Moina culture - CUBE Kolenchery

[center]Moina macrocopa culture at St Peter’s College, Kolenchery[/center]
How we did it:

  1. @Arunan sir brought us some Moina macrocopa when he visited our college for CUBE Meet at St Peter’s College, Kolenchery.
  2. We transferred the species into a plastic container and filled it with dechlorinated water.
  3. We plan to study histone remodelling in Moina via Haemoglobin protein expression through epigenetics and for this we must first increase culture of Moina.
  4. Rather than using milk for culturing, we decided to use a different method. We are currently culturing them using yeast and they seem to be multiplying very quickly.

[center]Close up of Moina macrocopa culture[/center]

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Short term objective: Largescale culture of Moina macrocopa
Long term objective: Histone remodelling in Moina macrocopa via Haemoglobin protein expression through epigenetics

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Protocol followed to grow Moina macrocopa using yeast:

  1. Take 50ml of dechlorinated water in a beaker.

    [center]50ml dc water[/center]
  2. Put 4-5 yeast granules into this 50ml of water. Yeast powder maybe used as well, but granules are easier to get an idea of the exact amount of yeast dissolved.

    [center]Yeast granules[/center]

    [center]Addition of 5 yeast granules into 50ml dc water[/center]
  3. Stir this really well until the yeast granules are completely dissolved in the water. You may heat the water to speed up this process, but ensure that the yeast solution has completely cooled down before adding into the Moina culture.

Unlike culturing of Moina macrocopa using milk, it isn’t necessary to add this yeast solution everyday. We observed that the Moina have stayed alive for 2 complete days without addition of yeast solution. We’ll proceed to check how long the Moina will survive without addition of yeast solution.

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These were already identified species of Moina which we recieved from CUBE Elphinstone via @Arunan sir. Hence, any identification attempts were unnecessary.

Moina contains the following species (those discovered so far):

  • Moina affinis
  • Moina australiensis
  • Moina belli
  • Moina brachiata
  • Moina brachycephala
  • Moina flexuosa
  • Moina hartwigi
  • Moina hutchinsoni
  • Moina macrocopa
  • Moina micrura
  • Moina mongolica
  • Moina rectirostris
  • Moina reticulata
  • Moina tenuicornis
  • Moina weismanni
  • Moina wierzejskii
    Reference: Moina - Wikipedia

Awaiting for your update :star_struck:even we want to know how many days can it survive without food. By the way how much yeast solution are you putting in the cultures ?

See I understand you are truly trusting on @arunan sir but you should confirm whether elphinstone college Moina macrocopa only and not any other species.

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What is the difference between yeast granules and yeast powder? By yeast powder are you referring to yeast extract powderr? In that case there won’t be live yeast as feed for Moina…

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Identifying features are necessary since your group at CUBE Kolanchery claims to have native Moina and Daphnia from a nearby pond!
Let’s face the question of how to
A. Identify Moina from Genus Daphnia
B. identify the local Moina species from Moina Macrocopa JSK 1 you got from HBCSE

IT IS ALSO IMPORTANT NOT TO MIX UP M Macrocopa JSK 1 WITH OTHERS! So, please inform what precautions you use while culturing both in the same lab at CUBE St Peter’s Kolenchery.

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Definitely sir.
A) Major difference between Daphnia and Moina are their size. All Moina are small or medium-sized cladocerans. The body length of adult parthenogenetic females varies between 0.5 and 1.8 mm, depending on the species. Daphnia females can grow up to 5 mm and the males about 2 mm.
We’ll compare the morphological features of Moina and Daphnia and add them to this thread.
B) We’ll also compare Moina macrocopa to the local moina species. College will remain closed till 8th so please wait till 9th for these updates :sweat_smile:

Precautions followed to prevent mixing of Moina cultures:
We have two Moina cultures. One is of the local unidentified Moina species and the other is Moina macrocopa.
In order to prevent mixing, we have made two seperate cultures. One, exclusively for Moina macrocopa and the other for the local Moina species.
We have also labelled both these containers to prevent any confusion.
Seperate droppers are used to prevent any mixing of the two Moina species, one for each culture.
We culture the local species as Hydra feed, and we want to increase the Moina macrocopa culture to study histone remodelling via Haemoglobin protein expression through epigenetics.

Yeast granules are larger sized granules. These are more easy to use as they are easier to measure. Yeast powder I was talking about is not yeast extract, rather these look more powdery than the granules but are actually very small sized granules :grinning:

I could only find so few articles on identification of the different Moina species and are quite difficult to distinguish between them as well. How did you perform identification? Is there any reference link I could use to do the same?
We’ll soon perform a morphological comparative study between the local Moina and Moina macrocopa. College is closed till 8th so please do wait for the updates :sweat_smile:

Can you please share those articles with us?

How about microphotographs with 4x and 10 x objectives, please!

And kindly see that the Moina Macrocopa JSK1 do not get mixed up.
Please ensure a protocol for that at your center and post that to all! @Akshitha @abhijith @Abhishek

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C. Key to Species in the Genus Moina
This identification key for morphological forms is derived from species descriptions in Goulden, who reported six species from N.A. Moina and Moinodaphnia are typically planktonic, but the majority of species are restricted to small
temporary ponds, saline, or alkaline lakes. The habitat is often ephemeral, turbid, and warm. Adult females reach 1–2mm, with one Moina species that is only about 0.5mm long.
1a. A simple distal tooth (next to the claw) in the row of teeth along the dorsal margin of the postabdomen; western N.A. saline lakes
Moina hutchinsoni
1b. A double (bifid) distal tooth (next to the claw) in the row of teeth along the dorsal margin of the postabdomen …2
2a (1b). A row of teeth along the side of the setae on the last and penultimate segments of the first thoracic leg; ephippium with two eggs; north
temperate, coast to coast … Moina Macrocopa
2b. Feathery setules along the side of the setae on the last and penultimate segments of the first thoracic leg; ephippium with one or two
eggs …3
3a (2b) Long hairs on head and carapace …4
3b. Hairs completely absent from head and carapace …5
4a (3a). Never over 1.2mm long, one egg per ephippium; an uncommon species found from midwestern and southeastern United States and west into Mexico … Moina affinis
4b. Some adults longer than 1.2mm; two eggs per ephippium; southwestern United States to Argentina … Moina wierzejski
5a (3b). Length less than 1.2mm, one egg per ephippium; temporary ponds to large lakes in the north temperate zone, coast to coast and south to
Paraguay; from temporary ponds to large lakes … Moina micrura
5b. Some adults longer than 1.2mm; two eggs per ephippium; one population known from southern California …Moina brachycephala

Reference: Ecology and Classification of North American Freshwater Invertebrates (Third Edition)
2010
Stanley L. Dodson, Carla E.Cáceres,D. Christopher Rogers
Chapter 20 - Cladocera and Other Branchiopoda
https://doi.org/10.1016/B978-0-12-374855-3.00020-0

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https://www.researchgate.net/publication/232709437_Moina_Crustacea_Anomopoda_Moinidae_in_the_Czech_Republic_a_review
This is yet another article I found, but it gives no information on the morphological features of the various species. It only talks about their habitat and behaviour :confused:

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7ml yeast solution in 4450ml dc water.

How is the yeast solution made?
Any standard protocol which you follow?

Looking for updates on your Moina cultures!! @Akshitha

We can address problems faced by the Moina Group of CUBE Kolenchery, discuss on the identification of your Moina species, the Protocol followed, the design and most importantly The Research Question!!

Update on Moina macrocopa brought from CUBE HBCSE, Mumbai @Akshitha