Densitaxis: Active particle motion in density gradients.

Organisms often swim through density-stratified fluids. Here, we investigate the dynamics of active particles swimming in fluid density gradients and report theoretical evidence of taxis as a result of these gradients (densitaxis). Specifically, we calculate the effect of density stratification on the dynamics of a force- and torque-free spherical squirmer and show that density gradients induce reorientation that tends to align swimming either parallel or normal to the gradient depending on the swimming gait. In particular, swimmers that propel by generating thrust in the front (pullers) rotate to swim parallel to gradients and hence display (positive or negative) densitaxis, while swimmers that propel by generating thrust in the back (pushers) rotate to swim normal to the gradients. This work could be useful to understand the motion of marine organisms in ocean or be leveraged to sort or organize a suspension of active particles by modulating density gradients.

pH-regulated single cell migration.

Over the last two decades, extra- and intracellular pH have emerged as fundamental regulators of cell motility. Fundamental physiological and pathological processes relying on appropriate cell migration, such as embryonic development, wound healing, and a proper immune defense on the one hand, and autoimmune diseases, metastatic cancer, and the progression of certain parasitic diseases on the other, depend on surrounding pH. In addition, migrating single cells create their own localized pH nanodomains at their surface and in the cytosol. By this means, the migrating cells locally modulate their adhesion to, and the re-arrangement and digestion of, the extracellular matrix. At the same time, the cytosolic nanodomains tune cytoskeletal dynamics along the direction of movement resulting in concerted lamellipodia protrusion and rear end retraction. Extracellular pH gradients as found in wounds, inflamed tissues, or the periphery of tumors stimulate directed cell migration, and long-term exposure to acidic conditions can engender a more migratory and invasive phenotype persisting for hours up to several generations of cells after they have left the acidic milieu. In the present review, the different variants of pH-dependent single cell migration are described. The underlying pH-dependent molecular mechanisms such as conformational changes of adhesion molecules, matrix protease activity, actin (de-)polymerization, and signaling events are explained, and molecular pH sensors stimulated by H+ signaling are presented.

The unexpected social cost of inguinal hernioplasty procedures derived from the COVID-19 pandemic: Surgical trend analysis based on an Italian hospital series.

BACKGROUND: The majority of inguinal hernias are usually paucisymptomatic, so are restored electively. The main purpose of this study is to assess the trends in hernia repair surgery before and during the pandemic period, analyzing an Italian hospital series of 390 patients, in an attempt to quantify the negative impact regarding social costs derived from the Covid-19 outbreak. Moreover, we want to focus on the concept of apparently minor pathology as hernioplasty which could represent a life-threatening condition for patients. METHODS: The study population consisted of all patients operated for inguinal hernia in a General Surgery Unit from 2019 to 2021, divided into a pre-pandemic and a pandemic period. RESULTS: The Covid-19 pandemic increased urgent operations in a complicated setting. A statistically significant difference was found regarding the trend of hospitalization length as well as a strong positive correlation between the severity of hernia and the hospitalization length. CONCLUSIONS: During the pandemic, it has been registered a mishandling of inguinal hernias to the detriment of both the healthcare system and patients, due to multifactorial issues and, in particular, to the restrictions imposed by the regional government that erroneously declassed hernia pathology as a minor problem for public health. We do believe that patients, after diagnosis of inguinal hernia, should learn the Taxis maneuver for its feasibility and ease of execution, in order to reduce access to emergencies in many cases and likewise to better pain and discomfort perceived, even in the event of unexpected worldwide healthcare scenario.

Redox properties and PAS domain structure of the Escherichia coli energy sensor Aer indicate a multistate sensing mechanism.

The Per-Arnt-Sim (PAS; named for the representative proteins: Period, Aryl hydrocarbon receptor nuclear translocator protein and Single-minded) domain of the dimeric Escherichia coli aerotaxis receptor Aer monitors cellular respiration through a redox-sensitive flavin adenine dinucleotide (FAD) cofactor. Conformational shifts in the PAS domain instigated by the oxidized FAD (FADOX)/FAD anionic semiquinone (FADASQ) redox couple traverse the HAMP (histidine kinases, adenylate cyclases, methyl-accepting chemotaxis proteins, and phosphatases) and kinase control domains of the Aer dimer to regulate CheA kinase activity. The PAS domain of Aer is unstable and has not been previously purified. Here, residue substitutions that rescue FAD binding in an FAD binding-deficient full-length Aer variant were used in combination to stabilize the Aer PAS domain. We solved the 2.4 Å resolution crystal structure of this variant, Aer-PAS-GVV, and revealed a PAS fold that contains distinct features associated with FAD-based redox sensing, such as a close contact between the Arg115 side chain and N5 of the isoalloxazine ring and interactions of the flavin with the side chains of His53 and Asn85 that are poised to convey conformational signals from the cofactor to the protein surface. In addition, we determined the FADox/FADASQ formal potentials of Aer-PAS-GVV and full-length Aer reconstituted into nanodiscs. The Aer redox couple is remarkably low at -289.6 ± 0.4 mV. In conclusion, we propose a model for Aer energy sensing based on the low potential of Aer-PAS-FADox/FADASQ couple and the inability of Aer-PAS to bind to the fully reduced FAD hydroquinone.

Trends in use of neoadjuvant systemic therapy in patients with clinically node-positive breast cancer in Europe: prospective TAXIS study (OPBC-03, SAKK 23/16, IBCSG 57-18, ABCSG-53, GBG 101).

PURPOSE: The aim of this study was to evaluate clinical practice heterogeneity in use of neoadjuvant systemic therapy (NST) for patients with clinically node-positive breast cancer in Europe. METHODS: The study was preplanned in the international multicenter phase-III OPBC-03/TAXIS trial (ClinicalTrials.gov Identifier: NCT03513614) to include the first 500 randomized patients with confirmed nodal disease at the time of surgery. The TAXIS study's pragmatic design allowed both the neoadjuvant and adjuvant setting according to the preferences of the local investigators who were encouraged to register eligible patients consecutively. RESULTS: A total of 500 patients were included at 44 breast centers in six European countries from August 2018 to June 2022, 165 (33%) of whom underwent NST. Median age was 57 years (interquartile range [IQR], 48-69). Most patients were postmenopausal (68.4%) with grade 2 and 3 hormonal receptor-positive and human epidermal growth factor receptor 2-negative breast cancer with a median tumor size of 28 mm (IQR 20-40). The use of NST varied significantly across the countries (p < 0.001). Austria (55.2%) and Switzerland (35.8%) had the highest percentage of patients undergoing NST and Hungary (18.2%) the lowest. The administration of NST increased significantly over the years (OR 1.42; p < 0.001) and more than doubled from 20 to 46.7% between 2018 and 2022. CONCLUSION: Substantial heterogeneity in the use of NST with HR+/HER2-breast cancer exists in Europe. While stringent guidelines are available for its use in triple-negative and HER2+ breast cancer, there is a need for the development of and adherence to well-defined recommendations for HR+/HER2-breast cancer.

Colloidal Active Matter Mimics the Behavior of Biological Microorganisms-An Overview.

This article provides a review of the recent development of biomimicking behaviors in active colloids. While the behavior of biological microswimmers is undoubtedly influenced by physics, it is frequently guided and manipulated by active sensing processes. Understanding the respective influences of the surrounding environment can help to engineering the desired response also in artificial swimmers. More often than not, the achievement of biomimicking behavior requires the understanding of both biological and artificial microswimmers swimming mechanisms and the parameters inducing mechanosensory responses. The comparison of both classes of microswimmers provides with analogies in their dependence on fuels, interaction with boundaries and stimuli induced motion, or taxis.

A Context-Aware, Computer-Vision-Based Approach for the Detection of Taxi Street-Hailing Scenes from Video Streams.

With the increasing deployment of autonomous taxis in different cities around the world, recent studies have stressed the importance of developing new methods, models and tools for intuitive human-autonomous taxis interactions (HATIs). Street hailing is one example, where passengers would hail an autonomous taxi by simply waving a hand, exactly like they do for manned taxis. However, automated taxi street-hailing recognition has been explored to a very limited extent. In order to address this gap, in this paper, we propose a new method for the detection of taxi street hailing based on computer vision techniques. Our method is inspired by a quantitative study that we conducted with 50 experienced taxi drivers in the city of Tunis (Tunisia) in order to understand how they recognize street-hailing cases. Based on the interviews with taxi drivers, we distinguish between explicit and implicit street-hailing cases. Given a traffic scene, explicit street hailing is detected using three elements of visual information: the hailing gesture, the person's relative position to the road and the person's head orientation. Any person who is standing close to the road, looking towards the taxi and making a hailing gesture is automatically recognized as a taxi-hailing passenger. If some elements of the visual information are not detected, we use contextual information (such as space, time and weather) in order to evaluate the existence of implicit street-hailing cases. For example, a person who is standing on the roadside in the heat, looking towards the taxi but not waving his hand is still considered a potential passenger. Hence, the new method that we propose integrates both visual and contextual information in a computer-vision pipeline that we designed to detect taxi street-hailing cases from video streams collected by capturing devices mounted on moving taxis. We tested our pipeline using a dataset that we collected with a taxi on the roads of Tunis. Considering both explicit and implicit hailing scenarios, our method yields satisfactory results in relatively realistic settings, with an accuracy of 80%, a precision of 84% and a recall of 84%.

COVID-19 and the Motorcycle Taxi Sector in Sub-Saharan African Cities: A Key Stakeholders' Perspective.

This article assesses the impact of the COVID-19 outbreak on the urban motorcycle taxi (MCT) sector in Sub-Saharan Africa (SSA). MCT operators in SSA provide essential transport services and have shown ingenuity and an ability to adapt and innovate when responding to different challenges, including health challenges. However, policymakers and regulators often remain somewhat hostile toward the sector. The article discusses the measures and restrictions put in place to reduce the spread of COVID-19 and key stakeholders' perspectives on these and on the sector's level of compliance. Primary data were collected in six SSA countries during the last quarter of 2020. Between 10 and 15 qualitative interviews with key stakeholders relevant to the urban MCT sector were conducted in each country. These interviews were conducted with stakeholders based in the capital city and a secondary city, to ensure a geographically broader understanding of the measures, restrictions, and perspectives. The impact of COVID-19 measures on the MCT and motor-tricycle taxi sector was significant and overwhelmingly negative. Lockdowns, restrictions on the maximum number of passengers allowed to be carried at once, and more generally, a COVID-19-induced reduction in demand, resulted in a drop in income for operators, according to the key stakeholders. However, some key stakeholders indicated an increase in MCT activity and income because of the motorcycles' ability to bypass police and army controls. In most study countries measures were formulated in a non-consultative manner. This, we argue, is symptomatic of governments' unwillingness to seriously engage with the sector.

Electrolocation? The evidence for redox-mediated taxis in Shewanella oneidensis.

Shewanella oneidensis is a dissimilatory metal reducing bacterium and model for extracellular electron transfer (EET), a respiratory mechanism in which electrons are transferred out of the cell. In the last 10 years, migration to insoluble electron acceptors for EET has been shown to be nonrandom and tactic, seemingly in the absence of molecular or energy gradients that typically allow for taxis. As the ability to sense, locate, and respire electrodes has applications in bioelectrochemical technology, a better understanding of taxis in S. oneidensis is needed. While the EET conduits of S. oneidensis have been studied extensively, its taxis pathways and their interplay with EET are not yet understood, making investigation into taxis phenomena nontrivial. Since S. oneidensis is a member of an EET-encoding clade, the genetic circuitry of taxis to insoluble acceptors may be conserved. We performed a bioinformatic analysis of Shewanella genomes to identify S. oneidensis chemotaxis orthologs conserved in the genus. In addition to the previously reported core chemotaxis gene cluster, we identify several other conserved proteins in the taxis signaling pathway. We present the current evidence for the two proposed models of EET taxis, "electrokinesis" and flavin-mediated taxis, and highlight key areas in need of further investigation.

Micro-Nano Motors with Taxis Behavior: Principles, Designs, and Biomedical Applications.

As a novel mobile nanodevice, micro-nano motors (MNMs) can convert the energy of the surrounding environment into mechanical motion. With this unique ability, they promise revolutionary potential in bio-applications including precise drug delivery, bio-sensing, and noninvasive surgery. Yet for practically reaching the target and fulfilling these tasks in dynamically changing bio-environment, environment adaptivity beyond propulsion is important yet challenging. MNMs with taxis behavior/autonomous target-seeking ability offer a desirable solution. These motors can adaptively move to the target location and complete the task. Thanks to the persistent efforts of researchers, tactic MNMs have shown automatic navigation to target under various energy fields, not only in static environments, but also in shear rheological conditions that simulate blood flow. Therefore, tactic motors with self-targeting capability lay a concrete foundation for targeted drug delivery, cell transplantation, and thrombus ablation. This review systematically presents the moving principle, design, and biological applications of tactic MNMs under different energy fields. Through in-depth analysis of state-of-art progress, the obstacles of the field and possible solutions are discussed. With the continuous innovation and breakthroughs of multi-disciplinary researchers, MNMs with taxis behavior are expected to provide a revolutionary solution for cancer and other major diseases in the biomedical field.

Microbial Volatile Organic Compounds from Tempered and Incubated Grain Mediate Attraction by a Primary but Not Secondary Stored Product Insect Pest in Wheat.

There has been a dearth of research elucidating the behavioral effect of microbially-produced volatile organic compounds on insects in postharvest agriculture. Demonstrating attraction to MVOC's by stored product insects would provide an additional source of unique behaviorally-relevant stimuli to protect postharvest commodities at food facilities. Here, we assessed the behavioral response of a primary (Rhyzopertha dominica) and secondary (Tribolium castaneum) grain pest to bouquets of volatiles produced by whole wheat that were untempered, or tempered to 12%, 15%, or 19% grain moisture and incubated for 9, 18, or 27 days. We hypothesized that MVOC's may be more important for the secondary feeder because they signal that otherwise unusable, intact grains have become susceptible by weakening of the bran. However, contrary to our expectations, we found that the primary feeder, R. dominica, but not T. castaneum was attracted to MVOC's in a wind tunnel experiment, and in a release-recapture assay using commercial traps baited with grain treatments. Increasing grain moisture resulted in elevated grain damage detected by near-infrared spectroscopy and resulted in small but significant differences in the blend of volatiles emitted by treatments detected by gas chromatography coupled with mass spectrometry (GC-MS). In sequencing the microbial community on the grain, we found a diversity of fungi, suggesting that an assemblage was responsible for emissions. We conclude that R. dominica is attracted to a broader suite of MVOC's than T. castaneum, and that our work highlights the importance of understanding insect-microbe interactions in the postharvest agricultural supply chain.

Changes in air quality in-taxis and in working conditions of taxi drivers pre- and post-lockdown, during the COVID-19 pandemic in the Paris area.

We evaluated the impact of the lockdown restriction measures in the Paris area on the variation of in-vehicle ultrafine particle (UFP) and black carbon (BC) concentrations between the pre- and post-lockdown period and professional drivers' working conditions and practices. The study was conducted with 33 taxi drivers. UFP and BC were measured inside their vehicles with DiSCmini® and microAeth® , respectively, on two typical working days pre- and post-lockdown. Job characteristics were self-reported. Our results showed that post-lockdown, both the number of clients and journey duration significantly decreased. Taxi drivers opened their windows significantly more and reduced the use of air recirculation. UFP decreased significantly by 32% and BC by 31% post-lockdown, with a weaker positive correlation compared to pre-lockdown. The reduction of in-vehicle UFP was due mainly to the reduction of traffic flow and ventilation settings, though the latter probably varied according to traffic conditions. The variation of in-vehicle BC also tended to be related to the decrease in traffic flow post-lockdown. We emphasize the role of traffic emissions on in-vehicle air pollution and that preventive measures such ventilation settings would help to minimize the exposure of professional drivers and passengers to air pollutants.

Caenorhabditis elegans exhibits positive gravitaxis.

BACKGROUND: Gravity plays an important role in most life forms on Earth. Yet, a complete molecular understanding of sensing and responding to gravity is lacking. While there are anatomical differences among animals, there is a remarkable conservation across phylogeny at the molecular level. Caenorhabditis elegans is suitable for gene discovery approaches that may help identify molecular mechanisms of gravity sensing. It is unknown whether C. elegans can sense the direction of gravity. RESULTS: In aqueous solutions, motile C. elegans nematodes align their swimming direction with the gravity vector direction while immobile worms do not. The worms orient downward regardless of whether they are suspended in a solution less dense (downward sedimentation) or denser (upward sedimentation) than themselves. Gravitaxis is minimally affected by the animals' gait but requires sensory cilia and dopamine neurotransmission, as well as motility; it does not require genes that function in the body touch response. CONCLUSIONS: Gravitaxis is not mediated by passive forces such as non-uniform mass distribution or hydrodynamic effects. Rather, it is mediated by active neural processes that involve sensory cilia and dopamine. C. elegans provides a genetically tractable system to study molecular and neural mechanisms of gravity sensing.

Navigation Along Windborne Plumes of Pheromone and Resource-Linked Odors.

Many insects locate resources such as a mate, a host, or food by flying upwind along the odor plumes that these resources emit to their source. A windborne plume has a turbulent structure comprised of odor filaments interspersed with clean air. As it propagates downwind, the plume becomes more dispersed and dilute, but filaments with concentrations above the threshold required to elicit a behavioral response from receiving organisms can persist for long distances. Flying insects orient along plumes by steering upwind, triggered by the optomotor reaction. Sequential measurements of differences in odor concentration are unreliable indicators of distance to or direction of the odor source. Plume intermittency and the plume's fine-scale structure can play a role in setting an insect's upwind course. The prowess of insects in navigating to odor sources has spawned bioinspired virtual models and even odor-seeking robots, although some of these approaches use mechanisms that are unnecessarily complex and probably exceed an insect's processing capabilities.

A model of the burglar alarm hypothesis of prey alarm calls.

When approached by a predator many prey species will emit an "alarm call" as a form of anti-predator behavior. One hypothesis for the function of alarm calls is the "burglar alarm" hypothesis whereby upon attack, a prey renders itself dangerous to a predator by generating an alarm call that attracts a predator at higher trophic levels in the food chain; that is, attracts a predator to the prey's own predator. This paper concerns a model incorporating a mechanism to test the burglar alarm hypothesis. We prove in one space dimension global existence, of positive bounded classical solutions, and establish existence of non-constant equilibrium solutions and assess their stability. We provide some representative numerical simulations to emphasize the nature of pattern formation for this model and demonstrate the benefit achieved by a signal inducing prey species under the burglar alarm hypothesis.

Aversive operant conditioning alters the phototactic orientation of the marbled crayfish.

Aversive learning was applied to affect the phototactic behaviour of the marbled crayfish. Animals initially showed negative phototaxis to white light and positive taxis to blue light. Using an aversive learning paradigm, we investigated the plasticity of innate behaviour following operant conditioning. The initial rate of choosing a blue-lit exit was analysed by a dual choice experiment between blue-lit and white-lit exits in pre-test conditions. During training, electrical shocks were applied to the animals when they oriented to the blue-lit exit. Memory tests were given to analyse the orientation rate to the blue-lit exit in trials 1 and 24 h after training and these rates were compared with the pre-test. In general, animals avoided the blue-lit exit in the memory tests. When training was carried out three times, the long-term memory was retained for at least 48 h, although a single bout of training was also enough to form a long-term memory. Cooling animals at 4°C or injection of cycloheximide immediately after training altered the formation of long-term memory, but had no effect on short-term memory formation. Administration of the adenylate cyclase inhibitor SQ22536, the PKA inhibitor H89 or the CREB inhibitor KG-501 immediately after training also blocked the formation of long-term memory, but had no effect on short-term memory formation. Thus, our pharmacological behavioural analyses showed that new protein synthesis was necessary to form long-term memories and that the cAMP/PKA/CREB pathway is the main signal cascade for long-term memory formation in the marbled crayfish.

Spatial drivers of instability in marine size-spectrum ecosystems.

Size-spectrum models are a recent class of models describing the dynamics of a whole community based on a description of individual organisms. The models are motivated by marine ecosystems where they cover the size range from multicellular plankton to the largest fish. We propose to extend the size-spectrum model with spatial components. The spatial dynamics is governed by a random motion and a directed movement in the direction of increased fitness, which we call 'fitness-taxis'. We use the model to explore whether spatial irregularities of marine communities can occur due to the internal dynamics of predator-prey interactions and spatial movements. This corresponds to a pattern-formation analysis generalized to an entire ecosystem but is not limited to one prey and one predator population. The analyses take the form of Fourier analysis and numerical experiments. Results show that diffusion always stabilizes the equilibrium but fitness-taxis destabilizes it, leading to non-stationary spatially inhomogeneous population densities, which are travelling in size. However, there is a strong asymmetry between fitness-induced destabilizing effects and diffusion-induced stabilizing effects with the latter dominating over the former. These findings reveal that fitness taxis acts as a possible mechanism behind pattern formations in ecosystems with high diversity of organism sizes, which can drive the emergence of spatial heterogeneity even in a spatially homogeneous environment.

Energy Taxis toward Redox-Active Surfaces Decreases the Transport of Electroactive Bacteria in Saturated Porous Media.

The fate and transport of bacteria in porous media are essential for bioremediation and water quality control. However, the influence of biological activities like extracellular electron transfer (EET) and swimming motility toward granular media on cell transport remains unknown. Here, electroactive bacteria with higher Fe(III) reduction abilities were found to demonstrate greater retention in ferrihydrite-coated sand. Increasing the concentrations of the electron donor (1-10 mM lactate), shuttle (0-50 µM anthraquinone-2,6-disulfonate), and acceptor (ferrihydrite, MnO2, or biochar) under flow conditions significantly reduced Shewanella oneidensis MR-1's mobility through redox-active porous media. The deficiency of EET ability or flagellar motion and inhibition of intracellular proton motive force, all of which are essential for energy taxis, enhanced MR-1's transport. It was proposed that EET could facilitate MR-1 to sense, tactically move toward, and attach on redox-active media surface, eventually improving its retention. Positive linear correlations were established among parameters describing MR-1's energy taxis ability (relative taxis index), cell transport behavior (dispersion coefficient and relative change of effluent percentage), and redox activity of media surface (reduction potential or electron-accepting rate), providing novel insights into the critical impacts of bacterial microscale motility on macroscale cell transport through porous media.

Multiscale modeling of glioma pseudopalisades: contributions from the tumor microenvironment.

Gliomas are primary brain tumors with a high invasive potential and infiltrative spread. Among them, glioblastoma multiforme (GBM) exhibits microvascular hyperplasia and pronounced necrosis triggered by hypoxia. Histological samples showing garland-like hypercellular structures (so-called pseudopalisades) centered around the occlusion site of a capillary are typical for GBM and hint on poor prognosis of patient survival. We propose a multiscale modeling approach in the kinetic theory of active particles framework and deduce by an upscaling process a reaction-diffusion model with repellent pH-taxis. We prove existence of a unique global bounded classical solution for a version of the obtained macroscopic system and investigate the asymptotic behavior of the solution. Moreover, we study two different types of scaling and compare the behavior of the obtained macroscopic PDEs by way of simulations. These show that patterns (not necessarily of Turing type), including pseudopalisades, can be formed for some parameter ranges, in accordance with the tumor grade. This is true when the PDEs are obtained via parabolic scaling (undirected tissue), while no such patterns are observed for the PDEs arising by a hyperbolic limit (directed tissue). This suggests that brain tissue might be undirected - at least as far as glioma migration is concerned. We also investigate two different ways of including cell level descriptions of response to hypoxia and the way they are related .

Mechanistic home range analysis reveals drivers of space use patterns for a non-territorial passerine.

Home ranging is a near-ubiquitous phenomenon in the animal kingdom. Understanding the behavioural mechanisms that give rise to observed home range patterns is thus an important general question, and mechanistic home range analysis (MHRA) provides the tools to address it. However, such analysis has hitherto been principally restricted to scent-marking territorial animals, so its potential breadth of application has not been tested. Here, we apply MHRA to a population of long-tailed tits Aegithalos caudatus, a non-territorial passerine, in the non-breeding season where there is no clear 'central place' near which birds need to remain. The aim is to uncover the principal movement mechanisms underlying observed home range formation. Our foundational models consist of memory-mediated conspecific avoidance between flocks, combined with attraction to woodland. These are then modified to incorporate the effects of flock size and relatedness (i.e. kinship), to uncover the effect of these on the mechanisms of home range formation. We found that a simple model of spatial avoidance, together with attraction to the central parts of woodland areas, accurately captures long-tailed tit home range patterns. Refining these models further, we show that the magnitude of spatial avoidance by a flock is negatively correlated to both the relative size of the flock (compared to its neighbour) and the relatedness of the flock with its neighbour. Our study applies MHRA beyond the confines of scent-marking, territorial animals, so paves the way for much broader taxonomic application. These could potentially help uncover general properties underlying the emergence of animal space use patterns. This is also the first study to apply MHRA to questions of relatedness and flock size, thus broadening the potential possible applications of this suite of analytic techniques.