Isolation of soil Nematodes &looking for C.elegans in Indian soil

Soil Nematode ,photo by @Jatin Joshi ,CHM College,Mumbai .

Cubists are interested in soil Nematodes especially 3 prize nobel prize winner C.elegans. But there are no report of isolation of C.elegans from Indian soil . Cubists are trying for that ,as you can see these are the certain experimental setup made by Cubists .

Cubists are using boiled potato slices as a medium and adding milk at the centre of potato slices,as a source of bacteria .

Setup by @shalinisharma98

Objective - Isolation of soil nematodes to look for the variations among the population.

Soil sample - I had collected the soil from two places, 1) Dolichos plant (hyacinth bean) from the potted plant 2) Brassica (Mustard) from the open field

Experiment design - This experiment was put on 18-01-2023 by putting soil sample at the periphery of the raw potato slice and a drop of milk at the centre.

Some questions may come in the mind of new cubists…

1) Why milk?
2) From where the nematodes will come?
3) What is the role of control here?
4) Why there are two test potato slice?

  1. Milk contain bacteria which will serve a purpose of food for nematodes.

  2. We are expecting nematodes to be there in soil, at certain time if much bacteria are not present in soil…so nematodes will come out of soil to search for their food which we are providing in the centre (bacterias from milk).

  3. In both control and test we will be able to see bacteria colonies as we are adding milk but nematodes we will only found in the case of test sample.

  4. I had put the replicate just to make sure if anyhow one test doesn’t work then i will have another test to proceed with an experiment.

Expectation - In each test and control sample from both the setup, I expected for some bacterial colonies to grow.

Observation - I could not see any bacterial colonies even after 8 days.

Interpretation - I assumed it may be because that time the temperature of Delhi/Noida was low (8-12°C) and bacteria requires optimal temperature for their growth.

So, the next time I put setup on 25-01-0233
And didn’t give giving clayey texture to the soil, and added buffalo milk.

On 29-01-2023, i observed the milk was solidifying and also at the periphery of the centre i could observe that it is degrading.

On 1st Feb, i could observe the some white spots were there in the centre and on 3rd Feb, i could observe the fungal growth is there.

:sparkles: Objective - Isolation of soil nematodes ( C. elegans or its relative).

:sparkles: Soil sample - I had collected the soil from hyacinth bean creeper from my home itself.

:sparkles: Experimental design - The experiment was put on 10-06-2023 at 6 pm.

First made the 6 potato slices using 3 raw potato.

  1. T1 and T2 - It is our test slices in which I have added a drop of milk (source of bacteria) at the centre and collected soil (source of nematodes) at the periphery.

  2. PC1 and PC2 (Positive Control) here i have added soil at the periphery and not the milk at the centre.

  3. NC1 and NC2 (Negative control) here i have added a drop of milk and but not the soil.

:sparkles: HYPOTHESIS
The expectations of ours after 3 days:-

PC1 and PC2 - We will be able to see the nematodes if the contamination of bacteria from air will be more than the bacteria in soil on the potato slice.

NC1 and NC2 - In this case, we will be able to see bacterial growth but not the nematodes.

T1 and T2 - Nematodes can be found on both the potato slices.

(T1, PC1, NC1 and T2, PC2, NC2 are replicate of each other respectively)

Detailed description of setup 1

Objective - Isolation of soil nematodes.
Soil sample was collected from the field of Brassica plant.

Data collector name - Shalini Sharma (CUBE, DSC)

Objective - Isolation of soil nematodes.
Setup 1 for soil sample 1 from Brassica (Mustard plant) open field.

  1. BS1 = Brassica Soil 1
  2. BS2 = Brassica Soil 2 (replicate)
  3. Control

Data collector name - Shalini Sharma (CUBE, DSC)

Objective - Isolation of soil nematodes.
Soil sample was collected from the potted plant of Dolichos. ( Setup -2 )

Data collector name - Shalini Sharma (CUBE, DSC)

Dolichos plant (Sem beans)

Objective - Isolation of soil nematodes.
Setup 2 for soil sample 2 from Dolichos (Sem beans) potted plant.

  1. DS1 = Dolichos Soil 1
  2. DS2 = Dolichos Soil 2 (replicate)
  3. Control

Data collector name - Shalini Sharma (CUBE, DSC).

Whole setup is covered with net.

Setup by @Swaha

AIM: To isolate soil nematodes
Name: Swaha
Place:Jalpaiguri, West Bengal

Setup :1 by @amritha

Data collector name :Amritha.k sn college alathur time :2:10 pm place :palakkad

Nematodes data collector name: Amritha.k sn college alathur place :2:10pm place :palakkad

Nematodes data collector name :Amritha.k sn college alathur. Time

Setup :2 by @Amritha

Data collector name: Amritha.k sn college alathur
Set up time:12:00
Time now:7:00
Experiment name:nematodes.

Data collector name: Amritha.k sn college alathur. How i do my nematode experiment in my home lab :point_up_2:place:palakkad date:2/2/23

Observations from Amritha .

@amritha says this is nematodes ,but according to sakshi this is not . @sakshiconsultant2002 says based on the size and the presence of segments in the worm . @sakshiconsultant2002 how can we prove or disprove your comment ? Any design of the experiments?

Photos & writeup about C.elegans by @Anshu_kadam_2813


C elegans culture back to CUBE
So I decided to do my Master’s dissertation on C elegans. And with help of my professors I got C elegans N2 strain culture yesterday.

Short term aim - Subculturing and Maintenance of C elegans

Main culture plate I received on 3/3/2023

Straight stoma

Observation under Compound microscope 40X magnification of Objective lens (4/3/23)

2 Pharyngeal bulbs are visible

Subculture plate made on 3/3/23. All the white structures are nematodes


Anatomical characteristics of C. Elegans that we use for identification are

  1. Straight stoma
  2. 2 Pharyngeal bulbs
  3. Vulva position in the middle
  4. Long tail

Please check @⁨Batul Ma’am⁩ @⁨~Shivam kumar Sriwas :man_wearing_turban:t2:⁩ @⁨Arunan sir⁩

This is a nematode we found out when I was in Ratnam college. It also had 2 bulbs but the vulva position was in the posterior end. That’s why we considered it as Caenorhabditis genus. ( I’ll check if I have the picture of vulva)

This is the picture I clicked yesterday. There is a small opening from where the eggs and some body fluids can out. So I’m guessing it should be the vulva. @⁨Arunan sir⁩ @⁨Batul Ma’am⁩ @⁨~Shivam kumar Sriwas :man_wearing_turban:t2:

Photo shared by @Arunan

From @⁨~Jatin Joshi⁩ on 17 February 2023 of local soil nematodes CUBE CHM college, Ullasnagar, Mumbai. @⁨~Jatin Joshi⁩ @⁨Anshu Kadam CUBE⁩ @⁨Batul Ma’am⁩ Please comment.

There was an interesting discussion in context curriculum Whatsapp group about the photo of Jatin Joshi on 21/02/2023

@magpie wrote

@⁨~Shivam kumar Sriwas :man_wearing_turban:t2:⁩ this is from Anshu’s group Can you now appreciate why I am saying Jatins pics are of Caenorhabditis and not Oschieus?

@⁨~Jatin Joshi⁩ nematode showing straight stoma,2 Middle bulb 3)Basal bulb

jatin’s nematodes could be of C.elegans !? What do you say @sakshiconsultant2002 @shalinisharma98 @Arunan & others

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Advisory_Services_for_Nematode_Pests.pdf (1.9 MB)

This is how i used to set up culture for isolation of soil nematodes but there were so many goof ups that i made while trying to culture nematodes! From @sakshiconsultant2002 's homelab .

@sakshiconsultant2002 wrote,

This was my 4th set up of nematode culturing and like amritha i also observed some flies on my potato. As amritha said in yesterday’s chatshala that how can fruitfly lay eggs on a potato slice?? I guess what she wanted to say was apart from fruitflies there must be other flies laying eggs on the potato slice is it @⁨Amirtha CUBE⁩. In this case the flies which i observed on the potato slice were not fruit flies, so the larvae amritha got could be of any flies. For identifying it amritha should wait for the larvae to turn in pupa and then when the fly comes out we can say if it a fruitfly or some other fly

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