Toxic cyanobacterial blooms are a growing environmental concern that affects freshwater ecosystems, drinking water supplies, and public health. The cyanobacterium Microcystis is among the most important bloom forming species. It often grows in large colonies, which enhances its flotation, reduces grazing, and improves nutrient regulation. Microcystis cells are held together by a matrix of extracellular polymeric substances (EPS), making colony mechanics crucial for bloom formation. However, an analysis of the biomechanical properties of cyanobacterial colonies, and how these properties relate to environmental conditions like nutrient availability, remains largely missing. Here, we use micropipette force sensors to quantify the linear and non-linear mechanical properties of individual colonies at single-cell resolution. Bulk shear rheology complements these measurements by probing macroscopic properties. The measured tensile strength and yield stress are broadly comparable to those of bacterial biofilms and are far greater than the hydrodynamic stresses typically found in wind-mixed lakes. This implies that cyanobacterial colonies are highly resistant to fragmentation by natural mix
The life history of an individual coral is archived within the accreting skeleton of the colony. While reef-forming coral colonies (e.g. massive $\textit{Porites}$ sp.) may live for hundreds of years and deposit calcareous structures many metres in height and width, their living tissue is a thin outer surface layer comprised of asexually-dividing polyps that only survive a few years. To understand the rate and timing of polyp division and the consequences for colony skeletal growth, scientists need to track the skeletal corallite deposited around each polyp. Here we propose CoralLite, an annotated $μ$CT scan dataset of entire calcareous skeletons and an associated, first corallite deep learning reconstruction baseline. CoralLite combines fully quantified volumetric segmentations with cross-slice linking for visualisations of 3D models for each corallite up to colony scale. For segmentation, we propose and evaluate in detail a hybrid V-Trans-UNet architecture applicable to segmenting tiled $μ$CT virtual slabs of $\textit{Porites}$ sp. colonies. The model is pre-trained on weakly annotated data and topology-aware fine-tuned using fully annotated slice sections with 8k+ manual coralli
The detection and classification of bacterial colonies in images of agar-plates is important in microbiology, but is hindered by the lack of labeled datasets. Therefore, we propose Colony Grounded SAM2, a zero-shot inference pipeline to detect and segment bacterial colonies in multiple settings without any further training. By utilizing the pre-trained foundation models Grounding DINO and Segment Anything Model 2, fine-tuned to the microbiological domain, we developed a model that is robust to data changes. Results showed a mean Average Precision of 93.1\% and a $Dice@detection$ score of 0.85, showing excellent detection and segmentation capabilities on out-of-distribution datasets. The entire pipeline with model weights are shared open access to aid with annotation- and classification purposes in microbiology.
Fluid flow has a major effect on the aggregation and fragmentation of bacterial colonies. Yet, a generic framework to understand and predict how hydrodynamics affects colony size remains elusive. This study investigates how fluid flow affects the formation and maintenance of large colonial structures in cyanobacteria, using an experimental technique that precisely controls hydrodynamic conditions. We performed experiments on laboratory cultures and lake samples of the cyanobacterium Microcystis, while their colony size distribution was measured simultaneously by direct microscopic imaging. We demonstrate that EPS-embedded cells formed by cell division exhibit significant mechanical resistance to shear forces. However, at elevated hydrodynamic stress levels (exceeding those typically generated by surface wind mixing) these colonies experience fragmentation through an erosion process. We also show that single cells can aggregate into small colonies due to fluid flow. However, the structural integrity of these flow-induced colonies is weaker than that of colonies formed by cell division. We provide a mathematical analysis to support the experiments and demonstrate that a population mo
Mechanical interactions among cells in a growing microbial colony can significantly influence the colony's spatial genetic structure and, thus, evolutionary outcomes such as the fates of rare mutations. Here, we computationally investigate how this spatial genetic structure changes as a result of heritable phenotypic variations in cell shape. By modeling rod-like bacterial cells as lengthening and dividing circo-rectangles in a 2D Brownian dynamics framework, we simulate the growth of a colony containing two populations with different aspect ratios. Compared to monodisperse colonies, such bidisperse colonies exhibit diminished intermixing between sub-populations when the less elongated cells are too short to nematically order, instead forming large clusters. We find that the cells with longer aspect ratio gradually segregate to the colony periphery. We present evidence that this demixing is related to nematic order in the bulk and to active nematic mixing dynamics near the periphery. These findings are qualitatively robust across different growth rate protocols and initial conditions. Because the periphery is often an advantageous position when nutrients are limited, our results su
Bacteria are prolific at colonizing diverse surfaces under a widerange of environmental conditions, and exhibit fascinating examples of self-organization across scales. Though it has recently attracted considerable interest, the role of mechanical forces in the collective behavior of bacterial colonies is not yet fully understood. Here, we construct a model of growing rod-like bacteria, such as Escherichia coli based purely on mechanical forces. We perform overdamped molecular dynamics simulations of the colony starting from a few cells in contact with a surface. As the colony grows, microdomains of strongly aligned cells grow and proliferate. Our model captures both the initial growth of a bacterial colony and also shows characteristic signs of capturing the experimentally observed transition to multilayered colonies over longer timescales. We compare our results with experiments on E. coli cells and analyze the statistics of microdomains.
Bacterial colonies growing on surfaces are shaped by mechanical stresses transmitted through the community, governed by the balance between cell growth and steric and cell-substrate interactions. Using overdamped dynamics simulations of nonmotile, stress-responsive bacteria, we examine how purely mechanical interactions determine colony morphology and internal organization. Growth-induced extensile stresses compete with steric constraints, giving rise to the spontaneous formation of microdomains composed of highly aligned cells. We characterize this self-organization through the distribution of microdomain areas and a nematic order parameter that quantifies colony-wide alignment. Mechanosensitivity does not systematically alter domain structure, but increasing substrate friction reduces the mean domain size and broadens the diversity of orientations. Shifting the balance toward steric interactions, by lengthening the cell division size, slows the relaxation of colony shape toward isotropy and broadens the distribution of contact forces, producing a slower exponential decay. In dense colonies, strong forces are transmitted anisotropically through chains of aligned neighbors within m
We study the genetic interfaces between two species of an expanding colony that consists of individual microorganisms that reproduce and undergo diffusion, both at the frontier and in the interior. Within the bulk of the colony, the genetic interface is controlled in a simple way via interspecies interactions. However, at the frontier of the colony, the genetic interface width saturates at finite values for long times, both for neutral strains and interspecies interactions such as antagonism. This finite width arises from geometric effects: genetic interfaces drift toward local minima at an undulating colony frontier, where a focusing mechanism induced by curvature impedes diffusive mixing. Numerical simulations support a logarithmic dependence of the genetic interface width on the strength of the number fluctuations.
Segregation of populations is a key question in evolution theory. One important aspect is the relation between spatial organization and the population's composition. Here we study a specific example -- sectors in expanding bacterial colonies. Such sectors are spatially segregated sub-populations of mutants. The sectors can be seen both in disk-shaped colonies and in branching colonies. We study the sectors using two models we have used in the past to study bacterial colonies -- a continuous reaction-diffusion model with non-linear diffusion and a discrete ``Communicating Walkers'' model. We find that in expanding colonies, and especially in branching colonies, segregation processes are more likely than in a spatially static population. One such process is the establishment of stable sub- population having neutral mutation. Another example is the maintenance of wild-type population along side with sub-population of advantageous mutants. Understanding such processes in bacterial colonies is an important subject by itself, as well as a model system for similar processes in other spreading populations.
Colonies of bacteria endowed with a pili-based self-propulsion machinery are ideal models for investigating the structure and dynamics of active many-particle systems. We study Neisseria gonorrhoeae colonies with a molecular-dynamics-based approach. A generic, adaptable simulation method for particle systems with fluctuating bond-like interactions is devised. The simulations are employed to investigate growth of bacterial colonies and the dependence of the colony structure on cell-cell interactions. In colonies, pilus retraction enhances local ordering. For colonies consisting of different types of cells, the simulations show a segregation depending on the pili-mediated interactions among different cells. These results agree with experimental observations. Next, we quantify the power-spectral density of colony-shape fluctuations in silico. Simulations predict a strong violation of the equilibrium fluctuation-response relation. Furthermore, we show that active force generation enables colonies to spread on surfaces and to invade narrow channels. The methodology can serve as a foundation for future studies of active many-particle systems at boundaries with complex shape.
Bacteria build multicellular communities termed biofilms, which are often encased in a self-secreted extracellular matrix that gives the community mechanical strength and protection against harsh chemicals. How bacteria assemble distinct multicellular structures in response to different environmental conditions remains incompletely understood. Here, we investigated the connection between bacteria colony mechanics and the colony growth substrate by measuring the oscillatory shear and compressive rheology of bacteria colonies grown on agar substrates. We found that bacteria colonies modify their own mechanical properties in response to shear and uniaxial compression with the increasing agar concentration of their growth substrate. These findings highlight that mechanical interactions between bacteria and their microenvironment are an important element in bacteria colony development, which can aid in developing strategies to disrupt or reduce biofilm growth.
We study the stochastic hydrodynamics of colonies of flagellated swimming cells, typified by multicellular choanoflagellates, which can form both rosette and chainlike shapes. The objective is to link cell-scale dynamics to colony-scale dynamics for various colonial morphologies. Via autoregressive stochastic models for the cycle-averaged flagellar force dynamics and statistical models for demographic cell-to-cell variability in flagellar properties and placement, we derive effective transport properties of the colonies, including cell-to-cell variability. We provide the most quantitative detail on disclike geometries to model rosettes, but also present formulas for the dynamics of general planar colony morphologies, which includes planar chain-like configurations.
The maintenance of the pluripotent state in human embryonic stem cells (hESCs) is critical for further application in regenerative medicine, drug testing and studies of fundamental biology. Currently, the selection of the best quality cells and colonies for propagation is typically performed by eye, in terms of the displayed morphological features, such as prominent/abundant nucleoli and a colony with a tightly packed appearance and a well-defined edge. Using image analysis and computational tools, we precisely quantify these properties using phase-contrast images of hESC colonies of different sizes (0.1 -- 1.1$\, \text{mm}^2$) during days 2, 3 and 4 after plating. Our analyses reveal noticeable differences in their structure influenced directly by the colony area $A$. Large colonies ($A > 0.6 \, \text{mm}^2$) have cells with smaller nuclei and a short intercellular distance when compared with small colonies ($A < 0.2 \, \text{mm}^2$). The gaps between the cells, which are present in small and medium sized colonies with $A \le 0.6 \, \text{mm}^2$, disappear in large colonies ($A > 0.6 \, \text{mm}^2$) due to the proliferation of the cells in the bulk. This increases the co
The dynamics of growth of bacterial populations has been extensively studied for planktonic cells in well-agitated liquid culture, in which all cells have equal access to nutrients. In the real world, bacteria are more likely to live in physically structured habitats as colonies, within which individual cells vary in their access to nutrients. The dynamics of bacterial growth in such conditions is poorly understood, and, unlike that for liquid culture, there is not a standard broadly used mathematical model for bacterial populations growing in colonies in three dimensions (3-d). By extending the classic Monod model of resource-limited population growth to allow for spatial heterogeneity in the bacterial access to nutrients, we develop a 3-d model of colonies, in which bacteria consume diffusing nutrients in their vicinity. By following the changes in density of E.coli in liquid and embedded in glucose-limited soft agar, we evaluate the fit of this model to experimental data. The model accounts for the experimentally observed presence of a sub-exponential, diffusion-limited growth regime in colonies, which is absent in liquid cultures. The model predicts and our experiments confirm
The in vitro clonogenic assay is a technique to study the ability of a cell to form a colony in a culture dish. By optical imaging, dishes with stained colonies can be scanned and assessed digitally. Identification, segmentation and counting of stained colonies play a vital part in high-throughput screening and quantitative assessment of biological assays. Image processing of such pictured/scanned assays can be affected by image/scan acquisition artifacts like background noise and spatially varying illumination, and contaminants in the suspension medium. Although existing approaches tackle these issues, the segmentation quality requires further improvement, particularly on noisy and low contrast images. In this work, we present an objective and versatile machine learning procedure to amend these issues by characterizing, extracting and segmenting inquired colonies using principal component analysis, k-means clustering and a modified watershed segmentation algorithm. The intention is to automatically identify visible colonies through spatial texture assessment and accordingly discriminate them from background in preparation for successive segmentation. The proposed segmentation algo
The emergent spatial patterns generated by growing bacterial colonies have been the focus of intense study in physics during the last twenty years. Both experimental and theoretical investigations have made possible a clear qualitative picture of the different structures that such colonies can exhibit, depending on the medium on which they are growing. However, there are relatively few quantitative descriptions of these patterns. In this paper, we use a mechanistically detailed simulation framework to measure the scaling exponents associated with the advancing fronts of bacterial colonies on hard agar substrata, aiming to discern the universality class to which the system belongs. We show that the universal behavior exhibited by the colonies can be much richer than previously reported, and we propose the possibility of up to four different sub-phases within the medium-to-high nutrient concentration regime. We hypothesize that the quenched disorder that characterizes one of these sub-phases is an emergent property of the growth and division of bacteria competing for limited space and nutrients.
We describe our optical and electron-microscopy observations of pearlite structures in eutectoid steels which seem to imply that the mechanisms of formation of pearlite colonies in these steels differ from those observed earlier for non-eutectoid steels. A simple theoretical model to study kinetics of pearlite transformations is suggested. Simulations of growth of pearlite colonies based on this model reveal that for the volume carbon diffusion mechanism usually-supposed such growth is always unstable, and the steady-state growth can be realized only via the interfacial carbon diffusion mechanism. A model of formation of pearlite colonies based on the assumption of a strong enhancement of carbon diffusion near grain boundaries is also suggested. The model can be applicable to the plastically deformed steels, and the results of simulations based on this model qualitatively agree with some microstructural features of formation of pearlite colonies observed in such steels.
We introduce and study a stochastic model for the dynamics of colonial species, which reproduce through fission or fragmentation. The fission rate depends on the relative sizes of colonies in the population, and the growth rate of colonies is influenced by intrinsic and environmental stochasticities. Our setting thus captures the effect of an external noise, correlating the trait dynamics of all colonies. In particular, we study the effect of the strength of this correlation on the distribution of resources between colonies. We then extend this model to a large class of structured branching processes with interactions in which the particle type evolves according to a diffusion. The branching and death rates are general functions of the whole population. In this framework, we derive a $ψ$-spine construction and a Many-to-One formula, extending previous works on interacting branching processes. Using this spinal construction, we also propose an alternative simulation method and illustrate its efficiency on the colonial population model. The extended framework we propose can model various ecological systems with interactions, and individual and environmental noises.
Honey bees face an increasing number of stressors that disrupt the natural behaviour of colonies and, in extreme cases, can lead to their collapse. Quantifying the status and resilience of colonies is essential to measure the impact of stressors and to identify colonies at risk. In this manuscript, we present and apply new methodologies to efficiently diagnose the status of a honey bee colony from widely available time series of hive and environmental temperature. Healthy hives have a remarkable ability to control temperature near the brood area. Our method exploits this fact and quantifies the status of a hive by measuring how resilient they are to extreme environmental temperatures, which act as natural stressors. Analysing 22 hives during different times of the year, including 3 hives that collapsed, we find the statistical signatures of stress that reveal whether honeybees are doing well or are at risk of failure. Based on these analyses, we propose a simple scale of hive status (stable, warning, and collapse) that can be determined based on a few temperature measurements. Our approach offers a lower-cost and practical bee-monitoring solution, providing a non-invasive way to tr
Bacterial colonies can form a wide variety of shapes and structures based on ambient and internal conditions. To help understand the mechanisms that determine the structure of and the diversity within these colonies, various numerical modeling techniques have been applied. The most commonly used ones are continuum models, agent-based models, and lattice models. Continuum models are usually computationally fast, but disregard information at the level of the individual, which can be crucial to understanding diversity in a colony. Agent-based models resolve local details to a greater level, but are computationally costly. Lattice-based approaches strike a balance between these two limiting cases. However, this is known to come at the price of introducing undesirable artifacts into the structure of the colonies. For instance, square lattices tend to produce square colonies even where an isotropic shape is expected. Here, we aim to overcome these limitations and therefore study lattice-induced orientational symmetry in a class of hybrid numerical methods that combine aspects of lattice-based and continuum descriptions. We characterize these artifacts and show that they can be circumvent