An Alternative Way of Culturing Chlorohydra :CUbists Proposal

Dhanraj and Sakshi @dhanraj7 @sakshiconsultant2002 have been culturing Chlorohydra for almost 3 years. Shakshi said that in her college, Sophia Mumbai, Artemia is a feed for Chlorohydra.

Sakshi introduced Moina to feed Chlorohydra, which she learned from CUBE. Parallely, Sakshi and Dhanraj had a hypothesis that marine organisms like Artemia can cause problems in Chlorohydra, a freshwater organism. It is good to replace Artemia with Moina, a freshwater organism.

Collaboration:

These are the photos of the workshop on Study and maintenance of Chlorohydra which was conducted at Ratnam College and @dhanraj7 was invited as a resource person.

Date - 8th of July 2022.

On the same day collaborator pritesh from Elphinstone college was also invited as a resource person for workshop on Chlorohydra by @⁨Prof.Farhaan Makba CUBE⁩ at royal college Mira road.
The photos are below.

Following are the photos from @sakshiconsultant2002

Moina culture maintenance done be me in the month of july 2022.


Long shot picture of hydra and moina culture maintained in ratnam college with labelling.


Video of moina culture showing moinas at the top of the bottle.
Date:27th july 2022
Name:sakshi
Location:ratnam college


Picture of chlorohydra culture maintained in the bottle!
Date of picture: 1st August 2022
Location: ratnam college
Name: sakshi


Moina culture and hydra culture shifted to homelab due to vacations!
2 bottles of moina culture and 1 bottle of hydra culture.
Date: 31st august 2022
Location:bhandup.
Name:sakshi.

Hydras maintained in transparent glasses.
Date:31st august 2022.


Video of moina culture maintained in the bottle.
Date: 9th September 2022
Location: bhandup
Name: sakshi


Subcultures of moina made on 13th September 2022.
Location: bhandup.
Name: sakshi


Video of hydra culture maintained in glass.
We can see green particles present at the surface of the glass and the moinas were not fed by hydra.
20 September 2022
Sakshi.

Number of moina’s increased also there is transfer of green particle from the chlorohydra glass to the bottle we can see some green algal growth at the bottom of the bottle.
Date: 9th October 2022
Location: bhandup.
Name:sakshi.


Made subculture of moina on 13th October 2022.
Bhandup.
Sakshi.


Approximately 400 moinas or more than that seen in this bottle culture along with green algae.
Lost hydra culture:(
Date: 27th November 2022
Location: bhandup
Name: sakshi.


Subcultures of moina with normal and algal growth.
Date:10th January 2023
Location:bhandup
Name:sakshi

Again got chlorohydra from dhanraj on 12th January 2023.
Location: somaiya college
Name:sakshi.


Transferred the chlorohydra in glass beaker the dots and dashes aare chlorohydra there was also presence of green algae at the bottom.
13th January 2023.
Bhandup.
Sakshi.


Done an experiment of light and dark conditions with hydra. Experiment went till 15 days.
Date: 20th January 2023
Location: ratnam college
Name:sakshi.


Lost the hydras again!
We were left with chlorella algae at the bottom of the beaker and moina’s feeding on that algae:(
Date: 1st February 2023.
Bhandup.
Sakshi.


Green algae seen in the intestine of moina.
2nd February 2023
Sakshi.


Moina culture with green algae maintained at home lab
9th june 2023
Bhandup.
Sakshi.


Accumulation of algae at the bottom of the bottle.
Date 21st july 2023.
Bhandup.
Sakshi.


Again, made subcultures of moina.
Date:25th july 2023.
Bhandup.
Sakshi.


Video of moinas in the subcultures.
1st august 2023
Bhandup.
Sakshi.


Video of moinas present in algal medium.
1st august 2023
Bhandup.
Sakshi.


Hydra culture in Sophia college.
Photo: Sakshi.

Here is the video of hydras fed with artemia.

This is the video of hydras feeding on moina.

Following are the updates from @dhanraj7

By @dhanraj7


Photo of the chlorohydra. These chlorohydras were cultured in a beaker and was given regular feed of moina.

11th March 2022 i.e two years ago when I started working on this model system.
Cube Elphinstone lab.


Photo of 11th March 2022, hydra was cultured ina beaker of volume 250 ml
Cube Elphinstone lab.


Moina culture in a plastic tank of 6 litres approx. Photo - 14th March 2022

Cube Elphinstone lab. 24 drops of milk was added everyday as a feed for moina.

Standard feeding protocol was 1 drop of milk in 250 ml of dechlorinated water

Whiteboard screenshots of Chatshaala:

Summary made by @dhanraj7 on 28/05/2024.

Yesterday we discussed about my journey with Chlorohydra, almost three years ago, when I was introduced to Chlorohydra by my seniors at Elphinstone college, later about how and what all we studied on Chlorohydra like particularly I have studied the regeneration in chlorohydra, about what all pathways must be included in this process of regeneration, about effect of light and dark conditions on symbiotic Chlorohydra, certain aspects of ageing, stem cell biology till my thesis work in somaiya which was to study the regeneration in Chlorohydra under hypoxic conditions. This whole period of more than two and half years almost three years helped me to learn many new things about this model system. The whiteboard shows about the journey of Sakshi’s experience with Chlorohydra where she was introduced with Chlorohydra in a workshop conducted at NES Ratnam College, Mumbai on 8th July 2022, where I was invited as a resource person. Sakshi who is currently a student of Sophia College, Mumbai is studying about hypoxia on hydra. There are many instances where Cubists from Kerala and Sapekhati, Assam tried to find hydra in the freshwater.

Later talking about the workshop, the Chlorohydra and the moina culture was handed over to Sakshi where they maintained these Chlorohydra as well as Moina in NES Ratnam College, Mumbai.

We discussed about how to culture, how do we maintain Chlorohydra in homelab for which we designed an experimental setup where a 250 ml of transparent plastic bottle was taken since it is easily available. And then adding around 200 ml of dechlorinated water to the plastic bottle. We discussed about obtaining dechlorinated water by taking some tap water in a vessel and then keeping it near the window for dechlorination. And keeping a muslin cloth or a cotton cloth on the vessel to avoid contamination. Since the chlorine evaporates in the form of gas at room temp. It is very easy to obtain dechlorinated water. Then next day, after 24 hours, we could use the dechlorinated water for culturing Chlorohydra. We discussed about the feeding of Chlorohydra done by adding moina to the plastic bottle. The feeding ratio will be 1:2. That is for each Chlorohydra we need to feed with two moina or daphnia.

We also discussed whether Paramecium can be given to Chlorohydra as a feed? Possibly it can be done. Chlorohydra can be fed on paramecium. Hydra are small, freshwater organisms belonging to the phylum Cnidaria, and they are known for their simple body structure and ability to capture prey using their tentacles which has specialized stinging cells called nematocysts. So, when Chlorohydra’s tentacles come in contact with paramecium or moina or daphnia, the nematocysts discharge, releasing some toxins, capturing and immobilizing the prey which is then brought to the Chlorohydra’s mouth for ingestion. This is known as the feeding behaviour of Chlorohydra.

Further we developed the discussion where I have seen some college facilities using artemia cyst as a feed for hydra. Artemia is a marine organism so wouldn’t it show any effect on freshwater Chlorohydra? @⁨Sakshi⁩ what do you think? The Artemia capsule or the eggs of the Artemia which are in dormant state are cultured and after hatching they are provided as a feed for hydra.

Sakshi mentioned about how her college facility used artemia as a feed for Chlorohydra. When Sakshi took Chlorohydra as a model system to work on in Sophia College. She told her story that how she used Moina for culturing Chlorohydra. Sakshi discussed that before feeding artemia to the Chlorohydra, the Artemia are given few washes in the normal water and then they are given to Chlorohydra as a feed.

This brought a change in method of doing science, where Sakshi used moina as an alternative for Artemia and challenged the ritual of culturing Chlorohydra with Artemia.

Apparently we also discussed about the journey of Cubists where we questioned about the strain of moina which Sakshi is having in Sophia College, is it the jsk1 or is it different strain.
I explained how we got moina macrocopa jsk1 from our seniors, Drishtant, Saida and Zahra, who then got it from CUBE lab in HBCSE and were working on their thesis on studying epigenetics in moina. Sakshi mentioned about the moina which she is having now, is the moina which was passed on by me during the Chlorohydra workshop at Ratnam College.

Through all these journey there are many goofups that I made, and Sakshi took. Like the self sustainable culture of moina which was an Alexander Fleming moment for all of us.

The problem was that we were afraid to fail in science and didn’t celebrate our goof ups like not showing consistency in sharing the photos which acts as a evidence in science. There are many goofups and TINKE moments during this journey. I think we could discuss it better by sharing photo as an evidence tk convince audience.

Group photographs of participants:30/05/2024