquito larva (Culex quinquefasciatus)

Detrended Fluctuation Analysis (DFA) is a method that has been frequently used to determine the presence of long-range correlations in human and animal behaviors. However, according to previous authors using statistical model systems, in order to correctly use DFA different aspects should be taken into account such as: (1) the establishment by hypothesis testing of the absence of short term correlation, (2) an accurate estimation of a straight line in the log–log plot of the fluctuation function, (3) the elimination of artificial crossovers in the fluctuation function, and (4) the length of the time series. Taking into consideration these factors, herein we evaluated the presence of long-range correlation in the temporal pattern of locomotor activity of Japanese quail (Coturnix coturnix) and mosquito larva (Culex quinquefasciatus). In our study, modeling the data with the general autoregressive integrated moving average (ARFIMA) model, we rejected the hypothesis of short-range correlations (d=0) in all cases. We also observed that DFA was able to distinguish between the artificial crossover observed in the temporal pattern of locomotion of Japanese quail and the crossovers in the correlation behavior observed in mosquito larvae locomotion. Although the test duration can slightly influence the parameter estimation, no qualitative differences were observed between different test durations.

Effect of the density of conspecifics on runway social reinstatement behavior of male Japanese quail genetically selected for contrasting adrenocortical responsiveness to stress

Runway tests are considered indicative of underlying sociality in birds and their ability to make social discriminations and establish interactions among conspecifics. Herein, social reinstatement behavior in male juvenile Japanese quail selected for a reduced (LS, low stress) or exaggerated (HS, high stress) adrenocortical response to brief mechanical restraint was evaluated. Individual males were given the choice to reinstate with either 2 (low density, LD) or 8 (high density, HD) unfamiliar randombred conspecifics placed in goal boxes at opposite ends of a two-choice runway (TCRW). Then, the same males were individually retested in a single goal box runway wherein they were exposed to a goal box containing either a LD or a HD of males. In the TCRW, a higher (P < 0.01) number of HS males started their ambulation toward the goal box containing the HD as opposed to LD of conspecifics. The HS males also spent more (P < 0.01) time in close proximity (within a 10-cm close zone; CZ) to the HD (218 s) rather than LD (57 s) of conspecifics. In contrast, the LS males did not differ in their initial direction of travel and they spent similar average amounts of times in the CZ of their stimulus LD (141 s) and HD (124 s) conspecifics. Similar to the TCRW results, in the single goal box runway, HS males spent more (P < 0.01) time in the CZ of HD rather than LD conspecifics, whereas LS quail spent similar amounts of time in the CZ of LD and HD males. Considering that runways are novel (and therefore frightening) environments, our findings suggest that HS quail may find better shelter (i.e., more comfort) in close proximity to a larger rather than smaller group of conspecifics, whereas LS birds find groups of varying conspecific density equally attractive. The results suggest that LS quail possess favorable social adaptive qualities because they appear to be better suited to cope with situations where the density of conspecifics is variable.

Temporal pattern of locomotor activity recuperation after administration of propofol in Japanese quail (Coturnix coturnix japonica)

The present study evaluates the organization and complexity of the temporal pattern of locomotion after an acute administration of propofol in Japanese quail by using traditional and fractal analysis. Birds were administered with propofol 0, 10, 20, 40 or 80 mg/kg. Ten min after administration, they were placed in an open-field apparatus and their locomotor activity was recorded during 45 min at a resolution of 0.5 s. A significant dose dependant increase in the latency to initiate ambulation was observed for doses of 20, 40 and 80 mg/kg when compared to the control group. A rapid recuperation of normal locomotor activity was observed after sedation with 20 mg/kg. Birds administered with propofol 40 mg/kg showed signs of recuperation of normal locomotion after 30 and 40 min (males and females, respectively) of propofol administration, that was not observed in quail treated with propofol 80 mg/kg. Our results suggest that depending on the dose, propofol administration in quail may allow full locomotor recovery of a sedative/anesthetic dose as early as 30 min post-administration.

Unexpected results when assessing underlying aggressiveness in Japanese quail using photocastrated stimulus birds

&NA; Aggressive behaviors can affect both animal welfare and productivity. Because the expression of aggressive behaviors is dependent on the quality of the opponent, they reflect relative rather than absolute levels of underlying aggressiveness. This study was aimed to characterize the aggressive responsiveness of photostimulated (14:10 h light:dark photoperiod) adult Japanese quail when interacting with a photocastrated (6:18 h light:dark photoperiod) counterpart in a novel test environment and to assesses interindividual variations. This was based on the assumption that photocastrated birds will not actively provoke an aggressive confrontation. Birds were reared in male‐female pairs. Frequencies of behaviors (i.e., pecks, threats, chases, grabs, mounts) were determined during 10 min social interactions in a novel environment. A first experiment evaluated 78 encounters between a photostimulated male or female with either a photocastrated male or female (photocastration of sexually mature birds started at 11 wk of age). High interindividual variability was observed and in general, highly aggressive birds (performing 20 or more aggressive interactions) received little or no aggression from their test counterpart. However, unexpectedly, we also found that 37% and 32% of photocastrated males and females, respectively, performed aggressions toward their photostimulated counterparts, and initiated the aggressive interactions in a similar proportion than photostimulated males. Aggressive photocastrated males did not perform reproductive‐type behaviors (i.e., grabs, mounts). Aggressiveness in the photocastrated birds was attributed to their social experience prior to photocastration. Therefore, a second experiment evaluated 106 encounters between a photostimulated male or female and a naive photocastrated male (photocastration started at 4 wk of age, prior to sexual development). Photocastrated males performed no aggressions toward their photostimulated counterparts. Consistently with previous studies, our findings show that naive photocastrated males can be used as a non‐aggressive stimulus during a social interaction aimed to assess expression of aggressiveness in photostimulated birds. However, caution should be taken when applying the photocastration protocol considering that prior fighting and sexual experience or other physiological changes related with maturation can interfere during subsequent aggressive testing.

High resolution, week-long, locomotion time series from Japanese quail in a home-box environment.

Temporal and spatial patterns of locomotion reflect both resting periods and the movement from one place to another to satisfy physiological and behavioural needs. Locomotion is studied in diverse areas of biology such as chronobiology and physiology, as well as in biomathematics. Herein, the locomotion of 24 visually-isolated Japanese quails in their home-box environment was recorded continuously over a 6.5 days at a 0.5 s sampling rate. Three time series are presented for each bird: (1) locomotor activity, (2) distance ambulated, and (3) zone of the box where the bird is located. These high resolution, week-long, time series consisting of 1.07×106 data points represent, to our knowledge, a unique data set in animal behavior, and are publically available on FigShare. The data obtained can be used for analyzing dynamic changes of daily or several day locomotion patterns, or for comparison with existing or future data sets or mathematical models across different taxa.

Cholesterol favors the emergence of a long-range autocorrelated fluctuation pattern in voltage-induced ionic currents through lipid bilayers.

The present paper was aimed at evaluating the effect of cholesterol (CHO) on the voltage-induced lipid pore formation in bilayer membranes through a global characterization of the temporal dynamics of the fluctuation pattern of ion currents. The bilayer model used was black lipid membranes (BLMs) of palmitoyloleoylphosphatidylethanolamine and palmitoyloleoylphosphatidylcholine (POPE:POPC) at a 7:3 molar ratio in the absence (BLM0) or in the presence of 30 (BLM30), 40 (BLM40) or 50(BLM50)mol% of cholesterol with respect to total phospholipids. Electrical current intensities (I) were measured in voltage (ΔV) clamped conditions at ΔV ranging between 0 and ±200mV. The autocorrelation parameter α derived from detrended fluctuation analysis (DFA) on temporal fluctuation patterns of electrical currents allowed discriminating between non-correlated (α=0.5, white noise) and long-range correlated (0.5<α<1) behaviors. The increase in |ΔV| as well as in cholesterol content increased the number of conductance states, the magnitude of conductance level, the capacitance of the bilayers and increased the tendency towards the development of long-range autocorrelated (fractal) processes (0.5<α<1) in lipid channel generation. Experiments were performed above the phase transition temperature of the lipid mixtures, but compositions used predicted a superlattice-like organization. This leads to the conclusion that structural defects other than phase coexistence may promote lipid channel formation under voltage clamped conditions. Furthermore, cholesterol controls the voltage threshold that allows the percolation of channel behavior where isolated channels become an interconnected network.