 Yeah, so we are beginning after the learn session, so the group which is now going to present is from, there is a nearby study center which is called as M-Ward Learning Center. So it is a study center like it has a library and a space. So we have started a CUBE program and they have set up a lab there. So they actually have now plenty of moina, they also have plenty of fruit fly, also algae. So they will be presenting about their moina work, how they have started, from nothing they have started a lab. So they will be talking about it. Hi friends, my name is Gul Sade and his name is Bindu, he is just 10 and his name is Isade. I am from M-Power and today we are going to be presenting about the moina, on which we have done our work, I would like to tell you all about it. First time in M-Power Library Center I got to know about CUBE, there I met Arunan sir, Jai sir, Mayur sir, Aarif, these people met me and we told them what CUBE is. We meet in two days in CUBE, in M-Power Library Center. What is CUBE? I would like to tell you about CUBE, collaboration, education, understanding, biology, education. This is a different way to learn about science. This is a simple model system moina, which was our second set up on 25th January. At that time we selected two bottles, which were 500 ml, and filled them with 250 ml DC water. And in that we transferred 500 ml to 50 ml DC water, and in both of them we put a label, one drop of milk, both of them were set at home, their feeding was milk, and in both of them we put one drop of milk. But before that I had done something, which I wanted to do, which I wanted to learn and which I wanted to learn. This is why our first set up was different from the date, which I did on 28th December, which was my first set up. At that time we selected two bottles, which were 500 ml, and filled with half bottle of DC water, and in that we transferred 500 ml to 50 ml. And in that we put a label, one drop of milk, and in the second we put five drops of milk. And we set it up at our home. Because we couldn't go to the library centre every time, so we set up that lab at our kitchen. And we used to take care of it. And every morning we would put milk in it. The purpose of doing this was that we wanted to know that they are moina colourless. And before that we wanted to know that there are five drops of one drop. We wanted to know which bottles have more oxygen in them. First one is drop, and the other is five drops of milk. After the first set up we observed something. It was that after three days moina increased. And after 20 days it became red. After 24 days it became dead, because it lacked oxygen. And my mother said that we should keep the bottle open. So we gave it to her, because she couldn't get oxygen to breathe. And five drops of milk was given in 15 days only. Moina organises itself increases the increase and percolation. And why did we learn something after observing it? It was that we feel that moina is in an organiser who doesn't need any other organiser to increase his or her progeny. He alone increases his or her organiser. Milk is its fruit. Milk is its fruit which is made of bacteria. And bacteria and moina both need oxygen. And due to lack of oxygen, when both of them need equal oxygen then due to lack of oxygen moina, it became red. After observing it, we understood that the way we need to breathe and eat, similarly small organisms also need it. Hello. This is the data. As they told me, it was the same model system as mine. But I found something different in it. Like when I did it, I found it in bottles. But I didn't put it in bottles. Maybe it was because the DC water was open for a long time or some other particles came in it. So I found rotifers in it. Moina was taken from the cube lab. I found rotifers in it. And I divided them all in it. So I found rotifers because of that. But I didn't find these people. And my progeny was increasing in two days. The amount of my progeny was increasing in two days. And as people told me, how many children can give a moina? How many progeny can a moina give? So I did that experiment. I saw that it was giving 5 to above and under than 10 progenies. So let me show you this video. Moina is a rotifer. Look at this. When I got this photo, I found it. Start. Look at this. This is a rotifer. And this is a moina. And this is a rotifer group. Thank you everyone. And I, Jai sir, Arunan sir, and the rest of the people wanted to thank us for coming here. And they told us what the cube is. So they want to know something. So Moina, Gulshan and Sipthin told us that moina. So I am working on moina. So you can see that moina. So I showed you how moina looks like without microsoft. And this moina is taken on T10x. And this is the diagram of moina. So Moina, as the king told us, moina is taken from here from the lab. So where did we get this moina from? So when we come to Vasi stations, look down there, there is a bridge. So maybe from there, from the place before, we got samples of moina from there. We also got catfish from there. So we got samples of moina from there. So after bringing moina from there, they identified what moina is. So moina is a living organism. We have seen it in Jhinga. So the relative size of Jhinga is here. So if you compare it with Jhinga, it is a prawn. So it looks similar. It is so big in size. And in microsoft, we will see the size of Jhinga as big as it is. So if we see it like this, we will see the size of around 2mm. So where will we get this moina from? If we go to find another moina from there. So I told you that for hydra culture, I told you that moina is important for hydra culture. If someone has hydra, we can also get hydra from the lake pond. There is water, there is algae. So we got moina from here in pond, dam and lake. We also got moina from the lake pond. I was with my team back there. It is not with me. We got moina from Gondi. So if you see here, there is water from the pond. So we got moina from here. And we also got moina from the learning centre of MR. So we got moina from rainy season. We also got moina from rainy season. After that, we have a ground where we go to play. So we also got moina from there. So if you see this stagnant water, it is not completely thick. And here, it is near the pond gutter. So moina can come from gutter if someone has done something. And here, the water is directly collected. And we got moina from it. So this question is, how did moina come from here? So we also identified some experiments that we can do that if it is filled with water, then after a few days, moina can come or not. So if we want moina from here, then in the rainy season, we can get it from stagnant water place. So we can get moina from here. And moina, how to culture moina? I have told you earlier that how to make moina in hydra. So we are using the same method. We take two bottles, two bottles are de-grounded water. De-grounded means DC water. I had told you that you have to keep it in a bucket after 24 hours, it becomes de-grounded. So if you take it in a bottle, then if you are doing it in 500 ml, if you are doing it in 500 ml bottle, then you have to fill half of it with water. And in that, you can see daily milk one drop, two drops. So the requirements, I have already told you once. This is two red moina. So red moina, what you see here is two drops, yellow moina, three days. So if we get three red moina, then what I have said is that I have made a set of bottles. In this, total eight bottles, six bottles, six bottles, one in two drops, one in four drops, one in six drops. Here, I have made a mistake. This is one drop milk and this is two drops milk. So I have made two bottles of one drop, two bottles of two drops, two bottles of four drops, two bottles of six drops. So I had to add different numbers in daily milk. So what observations I got in the microscope? So here you see, three days of yellow moina in one drop. Here in four drops, red moina is red. And in one drop, in six drops, red moina. If we want to culture red moina, then we will give six drops, six or four drops. So in six drops, in three days, red moina will take four days. So now, Hina and Chitra, we will tell you about the red moina. We will also tell you about the red moina. Chitra, we actually culture red moina in the lab. So basically, as I have explained, moina are something like Ginga only. They are insects from Crustinicious genus and they are found in lakes, pools, and basically wherever the water is tangled. So usually, the size of adult moina is from 0.7 to 1 millimeter. And the young moina size is 0.4 millimeter. Moina basically feeds on bacteria, yeast, fungi, but most people will be given bacteria and fungi. So moina has a head part and a trunk on the head. The antenna present is used for the locomotion. There is a brood pouch at the dorsal side of the moina. In the brood pouch, the embryos and the eggs are developed. Basically, in moina, the brood pouch is open whereas in Daphnia it's a closed pouch. Moina can reproduce sexually as well as asexually. Moina generally reproduce after four or seven ages and they can give 4 to 22 progeny at one go and they give 2 to 6 such brood pouches in their lifetime. As you can see, this is a visual diagram. So this means that when we brought this moina from the lab, there was a kajal in Delhi. Before we brought this moina, we used to call it Daphnia. In the search organization, they call it Daphnia. In the lab, it was called Daphnia. So the kajal found out that it was moina or Daphnia. They were also doing this experiment and they didn't even feel that it was Daphnia. So suddenly, the kajal observed something that it could see here. This is the mouth part. This is the antenna. This is the cuff. The point of the Daphnia is its mouth. So in this way, it identified that it was moina or Daphnia. The kajal found it in Delhi. It's a special quality that it can resist a wide range of temperature and it can also resist in a very low amount of oxygen content. Basically, moina, they are studying red moina, right? So red moinas are turned red because of the more production of hemoglobin, which as the oxygen level decreases, it produces more hemoglobin due to which they also appear red. No. Hello everyone. If you want to cultivate red moina, the requirements are you just need the five things that are easily available in every source. Like you can get DC water, you know, DC water has been explained, transparent bottle, a drop of milk and muslin clothes and five moinas you can start. Basically, we have started with the on 22nd January for cultivation of red moina. So procedure is we have started a project with a four, one little bottle. That is, we have taken just one bottle that is controlled and another three are for cultivating red moinas. So we have taken one liter bottle and we have added 500 ml DC water. In that, we have added 10 moinas. So after that, we have started with one drop of milk and of course, we have we have plugged it with the cotton plug. So after on the third day what happened actually, we were eagerly waiting for getting the red moina. So we thought on the third day, on the first day, we were having almost 15 to 20 moinas. After that, we were having 25 to 30 moinas. After on the third day, what happened we were like eagerly waiting for the red moina. We should get the red moina. We will get it quickly. So we thought, let's add more milk. So we started with one drop. In control, we have actually added one drop in control and in other three bottles, we have added three drops. So on the third day, in control, we have started with three drops and in the remaining we have added six drops. So after on the seventh day, we have found red moinas and so that was the goof up. So we have done that. This is the on your this colorless moina and this is red moina on the seventh day for see you can see here blue line is for control and the another three bottle having red, yellow. So the number has been increased after the red moina also. But at the on the 14 day, the moinas were dying. So main was short term objective was that to cultivate red moina. After that, our long term objective was that to find out whether this red moina are getting reversible. Are they getting again colorless or they are getting permanent red. So that was a thing. Thank you. One more thing, someone has a question here, which is moina culture. Someone has a question, why are we adding milk. Someone has a question, why are we adding milk to the bottle of moina. So the bottle of moina is that we are making the decolonized bottle. So there is bacteria in it. So to grow bacteria, we add milk. There are 14 present. If we take a little curd and add milk to it, then the milk is converted into curd. This is the growth of bacteria. So in the bottles, we have seen 1 drop, 2 drop, 3 drop. In the bottles we have 1 drop, 2 drop, 3rd drop, 4 drop, 6, then 10. We have increased the drop of milk to produce bacteria. So in the red, everyone knows that moina is red. So how is moina red? Why? Because in the red, there is a colorless moina. So in the 1 drop, there is 2 drop milk. So in the 2 drop, compared to 1 drop, there will be more bacteria. So the growth of bacteria will be more. So in the bacteria, we took 5 moinas and put 1 drop milk in it. If bacteria increase then bacteria will eat it. So in the 2nd bottle, we add the same number. We double the amount of milk. There are more bacteria in it. We said that in 7 days, they got red. In the 4 drop, they got red. So in that, we increased the amount of bacteria. So in the bacteria, in the living organism, moina is also in the living organism. So we need bacteria and moina. If we increase the amount of milk, bacteria will grow more. So compared, moina is less oxygen and bacteria is more oxygen. So in the moina, according to my information, there is a hemoglobin gene. So in our blood, there is a hemoglobin. So in the hemoglobin, there is no oxygen where there is no oxygen. So the hemoglobin gene is present in the moina. So in that, there are no red blood cells in the moina. There are no red blood cells present in the moina. So when moina gets oxygen, the hemoglobin gene produces it. So moina is red. That is why we increase the amount. We study the red moina because it is an epigenetic thing. So we try to study it. So we have a plan ahead. I also have something else. So moina is red. Thank you. If you put a fish in a tank where it gets oxygen. So I was thinking, suppose you gave oxygen, and then put milk in it. Will it not turn red because I am supplying oxygen in the water? Can you do this? Yes, we can do that. Because the food of moina is bacteria. He is saying that if we take a bottle and give it oxygen and feed the milk, bacteria will turn red. So in that, we had to give oxygen to air. So if we can try like algae, then we will have to buy it for oxygen. So we will have to buy it for air. So will my hypothesis change if I supply the oxygen? Will the redness will be less if I supply the oxygen? Will the concentration of milk be more? Even if I increase the milk concentration? If I get oxygen, I also think that it will not be red. So if we have red moina, after it gets red, we don't have anything. So we have to study genes. So what we have to do is that we have got the moina. So we will take the same moina in other bottles. The same red moina we got in 7 days, 5 days, 3 days. So the gene expression would differ in the case of the oxygen cycle. Yes. So can you study the regulation by supplying the oxygen from outside? We can't do it outside. We have to check the recieval. So for that, we will design another experiment. The same procedure. DC water 250 ml. So what we will do is that if we get the red moina in the milk, the same moina is not red in control. There is no growth of bacteria. So in that, the red moina, we will take out the moina and feed it with one drop of milk. So we will feed it with food. And in other bottles, we can control it so that there is no feeding. If there is no feeding, it will be alive. If we give bacteria, bacteria will grow in the water. If we give one drop of milk, bacteria will grow. So in control, the bacteria present in the water will stay in the DC water. So we can conclude that it is getting red or recieval. So this is the next experiment that we have to do. We have to check the recieval. How do you know if it is getting red? Have you tested the oxygen? No. How do you know if it is getting red? Normal... So you have seen it that it is getting red. So I took a red moina which was red before and I was putting it in a lot of milk drops. Now I have put it in a normal DC water where I have put only one drop of milk. So it is becoming colorless? Yes, it has become colorless. It happened in three days. So I confirmed that I had put it in a normal DC water. I did not do the experiment. So I put it in a side water. In one try? Yes. But you have not seen the oxygen, right? No. So they have said that if you provide the oxygen, if you give the red moina, it will happen. So you can do the same? Yes, you can do it. Yes, it tastes like dissolved oxygen. But if you don't do it, you can also do it. But if you can do another thing also that provides the oxygen, instead of showing that there is an oxygen decrease, you provide the oxygen and see that it is changing. Yes. It can be like if you are putting more drops of milk, there will be more bacterial growth. And what will be the bacteria? It will be lactobacilli. Lactobacilli is micro-aero-tolerant. It also needs some air. It needs some oxygen. It is not anaerobic. So because of that, it can consume the amount of oxygen in the bottle. So it will work for the moina. Oxygen is taking the bacteria. That is why moina is getting red. So if you are putting 6 drops of milk, there will be more bacteria than moina. So we can get a lot of bacteria in it. We can't even count the bacteria if we put them in one drop. If someone is here from chemistry or background or something, someone can suggest a test by which I can put some chemical one drop or one capsule of something into water and it changes colour according to the oxygen content. Because dissolved oxygen test is there. The compounds which will take up the oxygen. Actually milk also... Yeah. Yeah. Arif, you said that in moina, there are white blood cells. No, there are no white blood cells. I mean there is no RBC. There are no white blood cells. Sorry, there are no red blood cells. And when it gets red, it gets red because of hemoglobin. So it gets red because of red blood cells. Yeah. So hemoglobin is a protein that has hemoglobin gene and protein. It is present in every cell of moina. It doesn't function in it. It is present in every cell of moina. So it doesn't have red blood cells? No. It has red blood cells in humans. If it has blood, it will have more oxygen. No, it doesn't. We have an oxygen carrier to carry hemoglobin. RBC. So what is in it? Hemoglobin. The protein inside the RBC is responsible for taking oxygen. Hemoglobin. Like we have heard that hemoglobin is less, it has less oxygen. So we increase the amount of hemoglobin in red blood cells. The actual carrier is hemoglobin. Hemoglobin binds with the oxygen molecule and those hemoglobins are on the surface of red blood cells. So that's why our cells are we call it red blood cells. Because when they bind with oxygen, they look red. It's an oxide of iron. So iron oxides are red in colour. So in what he is saying is in moina, those proteins, those hemoglobins they are freely dissolved in the body fluid of moina. No special cell is there to carry it. So that's why the whole moina appears to be red. Anyone else? So I saw your pictures of the one drop and the two drop three drop, four drop, six drop moinas. So if you see the one which is colour less has a body structure that is not so swollen as the one which is red in colour. Can you tell me why is that difference? Which one is this? Yeah. White blood. Why is it so swollen? Yeah. Now, when you compare both of them then the colour less one looks smaller and the red moina looks bigger. Why is that? So in that the one which is visible is also big. This is big and this is also big. So this time the one which is bigger on the backside has a brood pouch. This means that the male has a sexual function. Both of them are in the same moina. The one which is visible on the brood pouch has a male function. The one which is visible on the brood pouch has a female function. It's not a small moina. It's not a small red. Red means it's progenies. So we can't see the red moina. So this is a very important point. If we take a small moina then it will take some time to culture it. Because the small moina is progeny. It will take some time to function so it will increase. So to start the culture we need to take a gravid moina. If we take a gravid moina then it will take at least 70 to 80 100 crosses. Take a big moina. If we take a small moina then it will take some time. If we take a small moina then it will produce in a third day. Like Sifthia said it will produce in 10 to 12 months. One moina in a day. So this red moina means the small moina has a one drop. If we take a red moina then it will take 3 drops. So the small moina is not a big moina. So we can experiment that the red moina we can put it separately and make it progeny. So the red moina is colorless. Or we have to see the progeny of the bottle. So the size is similar. The size of the bottle is small. So everything should be red. So I have seen that. So we will have to experiment that. So the progeny is colorless. The question is that the small moina is red. The small moina is red because the small moina is from birth. So whether it is born colorless or red that is the question. This can be an experiment. And the other day there were two visitors in cube lab and they were discussing. Some students were discussing there. I don't know whether you were there. Your graph pattern you were showing. If in so many days suppose in 5 days or 6 days colorless or red so if colorless there is 50. So all 50 and 50 together are red. On the 6th day. So they predicted that 40 are red and 10 or 20 are still yellow or colorless. So it will be interesting to see that graph pattern. I think Apurva can also become part of this project and I want people to take up this experiment. Of course along with that. We have to cut it. Yeah. Actually we did the experiment with red moinas also. We put the red moinas and we observed that they are progeny red colorless only. You don't have to ask. I don't know the answer. Yeah. Okay. So, you want to have. Yes. This is our problem. The way we take oxygen and leave carbon dioxide so will this system be there too? After doing this we have a question. Or the way we put milk for bacteria we want to see something else. And we want to do it. If you feel that we can do it then you can say it. The hemoglobin that they said is also in humans. We are also telling the same thing. But my point is that we take oxygen and leave carbon dioxide. So how do we leave it in water? The way we leave fish. Okay. Because if we take a glass and leave carbon dioxide then bubbles will come out of it. When the fish breath then they will observe it. Then bubbles will come out. For example, if we see fish tank then bubbles come out of the gills. So we feel that carbon dioxide is there. See, I want to divert your attention a little bit. Of course, first I wanted to just respond to it. This is normally the way how textbooks are written. We take in oxygen and give out oxygen. When we breathe, oxygen enters. When we breathe, air enters. When we breathe, air comes out. Oxygen and carbon dioxide we cannot take. Air enters, not oxygen. Air enters. So this is a general popular misconception for many people. But what actually happens is air enters in lungs where blood is more. When there is an exchange at that time, the dissolved carbon dioxide and dissolved oxygen both are there. But hemoglobin only takes oxygen. So if we leave it, both can come together. So oxygen will come out, carbon dioxide will come out. So that is one small important thing that you have to remember. Because in textbooks as it is written that leads to a lot of confusion. And this is something that we have to think about. Confusion creating issue for that the textbooks are also responsible teachers are also responsible. So at least we as Cubists we should try to keep a good eye and critical eye and all these things. Because we need evidence, right? Or that is why there are some ministers who said that cows release oxygen. They also take oxygen. Right? So now this is what is happening. People who are really ignorant are ruling us. We have to really do what? Cubists have to head on to say that we challenge you. We will do the experiment and we will find out which is that cow which will sort of you have to say the same thing. Cows also take air and they also take out air. Yes. And they also need oxygen. Daphnia also needs oxygen. Bacteria also needs oxygen. So having said that I want to throw a challenge today. Of course this is so if you look at the first two years of our conferences in cube and the presentations this slide would have been Daphnia. Not Moina. So we started with Daphnia because that is what is there in biology publications everywhere. So we started with Daphnia. Now today you haven't heard Daphnia as much as Moina because you heard only Moina today. That's a good sign. Now the challenge is if there is anybody anywhere in the country who will bring a native Daphnia they will get a big prize. There is one here. Earlier the presentation was there. Where did Hyderabad come from? No, we will not listen to anything. We have to show identify evidence it has to be Daphnia and not Moina. Now your question starts happening. What is the difference between the two? How do you find them all that? So I will leave that as an open challenge and the challenge sustains. I am going to give a big prize for people who are going to tell which area in India has Daphnia and which of course I am actually challenging that everywhere you get only Moina. In Delhi you get Moina and in Mumbai you get Moina and in Kerala you get Moina. This is Moina country. Which countries have Daphnia? Let's find out those things and maybe over a period of time we will come to know that the whole world has only Moina. So we will move on to the next presentation. You want to say something also? We will have the session or you can write to us. Sifthan said that one Moina gives 5 to 10 Pojinis and he said 5 to 20. Is there any difference between the two? Is it special? You can see. Both of them can be. Okay. So next group is what mosquito? To experiment we have taken 5 to 5 Moina in the sub-bottle. We are not sure whether they are male or female or they are generated by themselves. We will see one Moina in the set-up whether she does it herself or not. And how many do they do? Because there are two questions about how they are more or less. So we will take one and see whether she does it herself or not. What can be the difference between the two? Okay. So next is Ubaid and his group on mosquito. Mosquito mapping. Good afternoon everyone. Myself Rihanna, we are from Chamber Naga MPS we are collaborating in our school and we are in college in Kerala and also with Adarsh Vidyalam and Ubaid is going to tell you the seasonal cycle of mosquito. So we are doing according to seasonal cycle because in September if we got data in September and then we can compare to March means autumn to vernal equinox because the day and night time are same and if we got data in winter solstice and summer solstice then we can compare because in winter solstice days are shorter than the night and in June means summer solstice they are larger than the night. We are catching the mosquito by clap trap method. First we do by clap trap method but later on we we move to over trap method because we find the trapping method is not very effective like if we catch by clap trap method we can prevent the mosquitoes but it must be by someone but if we do by over trap method before the mosquito get adult we can like destroy them so they cannot regenerate. Ubaid is going to tell you how to make over trap. How to make over trap? Step 1 prepare plastic cup and step 2 paint it black because mosquito attract to dark color and step 3 fill the half cup with water step 4 walk white cloth on the scale step 5 label it step 6 keep it outside and check whether the mosquito has lay eggs or not. I found eggs on over trap. There are three different types of mosquito first anaphyl is second ad is enter culex. we are working on ad so we we have to find ad so it looks like this. it is different and anaphyl is anaphyl is in between white and in side black and culex all the eggs are joined. we found ad because you can see there is a dot on the side and these are different colors in different places. so we made a hypothesis in which this is Gorangi's hypothesis which was my colleague he said that in January to May there will be more non-ad compared to ad's and from from May to September the non-ad's will be lesser than the mosquitos will be more and the ad's will be more compared to non-ad's because in rainy season she thinks that if mosquitos lay eggs on the water then when rain comes it will flush out and in my hypothesis I used to think that from January to April the number of mosquitos will not be more because there will be no water and from May to September the mosquitos number will be more because rain will be coming and the water will be collected in a nutshell and some old shoes and in both the cases the similarities was that the ad's will be more compared to non-ad's and the results were really different because because in I used to think that in January to April the total number of mosquitos will be less but when I got mosquitos then the results were different because from January to May the total number of mosquitos was more compared to ad's how does the OVTRAP work? as you all know that mosquitos lay eggs on the water so that's why we fill it with a half of water and we wrap the scale by white cloth because the mosquitos eggs are black in color so that's why we can see it easily so if we keep outside then if we keep outside near the dark area mosquitos will come and lay eggs on the on the scale there so we can easily see and the benefit is that because we can get the data before the mosquitos get adult and before it it bites someone someone so we got this number of mosquitos then one time we got 125 mosquitos in a day so it's really good compared to this clap trap because when mosquitos get adult then we used to trap the mosquitos so in this one we can get before the mosquitos get adult so our future plan is that we will compare the data with the hypothesis that we make again for one year and we will map the mosquitos trap by the OV trap and differentiate the ADs and non-ADs eggs thank you any questions yes we got ADs more in July so I can say that in coming July you can get you can get dengu in more number you can get but in less number compared to July but you can get malaria or elephantiasis more because we got the non-ADs number more like 140 yes we can compare the data with the BMC data yes okay okay ADs because we got the ADs eggs we only got the ADs eggs every time in the second season you didn't get any OV trap or any mosquitos eggs we kept it in a very short time we will keep it in the future every day we will keep OV traps in every season and maybe you can get ADs because they lay eggs on fresh water and the non-ADs they are in dirty water they are in dirty type so in dirty type where there is dirty type water you can't keep your OV trap just scale and white cloth otherwise you can't keep some dirty water how do you know that your dirty water doesn't look dirty so if you keep OV trap in dirty type water you will get larvae in some kind of a drum and they were not ADs so you can try with water so this is your future plan why did you take this project why did you take this project because when we first came here we were not asked and we were put in a group and then later it was interesting for this project and most of all Daphne was there because he had friends and the teacher allowed him to go a few days ago you didn't get any ADs in his area Zube they were in his area they they didn't know about OV trap any more questions stop tell me if he came we would have killed him brother how will you identify male and female we can't do in X we need to be adult and we don't want that to be adult but if we get adult mosquito we can do that but how will you identify he is old he has more hair on his antenna and less hair on female antenna like there are moustaches in males and females don't have that's how it happens so you can see you can see on google if any mosquito is coming to kill it means he is female there is fibrenogen in blood and it helps and if females don't have then they spend time on plant juice what you were saying how can ADs cause any disease dengue yellow fever it means there is only one mosquito any other mosquito can cause any disease ADs and yellow fever one mosquito so which mosquito is dengue dengue sorry ADs and she had a question how can you use mosquito as a model first of all there is an ethical issue we are not supposed to rare mosquitoes so we can't use mosquito in our labs at least you know so just an experiment then to rare the mosquito is an ethical issue but you can use the mosquito to feed somebody else that is not an ethical issue otherwise you are killing the mosquito mayur vayakik some students feed him some chickens yes so if he had any harm then he would be banned from doing project so he can't use it as a model one more question what is ov trap this is ov trap ov means eggs and trap means trap net how do you make it how when can we take this type of half cut bottle we can paint it black or any dark color we can attract mosquitoes and we can wrap white cloth on it we can keep it don't take white tissue paper or paper because it will mix in water and you won't get anything this is how it is no question so we will wind up the session ok so next I will call abhijeet abhijeet mitesh and his group to present the work on snails snail is a wonderful model system for as we know in long time we are using it for learning and memory studies so they have some developed some essays and some kind of modifications in that experiment good evening to one and all present over here I am Rithvik he is Rohit he is Aniket and he is abhijeet and we are from Adarsh Vidyalaya chamber and our project is based on snails her title of the project is a study to develop methods to change the behavior pattern of snails by association conditioning abstract snails are destructive organisms that are detrimental to human and human concerns by using harmful chemical pesticides environment can be affected giant African land snails are one of the most dreaded crops raiders in the world this project is an attempt to develop a method to change a pestiferous giant African land snails into a non-pestiferous by associative conditioning our work plan is the project work timetable was started from 3rd July 2017 sample selected for observations the vegetable extract such as cucumber, tomato and cabbage vegetables experimentations and sample animal was giant African land snail snail culture was developed in the school laboratory in a closed ventilated basket which was kept in a tray containing water this prevents the hibernation of the snails by creating a scene like rainy season our hypothesis is our hypothesis is pestiferous giant African land snail can be turned into non-pestiferous by associative conditioning our objective immediate objective to find the food that attracts the snails and to study the response towards it to compare the response of snails is more towards samples or in direction in which they are kept short term objective to find does the Q9 has any or not to study the memory retention period of snails long term objective is to construct a method to change the behavioural pattern of snails by associative conditioning so as you can see that we took a circular region paper of 20 cm and on it we placed a glass plate on one side we kept sample and on the other side we kept control the snail was kept in the middle we used to change the direction of the snail after every run and we also kept a tank on it so that we could confirm that there is no other thing which attracts the snail actually here is a problem that this sample and control is both in one place only the sample is here and the control is placed on the other side so this we also calculated the old factor responsive index that is ORI so control versus control we used the formula CA-CB upon CA plus CB but when sample and control is kept on one side on both the sides sorry we used the formula C minus S upon C plus S now when we started our project so we took a snail actually we took four snails for our test and one was like control so here we took a test snail which was named named as interest we named it as snail's interest so here we did the experiments like control versus control ORI was calculated and like that to check that to quinine has in order or not so like this we have done and then based on this table we have made a graph and this graph what is quinine actually quinine is an additive which we have to so quinine is an additive which we are using for the means like when we add something in that sample which is cucumber extract so when we put quinine in that it is odorless and it is very bitter in taste so by this means when the snail go towards the sample assuming that it's a good smell is coming from it but it is having a bad taste so when it taste it it is getting we do back so that is we have to check that is quinine having some smell or not so that's why we placed control versus quinine also yeah plus point the plus shows the attraction but the it is not should be the ORI should be like attraction on repulsion it should be also like equal distribution means there is like if you control versus control also point if you go back to your thing this is the last column the control versus control it is also point two so I would like to so like this this circular region we have used so here we can see that when control is kept on both the sides means let's assume that here also control is there and here also control is there means there is no odor in the in this particular region and we have placed it it is water it is still water still water we are using as a control so when control is placed there not only the ORI should become zero but it should have equal distribution means there is no smell so it can go anywhere it wants to so it's also like so it was like equal distribution only yeah the figure was not going that's why we put like this diagram type of thing yeah but photographer was not going so some technical problem was there ORI zero and the last one is also control versus control ORI plus zero two why do you put like that what is this why why did you do again control versus control which is very different from zero yeah we started with control versus control so we have to check that in last role so that there is a experimental setup is good till the last we did till the last experiment what is that could you just explain that so is it only once you did it that you got zero the control versus control that left side control versus control once you did it and you got an ORI of zero then you did control versus quinine you got an ORI of plus two is it once is it a no no with each nail we did it 20 times so then there should be a mean and a standard deviation or something yeah only one trial yeah sorry one one set of 20 runs of one one set is of 20 runs so only once you have run the 20 times and the reading is 20 times when you ran it in order to get zero you should get 10 intercepts one side and 10 intercepts one side and that's all you got next time if you did it 0.5 plus 0.5 how will you how will you roll out that possibility you do it once and then say that that's the fact that's not real yeah force nails so this is a particular stale called interest is it so you did control versus control you got 0 and here you got plus 0.2 if you did it so twice you did it control versus control if you did it five times you would have got probably not zero you would have got 0.5 sorry 0.4 something like that so how many times you did it and how many times if you did it only once how many times you should be doing it in order for you to get convinced about it after each experiment like first we had control versus control then we did control versus queen island again we should do control versus why why didn't you tested all your force nails we have tested but here we have this nail car what you wanted this nail goes according to you other snails what like they contradicted interest your interest no no it was something nearby this only means what should we expect so if you if you have put that like one interest one non-interest maybe interest so all those four snails and you do for all five times you will get a bigger data set from which you can like easily more convincing you can find the standard deviation in the mean yeah we have but logbook is in school yeah I have idea of two snails they were having like like in first time we did then it was like this is interest so we got zero and there is another snail called as so this okay this interest so it was giving the what as something like plus zero point one and some was yeah then it should be like equal distribution and the near to zero near to zero minus if you put control and control on both the sides what you expect normally from a snail like all the snails you test you you what you hypothesize that the snail cannot differentiate between water and water right it should not show any preference towards water versus water it should be like that's why it's zero it's not going anywhere else repeatedly so that you can only find out if you test all the four snails five times each so you get five for the 20 set 20 points that 20 times iron different snails and they move towards this or that thing the same thing is with Q9 and your sample which you are putting if a snail if you run only once and the snail is going by chance it is going towards your Q9 or your sample you will see that the ORI is 0.9 and I am very sure that this is the hypothesis so might be it's wrong right by chance it got there but four snails each running five times then it cannot be by chance right every time it cannot move that side by chance yeah so according to that we have made a graph of the snail interest again so here like first we did control versus control so it was like zero and then when we did control versus Q9 that is you have to check that is Q9 having some order or not so we did so it was get 0.2 and then we did control versus sample so control versus sample sample was a cucumber extract we use cucumber extract for the this test and then it shows the plus 0.7 and then we did with in the next run we took sample and we added Q9 before doing the experiment we took a glass slide and on that we mixed cucumber extract plus Q9 and in that we run the snail on the slide and it went towards the cucumber extract and it tasted it and after that when we do back to the shell and after that we did the experiment so we got the ORI which was minus which was minus 0.4 minus 0.5 sorry so this is a very good thing which we expected so from control versus sample and control versus sample plus Q9 so difference is in control versus sample plus Q9 you are actually letting the snail expose to this thing first then you are taking the reading so because once it's exposed and it knows it's a bitter taste now it will go or not so what time frame you are selecting as like after exposure immediately you are doing it after 15 minutes something so after 15 minutes so like you hypothesize that after 15 minutes also the snail 15 or 5 0 so after 15 minutes also the snail knows that this side is having sample plus Q9 I should not go there we assume like that and for the sample also you let the snail expose for the first time then you took the reading immediately then how it makes sense see for the samples the snail was not knowing what is there so you didn't let the snail taste it and then because if you are adding Q9 what is your hypothesis that Q9 is having some order or not but because the graph goes negative because they are saying that when they tested with sample plus Q9 they first let the snail taste it it is bitter and then they run they did the essay no they already mixed sample with Q9 and they let the snail taste it obviously the snail will taste a bitter thing then after 15 minutes they are running this control versus this mixture essay yes and if you are doing that why didn't you do that with your control versus sample for sample also for sample also let the snail first taste it then after 15 minutes you should do you should keep the methodology same otherwise in one case you have not let the snail taste the sample so it is only sample control and you are taking reading on the other hand when you are putting Q9 you are letting it taste it and then you are taking the reading so we did like this because when if we did directly control versus Q9 so first time in the first run suppose we are taking 20 runs so in the first run it will go definitely towards the sample because it is not experienced that thing do you have not run anywhere sample versus Q9 have you run sample versus Q9 at one side only Q9 at one side only sample have you done that no like that we did no where sample versus Q9 this control versus Q9 so we have done control versus sample plus Q9 what I am saying is don't make the snail taste the mixture first and then only put control at one side and sample versus Q9 on other side and see what happens like you did for control versus sample I am asking why you exposed the snail in the first place to this mixture before taking the reading what was the thinking behind that we think that in the first runs the snail will go towards the sample and it will experience the bitter taste of the Q9 with the sample and then it will start rippling towards the sample rippling it will start repulsion towards the sample from the second run you should have done a sample at one side cucumber extract put cucumber extract at one side and Q9 alone at other side then you can what you can see if it goes first time towards the Q9 next time it is always going towards the sample or not this will suggest that it got the taste of a bitter Q9 so it is always going towards the sample but here your sample and Q9 is mixed and once you are pre-exposing it to it before taking the reading from next time it will always go towards control is it not happening from next time it is always going towards control that is why it is coming negative that is control versus sample so after one day one second Rahul you suggested that we do sample versus Q9 right but Q9 is odorless so if you do Q9 is odorless why they are getting 0.2 he wants to prove that only see in control versus Q9 they are getting 0.2 if it is odorless it should be 0 but it is getting plus 0.2 and they are like on that if we could tell the number of intercepts then we could say that if it is significant or insignificant that is what I am saying with all the four snails run it five times each snail control versus sample again because the snail was in starvation starvation periods because so before doing all these experiments like control versus control versus sample we starve the snails for 18 hours so means like if we are not hungry then why we should respond to the suppose Biryani given to us what he is asking is you starve the snail for 18 hours now when you are starting your experiment you should probably make six this essay boxes or slides what you have like six arenas one with control versus control control versus Q9 control versus sample then again control versus sample and if possible sample versus Q9 you can also test but the time period in which you are doing each essay the in between time period should be fixed like for from going from second to third you are taking 15 minutes of gap from mixture to again control versus sample you are taking one day of gap so why is it you are again starving it yeah so it again goes to the again shows the attraction towards the sample it is giving the ORI plus 0.4 what do you mean by that so it is going towards its memory is gone its memory is gone means he get experience on sample versus control versus sample versus Q9 that it is having a bad taste but yeah because your data suggest both the things see first control versus sample look at your slide the first time you put control versus sample what is the ORI plus 0.7 you let it taste the bitter things sample versus Q9 now next day again control versus sample but next day next day but look at the ORI yeah if it has forgotten the bitter taste why the ORI has reduced so maybe one time he went to the sample side and he get the new memory that but next time there is no Q9 to remember yeah so maybe it has not forgotten in one day ok maybe because your numbers are very less so that is why we cannot conclude so if you do it like what we have suggested with all the four snails in multiple times you can get to a better data and better graph the design is good you need to make some changes and you need to increase the N because the question is very good question is good design is not good this was the summary of the graph so the control versus control shows that aroma was the setup was aroma free no no no no yeah yeah the conclusion was yeah yeah we will be moving to next presentation that is of cardamine that is by Preeti Preeti and Arjun yeah ok yeah and Abhijeet ok Abhijeet thank you good evening everyone I am Arjun Jaysmani from Bombay College of Pharmacy this is Preeti and this is Abhijeet and we are going to present about plant which is called as cardamine cardamine now is a very simple and small plant which can grow anywhere and is easily available so cardamine is a small plant which is easily available throughout the country and it can be easily grown in favorable conditions as we can see in the PPT it's quite easily grown and as it says it's a tabletop farm and it can be grown in labs, houses or anywhere it's easily grown now coming to why cardamine cardamine basically looks like a simple plant but it is not so it has a very complex structure and its anatomy and morphology is quite different and compared to other plants as we can see the parts of the flower and the bud are very complicated it has a stamen, an ovary, a piston and hence it is so complex in its development hence we study cardamine Arabidopsis thaliana is a relative of cardamine hirsuta and has a lot of differences between them but belongs to the same family there are quite a few differences between them Arabidopsis thaliana the leaf shape is simple whereas cardamine is compound there are 6 statements in Arabidopsis while 4 in cardamine there are 4 petals while in cardamine it ranges from 0 to 4 the pod-shatter pattern ranges from is non-explosive in Arabidopsis while in cardamine it's explosive the seed shape is oblong in Arabidopsis while in cardamine it's disc-shaped and the seed surface is bumpy in Arabidopsis while in cardamine it's got indented dimples in cardamine we can have different types of studies this flowchart was made last year during march march meet so using cardamine we can study what is the life cycle and how it varies the physiology according to the place to place what is the pollen variability organ size if it's differing if the amount of time required to form one organ and to develop into the next organ is same or different in different conditions the flowering time seed production depending on what is the photo period how does it affects the plant depending on different growth conditions like ecology soil water, temperature, humidity how the plant is differing and it becomes more easier to generate because this plant requires only 60 days from seed to seed so this becomes easier to do replicate studies in one year we can also study different types of conditions but if you have different types of soil so how it is affecting the plant and the question which we were and also the embryology and stuff we were interested in is gene transfer like what we were interested in we wanted to perform the gene transfer from one plant to another like in cardamine like by a simple method called as floral dip method so what we can do is we take the gene this is what was our long term objective transfer it via agrobacterium into the plant using a marker gene I will describe a little detail about it in next slides this is a work done by Angela Hay Etel they are from Max Planck Institute Germany they have been working on cardamine as a model since a long time they have done a comparative studies between Arabidopsis and cardamine and out of this we are interested mainly in the flower since it will be used in the floral dip method so Arabidopsis flower we can see it has 4 petals cardamine it can have 0 to 4 petals it is variable I am extremely sorry there are some few images which I am missing because of some technical error this is a cardamine and this is the Arabidopsis cultivated by Shruti from NCBS this is a this is a pick from cardamine from Ranchi Benita another cubist who was working on it sorry for the missing image this is a cardamine from which was done 2-3 years back by Dale Rodricks and here were 2 picks of cardamine from Manipur which is near Asim and this is from cube Mumbai it is missing I am sorry what was our immediate objective the immediate objective was to understand what is the life cycle of our cardamine so that if we know it then only we can perform some experiment on it so we started with culturing of cardamine so to understand morphology and once we started with understanding what is the life cycle because the cardamine has a compound leaf it means that it if you see this it has got a this whole thing is a leaf and it has got 3 leaflets 1, 2 and 3 leaflets so this is called a compound leaf how it has got a compound leaf and how one second sorry so here you can see there are 3 compound leaflets 3 leaflets this it has got 7 leaflets so the number of leaflets is also variable so we started with counting how much leaflets it has like the first leaf, second leaf how much leaflets it has then sorry we went to understand what is the morphology and anatomy for flower and what are the different stages of flower development with respect to time how much time it requires from green bud to a whole flower and then to a mature pod the reason behind this is our short term objective was to perform floral dip for gene transfer using some marker gene like if we have to understand whether the gene transfer has taken place or not we need to have some signal that signal can be any antibiotic resistant gene like for example gene which we will add along with our whatever agrobacterium gene the gene in the agrobacterium and when if the genes is transferred we can select those seeds which are gene transform whereas we will not select those seeds which are not individual gene transfer is not taken place and the long term objective was to carry out gene transfer for altering the leaf shape of cardamine I am sorry for the missing image we were interested in stages of flower development so this is the green floral bud with green sepals you can see this was a leaf and here a new leaf is getting initiated along with it you can see there is a green bud a bud with green sepals the bud now it was earlier only green in color but you can see at the tip there is a little white dish part so here actually there was an image which was actually a green bud with white sepals in between now here we can see the white petals will become more prominent just show you the image of a flower I am sorry for the inconvenience actually here we had a very beautiful image of a cardamine flower and when it is bloomed how it looks with its stigma at the bottom and how its anther are protruding like four anthers this is the next stage where the stigma has now already come up the anthers would be below it so the stigma has started coming up now it is developing into a pod so this you can see the stigma along with its growing and we can see stigma at the top so this ovary is developing into a pod further we can see how it is elongating and earlier we had green sepals which are looking prominent and also the white petals then we can see the green sepals are not there they are absent we can see only white petals are present this is an image where we can see the pod is almost like maturing and the white petals have loosened up here the white petals have disappeared like this is the inflorescence how a cardamines it is called the rassimus inflorescence where the older flowers will be at the base and the younger ones are at the top so after after this pod is matured you just need to touch it so if you touch it it will burst open this is how the seeds look there was a video of how to collect the seeds I am sorry it won't be able to play now what inspired us to take up this long-term objective was the work then by Angela et al what they had done was they took this cardamine and they had this arabidopsis arabidopsis has a simple leaf you can see it is only one leaf and cardamine it has different leaflets so it is a compound leaf so they introduced this arseo gene into cardamine they introduced this gene in this because of which it started developing a little bit like a simple leaf and they inhibited one gene in this arabidopsis because of which it is becoming a little curved shape similar to what is happening in compound leaf but not exactly because it is not only one gene which is playing role in the formation of a leaf shape the summary will be told by abhiji from all the observations we could say that plant take an average to 6 days to develop a new leaf it means that average 6 days it take to develop a new leaf generally the leaf pattern which we observe is that we expect the leaf pattern should be like 1 1 3 3 5 5 7 7 like this but there should be there can be variations in this but is generally observed with the initiation of 7th leaf average on 6 weeks post germination type of influences race most current hypothesis will be explained by Priti whatever the number of samples we have taken accordingly we could see that the time taken from a green bird with green sepals to a fully bloomed flower is 2 days but we didn't have the enough number of observations and also we didn't like because of our not proper follow up we couldn't observe it properly for one plant the whole development from a bud to a whole mature pod so whatever pictures we shown it was like capture at different stages of different flowers so we need to do a proper follow up then only we can say with confirmation that yes bud to flower takes only 2 days and for flower to mature from becoming a fully bloomed flower to mature pod it requires approximately 10 to 12 12 days which needs to be validated with appropriate number of samples the learning outcome is that this is a very easily available plant model to study plant morphology like whenever we study in text books how a plant like flower develops into fruit we just have it given okay there's a flower there's this pollination takes place and then it develops into a fruit but you can actually observe it within a short span of time so it is a very good model to show it in schools and colleges it is a good model for carrying out molecular biology experiments at a college level like as we said gene transfer using floral dip method the goof ups like we made many goof ups like we didn't write proper details in log books like we didn't mention what is the watering regimen so if I'm having a certain kind of I'm watering some amount for some for the plants it won't be followed by all my team members so then we came to know okay we don't have the same idea it should be the watering regimen then we we had a cossery and then we designed how our watering regimen should be for each type of cultures like we can see we have tray cultures also we had some lawn cultures near g6 lab so each thing needs to be devised how what it should be ordered so even if any one of us is not present our cultures won't cultures would be taken care of by someone else so that was one thing then while writing in lab record book we used to just mention all the details we used to just write okay this many number of leaves bird flower everything but what is the take away message from the whole page this way our whole log book is getting filled but what is the take away important message from that whole page if somebody just picks up our log book how will they come to know what is the most important thing to look for in this page so we should at least make that important highlight point with some other color we should have a remark column where we should write what is the important thing in the whole page then we need to like we were not initially interested in knowing the whole anatomy of a plant we were like interested okay so we have a long term objective we should focus only on that but no then we had during Kaasari sir and everyone like when they discussed we got to know the know before we step into anything like a short term or long term objective first important step is knowing our organism if you don't know our organism we cannot step into the further plans then or if we have a design like we have to plan everything a day prior to it if we have designed it then we should start posted on the groups or emails and then discuss with others so they may point out okay this is a problem in your design like in the snail group they had a certain design but then somebody said okay you need to do a number of replicates need to be there this this is supposed to be amended so when you discuss that time you start understanding how you should design it properly and your design gets optimized so that we started learning during this project number of replicates like earlier we just had one tree like this is our tree a which we started with and I thought it is enough we have many plans but then what if the tree gets damaged some by somehow something happens and it is damaged then my whole culture is lost so we should not just have one replicate I mean we should have a number of replicates not just at one place but at different places as well we should be a good observer and we should report whatever our findings with good photos and considering the audience we should label what should be looked in the photos and the last and but the most important thing is following up do you have any questions Preeti how this plant can be linked to co-dominants because the leaves are showing two dimorphic shapes at a time so can this I don't know but can this be linked to co-dominants is it always only two at a time like two types of leaves are there in one plant because I saw more than two we have one leaflet stage three leaflet in the same plant five leaflet and seven leaflet stage then it is not co-dominants it is just the compound leaf it is not a different type of leaf in the same plant like it could be there but we have no idea about it like why don't you go to the picture back it has different number of leaflets but it is actually a compound leaf for me the shape is also different like if you observe it for the first initial two leaves which are one and one leaflet leaves it is the leaflet shape is quite different as compared to the leaves which will come later for seven leaflet stage this is the picture of a fully grown plant can you explain that that answers what she is asking see like the here we can see we have five leaflets instead of that if we have only one leaflet so that time it will be only one broad leaf then when it has three leaves we can see a leaf is similar to this and we have two leaflets and as in when leaflets start increasing like in this picture that is the gene responsible for this leaf shape the compound leaf shape and like if you see in the seven leaflet stage like here we can see the leaf shape is quite different as compared to this this is a broader leaf which is at the base but the upper leaf as it the stem grows up the upper leaves which are having more number of leaflets it is becoming more narrower yes could be possible regarding differential like like what we observed for our sample almost similar yes but for literature studies we didn't look into this detail in this like we can see we have this five leaflet stage now here it is almost like what they did was they mutated this gene in cardamom they must have carried out some either a gene transfer or some mutation because of which the leaf shape has yeah they have done because we have a little bit only now we have we had read yeah they had done like in I don't exactly recall which paper it was Arribidopsis thaliana Arsinoe transgenics shows that it becomes something similar to Arribidopsis thaliana when Arsinoe is introduced into that it looks like the third one Lyraton Lyraton Lyraton Lyraton Lyraton Lyraton Lyraton Lyraton they have also performed some similar gene transfer because we have less time you can look at the things which he said that you know whether the whether the leaves differentiation increases as if it grows grows yeah in their own setup regarding differential gene expression yeah you may not have the answer now but that what you should be looking for yeah okay so I think we should proceed for the next present that is of Shreyas and he will be presenting on yeah we know the pagala post thing and he has worked on rotifers specifically so he will be talking about the rotifers and some yeah so hi good evening everyone I have worked on pagala post and as I mentioned here life at your doorstep now this particular line life at your doorstep is very very important because when you look at this word pagala post it won't make any sense if you have heard it for the first time basically it comes from this famous islands that is Galapagos islands now as we have been talking since the morning about Darwin so Darwin was the person who visited Galapagos island and he studied evolution by looking at the finches finches a bird who used to like finches were there on the Galapagos island now in this Galapagos island you can see it is not just one single island but it is a series of islands and you can see there is a land over here surrounded by water now why we call this something like this how can this be connected so it is very simple here it is land and surrounded by water all over here in this two small holes the sewer lead the two small holes of the sewer lead they have water in them most of the time of the year and there is land all around so it is just an exact replica of it so what Darwin actually did at this Galapagos island so you can see these are the finches which were there on the Galapagos island on different island there were different kinds of finches so like suppose let's consider island number one two three four five and six okay so in in an island there were finches like this and when Darwin observed like what is the vegetation over here so he found that mostly there were plants which were having a lot of leaves and he said that maybe this finches are eating leaves he went to another island and he saw a slight difference in the shape of the beak a slight difference okay and you can see that this kind of beak will help the finch eat the seeds same you can see these are for the birds and fruits these are for growth these are for insects so what is the importance okay we see in our everyday life we see there are differences between the animals between the birds but we don't pay attention to it but the important thing over here was that all of these were the same bird they were the finches but they were showing difference in the shape of the beak and how did that particular population of finches increased on that particular island so that is how the natural selection works like the the finches which were the most fit to survive on that particular island with that kind of food were in abundance on that particular island and that is how we also want now if you want to study evolution just like this we don't have resources to go to the Galapagos island or and we don't even need it okay we don't even need to visit such far away places we can just go out on our footpaths and just have a look at how this seaward leads and these are pretty much just like our Galapagos islands just the only thing is that you won't be able to see big animals and finches but you'll have to see small microorganisms and study the same evolution now well as we saw the Galapagos island the the person Darwin he choose islands having different kind of vegetations okay now in Paglopos we will choose different kinds of wells now I have termed it as wet semi wet and dry let me just brief what will be wet semi wet and dry many times you make a confusion wet will be that which stays wet throughout the year okay not just in the rainy season but throughout the year it means that it should be near some leaking tap or some fountain or something like that the semi wet should be the one where it usually gets wet and dry wet and dry wet and dry we can also call it ephemeral which is constantly changing so it can be somewhere near a garden where the gardener every day waters in the morning but by the afternoon it again gets dried up but the next morning again it will get wet because the gardener will do his routine work and the next one will be dry the dry will be the one which will be dry for long time and I would prefer to say or if you can find such well which will not even get during the monsoon which will be some under some shade or under like there are tea stalls or something and under that you can find such kind of wells so method to collect the sample how are we going to collect samples I am extremely sorry for not having the photographs that's a huge goof up of me so to collect the samples we just need simple tools like droppers we need sterile water, sterile test tube, cotton plugs and log books why sterile because we want the samples only from those wells we don't want any microbial load from our water the DC water which we will take we don't want anything from it so sterile water, sterile means just simply pressure cook it and when we are taking the observation some things which we should consider which are very important are the time of observation we should note down at what time we are taking the observation because when we take the dry sample we can just observe the dry sample under the microscope obviously add some sterile water to the sample and then observe it so when you look at such sample it is very crucial to note down the time because at zero hours sometimes the microorganism won't be present but after 24 hours or 48 hours some microorganisms would just emerge and it is not just jadu but they are just coming out of the dormant stage the next thing is magnification is very important so that we can know the size of the organism and it becomes easy for us to recognize to identify which organism it is and size of the sample to observe it is very necessary that you don't always observe just one drop of one sample but we take multiple drops that is what we are saying since morning duplicates replicates that is how so you should determine that what size of sample you should use I prefer three drops of every test tube to observe now when I did this I observed many different organisms I was able to identify this many there were many others which I couldn't identify so amoeba was there volticella was there there were many diatoms there were rotifers and there were parameshim these are some of the poor images but I hope you can make out these are rotifers over here this is again a rotifer over here he is just exiting the the anterior part is out and over here if you can see this is a volticella this this over here it is and this is its body so while working on the paglopos as I told you I came out all of these many organisms and I further went ahead to study about rotifer now as you can see it's deloid rotifer deloid is just the class of that rotifer and I have said it is a model for HGT that is horizontal gene transfer now a very simple question can an organism survive without sexual reproduction that is without any kind of variation over a long span as long as 80 million years because the last fossil like the oldest fossil found of deloids is 80 million years old so obviously many of you would say that it's not very feasible it won't it won't agree with the theory of evolution which we have sorry yeah so why that okay so if there is no variation in an organism all of them would be the same there won't be any difference so because of that if there is any any like any chemical compound coming which is a poison to them or any change in the environment which is not feasible for them not suitable for them all of the population will die at once but when there is variation happening in the population some of them will get the ability to survive that kind of maybe climate change or as I am saying a toxic chemical maybe so at least few of them will survive and then when the few of them will reproduce then the population will again be surviving that is what we call natural selection but the question is has their habitat changed over this 80 million years significantly like they were aquatic they are still aquatic yes so has the environment changed sorry I couldn't get you like why is adaptation required to deal with the changes to deal with the changes which are happening around us exactly if the environment is constant and favorable for a longer period of time we can't say for this many long years the environment hasn't changed at all even if you are taking just the aquatic environment as you said they are living in so this rotifers are found almost in every plant body whichever we see in the lakes in the rivers and I can't like even we would agree on this particular point that river 50 years back and river today is not the same if we consider all the chemicals in it yeah so there has been many changes even if they are still aquatic so but they are surviving this rotifer is surviving so how there is one answer that is horizontal gene transfer now let me just brief you of how what is deloid how is a deloid rotifer and I will move ahead how HGT happens in this so obviously it is a kingdom animalia it's a heterotroph it's a multicellular its phylum is rotifera now many of us have not heard of this phylum but yeah it's a phylum and how we identify the phylum as rotifera this small corona of cilia this rotating cilia so which are organism bears this it is under the phylum rotifera and this is the class deloidia okay I am extremely sorry again for the images I will put forward on the whatsapp groups this is just you can easily make it out by the shape the other classes are cisnoidia which is marine so don't even think that it can be cisnoidia and the other one is monogonata it is a freshwater but it doesn't resemble at all with the shape it is much more circular whereas you can see it is much more elongated elongated so why I am talking about deloid because this deloids are the only one which are asexual since this long time the other class which was monogonata they are not exclusively asexual they become sexual at times and the size is around 100 to 200 micrometers it means we can see it with our naked eye by that I mean you can't see the image properly you can't make out the shape but you can just see it like a dot in your bottle or in your culture it can be in a tank it can be in anything I have it in bottle in a tank as well in the lab and it is a freshwater organism now my objective is the long term objective is to understand the mechanism for deloid rotifers the short term is to transfer the gene of my own interest into a deloid rotifer and immediate objective is to determine the concentration and type of antibiotic sensitive to deloids now why I am talking about antibiotics I will come back to it in few moments so now my hypothesis regarding the horizontal gene transfer how horizontal gene transfer takes place now my hypothesis is based on there is a group of researchers Iris and group in the BMC biology and it is based something on them so according to that the undigested DNA from the food of deloid so whatever deloid will eat and when it will eat bacteria or maybe algae there will be the DNA of that bacteria that algae and this rotifers get into the dormant stage on desiccation that is a very important point that they whenever the environment desiccates around them they go into a dormant stage that dormant stage is termed as tun t-u-n-tun so that is a very important stage so whenever they go into the dormant stage at that time there will be some part of the DNA of the bacteria of the algae which will be undigested which means that DNA part will not be broken so during that time when the rotifer shrinks the food is over here the food is in the stomach and these are the ovaries of the rotifer ok am I confusing you going on starvation just go ahead not starvation I am saying into a dormant stage because of desiccation ok the water has dried up around it so it has to go into a dormant stage so while going into the dormant stage it will shrink its body it will slow down its metabolism so during that time whatever food will be there in its stomach it is not necessary that all of the food will be undigested or all will be digested there can be some part which is digested some is undigested so the undigested DNA which means the DNA which is not broken of the food of the bacteria which it has eaten of the bacterial DNA that DNA will go and get I am sorry it will go and fuse with its own genome in its ovaries ok which are just beside the stomach progenic genome which is the progenic genome obviously how can it fuse with all the cells ok so after that when the rotifer will again rehydrate means whenever the conditions will become favorable when again the water will be available in the surrounding at that time the rotifer will again rehydrate it will again move around at that time again the cell membranes will close and by that time the bacterial DNA is now fused with the rotifer's DNA ok so now the progeny which is born it won't be the same as its parents but it will be different not all progenies right all progenies maybe we can look at it but we haven't looked at it yet because it must be having like the alien DNA which is the bacterial DNA which is there it must be like only few in numbers as compared to the X which are there in the ovary right so one DNA we can't be pretty sure that it can be very less or very more because we don't know while going to the desiccating state the rotifer has just fed or it has eaten a long time ago is there any criteria that one fragment will go to each of the eggs very very little about this is known the initial study which was done by Missilsen in Harvard University in 2008 they have done some bioinformatic assay and because of that they say that it is massive horizontal gene transfer because while comparing they saw that most of the genome of the rotifer was belonging from the bacterial class and all such things so very little about this is known it is pretty interesting then over 80 million 80 million years of lifespan it is still a rotifer while incorporating so much of DNA from other organisms why it is still a rotifer because you are talking about the evolution yeah so this has got a bit distorted because it was in the windows and now we are on the annex okay so how did I isolate the deloids from the pagolapos sample which I got I used two different methods initially I used the serial dilution method when I was along with Himanshu doing it Himanshu is also from Elphinstone college so we used a microtiter plate the 96 well plate which we have and in the first plate we put the three drops of the sample without looking anything under it because once we made sure that every drop which we are taking under the microscope is having rotifer in it is having a lot of deloids in it we just took randomly one drop in the first 12 of the 96 well plate and we just did serial dilution we used to take one drop in the 96 we just used to take one drop from the adjacent plate and add other two drops of sterile water and in the same way we just did serial dilution and after 10 such dilutions we observed it under the microscope to see if there is one single rotifer present and once we got one single rotifer present initially we believed that okay we have got a single line culture but after as we all know the rosaries in the cube lab I got to know that okay maybe there are some eggs coming along with it which I might have missed and this is not enough to be called a single line culture then along with the help of Arif Arif is an expert looking at very minute things with the naked eyes so he was the one who said that okay I can just see one rotifer in a dropper and I can help you so along with his help he helped me to pick up one single rotifer from my culture into a 24 well plate and as you can see just one single rotifer means hardly a drop and one drop of rotifer culture just one single rotifer and I diluted it with around 2 ml of sterile water and such way I made 5 dilutions and thus I made sure that when it comes into the last well that is that was not I would not call it serial dilution but I would call it washing washing my rotifer so that all the eggs of the rotifer come out or stay in the initial wells and when it was in the fifth well it was pretty much sure that it doesn't have any eggs in it and now it can be called a single line culture and after that I was just feeding it with again the same moina method filled it with milk because it needs bacteria but now as it was just one single rotifer in a 24 well plate I can feed it with a large amount of milk so simple thing I just took 500 ml water sterile water I added one drop of milk to it to make it the same concentration and of this solution I added one drop to the 24 well plate and that is how the culture increased in a 24 well plate initially then in a test tube from a test tube to the bottle from bottle to the tank so this is just how our media works the milk it will lead to growth of bacteria and that bacteria will be the feed for our rotifers so now as I talked initially my immediate objective was to know which antibiotics which the rotifer is sensitive to which antibiotic now why antibiotic what I think is that as I said I want to put my gene of my interest into the rotifer so I thought why not put antibiotic resistant gene in it so first for that I wanted to initially observe that which antibiotic 2 are this rotifer sensitive so I checked with this 4 different antibiotics that is chloramphenicol tetracycline gentamicin streptomycin and ampicillin as a control why ampicillin as a control it is because ampicillin as you all know it disrupts the cell wall formation and rotifer doesn't have a cell wall so yeah that was the initial goof up initially I thought that ok ampicillin work and ampicillin resistant bacteria is very easy to find so I just added different concentrations of ampicillin to the rotifer culture and nothing happened I added 100mg 100 micrograms per ml 200, 300, 400, 500 nothing happened at 500 I added 1000, 2000, 3000, 4000, 5000 nothing happened then I had a stock I just added a stock and I was like what should I die but nothing happened then I went then I was like why it is not happening and I just googled a very small thing the mechanism of ampicillin how ampicillin works and I was like oh man I am so stupid so that is why I chose ampicillin as a control because ampicillin won't have any effect on the rotifer as it just obstructs the cell wall formation and rotifer being an animal doesn't have a cell wall so after adding this again why 60 micrograms because in our college lab we have the discs which are of 30 micrograms so I thought 30 is very less so I add 2 I couldn't make it 50 because there were 30 microgram discs so looking at the results of this experiment I don't think they are very reliable because I just did it once but I did it just few days ago so I am ampicillin as it was control it was showing growth in the ampicillin gentamicin no rotifers were found they were all dead in chloramphilicol they were all dead but in tetracycline and streptomycin they were still present but as compared to the control the amount of or the number of rotifers in the tetracycline and streptomycin was less but I can't say that it just like they were not very sensitive they were I can say they were bit sensitive to it one more thing what I did was desiccation tolerance and revival as I said that rotifers undergo desiccation and then they again revive back so I wanted to check that the culture which I am having how many of them like what is the percentage of rotifers can revive back ok so again I took a 96 well plate and I had that that time a good tank of rotifers so I can use as many as I want so I just took 10 rotifers in each of the 96 well plate and I just kept the well plate open for around 2 days no around 4 days I guess ok I am entertained here I just kept the well plate open so that all the water will evaporate and then again I added 2 drops of water to each of this well and I closed it with a lid and I kept it like that and I observed it after every hour then after observing every hour I couldn't find any kind of revival I am still very much confused regarding it that why I couldn't see any revival because initially from where I got this rotifers they were from a dry paglipos and when I put water they came back to life so I think they should have come back to life even in while performing this experiment in a 96 well plate but it didn't happen I think the reason might be for this when I looked at when I looked at how people did it initially Claudia Ricci she was the one who did many experiments with the Deloitte rotifers and when I looked at her methods in the literature what she always used to do is she used to put some filter paper or something like that in the well plate so that when the rotifers desiccate they will adhere to that filter paper or something and the toon the cyst of this rotifers won't just fly off with the air so maybe that was the goof up why I couldn't find the revival of this rotifers but if I do it again maybe I can find it 10 rotifers per well so 96 in around 1000 rotifers um no I couldn't find it anywhere it is very less likely but I don't think that it can happen because I initially got it from the dry itself so they should revive that I don't think that would be the case because sometimes it just happened by chance we didn't do it deliberately but while looking at the rotifer on our slide sometimes we just went out for a lunch and while coming back it was all dried up and it was just kept there and after sometime someone thought that okay let's just put some water and maybe it can revive and he would just put some water and while the cause he was going on he would just make sure that the water don't dry up again and he was to put water and by the time we are leaving again around 6-7 he would see the rotifers are again moving so I kept checking the first day I kept checking every hour because I was not knowing then the second day again I kept I didn't check it every hour the second day but in intervals so it's not like after a gap of 4 days I checked it but I was checking it in between it maybe I have all flown away because as I said I just kept it open to dry I didn't cover it with any lead or anything after that sorry for the dormancy so yeah they mostly revive back within 4 hours and the revival rate is around 70-80% in deloids mostly sorry they just just used to keep the well plates open by keeping a filter paper below it so and she stated that so that the tones will adhere to the filter paper and they won't fly away yeah so that's it and the goof ups which the goof ups which I had done they were obviously lack of maintenance I didn't maintain my culture very properly I wasn't very regular coming to the lab there was there is a lack of collaboration as you can see no one is standing here besides me yeah there were a few there were a few from Chennai Krishnamurti was from Chennai Adarsh was from Raji a colleague of me from Elphinstone College himself Himanshu was there working with me but okay and irregularity in the work okay that's it I'm open for your questions any questions please okay I have one question yeah because you said that the rotifers they don't reproduce sexually yeah most of the time they reproduce sexually and you said so variation how does it come by taking the yeah you're taking the this thing the DNA of the other other species so the question is that you know if they are taking the DNA you know they should be changing very fast because they are directly taking the chunks of the DNA and putting incorporating into their genome yeah you know that is not how no random mutation happens very slowly one mutation at a time you know but here that taking lot of lot of DNA at a time you know but so they should be they should be evolving very fast all the time mutation does not add up to the genome size here it's every time it's adding up to the genome okay so what about the genome size itself yeah okay you have a clock clock timepiece or a clock working and you poke it what is the probability of that running better you understand what I am trying to say a clock is running reasonably okay and you just go on poking it what's the probability of it running more efficiently what's the probability of it conging off yeah more so to me it's something like poking with the finger it's not that it is picking and choosing the right type of DNA it is just a random thing happening so most of the random thing is like you are poking into the machinery of the clock so now the point is that what is it that is becoming sometimes once in ten million times or something you poke it to the clock and probably clock is working better so that doesn't mean that you will be going on clock my question is different my question is that if it is taking lot of DNA it should be evolving very fast okay at least it should happen something very fast something very you know not should be happening unlike what is happening in the random variation so in that case you should be can you show one organism evolving into another like one rotifer it is taking some DNA and it is becoming another for example another species of a rotifer species of a rotifer antibiotic resistance is okay I don't care about rotifer I am saying that can you show one species evolving into another species that's what you said about okay this is a question so this is a scientific question people scientists are asking this question no as you are talking as the Misselson's paper said according to their bioinformatics study what they say is that most of the transfer of the gene which is taking place it is mostly with the genes related to the enzymes okay I don't know how much I am answering to your question but what they say is that it is mostly the parts which are getting changed they are the parts which codes for some enzymes in those rotifers so maybe the phenotypic changes we are not seeing because of that can I function did you finish he was referring to something very casually a Misselson I think he said the same Misselson stall yes he is the one who designed a very famous Misselson and stall experiment okay so if you look at from that serious point of view it is the same serious Misselson talking about it so what is this I think you are also talking about enzymes genes responsible genes related to enzymes are being modified so what could be possibly happening one of the easiest to think that could be possibly happening is that there will be a homologous recombination that is possible homologous recombination that means some homologous sequences that is coming from a bag some other foreign source it has got much better probability of getting incorporated into an existing gene which has got homology with those sequences so those are the ones that is getting incorporated so you have a nice amylase gene a gene for amylase which is doing very well for the animal I am just using and a homologous sequence another sequence for foreign DNA has got a homologous sequence is getting now incorporated into this very finely functioning amylase gene so what is the possibility of getting conging off even if it is a homologous transfer there is a good possibility of the existing amylase gene will be counter there is a rare case where this amylase gene will become extremely fantastic I am just using a word amylase so the probability you will just see that my earlier analogy of the clock so here we are also explaining a mechanism of how homologous recombination can take place even in a homologous recombination the probability of it becoming great for the organism to reproducibly multiply better reproduction will be much less but once it happens and if it is aiding it that will go proliferating in a particularly in a non conducive atmosphere where all other fellows are very very meekly functioning where this fellow will be doing so the probability is what we need to worry about so the probability of it is getting becoming dull and inefficient is much more than the probability of getting a very very nice nicely functioning enzyme coming in the nicely functioning enzyme also will not be nicely functioning in the given situation in a changed situation probably that will be the best one yeah it can happen is that make sense also the question rises is there any mechanism of like horizontal gene transfer so is it there something horizontal gene elimination because it has to maintain its genome size as it is right it cannot sustain more than a limit of a genome size in that rotifer rotifer is not a higher organism so have you come across something like is it or how frequent is it is it not more is it frequently it is very frequent that is why they call it massive then within ten years of the time from one generation to the fiftieth or the hundred there should be a large genome expansion okay so it's a very interesting thing so I think you should send us all these some interesting papers so the papers must have used some markers I think you were using a marker antibiotic resistance so they must have said they must have concluded it is massive gene transfer maybe because they have found out several markers when they used it must have been all getting incorporated and many of them are not really becoming deleterious for the organism so those papers are very good it's a fantastic area so I think we will be breaking for the tea now so tea and snacks after the tea we actually are passing you a small paper sheet of paper we will be after this we will be showing after one presentation