BacFighT6: Simulation of T6SS-mediated Bacterial Interactions

Version: 10.0 (27.6.2026)

Developed by: Marek Basler (University of Basel, Biozentrum) with assistance from Gemini 2.5 - 3.5.

Inspired by:

• Pukatzki et al. 2006 (https://doi.org/10.1073/pnas.0510322103)

• Russell et al. 2011 (https://doi.org/10.1038/nature10244)

• Basler et al. 2012 (https://doi.org/10.1038/nature10846)

• Alteri et al. 2013 (https://doi.org/10.1371/journal.ppat.1003608)

• Basler et al. 2013 (https://doi.org/10.1016/j.cell.2013.01.042)

• Borgeaud et al. 2015 (https://doi.org/10.1126/science.1260064)

• Borenstein et al. 2015 (https://doi.org/10.1371/journal.pcbi.1004520)

• Ringel et al. 2017 (https://doi.org/10.1016/j.celrep.2017.12.020)

• Mavridou et al. 2018 (https://doi.org/10.1016/j.cub.2017.12.030)

• Smith and Vettiger et al. 2020 (https://doi.org/10.1371/journal.pbio.3000720)

• Smith et al. 2020 (https://doi.org/10.1038/s41467-020-19017-z)

• Lin et al. 2022 (https://doi.org/10.15252/embj.2021108595)

• Mashruwala et al. 2022 (https://doi.org/10.1016/j.cell.2022.09.003)

• Flaugnatti et al. 2025 (https://doi.org/10.7554/eLife.101032)

• Brüderlin et al. 2025 (https://doi.org/10.1126/sciadv.adr1713)

• Stubbusch et al. 2025 (https://doi.org/10.1126/science.adr8286)

Reviews on various aspects of the Type VI Secretion System:

• Wang et al. 2019 (https://doi.org/10.1146/annurev-micro-020518-115420)

• Hespanhol et al. 2023 (https://doi.org/10.1099/mic.0.001376)

• Wohlfarth et al. 2025 (https://doi.org/10.1038/s41579-025-01256-w)

1. Overview

This simulation models the dynamic interactions between up to three distinct bacterial cell types—Attackers, Prey, and Defenders—within a shared environment. The primary mechanism of interaction is the Type VI Secretion System (T6SS), a sophisticated molecular weapon that bacteria use to inject toxins into neighboring cells upon direct contact. Additionally, Prey cells are not entirely defenseless; they can produce and release their own bacteriocin toxins into the environment, allowing them to kill or inhibit rival Attackers and Defenders.

Cells inhabit a two-dimensional hexagonal grid and compete for space. The simulation progresses in discrete time steps, where each step typically represents one minute of real time. During each step, cells can undergo several actions:

• Replication: Cells can divide to produce offspring if space is available and their internal replication cooldown has completed.

• Replication Reward (Lysis-for-Replication): Attacker and Defender cells can be rewarded for lytic activity, earning an accelerated or entirely new replication cycle for successfully lysing a set number of targets. This creates a powerful fitness feedback loop where combat success directly drives proliferation.

• T6SS Firing: Attacker and Defender cells can fire their T6SS, targeting adjacent cells. Attacker firing is regulated by a cell density-dependent Quorum Sensing system.

• Toxin Effects: Cells hit by T6SS toxins accumulate damage. This can lead to cell death (growth arrest) or lysis (cell bursting), depending on the type and amount of toxins received.

• Retaliation: Defender cells can retaliate specifically against attackers.

• Cell Movement: Cells can move to adjacent empty hexes based on configurable cooldowns, probabilities, and directional preferences.

• Attacker Chemotaxis: Attacker cells can exhibit chemotactic behavior, preferentially moving towards higher concentrations of autoinducers released by Prey cells.

• Prey Capsule Formation: Prey cells can develop protective capsule layers in response to their own local cell density (via Prey autoinducer signals), offering a chance to resist T6SS attacks.

• Prey Toxin Production & Release: Prey cells can synthesize and release their own toxins (bacteriocins) to inhibit or lyse Attacker and Defender cells, either continuously or via a lysis-dependent release mechanism.

• Reporter System: Lysis of Prey cells releases a reporter enzyme (LacZ), which converts a substrate (CPRG) in the environment. This conversion is visualized by a change in the arena's background color, providing a proxy for overall Prey lysis.

The simulation allows users to configure a vast array of parameters, offering a flexible platform to explore complex ecological dynamics, the evolution of T6SS strategies, and the outcomes of bacterial warfare under different conditions. It is inspired by numerous experimental findings and theoretical models in microbial ecology and T6SS research. 🎮 Click here to launch "Battle Royale" demo

2. Getting Started & User Interface

The simulation interface consists of four main parts:

1. Control Panel (Left): Contains all settings, setup options, and simulation controls. This panel now includes the Time-Travel Control section for history navigation.

2. Simulation Arena (Center): The hexagonal grid where bacterial interactions are visualized.

3. Simulation Statistics (Right): Located on the right side of the arena, above the Hex Inspector, this panel displays real-time statistics. Cell-type specific data (live, dead/lysing, kills, lyses) are shown in a table format, while general simulation metrics (time, total cells, firings, CPRG conversion) are listed below it.

4. Hex Inspector (Right): A panel that displays detailed real-time information about the hexagonal cell your mouse is currently hovering over.

Basic Workflow:

1. Configure Arena & Initial Population:

   • The simulation starts with an empty arena in manual placement mode by default.

   • Select a tool from the "Setup Arena" panel (e.g., 'Att', 'Prey', 'Def', 'Barr', or 'Rem'). The active tool is highlighted with a colored ring.

   • Click and drag on the arena to place or remove cells.

   • For quick setups, you can use:

     • Place Cells Randomly: Fill the arena randomly based on the "Initial Count" specified for each type under "Cell-Specific Settings". The positions are defined by seed.

     • Clear All Placed: Empties the arena.

     • Import Arena (TSV): Load a predefined spatial arrangement from a TSV file.

     • Export Arena (TSV): Save your current manual layout.

   • The simulation is ready to run as soon as you have placed cells.

2. Adjust Parameters:

   • Navigate through the control panel sections: "Simulation Control," "General Settings," "Exports," "Cell-Specific Settings" (for Attacker, Prey, and Defender), and "CPRG Reporter Settings."

   • Modify parameters as needed for your experiment. You can also "Load Preset Scenario" for pre-configured setups.

3. Run the Simulation:

   • Use the main control buttons. These can also be activated via keyboard shortcuts:

     • Start [S]: Begins or resumes the simulation.

     • Pause [P]: Temporarily halts the simulation.

     • One Step [O]: Advances the simulation by a single time step (one minute).

     • Stop & Report [R]: Ends the simulation run and displays the results report.

     • Step Back [Left Arrow] (when paused): Navigates back one step in the recorded history.

     • Step Forward [Right Arrow] (when paused): Navigates forward one step in the recorded history. Note: For better performance on large arenas and long runs, increase the Render Rate in the 'Simulation Control' panel to draw the arena less frequently.

4. Analyze Results:

   • Upon stopping, a Report Modal appears, summarizing the outcome and key statistics.

   • From the report, you can:

     • View Graph: Visualize population dynamics and CPRG conversion over time.

     • Save All Data: This is the main button to save archives of all currently buffered data (Images as ZIP, Arena States as ZIP, Full Session History as .bft6) and a TSV file with the data for the population dynamics graph.

     • Load Step to Arena: If Arena Layouts were saved, you can initialize the arena from a specific past simulation step.

3. The Simulation Arena

The arena is a hexagonal grid. Each hexagon represents a potential space a single bacterium can occupy.

• Coordinates: Cells are located using axial coordinates (q, r).

• Arena Radius: Set in the "Setup Arena" panel. This determines the size of the grid. This can be changed at any time while the simulation is paused. Changing the radius is non-destructive; it will not remove any cells you have placed, but cells outside the new radius will not be visible or interactive.

• Cell Placement: Cells are placed either randomly or manually. Replication results in new cells appearing in adjacent empty hexes. Additionally, cells can actively move to adjacent hexes based on defined movement parameters (see Section 6.1).

4. Visualizations

The simulation provides real-time visual feedback:

• Attacker Cells: Rendered as red hexagons.

• Prey Cells: Rendered as blue hexagons.

• Defender Cells: Rendered as yellow-orange hexagons.

• Barrier Cells: Rendered as dark brown hexagons. These are static, impassable obstacles.

• Cell States:

  • Live Cells: Displayed in their solid type-specific color.

  • Dead Cells: Appear as dark gray hexagons. They also feature a smaller, faint inner dot matching their original cell type color (e.g., a faint red dot for a dead attacker). Dead cells are metabolically inactive and cannot replicate or fire, but they continue to occupy space until they lyse.

  • Lysing Cells: Appear as light gray hexagons, also with a faint inner dot of their original color. These cells have accumulated enough lytic toxins to burst and are on a timer to be removed from the grid.

  • Prey Capsule Indication: A Prey cell with a capsule is indicated by a thicker, purple inner outline. This inner border is drawn just inside the cell's main outline. The thickness and darkness of this inner outline correspond to the number of capsule layers, providing a visual cue of its increased protection.

  • Prey Toxin Accumulation Indication (Lysis Mode): Prey cells actively producing and accumulating internal toxins feature a green inner circle that darkens as the cell approaches its self-induced lysis threshold.

• T6SS Firing Events:

  • A bright green sector emanating from an attacking cell indicates a "precise hit" that can deliver toxins.

  • A darker, semi-transparent green sector (Attacker cells only) indicates a "missed" T6SS firing attempt that resets the cooldown but delivers no toxins.

• CPRG Reporter (Arena Background Color): The background color changes from white towards magenta as the amount of converted CPRG (released from lysed Prey) increases.

• Prey AI Field: A faint, transparent blue overlay on empty hexes visualizes the concentration of the Prey autoinducer signal. The overlay becomes more opaque as the concentration crosses key thresholds: Attacker's attraction limit and a fraction of the theoretical maximum AI level. This helps visualize areas attractive to Attackers.

• Prey Toxin Field: A transparent green overlay on empty hexes visualizes the concentration of the Prey-derived toxins (bacteriocins NL and L) in the environment. The overlay becomes more opaque as the local toxin concentration increases, helping visualize the chemical defensive barrier created by Prey cells.

• Empty Spaces: Unoccupied hexes are rendered with a light gray outline.

• Hex Inspector: This panel provides detailed, real-time data for the hex under the mouse cursor, including coordinates, local AI concentrations, cell ID and type, cooldowns, toxin levels, and special states like QS P(active) (for Attackers) or Capsule Layers (for Prey).

5. Simulation Controls & Settings Detailed

5.1. Setup Arena (Panel)

• Arena Radius: An integer defining the radius of the hexagonal grid.

• Placement Buttons: (Att, Prey, Def, Barr, Rem): Selects the action performed when clicking on a hex in the arena. The active tool is highlighted.

• Place Cells Randomly: Clears the arena and populates it with a random spatial distribution of cells based on their "Initial Count" settings.

• Clear All Placed: Removes all cells from the arena.

• Import Arena (TSV): Loads an initial cell configuration from a user-provided Tab-Separated Values (TSV) file (q\tr\ttype).

• Export Arena (TSV): Saves the current manually placed cell configuration to a TSV file.

5.2. Simulation Control (Panel)

This panel features the main simulation parameters as well as advanced controls for reproducibility.

• Step Delay (ms): The delay in milliseconds between consecutive rendering steps. Lower values result in faster animation.

• Render Rate: Render the arena only every N steps. '1' renders every step. '10' renders on steps 0, 10, 20, etc. Higher numbers improve performance.

• Duration (min): The maximum number of simulation steps (minutes) the simulation will run before automatically pausing.

• Seed: The input field for the random number generator (RNG) seed. The simulation's outcome is determined by this seed.

• New Seed Button (🔄): Generates a new, random seed for starting a completely new experiment.

• RNG Reset Button (🡄): This button appears (in green) when the RNG has been used. Clicking it resets the random number sequence to the beginning for the current seed, without changing the seed itself. This is essential for precisely replicating starting conditions.

• Cell Sync Button (🔄): This button appears (in red) at the top of the "Setup Arena" panel whenever placed cells become de-synchronized from the RNG (e.g., if you change the seed after placing cells). Clicking the red 🔄 button will iterate through every cell on the grid and re-calculate all of its initial, randomly-determined properties using the currently active seed, bringing the population back into a valid, reproducible state with the current seed.

• Reset Simulation Button: This performs a hard reset. It clears the arena, erases all history, and generates a new random seed.

5.3. Main Control Buttons

• Start [S]: Begins the simulation from step 0 or resumes a paused simulation.

• Pause [P]: Halts the simulation at the current step.

• One Step [O]: Advances the simulation by exactly one step. Useful for detailed observation.

• Stop & Report [R]: Terminates the current simulation run and displays the end-of-simulation report modal.

5.4. Time-Travel Control (Panel)

• Scrub Through History Slider: This slider becomes active if 'Full State History' is enabled. When the simulation is paused, you can drag the slider (or use the step buttons/arrow keys) to view the state of the arena at any past time step.

• Step Back (◀) & Step Forward (▶) Buttons: When paused, click these to step backward or forward through the history one step at a time, based on the current Render Rate.

• Play & Stop Autoplay Buttons: When paused, click Play to automatically replay the simulation history forward step-by-step. The playback speed is controlled by the Step Delay (ms) setting. Click Stop to halt the autoplay.

• Resume Simulation from this State: When scrubbing to a past point, this button becomes active. It is one of two ways to create a new timeline. Clicking it will discard all future history and immediately start the simulation from the selected point. Alternatively, you can simply start modifying the arena (e.g., placing a cell), which will also automatically branch the timeline, after which you can use the main "Start" or "One Step" buttons to proceed.

• Import Session (.bft6): Loads a complete, previously saved simulation session, including all settings and the full step-by-step history. During the import process, a Stop & Save History Loaded button is available. Clicking it terminates the file reading immediately, commits the loaded frames to the database, and sets the last successfully read step as the end of the history.

• Import Step (JSON): Loads a complete simulation state from a previously exported JSON file. This overwrites all current settings and cell placements to perfectly replicate the conditions of the saved step. After importing, you can either resume the simulation to continue the original timeline, or click "New Seed" to branch off and run a new experiment from that exact starting point.

• Export Step (JSON): Exports the complete state of the currently viewed time step (live or historical) into a single, human-readable JSON file. This file contains all settings, cell properties, AI grid concentrations, and the precise RNG state, making it ideal for debugging or for use with the "Import Step" feature.

5.5. General Settings (Panel)

• Load Preset Scenario: Opens a modal allowing selection from various pre-defined parameter configurations. When you click a preset box to select it, you can adjust specific interactive sliders (e.g. fill percentage, cell ratios, AI levels) or selectors (e.g. selectivity, lysis/release mode, trigger mechanism) directly within that preset's panel before running it. Clicking "Apply Selected Preset & Close" will apply these customized overrides, reset the arena and RNG, and randomly populate the cells.

• Import/Export Settings (TSV): Loads or saves a full set of simulation parameters from a TSV file.

• Copy Shareable Link 🔗: Generates and copies a URL to your clipboard that reproduces your current simulation state.

  • What it saves: The current RNG Seed and every Parameter that differs from the default values.

  • What it doesn't save: Specific positions of manually placed cells. (It relies on the Seed + Initial Counts to reproduce random populations).

• Reset Settings to Defaults: Resets all simulation parameters in the control panel to their original default values. This action does not clear the arena or change the current simulation seed.

5.6. Exports & Buffers (Panel)

• Arena Layout: Checkbox. If enabled, the q,r,type of all active cells is recorded at each step. This data can be downloaded as a ZIP of TSV files. This also enables the "Load Step to Arena" feature in the report modal.

• Full State History: Checkbox. If enabled, the entire simulation state (all cell properties, AI grids, stats) is saved at each step. This is required for the Time-Travel feature and for saving/loading complete sessions (.bft6 files). ⚠️ This is a memory-intensive option.

• Arena Images: Checkbox. If enabled, a PNG image of the arena is captured at each step.

  • ⚠️ Warning: Saving states or images can consume very significant browser memory for long simulations.

• Image Size (px): Defines the width and height of the saved square PNG images.

• History Buffer Size Limit (MB): Sets the maximum memory threshold (RAM) for full state history frames. Once exceeded, the simulation automatically offloads older frames to the browser's local database (IndexedDB) to prevent crashes.

• Arena Layouts Buffer Size Limit (MB): Sets the maximum RAM threshold for caching cell coordinates and types (q,r,type). Exceeding this limit triggers automatic offloading to IndexedDB.

• Images Buffer Size Limit (MB): Sets the maximum RAM threshold for storing step-by-step PNG screenshots. When exceeded, images are offloaded to IndexedDB to protect memory.

5.7. CPRG Reporter Settings

• Initial CPRG Substrate: The total amount of CPRG substrate units available at the beginning of the simulation.

• LacZ kcat: The catalytic rate constant (turnover number) for LacZ.

• LacZ Km: The Michaelis constant for LacZ with CPRG.

5.8. Cell-Specific Settings (Panel)

This section of the control panel is tabbed for Attacker, Prey, and Defender types. Each tab contains parameters unique to that cell's behavior.

Common Properties (Across Tabs)

• Initial Count: The number of cells of this type to place when using the Place Cells Randomly function.

• Replication (min): The mean division time and the range (± variation) in minutes. The division cooldown is randomized per cell within this range. Setting the mean to -1 completely disables standard cell division for this cell type.

• Movement Behavior:

  • Cooldown Min/Max (min): The time range (in minutes/steps) a cell waits before trying to move again.

  • Probability (%): The chance (0-100%) that the cell will actually attempt to move when its cooldown reaches zero. Set to 0% for non-motile cells.

  • Directionality (%): Influences target hex selection. 100% means it looks only at empty adjacent hexes; 0% means it picks a random adjacent hex first and moves only if it happens to be empty. Intermediate values blend these two strategies.

1. Attacker Tab

• Movement Behavior (AI-Guided Attraction):

  • Prey AI Attraction (%): The probability that the Attacker will move toward the adjacent empty hex with the highest concentration of Prey-derived autoinducer (AI) signal instead of a random empty neighbor.

  • Prey AI Attraction Threshold: The minimum local concentration of Prey AI required to trigger guided chemotactic attraction. If all empty adjacent spots are below this, the cell falls back to random movement.

• T6SS Firing:

  • Fire Cooldown Min/Max (min): The randomized cooldown range (in minutes) after a T6SS firing event.

  • Precision (%): The chance that a firing event results in a precise hit (delivers toxins). The remaining percentage represents "misses" (resets cooldown but does no damage).

  • Contact Sensing Bias (%): The chance that the Attacker commits to firing only at a physical neighbor (another cell or barrier). If no contacts are adjacent, the firing is aborted. The remaining percentage represents a random firing direction attempt (which may hit empty space).

  • Kin Exclusion (%): The probability that the cell's self-recognition mechanism activates when targeting another Attacker cell.

  • Kin Exclusion Penalty (min): If kin recognition activates, a value $\ge 0$ cancels the shot and sets a cooldown penalty (in minutes); a value of -1 enables Smart Re-targeting, forcing the cell to pick a new, non-kin target from its neighbors.

• T6SS Effectors (vs Prey/Defenders):

  • NL Units/Hit & L Units/Hit: The number of Non-Lytic (NL) and Lytic (L) toxin units delivered per precise hit.

  • NL / L Delivery Chance (%): The individual probabilities that the NL or L toxin component is successfully injected during a hit.

• Quorum Sensing (QS) (T6SS Activation):

  • AI Prod. Rate (U/min): The units of Attacker autoinducer (AI) secreted by each live Attacker cell per step.

  • AI Degrad. Rate (%/min) & AI Diffusion Rate: The degradation percentage and spatial diffusion speed of the Attacker AI signal grid.

  • Activation Midpoint (K): The local AI concentration at which T6SS activation reaches 50% (Hill equation midpoint). Set to -1 for constitutive (always-active) T6SS.

  • Cooperativity (n): The Hill coefficient. A higher value makes the density-dependent activation switch sharper and more binary.

• Replication Reward:

  • Lyses per Reward: The number of successful prey or defender lyses this cell must cause to earn a reward. Set to 0 to disable.

  • Reward Repl. CD (min) [Mean/Range]: The division cooldown (or reduction) applied upon earning a reward. A mean of -1 triggers immediate replication.

• Prey Toxin Sensitivity:

  • NL Resist. (%) & L Resist. (%): The chance to completely ignore environmental Prey toxins absorbed from the grid.

  • NL / L Absorp. (%): The percentage of local grid-based Prey toxins absorbed by the cell in each step.

  • NL Die / L Lyse Thresh.: Cumulative absorbed Prey toxin units required to kill (growth arrest) or lyse (burst) the Attacker cell.

2. Prey Tab

• Sensitivity vs Attackers / Defenders:

  • NL Die Thresh. & L Lyse Thresh.: The cumulative toxin units received from Attacker/Defender hits required to cause cell death (growth arrest) or lysis.

  • Base Lysis Delay (min): The baseline delay before a lysing cell bursts. The active delay is calculated as ceil(Base Lysis Delay / effectiveLyticUnits).

  • NL / L Resist. (%): The chance to completely negate incoming Attacker or Defender toxins per hit.

• LacZ Units/Lysis: The units of LacZ enzyme released into the environment when this Prey cell lyses, driving the CPRG reporter reaction.

• Quorum Sensing (QS) (Autoinducer):

  • AI Prod. Rate (U/min): The units of Prey autoinducer (AI) secreted per minute.

  • AI Degradation & Diffusion: Rates governing the degradation and spread of Prey AI across the hex grid.

• Toxin Production & Release:

  • NL / L Prod. Rate (U/min): The units of Bacteriocin NL and L produced per minute.

  • NL / L Degradation & Diffusion: Rates governing the environmental spread and decay of Prey-derived bacteriocins.

  • Trigger Mode: The mechanism for activating toxin production. Can be:

    • Standard: Stochastic activation check based on the start probability.

    • QS: Activation probability scaled by local Prey AI density (Hill equation).

    • Attacker NL: Production is triggered once the Prey accumulates attacker NL toxins $\ge$ NL Init. Threshold.

  • Stochastic Start Prob. (%/min): The active chance (or scaling probability) for initiating toxin production. Must be $>0$ for any trigger mode to activate.

  • QS Derepression Midpoint (K) & QS Cooperativity (n): Activation parameters for the QS trigger mode.

  • NL Init. Threshold: The minimum attacker NL toxin damage required to trigger production in the Attacker NL mode.

  • Release On Lysis: If checked (true), toxins accumulate internally and division is arrested until the cell crosses a random lysis threshold (between the configured Min and Max limits), at which point it lyses and dumps all internal toxins onto the grid. If unchecked (false), toxins are continuously secreted into the grid, and cells return to normal growth once their production target is reached.

• Capsule Synthesis (Defense):

  • Enable Capsule Synthesis: Master switch to activate capsule protection.

  • Max Protection (%): The maximum protection chance provided by a full 5-layer capsule. Each layer provides 1/5th of this value.

  • Derepression Midpoint (K) & Cooperativity (n): Controls for the derepression switch that responds to local Prey AI concentration. Setting K to -1 makes capsule synthesis always-on (constitutive).

  • Capsule Cooldown Min/Max (min): The randomized time range required to synthesize one protective capsule layer.

3. Defender Tab

• Retaliation Behavior:

  • Sense Chance (%): The probability that being hit by a T6SS attack will trigger Defender retaliation.

  • Max Retaliations: The maximum number of counter-attack shots fired in a single burst (one shot per step) at the attacking cell.

• Random Firing:

  • Rand. Fire CD (min-max): The cooldown range (in steps) between proactive random firing attempts.

  • Rand. Fire Chance (%): The probability per step (when not on cooldown or retaliating) to fire a single T6SS shot at a random adjacent cell.

• T6SS Effectors (vs Attackers/Prey):

  • NL / L Units/Hit and NL / L Delivery Chance (%): Toxins delivered and injection chances when attacking other cells.

• Replication Reward:

  • Lyses per Reward & Reward Repl. CD (min) [Mean/Range]: Lytic reward settings identical in function to the Attacker's reward system.

• Sensitivity vs Attackers:

  • NL Die Thresh., L Lyse Thresh., Base Lysis Delay, and NL / L Resist. (%): Toxin thresholds and resistance chances against direct Attacker T6SS hits.

• Prey Toxin Sensitivity:

  • NL / L Resist. (%), NL / L Absorp. (%), and NL Die / L Lyse Thresh.: Configurable parameters for environmental Prey toxin absorption, resistance, and survival thresholds.

6. Cell Types & Detailed Mechanics ("Under the Hood")

Each cell in the simulation is an object with properties tracking its state and behavior.

6.1. General Cell Properties & Processes

• Coordinates & ID: Each cell has q and r axial coordinates and a unique id.

• Type: attacker, prey, or defender.

• Replication:

  • A cell becomes eligible to replicate when its replicationCooldown timer reaches zero. This timer is initialized randomly for new cells and reset after each successful division based on "Mean" and "Range (±)" settings for its type (e.g., time = mean ± random_value_within_range).

  • Replication for any cell type can be disabled by setting its 'Mean' replication time to -1 in the settings.

  • For replication to occur, an empty, valid hexagonal space must be adjacent to the parent cell.

  • If multiple empty neighbors exist, one is chosen randomly for the daughter cell.

  • Both parent and new daughter cell then get their replicationCooldown reset.

  • If no empty space is available when a cell is eligible to divide, its replicationCooldown is simply reset (simulating a missed division opportunity due to crowding).

  • Example: A Prey cell has replication settings: Mean=20min, Range=5min. Its next division time might be randomly set to, say, 18 minutes. After 18 simulation steps, if an adjacent hex is empty, it divides. If not, its cooldown resets, and it tries again later.

• Replication Reward (via Lysis):

  • To simulate a fitness advantage for successful killers, both Attacker and Defender cells can earn a replication reward for their lytic activity. This system provides a direct feedback loop where effectiveness in combat translates to faster proliferation.

  • Triggering Mechanism: Each Attacker and Defender maintains a personal counter for the number of cells it has successfully lysed (lyses). A reward is triggered when this counter reaches a multiple of the value set in Lyses per Reward. For example, if this is set to 3, a cell will receive a reward when its lyses count reaches 3, then again at 6, 9, and so on. The system internally tracks "claimed" rewards to ensure that a reward is given only once per threshold crossing. This feature is disabled if Lyses per Reward is set to 0.

  • Reward Action: When a reward is triggered, the cell's replicationCooldown is modified based on the Reward Repl. CD (min) settings:

    • Accelerating an Existing Cycle: If the cell is already counting down to a normal replication, the reward value is subtracted from this cooldown, accelerating the division.

    • Initiating a New Cycle: If the cell is not currently in a replication cycle (e.g., because normal replication is disabled), this reward initiates a new, fast replication cycle, setting the replicationCooldown to the reward value.

  • If the Reward Repl. CD (min) mean value is set to -1, the reward is immediate, setting the replicationCooldown to 0.

  • Example: A Attacker has its normal replication disabled (Mean = -1). Its reward settings are Lyses per Reward = 2 and Reward Repl. CD = 10 ± 2 min. The Attacker lyses its first target; its lyses count is now 1. Later, it lyses a second target, bringing its count to 2. This meets the threshold and triggers a reward. Since its replicationCooldown was Infinity, a new cycle is initiated with a cooldown between 8 and 12 minutes.

• Toxin Accumulation & Cell Fate:

  • Cells accumulate damage from two distinct sources: direct Type VI Secretion System (T6SS) contact attacks (from Attackers or Defenders) and environmental toxin absorption (from Prey-derived bacteriocins in the grid).

  • T6SS Contact Attacks (Attacker & Defender T6SS):

    • Prey Capsule Resistance (Prey Only): If the capsule system is enabled and the target is a Prey cell with active capsule layers, there's a chance the attack is rendered harmless. The maximum possible protection (e.g., 100%, 80%, etc.) is user-definable via the 'Max Protection (%)' setting. Each of the 5 possible layers provides 1/5th of this user-defined maximum. If the attack is negated by the capsule, subsequent resistance and toxin accumulation steps are skipped for that hit.

    • Innate Resistance Check (Prey & Defenders): If the attack was not negated by a capsule, Prey and Defenders then have a chance to resist NL and L toxins independently (based on % Resistance settings against the specific attacker type). If resisted, no toxins of that type are added from that hit component. Attackers do not have a configurable % Resistance setting against Defender attacks in the current model; they rely solely on their fixed sensitivity thresholds to determine the effects of accumulated toxins.

    • Toxin Accumulation: If not resisted by a capsule or innate resistance, the delivered toxin units are added to the cell's accumulators.

  • Environmental Toxin Absorption (Prey Bacteriocins):

    • Live Attacker and Defender cells absorb a percentage of Prey-derived toxins (NL and L) from their local grid cell at each step (defined by % Absorption settings).

    • They check their specific % Resistance against these absorbed toxins. If not resisted, the toxins accumulate in the cell.

  • State Update (updateStateBasedOnToxins):

    • Non-Lytic Death: If accumulated NL toxins (from either contact hits or environmental absorption) $\ge$ NL Die Thresh., the cell becomes isDead = true. It stops replicating and firing but remains on the grid.

    • Lytic Lysis: If accumulated L toxins $\ge$ L Lyse Thresh., the cell becomes isDead = true and isLysing = true. A lysisTimer is initiated. The duration of this timer is calculated as ceil(baseLysisDelay / effectiveLyticUnits), where effectiveLyticUnits is the actual accumulated lytic toxin (ensuring more toxin leads to faster lysis).

    • When lysisTimer reaches 0, or if lysis is immediate (e.g., delay calculates to 0), the cell becomes isEffectivelyGone. At this point, if it's a Prey cell, it releases its lacZPerPrey units. isEffectivelyGone cells are removed from active simulation and counts in the next step's cleanup phase.

  • Example: A Prey cell (L Lyse Thresh: 5, Base Lysis Delay: 20min) is hit by a Attacker delivering 3 Lytic units. Prey accumulates 3 L units. Later, another hit delivers 3 L units. Total L units = 6. Since 6 ≥ 5, Prey becomes isLysing. lysisTimer starts, e.g., ceil(20 / 6) = 4 minutes. After 4 steps, it becomes isEffectivelyGone.

Cell Movement

For each of the three cell types (Attacker, Prey, Defender), you can define their movement behavior using the following parameters, found in their respective "Cell-Specific Settings" sections under "Movement Behavior":

• Movement Cooldown Min (min) and Movement Cooldown Max (min):

  • This defines the time range (in simulation minutes/steps) a cell waits before it can attempt to move again.

  • When a cell needs a new movement cooldown (e.g., after an attempted move, or after deciding not to move due to probability), a random duration is chosen from this range (inclusive).

  • Example: If Min is 5 and Max is 10, the cell will wait between 5 and 10 minutes before its next movement attempt becomes possible.

• Movement Probability (%):

  • When a cell's movement cooldown reaches zero, this parameter (0-100%) determines the chance that the cell will actually attempt to make a move in the current simulation step.

  • If set to 0%, the cell is non-motile and will never attempt to move, even if its cooldown is zero.

  • If set to 100%, the cell will always attempt to move as soon as its cooldown is zero.

  • If a cell's cooldown is zero but it does not attempt to move due to this probability (e.g., a 30% probability means a 70% chance of not attempting the move right now), its movement cooldown will still be reset for the next attempt.

• Movement Directionality (%):

  • This parameter (0-100%) influences how a cell chooses its target square if it decides to attempt a move (i.e., its cooldown is zero AND it passed the "Movement Probability" check).

  • 100% Directionality: The cell will only look for adjacent empty squares. If it finds one or more, it will randomly pick one to move into. If there are no empty adjacent squares, the cell will not move in this attempt (but its cooldown will be reset).

  • 0% Directionality: The cell will pick one of its 6 adjacent squares completely at random, without regard for whether it's empty or occupied. However, the cell will only successfully move if this randomly chosen square is actually empty. If it picks an occupied square (another cell or a barrier), the move is cancelled for this attempt (and its cooldown is reset).

  • Between 0% and 100% (e.g., 75%): The cell has a Directionality % chance of using the "100% Directionality" strategy (look for empty spots only). If it doesn't use that strategy (i.e., with 100 - Directionality % chance), it will use the "0% Directionality" strategy (pick a random direction, then check if it's empty).

General Movement Rules:

• Cells attempt to move one hexagonal space at a time.

• Cells cannot move into squares already occupied by other cells or by barriers.

• If multiple cells attempt to move to the exact same empty square in the same simulation step, only one cell (chosen randomly from the contenders) will successfully occupy the square. The other cells' moves to that specific square will fail for that step.

• Regardless of whether a move attempt is successful, fails due to an occupied target, or fails due to conflict with another moving cell, the cell's movement cooldown is reset.

• In each simulation step, movement calculations occur after other actions like T6SS firing and cell replication. The state of the arena after movement becomes the state for the beginning of the next simulation minute.

State Synchronization & Reproducibility

For a simulation to be perfectly reproducible, two conditions must be met:

1. The Seed must be the same.

2. The sequence of random numbers drawn from that seed must be the same. A cell's initial random state (e.g., its starting replicationCooldown) is set the moment it is created, using the next available number from the RNG sequence. If you place a cell and then change the seed or reset the RNG sequence, that cell's state becomes "out of sync"—it was created with a random number that no longer corresponds to the current state of the generator. The simulation tracks this and provides a tool to correct it.

• The Cell Sync Button (🔄): This button appears (in red) at the top of the "Setup Arena" panel whenever a de-synchronization event occurs. This happens under two specific conditions:

  1. The Seed is changed (manually, via settings import, or with the 🔄 button) while there are already cells on the arena.

  2. The RNG sequence is reset to the beginning (using the 🡄 button) while there are already cells on the arena. Clicking the red 🔄 button will iterate through every cell currently on the grid and re-calculate all of its initial, randomly-determined properties. This action brings the entire population of cells back into a valid, reproducible state with the current seed and RNG sequence.

6.2. Attacker Cells

• Role: Actively hunt and attack Prey and Defender cells.

• Movement Behavior (AI-Guided): Attackers can use AI signals produced by Prey to guide their movement.

  • Prey AI Attraction (%): If the Attacker decides to move into an empty space (based on its 'Movement Directionality' setting), this is the chance (%) it will attempt to pick the empty neighboring hex that has the highest concentration of Prey-derived Autoinducer (Prey AI). A 0% chance means it will pick a random empty neighbor. A 100% chance means it will always try to pick the highest Prey AI spot.

  • Prey AI Attraction Threshold: This is the minimum concentration of Prey AI that must be present in an empty neighboring hex for that hex to be considered a candidate for the 'highest concentration' selection during Prey AI-attracted movement. If no empty neighbors meet this threshold, the Attacker will revert to choosing a random empty neighbor (if 'Movement Directionality' allows movement into empty hexes).

• Quorum Sensing (QS) Behavior (for T6SS Activation): These parameters control a cell-density-dependent communication system using the Attacker's own Autoinducer (AI). Attackers produce this AI signal, and when its local concentration is high enough, Attackers become "activated" and can fire their T6SS.

  • AI Prod. Rate (U/min): The number of AI signal units each live Attacker cell produces per minute.

  • AI Degrad. Rate (%/min): The percentage of AI signal that degrades or disappears from each hex per minute, simulating signal instability.

  • AI Diffusion Rate: A coefficient (0-0.166) that determines how fast AI spreads to adjacent hexes. It is based on the concentration gradient, meaning AI flows from areas of high concentration to low concentration. A higher value means faster diffusion.

  • Activation Midpoint (K): The threshold for T6SS activation, functioning as an EC50. If set to a value less than 0 (e.g., -1), QS is always active and T6SS firing is constitutive (not regulated by cell density). If set to 0, T6SS is active as long as any AI is present. If set to a value greater than 0, it acts as the midpoint of a Hill function, where T6SS activation is at 50% when the local AI concentration equals K.

  • Cooperativity (n): The Hill coefficient for the QS response. This controls the steepness and sensitivity of the switch. A value of 1 gives a gradual, graded response to AI concentration. Higher values (e.g., 4 or more) create a much sharper, more "all-or-nothing" switch.

• T6SS Firing Logic (runSimulationStep): The process for a Attacker attempting to fire follows a strict decision tree each simulation cycle:

  • Activation Check: A Attacker can only attempt to fire if it first becomes "active" based on the Quorum Sensing system and its t6ssFireCooldownTimer is 0.

  • Strategy Selection (Contact vs. Random): The simulation first determines the firing strategy based on the Contact Sensing Bias %.

    • If the random check passes (e.g., a 70% bias gives a 70% chance to pass), the Attacker commits to the "Contact-Biased" strategy.

    • Otherwise, it commits to the "Random" strategy.

  • Preliminary Target Selection:

    • If using the "Contact-Biased" strategy: The Attacker scans its adjacent hexes for contacts (any other cell or barrier). If one or more contacts are found, it randomly picks one as its preliminary target. If no contacts are found, the firing sequence is aborted for this cycle. The Attacker will not fire into an empty space.

    • If using the "Random" strategy: The Attacker picks one of the six possible directions at random and sets that hex as its preliminary target, regardless of whether it is empty or occupied.

  • Kin Exclusion Check: If a preliminary target was successfully selected in the previous step, the code now checks if that target is another Attacker.

    • If it is, the Kin Exclusion % determines if the recognition system activates.

    • If the system activates, the Kin Exclusion Penalty value dictates the outcome:

      • Penalty ≥ 0 ("Cancel & Penalize" mode): The shot is cancelled. If the penalty value is greater than 0, a special cooldown is applied.

      • Penalty = -1 ("Smart Re-targeting" mode): The shot is not cancelled. Instead, the Attacker attempts to find a new target by running the targeting logic again, but from a pool of neighbors that has been pre-filtered to exclude all kin. This new selection still respects the original Contact Sensing Bias strategy. If no valid non-kin targets can be found, the shot is aborted.

  • Final Firing Event: If the shot has not been aborted or cancelled, the firing event is visualized and counted. The simulation then checks the Precision % to determine if the shot was a "precise hit" (delivers toxins) or a "miss" (no damage).

    • Example of the firing logic: A Attacker cell is ready to fire, with the following setup:

    • Parameters: Contact Sensing Bias = 80%, Kin Exclusion = 90%, Kin Exclusion Penalty = -1 ("Smart Targeting" mode).

    • Environment: The Attacker is adjacent to another Attacker (kin), a Barrier (a non-kin contact), and an empty hex. In this scenario, the Attacker's decision process is as follows: With an 80% chance, it commits to a "Contact-Biased" strategy and selects an adjacent contact (e.g., the kin cell). The 90% kin exclusion check then succeeds and triggers "Smart Re-targeting" due to the -1 penalty. A new pool of non-kin targets is created ([Barrier, empty_hex]). Because the original strategy was "Contact-Biased", the simulation filters this new pool for contacts, leaving only the [Barrier]. The final shot is therefore correctly fired at the Barrier. This logic ensures the Contact Sensing Bias has priority, preventing the Attacker from firing into empty space when its intent is to hit a contact, even after a complex re-targeting event.

• T6SS Effectors (vs Prey/Defenders): Parameters define NL Units/Hit, NL Delivery Chance %, L Units/Hit, and L Delivery Chance % when attacking Prey or Defenders.

• Sensitivity (to Defender Attack): Attackers have their own thresholds (NL Die Thresh., L Lyse Thresh., Base Lysis Delay) for damage received from Defender attacks. They do not have % resistance values like Prey/Defenders.

• Sensitivity & Resistance vs. Prey Toxin:

  • NL Resist. (%) and L Resist. (%): Chance (0-100%) to completely ignore NL or L toxin absorbed from a local grid cell in a step.

  • NL Absorp. (%) and L Absorp. (%): Percentage of local grid-based Prey toxin absorbed by the Attacker cell in each step.

  • NL Die Thresh. and L Lyse Thresh.: The cumulative Prey toxin units (absorbed from the grid) required to kill (growth arrest) or lyse the Attacker cell.

6.3. Prey Cells

• Role: Primary targets for Attackers and Defenders. They can replicate, protect themselves via a capsule system, and actively synthesize/release their own bacteriocin toxins to damage or compete against rival cell types.

• Sensitivity & Resistance: Have distinct sets of parameters for damage from Attackers and damage from Defenders:

  • NL Die Thresh., L Lyse Thresh., Base Lysis Delay.

  • NL Resist. %, L Resist. %: Chance to completely negate incoming NL or L toxins from a specific hit.

• Quorum Sensing (QS) Behavior: Prey cells can also produce their own Autoinducer (AI) signal, which can be detected by Attackers (if Attacker Prey AI Attraction is enabled). This system is simpler than the Attacker's QS and does not directly control Prey behavior but contributes to the AI landscape.

  • AI Prod. Rate (U/min): The number of AI signal units each live Prey cell produces per minute.

  • AI Degrad. Rate (%/min): The percentage of Prey AI signal that degrades or disappears from each hex per minute.

  • AI Diffusion Rate: A coefficient (0-0.166) determining how fast Prey AI spreads to adjacent hexes, based on concentration gradients.

• Capsule Formation: Prey cells can develop a protective polysaccharide capsule as a defense mechanism.

  • Master Switch: The entire feature can be enabled or disabled globally using the "Enable Capsule Synthesis" checkbox. By default, this feature is off.

  • Regulation: When enabled, synthesis is controlled by a derepression switch that responds to local Prey AI concentration. High AI levels relieve a default state of repression, increasing the chance of synthesis.

  • Derepression Midpoint (K): This control works identically to the Attacker's T6SS activation control:

    • If K > 0: This is the standard mode. K acts as the midpoint concentration (EC50) for derepression, and the chance of synthesis increases with AI.

    • If K = 0: Synthesis is triggered if any AI is present (AI > 0).

    • If K < 0: Synthesis is Always ON (constitutively derepressed), regardless of the AI level.

  • Protection Effect: When an attack hits a prey with a capsule, there is a chance the attack is rendered harmless. The maximum protection offered by a full 5-layer capsule is set by the user via the "Max Protection (%)" setting. Each layer provides 1/5th of this total protection (e.g., if Max Protection is 80%, each layer gives a 16% chance to block an attack).

• Toxin Production & Release: Prey cells can synthesize their own bacteriocin toxins (Bacteriocin NL and L) to damage or compete against rival cell types.

  • Release Modes:

    • Continuous (Standard) Release: Toxins are secreted directly and continuously into the local grids (preyToxinNLGrid and preyToxinLGrid) at each step. Cells only start toxin production stochastically based on the Stochastic Start Prob. (which defaults to 0). Upon starting production, each cell chooses a target limit (max production) randomly from the configured range (200-1000). While producing, cell division is arrested, and the cell is visually marked with a small mid-green center dot. Once the cumulative toxin produced reaches its target limit, production is disabled and the cell returns to normal growth and division.

    • Lysis-Dependent Release: Toxins are accumulated internally (internalPreyToxinNL and internalPreyToxinL). Once the sum of accumulated internal toxins reaches the cell's individual Lysis Threshold (a random value chosen within a configured range exactly once when the cell is first activated as a producer), the cell lyses (dies and bursts), dumping its accumulated internal toxins onto the hex grid. Toxin-producing cells halt cell division while producing. If a cell divides (if production is inactive), any accumulated internal toxins are split equally (halved and rounded down to the nearest integer) between the parent and daughter cells. If a cell is lysed by attackers' T6SS before hitting its own threshold, its accumulated internal toxins are still successfully released.

    • Lysis Visualization: Prey cells with accumulated internal toxins (in lysis-dependent mode) are drawn with a green inner circle that darkens as they approach the lysis threshold. Continuous producers are drawn with a small, static mid-green center dot.

  • Activation Mechanisms (Trigger Modes):

    • Standard / Stochastic: Prey cells start with toxin production turned OFF by default. At each simulation minute, live cells have a chance to activate toxin synthesis governed stochastically by the Stochastic Start Prob. (%/min). Note: If this probability is left at 0%, no toxins will ever be synthesized under this mode.

    • QS Triggered: Activated based on local Prey AI concentration, modeled by a Hill equation with midpoint $K$ and cooperativity $n$. The QS-induced chance is scaled by the Stochastic Start Prob. (%/min). Once activated, the transition is permanent until completion (lysis or reaching the production target).

    • Attacker NL Toxin Triggered: Activated once the cell has been hit by attacker non-lytic toxins such that the accumulated units are $\ge$ NL Init. Threshold. Once the threshold is crossed, the activation chance per step is governed by the Stochastic Start Prob. (%/min). This is a permanent one-way transition once activated.

• Reporter System: Upon lysis, each Prey cell releases lacZPerPrey units of LacZ enzyme.

6.4. Defender Cells

• Role: Can attack Attackers and Prey, and importantly, can retaliate against attackers.

• Retaliation Behavior:

  • When a Defender is hit by a T6SS attack (from a Attacker or another Defender), it "senses" the attack.

  • Sense Chance %: The probability that this sensed attack triggers a retaliation state.

  • If retaliation is triggered:

    • The Defender locks onto the original attacker.

    • It fires a burst of shots, with the number of shots chosen randomly between 1 and Max Retaliations.

    • One shot from this burst is fired per simulation step (minute) until the burst is complete.

  • Example: Defender X is hit by Attacker Y. Defender X's Sense Chance is 60%. If the 60% chance succeeds, Defender X will retaliate against Attacker Y. If Max Retaliations is 7, it might fire (e.g.) 4 shots, one per step for the next 4 steps, at Attacker Y.

• Random Firing (attemptDefenderRandomFire):

  • Defenders can also fire proactively if their t6ssRandomFireCooldownTimer is 0.

  • Rand. Fire CD (min-max): Cooldown range after a random firing event.

  • Rand. Fire %: Chance per step (if not on cooldown) to fire one shot at a randomly chosen adjacent cell (Attacker, Prey, or another Defender).

• T6SS Effectors (vs Attackers/Prey): Parameters define NL Units/Hit, NL Delivery Chance %, L Units/Hit, and L Delivery Chance % for their attacks.

• Sensitivity (to Attacker Attack): Defenders have their own thresholds and resistance values against Attacker toxins.

• Sensitivity & Resistance vs. Prey Toxin:

  • NL Resist. (%) and L Resist. (%): Chance (0-100%) to completely ignore NL or L toxin absorbed from a local grid cell in a step.

  • NL Absorp. (%) and L Absorp. (%): Percentage of local grid-based Prey toxin absorbed by the Defender cell in each step.

  • NL Die Thresh. and L Lyse Thresh.: The cumulative Prey toxin units (absorbed from the grid) required to kill (growth arrest) or lyse the Defender cell.

• Defender vs. Defender Interaction: Defenders are immune to toxin damage from other Defender cells (they don't accumulate NL/L units from them). However, they do sense these hits and can initiate retaliation against the attacking Defender based on their Sense Chance.

6.5. Barriers

• Role: Static, impassable obstacles within the arena.

• Properties:

  • Barriers are placed manually or via arena import.

  • They do not replicate, do not fire T6SS, and are unaffected by toxins (they do not accumulate damage, die, or lyse).

  • They occupy a hex space, preventing other cells from moving into or replicating into that space.

  • Other cells (Attackers, Defenders) can target Barriers with their T6SS, but these firings will have no effect on the Barrier itself. The firing cell will still undergo its usual cooldown.

  • Barriers remain in place for the entire simulation unless explicitly removed during manual setup.

7. Simulation Loop / Step Logic (runSimulationStep)

Each simulation step (representing one minute) executes the following sequence of events:

1. Initialization & Cleanup:

   • Increment simulationStepCount.

   • Clear per-step counters (firings, kills, lyses for this step).

   • Clear activeFiringsThisStep map (used for visualization).

   • Remove any cells marked isEffectivelyGone from the main simState.cells map.

2. Cooldowns & Lysis Progression:

   • Iterate through all active cells and decrement their cooldown timers:

     • replicationCooldown

     • movementCooldown

     • t6ssFireCooldownTimer (Attackers)

     • t6ssRandomFireCooldownTimer (Defenders)

     • lysisTimer (for cells that isLysing). If lysisTimer reaches 0:

       • Mark the cell as isEffectivelyGone.

       • Increment per-step and cumulative lysis counters for its type.

       • If the cell is a Prey, add its lacZPerPrey to totalActiveLacZReleased.

3. Attacker Firing Phase:

   • Iterate through all active Attacker cells that are not dead/lysing.

   • A Attacker's ability to fire is first gated by Quorum Sensing. It calculates its probability of being "active" (P(active)) based on local autoinducer concentration.

   • If the Attacker passes this activation check and its t6ssFireCooldownTimer is 0, it attempts to fire (attemptAttackerT6SSFire):

     • Determine target hex based on contact sensing bias or random direction.

     • Check for Kin Exclusion if the target is another Attacker. If excluded, apply penalty cooldown.

     • If not excluded, determine if the shot is a "precise hit" based on AttackerPrecisionInput.

     • Record the firing event in activeFiringsThisStep for visualization (precise or miss). Increment firingsThisStep.

     • Reset the Attacker's t6ssFireCooldownTimer.

     • If it was a precise hit and a valid target cell exists in the target hex (and is not effectively gone):

       • The target cell (Prey or Defender) receives toxins (receiveHit('attacker')). This involves resistance checks and toxin accumulation.

       • The target cell's state is updated based on accumulated toxins (potentially becoming dead or lysing).

       • If the target is a Defender, it records the attacker's key (sensedAttackFromKey) for potential retaliation.

4. Defender Retaliation Triggering Phase:

   • Iterate through all active Defender cells that are not dead/lysing.

   • If a Defender sensedAttackFromKey (i.e., was hit in the Attacker phase or by another Defender in a previous step/phase) and is not already retaliating:

     • Check defenderSenseChanceInput. If successful, the Defender enters retaliation mode:

       • Sets isRetaliating = true.

       • Sets retaliationTargetKey to the key of the attacker.

       • Determines retaliationsRemainingThisBurst (randomly, 1 to defenderMaxRetaliationsInput).

     • Clears sensedAttackFromKey.

5. Defender Firing Phase (Retaliation & Random):

   • Iterate through all active Defender cells that are not dead/lysing.

   • If Retaliating:

     • If retaliationsRemainingThisBurst > 0, the Defender fires one shot at retaliationTargetKey (attemptRetaliationFire).

     • Record firing in activeFiringsThisStep. Increment firingsThisStep.

     • If a valid target cell exists:

       • Target cell (Attacker, Prey, or another Defender) receives toxins (receiveHit('defender')).

       • If the target is another Defender, it records sensedAttackFromKey (but is immune to damage).

     • Decrement retaliationsRemainingThisBurst. If it reaches 0, clear isRetaliating state.

   • Else (Not Retaliating or Burst Ended), Attempt Random Fire:

     • If t6ssRandomFireCooldownTimer is 0, check defenderRandomFireChanceInput.

     • If successful, the Defender fires one shot at a random adjacent hex (attemptDefenderRandomFire).

     • Record firing. Increment firingsThisStep. Reset t6ssRandomFireCooldownTimer.

     • If a valid target cell exists, it receives toxins as above.

6. Replication Phase:

   • Create a list of all cells eligible for replication (canReplicate() is true: not dead/lysing, replicationCooldown is 0). This list is shuffled.

   • Iterate through these eligible cells:

     • Find valid empty adjacent hexes.

     • If empty spots exist:

       • Choose one random empty spot.

       • Create a new daughter cell of the same type in that spot. Assign it a new ID and an initial (randomized) replicationCooldown. Add to simState.cells.

       • Reset the parent cell's replicationCooldown.

     • If no empty spots exist, simply reset the parent cell's replicationCooldown (missed opportunity).

7. Cell Movement Phase:

   • Iterate through all active, non-barrier cells (Attackers, Prey, Defenders) that are not dead/lysing.

   • For each eligible cell:

     • If its movementCooldown is 0, check its Movement Probability %.

     • If the cell attempts to move (passes probability check):

       • Determine a target adjacent hex based on Movement Directionality % (preferring empty hexes if directionality is high, or choosing randomly and then checking if directionality is low).

       • If a valid, empty, and non-barrier target hex is found, the cell is added to a list of pending movements.

     • The cell's movementCooldown is reset, regardless of whether a move was attempted or successful in this phase.

   • Process all pending movements:

     • Shuffle pending movements to handle conflicts randomly.

     • For each pending move, if the target hex is still empty (not taken by another cell in this step's movement phase), the cell is moved.

     • Cells cannot move into hexes occupied by other cells (including barriers) or hexes that become occupied by another moving cell in the same step.

   • Update cell positions in the main simulation state (simState.cells).

8. Quorum Sensing & Prey Toxin Update Phase:

   • Production:

     • Live Attacker cells add autoinducer (AI) to the attackerAiGrid.

     • Live Prey cells add autoinducer (AI) to the preyAiGrid.

     • For Prey cells in Continuous Release mode, toxins are added to preyToxinNLGrid and preyToxinLGrid at each step. (For Lysis-Dependent Release, toxins are stored internally and dumped onto the grids upon cell lysis).

   • Diffusion & Degradation:

     • New grids (attackerAiGrid, preyAiGrid, preyToxinNLGrid, and preyToxinLGrid) are calculated for the next step.

     • For each grid, the respective signal/toxin diffuses to/from neighboring hexes based on concentration gradients and the configured Diffusion Rate.

     • The concentrations are then reduced by the respective Degradation Rate %.

9. Update Statistics & Reporter:

   • Update cumulative counters for firings, kills, and lyses using the per-step values.

   • CPRG Conversion:

     • Calculate Vmax_current = simState.config.cprg.k_cat * totalActiveLacZReleased.

     • Calculate convertedThisStep = (Vmax_current * remainingCPRGSubstrate) / (Km + remainingCPRGSubstrate).

     • Ensure convertedThisStep does not exceed remainingCPRGSubstrate.

     • Add convertedThisStep to totalCPRGConverted and subtract from remainingCPRGSubstrate.

10. Record History & Capture Data: * Store current cell counts, cumulative stats, and CPRG converted in historicalData array (for graphing). * If saveImagesEnabled is true, capture an image of the arena. * If saveArenaStatesEnabled is true, capture the current cell layout as TSV.

11. Render & Schedule Next Step:

    • Redraw the simulation grid (drawGrid()).

    • Update the statistics display panel (updateStats()).

    • If simState.isRunning (not paused or just stepping), use setTimeout(runSimulationStep, simState.config.simulationControl.simulationSpeedMs) to schedule the next iteration.

    • If simState.isStepping was true, set it to false and update button states.

12. Termination & Database Offloading Checks:

    • The simulation checks if the time limit has been reached.

    • It also checks if any of the buffer limits (history, images, or arena states) have been reached, silently offloading the buffered data to the browser's local database (IndexedDB) to protect system memory.

8. CPRG Reporter System Details

The CPRG reporter system simulates a common laboratory assay.

• When Prey cells lyse, they release a fixed amount of LacZ enzyme (lacZPerPrey) into the environment. totalActiveLacZReleased tracks the cumulative active enzyme.

• This enzyme converts the available remainingCPRGSubstrate into a colored product.

• The rate of conversion follows Michaelis-Menten kinetics: Velocity = (Vmax * [S]) / (Km + [S]) Where:

  • [S] is remainingCPRGSubstrate.

  • Km is simState.config.cprg.Km.

  • Vmax (for the current total enzyme) is simState.config.cprg.k_cat * totalActiveLacZReleased.

• The amount converted in one step (convertedThisStep) is this velocity (since each step is one minute).

• The total CPRG converted (totalCPRGConverted) dictates the intensity of the magenta background color of the arena, scaled relative to initialCPRGSubstrateInput.

9. Preset Scenarios

The "Load Preset Scenario" button opens an interactive modal presenting several pre-configured simulation layouts.

• Interactive Customization: To select a preset, click anywhere inside its box (the border will highlight in blue). When a preset is selected, you can adjust its custom sliders and options (such as fill percent, attacker-to-prey ratios, trigger modes, and specific strategy toggles) directly within that preset's panel.

• Applying the Preset: Once you have customized the settings to your liking, click the "Apply Selected Preset & Close" button at the bottom of the modal. The system will:

  1. Apply all specific overrides and your custom slider configurations to the simulation settings.

  2. Reset the simulation arena and the RNG sequence.

  3. Populated the hexagonal grid randomly using your specified fill percentage and cell ratios.

The available preset scenarios are:

• 1. Role of Initial Density: Focuses on how varying initial cell densities and ratios between Attackers and Prey affect outcomes. Defenders are typically disabled.

• 2. Prey Sensitivity / Effector Type: Explores how different Prey sensitivities (to lytic vs. non-lytic toxins, or both) influence interactions. Defenders are typically disabled.

• 3. Contact Sensing & Kin Exclusion: Investigates the impact of Attacker T6SS firing strategies like biased targeting of adjacent cells and avoiding self-attack. Defenders are typically disabled.

• 4. Tit-for-Tat Dynamics: Sets up scenarios with all three cell types to explore retaliatory strategies, where Defenders respond to Attacker attacks, and Prey might have differential susceptibility. Parameters often include varying Defender retaliation accuracy vs. random firing.

• 5. Role of Capsule in Protection: Disables defenders and sets the prey capsule to be constitutively active, allowing exploration of capsule protection levels and synthesis time.

• 6. Predation by T6SS Activity: Disables defenders and normal attacker replication, forcing attackers to rely on the "Replication Reward" system for proliferation, linking killing to fitness.

• 7. Role of Movement in Killing: Disables defenders and provides sliders to configure prey AI production and attacker movement (probability, directionality), allowing exploration of chemotaxis and mobility in predation.

• 8. Quorum Sensing Dynamics: Disables defenders and enables the Prey capsule system. This preset provides a two-column layout to configure two separate QS systems: Attacker T6SS activation (via Attacker AI) and Prey capsule synthesis (via Prey AI). Sliders are available for AI production, activation/derepression midpoint (K), and cooperativity (n) for both systems, in addition to shared controls for cell density and ratio.

• 9. Prey Toxin Production: Focuses on Prey-derived diffusible toxins. It provides options for Arena Radius, Total Fill %, Attacker:Prey ratio, Stochastic Start Probability, and NL/L Toxin Production rates, plus buttons to toggle lysis-dependent release (Lysis Yes/No) and trigger activation modes (Standard, QS, Attacker NL).

• 10. Battle Royale: A comprehensive "sandbox" preset to quickly configure complex three-population scenarios. It provides high-level strategy switches instead of granular parameter inputs:

  • General: Sliders for Arena Radius and Total Fill %.

  • Population Mix: Two linked sliders control the Attacker % and Defender %, with the remaining population automatically assigned as Prey.

  • Strategy Switches:

    • Attacker: Sliders/checkboxes for Movement (No/Yes/AI), QS Regulated T6SS, Kin Exclusion, Contact Sensing, and Predation.

    • Prey: Checkboxes for Movement, AI Production, Capsule (capsule logic is linked to AI production), and Toxin Production.

    • Defender: Checkboxes/sliders for Movement, Selectivity (Low/Med/High), and Predation.

Applying a preset will only override the specific settings relevant to that scenario, leaving all other parameters (like Arena Radius or any settings you have manually changed) untouched. You can use the "Reset Settings to Defaults" button if you wish to return to a baseline state before applying a preset. You can further tweak any settings after applying a preset.

10. Data Export & Import

The simulation provides robust options for data management:

• Simulation Data (TSV): Time-series data of population counts and cumulative statistics.

• Settings (TSV): A file containing all simulation parameters, which can be saved and re-imported.

• Arena Layout (TSV - Manual Mode Only): Saves or loads a specific spatial arrangement of cells.

• Per-Step Arena Images (ZIP of PNGs): A ZIP archive of arena images, one for each step.

• Per-Step Arena States (ZIP of TSVs): A ZIP archive of cell layout files, one for each step.

• Single Step State (JSON):

  • Export: From the Time-Travel Control panel, you can export the complete state of any single step into a detailed, human-readable JSON file.

  • Import: This file can be re-imported to perfectly reset the simulation to that specific state, including all parameters and the precise state of the random number generator. This allows you to create exact replicas of a simulation state to run new experimental branches.

• Full Simulation Session (.bft6):

  • Export: If Full State History was enabled, the Save Full Session (.bft6) button appears in the report modal. This saves the entire simulation—all settings and the complete step-by-step history—into a single, highly compressed binary file. This is the definitive way to archive an experiment.

  • Import: From the Time-Travel Control panel on the main page, click Import Session (.bft6) and select a .bft6 file.

  • If loading the first file: This will load the entire saved session, including all settings and the full history.

  • Additive Loading (Advanced): If you already have a history loaded, you can import another .bft6 file to merge its history with the existing one. The application will validate that the Seed and Arena Radius match before merging. This powerful feature allows you to piece together very long simulation runs that were saved in multiple segments. The import logic correctly handles overwriting steps if there is an overlap, and can manage non-contiguous data (e.g., loading steps 0-100 and then 500-600).

• Retroactive Range Rendering ("Render from History")

When full state history is enabled, you can retroactively generate high-resolution PNG image sequences for any specific window of past time steps without re-running the live environment: * Custom Range Bounds & Rates: Input target values into the "From:" and "To:" step fields to isolate specific milestones. You can also specify a "Rate" to sample frames at regular intervals (e.g., setting Rate to 100 will render steps 5, 105, 205, etc.), which is useful for generating timelapses of very long simulations without rendering thousands of images. The final step in your range is always included. * Low-Memory Batch Generation: The engine sequentially loads frames out of IndexedDB, captures an offscreen canvas rendering, and pipes the output into memory-safe, chunked ZIP archives compiled via JSZip. * Dynamic Cancellation: Clicking the "Stop & Save Rendered" button instantly terminates active rendering loops, safely packages all frames processed up to that exact millisecond into a valid ZIP archive, and restores the live control interface.

#### Advanced Memory Management (IndexedDB Database Offloading)

Saving per-step data is memory-intensive. To prevent browser crashes during very long runs, the simulation includes buffer limits for images, arena states, and history, which can be configured in the "Exports & Buffers" panel.

• Silent Database Offloading: When any data buffer exceeds its defined size limit in memory (RAM), the simulation automatically offloads the heavy data (e.g. detailed frame objects, captured base64 images, or arena TSV layouts) to the browser's local database (IndexedDB).

• Multi-Tab Isolation: Each simulation instance dynamically generates a unique Session ID for its IndexedDB storage. This ensures you can safely run multiple independent simulations in separate browser tabs or windows simultaneously without their databases interfering with or corrupting each other. Orphaned session databases are automatically cleaned up when starting new runs.

• RAM Protection: The offloaded data is cleared from RAM, leaving only lightweight references in memory. This happens seamlessly in the background, allowing the simulation to run uninterrupted without needing user interaction, pausing, or intermediate file downloads.

• On-Demand Retrieval: When you click the download or save options (such as downloading ZIPs of images/arena layouts, or saving the full .bft6 session), the application automatically retrieves the offloaded frames and files from the database to build the download archive.
  Native Directory Access (Single Prompt Workspace)

To simplify high-frequency data logging, the application takes advantage of modern browser File System Access APIs:

• One-Time Workspace Permission: When clicking "Save All Data" or generating batch imagery, you will be prompted to select a local directory workspace.

• Silent Background Streaming: Once approved, all subsequent .bft6 history files, data tables, and image ZIP chunks write directly and silently to your local drive without triggering repeated browser download popups or confirmation dialogs.

10.1 Configuring Simulation via URL Parameters (Advanced)

You can pre-configure many aspects of the "BacFighT6" simulation by adding parameters directly to the browser's URL (the web address displayed in your browser, e.g., https://baslerlab.github.io/BacFighT6/index.html). This is a powerful feature for saving specific setups, sharing them with collaborators, or scripting different starting conditions for experiments.

How it Works:

• Parameters are appended to the main page URL, starting with a ?.

• Each parameter consists of a name=value pair.

• Multiple parameters are separated by an &.

  1. Example: https://baslerlab.github.io/BacFighT6/index.html?param1=value1&param2=value2

• Order Matters: The simulation applies settings in a specific hierarchy:

  1. System Defaults are loaded first.

  2. Presets (if ?preset= is present) are applied next, establishing a baseline scenario.

  3. URL Overrides are applied last. These are processed left-to-right and will override both the defaults and the preset values. This is crucial because later parameters can override settings or modify the arena state established by earlier ones.

🚀 Pro Tip: You don't need to write these URLs manually! Configure the simulation exactly how you want it in the UI, then click the "Copy Shareable Link 🔗" button in the General Settings panel. This will automatically generate the optimized URL for you.

Available Parameter Types:

1. Loading a Full Simulation Session from a URL (Highest Priority):

   • Parameter: sessionFileURL

   • Value: The full, URL-encoded web address of a .bft6 session file.

   • Behavior: This parameter takes highest priority. If the sessionFileURL is present in the URL, the simulation will immediately attempt to load the complete session from that file. All other configuration parameters in the URL will be ignored. This provides a direct, one-click way to load and view a specific archived experiment.

   • Example: ...?sessionFileURL=https://yourserver.com/sessions/tit_for_tat_run_01.bft6

   • 🔴 Important (File Access & CORS): See "Important General Notes" below.

2. Loading a Preset Scenario (Baseline Configuration):

   • Parameter: preset

   • Value: The ID of the preset scenario.

   • Valid Values: density, sensitivity, contactkin, titfortat, capsule, predation, movement, qs, preytoxin, battleroyale.

   • Behavior: Loads the default configuration for the chosen scenario (e.g., enabling specific rules, setting specific ratios). This is applied before other individual parameter overrides, making it the perfect "base" for custom links.

   • Example: ...?preset=battleroyale

3. Individual Settings Overrides:

   • You can change most simulation settings found in the control panel.

   • Parameter Names: These names must exactly match those listed in the first column of a "Settings (TSV)" file that you can export from the simulation's report modal (e.g., Arena_Radius, Attacker_Initial_Count, Attacker_T6SS_Precision_Percent, Image_Export_Enabled).

   • Behavior:

     • Changing Arena_Radius via a URL parameter is non-destructive. It will set the size of the arena boundary but will not clear any cells that have been placed by other preceding or subsequent parameters.

     • Other individual settings will update the simulation's configuration parameters. If these settings don't inherently trigger a full arena reset through their own event handlers, the current cell layout will be preserved.

   • Examples:

     • Set arena radius to 10 and Attacker initial count (for random seeding if no specific arena file/data is given later): ...?Arena_Radius=10&Attacker_Initial_Count=50

     • Enable image saving (for checkbox settings, use true or false): ...?Image_Export_Enabled=true

     • Set a specific T6SS parameter and disable Prey replication: ...?Attacker_T6SS_Kin_Exclusion_Percent=80&Prey_Replication_Mean_min=-1

     • Enable the capsule system and set its max protection to 50%: ...?Prey_Capsule_System_Enabled=true&Prey_Capsule_Max_Protection_Percent=50

4. Loading All Settings from a File URL:

   • Parameter: settingsFileURL

   • Value: The full, URL-encoded web address of a settings TSV file. This file must be in the same format as the one exported by the "Download Settings (TSV)" button.

   • Behavior: This loads all parameters from the specified file.

     • If the file contains an Arena_Radius setting, its value will be applied non-destructively. It will change the arena boundary size but will not clear any existing cells on the grid.

     • Settings from this file can be overridden by individual parameters that appear after it in the URL.

   • Example: ...?settingsFileURL=https://yourserver.com/path/my_experiment_settings.tsv

   • 🔴 Important (File Access & CORS): See "Important General Notes" below.

5. Loading/Modifying Arena Cell Layout from a File URL (TSV Format):

   • Parameter: arenaFileURL

   • Value: The full, URL-encoded web address of an arena TSV file (must be q\tr\ttype format).

   • Behavior (Additive/Overwriting): This parameter adds cells to the current arena or overwrites cells at the same coordinates. It does not automatically clear the entire arena before loading. Cells are placed onto the grid whose size is determined by the Arena_Radius active at the moment this parameter is processed (which itself could have been set by a preceding parameter).

   • Example: ...?Arena_Radius=12&arenaFileURL=https://yourserver.com/arenas/base_colony.tsv

   • 🔴 Important (File Access & CORS): See "Important General Notes" below.

6. Loading/Modifying Arena Cell Layout with Compact cellsData String:

   • Parameter: cellsData

   • Value: A continuous string where each cell operation is q<number>r<number><Type>. No spaces or other separators.

     • q<number>: Axial q coordinate (e.g., q-5, q0, q12).

     • r<number>: Axial r coordinate (e.g., r3, r0, r-7).

     • <Type>: A single character:

       • A: Place/overwrite with Attacker.

       • P: Place/overwrite with Prey.

       • D: Place/overwrite with Defender.

       • B: Place/overwrite with Barrier.

       • E: Empty/Remove cell at the specified (q,r).

   • Behavior (Additive/Overwriting/Removing): This parameter modifies the current arena state. It adds new cells, overwrites existing cells if coordinates match, or removes cells if E is used. It does not clear the entire arena by itself. Operations are validated against the Arena_Radius active when this parameter is processed.

   • Examples:

     • Place Attacker at (0,0) and Prey at (1,-1): ...?cellsData=q0r0Aq1r-1P

     • Place a Defender at (2,0), then remove any cell at (0,0): ...?cellsData=q2r0Dq0r0E (Result: Defender at (2,0), (0,0) is empty)

Combined Examples:

1. The "Challenge" Link: Load the Battle Royale preset, but force a specific Seed and extend the Duration to 10,000 steps. ...?preset=battleroyale&Simulation_Seed=12345&Simulation_Duration_Minutes=10000 🚀 Launch Battle Royale Challenge

2. The "High-Res" Run: Load the Tit-For-Tat preset, but increase the Arena Radius to 50 and enable Image Export. ...?preset=titfortat&Arena_Radius=50&Image_Export_Enabled=true 📸 Launch High-Res Simulation

3. Specific Arena Configuration: Set the radius to 10, set specific initial counts (for random seeding), and force a specific seed. ...?Arena_Radius=10&Attacker_Initial_Count=50&Prey_Initial_Count=50&Simulation_Seed=888 🔬 Launch Small Colony Setup

Understanding Order and Arena State Interactions:

The sequence of these parameters in your URL dictates the final setup.

• Arena Sizing (Non-Destructive):

  1. The simulation page initially loads with the default radius from the HTML input.

  2. The Arena_Radius can be set by an individual parameter (e.g., ?Arena_Radius=25) or from within a loaded settingsFileURL.

  3. Changing the Arena_Radius via URL does not clear the arena or reset the simulation. It simply defines the grid size. Cells placed by other URL parameters will be preserved, even if the radius is defined after them in the URL.

• Cell Placement/Modification (Additive/Overwriting/Removing):

  1. Both arenaFileURL and cellsData operate on the arena state as it exists when they are processed. They add new cells, or if a cell already exists at a specified coordinate, the new definition from the currently processing parameter will replace it. cellsData with type E will remove a cell. These two parameters do not call initializeBlankSlate() themselves.

• Combined Examples:

  1. Settings File (no radius change), then cellsData: ...?settingsFileURL=path/config_no_radius_change.tsv&cellsData=q0r0A

     • Initial blank state.

     • config_no_radius_change.tsv loads. It does not change Arena_Radius. Other settings are applied. The grid remains empty (or as it was if modified by a preceding parameter not shown here).

     • cellsData then places a Attacker at (0,0) on the current grid.

     • Outcome: Grid has q0r0A, and simulation parameters are from the file.

  2. Arena File for base layout, cellsData for specific additions/changes: ...?Arena_Radius=10&arenaFileURL=https://example.com/base_arena.tsv&cellsData=q0r0Aq5r5Pq-1r-1E

     • Initial blank state.

     • Arena_Radius=10: Sets radius to 10. Arena is re-initialized (empty, radius 10).

     • arenaFileURL: Cells from base_arena.tsv are loaded onto the radius 10 grid. (e.g., file has a Defender D at q0r0).

     • cellsData:

       • The Attacker q0r0A replaces the Defender from the file at (0,0).

       • The Prey q5r5P is added to an empty spot (if within radius 10 and empty).

       • Any cell at q-1r-1E is removed.

     • Outcome: Arena is primarily from base_arena.tsv, but specifically modified by cellsData.*

  3. cellsData first, then arenaFileURL (both additive/overwriting): ...?Arena_Radius=10&cellsData=q0r0Pq1r1A&arenaFileURL=https://example.com/main_arena.tsv

     • Initial blank state.

     • Arena_Radius=10: Arena set to radius 10, empty.

     • cellsData: Places Prey P at (0,0) and Attacker A at (1,1).

     • arenaFileURL: Loads cells from main_arena.tsv.

       • If main_arena.tsv defines a cell at (0,0), it will overwrite the Prey.

       • If main_arena.tsv defines a cell at (1,1), it will overwrite the Attacker.

       • Other cells from main_arena.tsv are added.

       • The cells from cellsData remain if their coordinates are not specified in main_arena.tsv.

     • Outcome: A merged arena. Cells from main_arena.tsv take precedence at shared coordinates.

  4. Revisiting Order: Arena_Radius change after placing cells: ...?cellsData=q0r0A&Arena_Radius=5

     • Initial blank state (default radius).

     • cellsData places a Attacker at (0,0).

     • Arena_Radius=5 is processed. The arena boundary is now set to a radius of 5. This does not clear the cell. The Attacker at (0,0) is preserved.

     • Outcome: An arena of radius 5 with a Attacker at (0,0).

Important General Notes for URL Configuration:

• Parameter Names: For individual settings, always use the exact names from an exported "Settings (TSV)" file (first column).

• File Access (CORS): For settingsFileURL and arenaFileURL, the linked files must be publicly accessible on the internet. If they are on a different domain than your simulation page, the server hosting them must have Cross-Origin Resource Sharing (CORS) enabled (e.g., by setting Access-Control-Allow-Origin: * or to your specific domain). For local testing with your own files, running a simple local web server (e.g., Python's http.server) to serve your HTML and TSV files via http://localhost/... URLs is recommended.

• URL Encoding: Values in URLs, especially if they are other URLs or strings with special characters (like spaces, &, ?, =), must be properly URL-encoded. Browsers often handle this if you copy-paste a full URL, but be mindful if constructing links manually or programmatically (e.g., JavaScript's encodeURIComponent()).

• Debugging: Open your browser's JavaScript console (usually F12). The simulation logs messages about which URL parameters it's processing and any errors encountered (e.g., file not found, CORS issues, malformed data). This is very helpful for troubleshooting your URLs.

11. Technical Notes & Considerations

• Browser: Best viewed on modern desktop browsers (Chrome, Firefox, Edge, Safari) that fully support HTML5 Canvas and modern JavaScript (ES6+).

• Performance: Very large arena radii (e.g., >40-50) combined with high cell densities and long simulation durations can become computationally intensive and slow down the browser, especially if per-step image/state saving is enabled.

• Current Limitations: The model is grid-based and does not include nutrient diffusion, complex spatial structures beyond hex occupation, or evolutionary changes within a single run (parameters are fixed once a simulation starts).

License and Attribution

This work (BacFighT6: Simulation of T6SS-mediated Bacterial Interactions), available at GitHub, is licensed under the Creative Commons Attribution 4.0 International License (CC BY 4.0).

Copyright (c) 2025 Marek Basler

You are free to:

• Share — copy and redistribute the material in any medium or format for any purpose, even commercially.

• Adapt — remix, transform, and build upon the material for any purpose, even commercially.

The licensor cannot revoke these freedoms as long as you follow the license terms.

Under the following terms:

• Attribution — You must give appropriate credit to Marek Basler - University of Basel - Biozentrum, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.

For the full license text, please visit: https://creativecommons.org/licenses/by/4.0/

Support

Development of BacFighT6 runs on coffee! If you find it useful, please consider a small donation. Thank you!