Establishing moina culture in smt.C.H.M.College 2

[Siddhesh]

2019-08-02T18:30:00Z
On 01/08/2019 we collected moina from CUBE lab HBCSE,we carried them to Ulhas nagar at my home

along with aprox 400 ml dc(dechlorinated water) from cube lab .
After reaching at home I transferred aprrox 10 - 10 moinas in both the bottles on same day .
02/08/2019

in each bottle number was observed around 15-20
then I kept 3 glass bottles 2(approx 250ml) 1 (500ml approx) for dechlorination of water .

Today 03/08/2019
I’ve transferred 5 moina in 2 glass bottles of 250 ml
10 moina in 1 glass bottle of 500 ml
Feeding
250ml bottles with 1 drop milk
500ml bottles with 2 drop milk
Yet moina culture is not established in college till the time I’ll maintain it at my home today I tried to take some photographs with help oh available magnifying glass photos are not as distinct but still observable.

Hope my writing and observation may increase and improve day by day

[Arunan]

Why not take a short video? Movement of Moina
can be seen easily!

[Siddhesh]

Uploading: VID_20190803_161235.mp4… https://stemgames.metastudio.org/uploads/default/original/2X/4/4ae520589e8409e22fceb483592e4796d6016db2.mp4
Video is taken with only mobile camera without any magnification but still we can observe natural size of moina ie around 0.5-2mm
Sorry for background noise it’s raining heavy here.
https://stemgames.metastudio.org/uploads/default/original/2X/7/7bb8b83b32e4d39c26f1476ffe329e5491a97279.mp4

[Siddhesh]

Number of moina increased from 5 to 15and 7 in
10 to 13

[jaikishan]

Please share pictures of moina under microscope. @Siddhesh

[Siddhesh]

Sorry college will be open tomorrow so i can get good images tomorrow till the time I’m sending the photo that I’ve clicked in cube lab

[dialecticalbiologist (dilecticalbiologist)]

Looking for updates on Moina culture at CUBE CHM college.

[Rechel_tirkey]

@Siddhesh do you keep moina container capped.

[jaikishan]

@Siddhesh are all the moina of same age in the picture? How to find out their age? looking forward to the status of moina culture? How many bottles do you have? Have they increased in numbers?
How much time it takes to give one generation?

[Siddhesh]

@Rechel_tirkey yes I’ve kept lid on the container and bottle cap to avoid contamination due to entry of mosquito and other insects but they are not tightly packed.

[Siddhesh]

Currently moina in th 3 glass jar are died
Other 2 pastic bottles collectively have around 150 moina
Reason may be excess milk drops in the glass jars

These are few survivals of the 500ml glass jars

All are red due to haemoglobin
moina belongs to phyllum arthropods as in photograph we can observe that they have jointed appendages (legs)

this is the same photograph taken in cube lab hbcse .
Arthropods have haemolymph as circulatory fluid as blood in humans haemolymph moves or circulates in open circulatory system haemocoel.but in unfavourable condition in hypoxia moina develops haemoglobin which is common pigment in higher organisms so the point in producing haemoglobin is it have higher affinity towards oxygen and thus moina becomes able to survive in hypoxic condition.

About male and female I’ve only idea that females have comparitively larger size than males and can be easily identified with their brood pouch
College lab is undergoing renovation till the time I’ll continue to culture moina at home

[dialecticalbiologist (dilecticalbiologist)]

Please tell us the whole story as of today ( in summary) of the Moina culture @Siddhesh

[Siddhesh]

01/08/2019
we collected moina(approx 15-20 in 13ml centrifuge tubes) from CUBE lab HBCSE,we carried them to Ulhas nagar at my home along with aprox 400 ml dc(dechlorinated water) in 2 plastic bottles from cube lab .
After reaching at home I transferred aprrox 10 - 10 moinas in both the bottles on same day .

02/08/2019
in each bottle number was observed around 15-20
then I kept 3 glass bottles 2(approx 250ml) 1 (500ml approx) for dechlorination of water .

03/08/2019
I transferred 5 moina in 2 glass bottles of 250 ml
10 moina in 1 glass bottle of 500 ml
Feeding
250ml bottles with 1 drop milk
500ml bottles with 2 drop milk
Plastic bottles with 2 drop milk

03/08/2019
Moina count bottle1 bottle 2

250ml bottles 7 10
500ml bottles 12
Plastic bottles 21 18

04/08/2019
Moina count bottle1 bottle 2

250ml bottles 15 18
500ml bottles 20
Plastic bottles 30 21

05/08/2019
Moina count bottle1 bottle 2

250ml bottles 13 22
500ml bottles 24
Plastic bottles 30 21

06/08/2019
Moina count bottle1 bottle 2

250ml bottles 26 28
500ml bottles 41
Plastic bottles 45 32

I Increased feeding by double

07/08/2019
Moina count bottle1 bottle 2

250ml bottles 30 20
500ml bottles 50
Plastic bottles 53 35

08/08/2019 (due to excess milk feeding water becomes turbid)
Moina count bottle1 bottle 2

250ml bottles 10 25
500ml bottles 45
Plastic bottles 40 28

09/08/2019
Moina count bottle1 bottle 2

250ml bottles 15 18
500ml bottles 20
Plastic bottles 30 21

10/08/2019
Moina count bottle 1 bottle2

250ml bottles 0 10
500ml bottles 25
Plastic bottles 60 50

11/08/2019
Now I’m again starting culture in
2 glass jasrs (500ml)
3 glass bottles (250ml)
This time I’ve added around 30-40ml dc water in all bottles + 1drop milk

In stock culture we can observe read moina clearly
Probably the count is also more than
https://photos.app.goo.gl/seZk3Gccr7MUdA5h6

[drishtantmkawale (Drishtant Maruti)]

This looks great!! You’ve cultured red moinas successfully. So why don’t you establish a short or long term objective regarding these moinas?
And if you want culture moinas, then you can take a large plastic / container and culture in it.
In future it would be easier to take out cultured moinas as per your convenience.
Keep us updated about your culture!!

[Siddhesh]

It is my observation that moina usually settle at bottom so according to me if I’m using vertical container I’ve to arrange pippate or kind of instrument to transfer moina so if I culture them in flat flask it’ll more beneficial for transfer with help of dropper (3ml plastic dropper) which I’m using also currently I’ve set again 5 bottles with low water level as it also allows more concentration of milk in high amount of water same drop will give low concentration
Also as population increases I’ll add water in it or else I’m planning to transfer them in 2 liters plastic bottles

Currently
Short term objective is to maintain stock culture
Long term ie for 2-4 weeks this is 2nd to learn techniques of culturing so that I could start doing some experiments on it
Because if we successfully standardize the amount of water and milk drops in small amount we can multiply the variables according to our convenience or requirements and also can switch red moina to pale colour and vice versa in short time

[drishtantmkawale (Drishtant Maruti)]

Ok cool.
You used the word ‘standardize’, may I know the meaning of it or what is standardizing in this case?? And yes,
Do you know the mechanism behind the moinas turning into red and vice versa?

[Siddhesh]

Yes
Standardizing technique of culturing means if i got results in 30ml of water 1 drop of milk if yield of moina is around 300 in 8-9 days from just 5 moinad at day 0 this result should be repeated in each and every trial with same amount of water and milk and number of moina as we can’t gete exact results all the time but they must be identical and have very less difference.

Arthropods have haemolymph as circulatory fluid as blood in humans . haemolymph moves or circulates in open circulatory system haemocoel.but in unfavourable condition in hypoxia ie low amount of oxygen moina develops haemoglobin which is common pigment in higher organisms . haemoglobin is having higher affinity towards oxygen and hence moina becomes able to survive in hypoxic condition . exact mechanism about the process of development of hemoglobin ive not idea I’m searching for it as iget it I’ll definitely update here .

[drishtantmkawale (Drishtant Maruti)]

So about the mechanism.
The genome of Moina macrocopa have at least four globin genes. In normoxic conditions, only two of them are activated or only two express themselves. All four respond differently to different conditions (hypoxic and normoxic). The haemoglobin gene is tightly bound by histone protein but when there is less oxygen, moinas need more haemoglobin to survive. So, when there is hypoxic condition (less oxygen), the cell introduces or calls for HAT (histone acetyl transferase) enzyme. As DNA is -vely charged and histone is +vely charged, they are bound together, so RNA polymerase is not able to read and proceed to form RNA and protein. HAT adds acetyl group (-COO which is -vely charged) removes histone protein so DNA gets free and RNA polymerase reads DNA and makes RNA and proteins hence more haemoglobin protein is made and moinas turn red in colour. On the other hand, if we keep the red moinas in normoxic conditions, the cells recruits HDAC (histone deacetylase) enzyme which removes acetyl group so once again histone comes and binds with DNA and hence making it unreadable and inactive.

[GN (Nagarjuna G)]

If you want red moina, narrow bottle is better than wider container with low level of water. What do you think is my reason?

[Siddhesh]

Can you please send reference for it because now igot the mechanism still few questions arises like

1. what inhibits HAT to come in action?
2. what is the structure of HAT?
3. and as my observation progeny’s of red moina is whitish or what we observe in normoxic condition so the expression of allele is due to environmental factor which is clear in our results so why this expression is not inherited ?
   Is this phenomenon is common in all lower organisms having haemolymph ?
   4)does haemoglobin remains as suspended particle because they don’t have rbcs to bind