Experiment: Octopus Fighting

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Background

Octopuses, or Octopodes, are some of the sea's most fascinating creatures. Just like cuttlefish and squid, they are Cephalopods, which is a surprisingly good descriptor for them as "Cephalopod" is a combination of the Greek "kephale" meaning head, and “pod” meaning feet; they are heads on feet with no body in the middle to speak of.

Though undoubtedly anti-social animals, doing their best to live as far from their brothers as possible and going as far as to eat ones that come too close, this doesn't meant that they are completely devoid of inter-octopus interaction. Quite the opposite is true, they exhibit fascinating behaviours when put in close quarters with one another. They dance around each other, flash colouration, poke at one another with their outstretched arms, and given a good opportunity they will begin a friendly match of octopus wrestling!

The octopus of interest for this experiment is the California two-spot (Octopus bimaculoides), named after the two blue spots found on the sides of it's head. (Read more info here) They're a great candidate for experimentation since they are an extremely hardy and easy to keep animal, living comfortably in temperatures all the way from 60-80°F. They're diurnal (they sleep at night), which makes for very convenient testing since they'll actually be awake during the same hours as you.

The most difficult problem with them is their pickiness when it comes to food; these octopuses aren't opportunistic hunters so if the food doesn't move they don't want it (but that's fine, just keep a bucket of live crabs to throw them occasionally).

DIY Neuroscience Episode of Experiment

Materials

For this experiment you will need:

• Octopus chamber (Laser cut and assembled) - Source File: Octopus_Chamber.ai
• Go Pro camera
• Mini-USB cable
• Laptop
• Aquarium with sea water
• Two California two-spot octopuses
• Empty, cleaned out milk jug for sea water
• Lamp with a diffusive filter/cover
• Paper towels (lots)
• Small plastic cup to carry octopus

Hardware Setup

You can assemble our laser cut octopus chamber using preferably non-toxic glues. Use hot glue and silicon to seal. Make sure the bottom is as watertight as possible, but still keep some paper towels under it.

You can also use your own container, but just be sure the camera is steady above the small "arena".

Software Setup

You can find the software referenced on our Github.

You are going to need quite a lot of software to be able to run the provided Jupyter notebook; and you will very likely run into bugs. Don't panic... this kind of problem-solving is an essential experience for anyone looking to delve into computer science.

If you are on a Windows machine, we highly recommend Git Bash as a pseudo-bash console: Git

You will need, first and foremost, PIP, the Jupyter notebook itself, and OpenCV:

PIP: https://packaging.python.org/tutorials/installing-packages/

Jupyter: http://jupyter.readthedocs.io/en/latest/install.html (We recommend Anaconda)

OpenCV: If all goes well all you need to do is "pip install opencv-contrib-python" Remember to install opencv-contrib specifically as it contains packages that we need.

Octopus Filming

1. Fill jug about a quarter of the way with sea water and shake it to aerate.
2. Fill the octopus chamber until there's at least an inch of water.

3. Set up Go Pro on top of the chamber.

4. Set up lamp so that the chamber is lit as evenly as possibly, meaning the walls don't cast major shadows, but also such that the lamp is not facing directly downwards causing reflection in the water. (Take Go Pro footage and check multiple times to see that the lighting looks good)
5. Scoop up one octopus with the plastic cup, be careful not to hurt their arms.

6. Gently let the octopus leave the cup into the chamber.
7. Scoop up second octopus.
8. Press record.
9. Place second octopus into chamber and record for 15-25 minutes.
10. Stop recording, and place octopuses back into their respective tanks.
11. Repeat this filming up to 3 times per day, giving the octopuses at least 2 hours of break in between.
12. Move the video recordings from the Go Pro onto a computer, and use any preferred video-editing software to cut it into 15-20 second clips surrounding the observed interactions (i.e. starting 10 seconds before a fight and ending 5 seconds after). Place the behaviour videoclips in the same folder as the Octopus_Optical_Flow.ipynb. Important: Video clips must be 720x1280, anything else requires significant code changes and you're on your own. 60fps is recommended.
13. Press Shift and Tab at the same time to run a cell of the Optical Flow notebook. Start by running the cell with all the import statements.
14. Change the filepath to match that of the videoclip you want to analyse
    
15. Leave one of the two options uncommented: top is full-colour, bottom is black and white with the mask applied, exactly what the computer sees:
    
16. Run the optical flow codeblock. A GUI will pop up if the video is loaded correctly. Since the corners will be selected manually the only important settings are in the top half of the GUI:
    

    The top switch is to start the corner selection, flip this when finished with the four settings below it.

    • Winsize: Size of the search window at each pyramid level
    • maxLevel: Height of image pyramid created
    • criteria: Parameter, specifying the termination criteria of the iterative search algorithm
    • liveCutoff: Maximum brightness (0-255) that the algorithm will look at (anything above is ignored)

    Best way to really figure out what these do is via trial and error.

17. After the switch is flipped, select two points for tracking on one octopus (the eyes are recommended) by clicking on them.
    

    Press escape to run the algorithm.

18. The code will now run and trace paths of the octopus, if one of the points loses contact with the octopus, terminate the code by pressing escape and try again. Try changing settings or clicking different points on the octopus.
    
19. Upon completion, the codeblock will print out an image like this, this is the traced path:
    
20. Run first pointcopy block:
    
21. Run steps 17-19 for second octopus. Save by running only pointcopy2 block. Don't run the first pointcopy block again, this will overwrite the data.
    
22. Change number to be a name you'll remember (probably the current videoclip name)
    
23. Save the data. This will create two files, a for the first octopus and b for the second.
    
24. Repeat steps 14-23 for as many videoclips as needed.

Experiment: Angle and Distance after Contact

1. First in Octopus_Optical_Flow load the a and b point data saved from the videoclips analyzed in the methods. To do this define number as the name of the saved points (the .npy file without the a or b at the end):
   
2. Run the two np.load codeblocks to load and display the a and b paths.
   
3. Run the next codeblock to make sure both arrows go from the octopus' mantle to its head. If it is incorrect such as this:
   

   Use the codeblocks below to flip the points:

   

   Until both arrow look like this:

   
4. Run the last 2 codeblocks to calculate and save the distance and angle data:
   
5. For all the videoclips analyzed for distance/angle data it is now necessary to find contour gradient data so that all the points of contact can be aligned. Open up Octopus_Contact and change the filename for the desired videoclip:
   
6. Run the codeblock and adjust the sliders (mostly v-high) until the green lines draw a nice contour of the octopus:
   
7. Flip the Off-On switch and let the video run.
8. Upon completion of videoclip, run the codeblock below it:
   
9. Save the data and repeat the procedure for the rest of the videoclips:
   
10. Now it's time to write some code. Create a new Jupyter notebook and write a sequence of codeblocks that will:
    1. Load all the distance, angle, and gradient files.
    2. Cut out a 200 frame width window from the distance and angle vectors that is +100 frames and -100 frames of the frame where the respective gradient vector has its peak.
    3. Normalize and add together all these windows.
    4. Plot the results.
    5. Add some extra features like labels, lines of best fit, coloration!
    The end result shoud look like this:
    

Science Fair Project Ideas

• Is there a heirarchy formed amungst octopodes?
• What other patterns do you observe in their behaviours?
• Do the octopodes always approach on the same side? Can you write code to determine this?