Fruitfly culture in homelab: Kolkata. @magpie
Trapped one gravid female from the fruitflies hovering over banana in the kitchen.30/12/23Homelab Kolkata BatulP.
I succeeded in putting a piece of banana inside the plastic cup without losing the fly.After 4days I expect to see fruitfly larvae.Homelab Kolkata BatulP 30/12/23
[30/12/2023, 1:32 pm] Batul Ma’am: My fruitfly seems to be laying eggs on the banana.please comment.Homelab Kolkata BatulP 30/12/23
[30/12/2023, 1:33 pm] Batul Ma’am: The eggs if laid today should form larvae tomorrow. But they maybe difficult to see till they are moving around in the the 4th instar larvae stage so I said four days.
[30/12/2023, 1:39 pm] Batul Ma’am: We can see the fly abdomen more clearly here.
I succeeded in putting a piece of banana inside the plastic cup without losing the fly.After 4days I expect to see fruitfly larvae.Homelab Kolkata BatulP 30/12/23
Life cycle of fruitfly Drosophila.
A close up pic showing the fruitfly I trapped is gravid.30/12/23Homelab Kolkata BatulP
Are these eggs of fruitfly @Kiran Ma’am and others
I can see three larvae actively moving in the cup at 5pm on 3/1/24 and one larva is at the top so total four larvae in the cup of 4mm size four days after trapping a gravid fruitfly.Homelab Kolkata BatulP
Today is 8days after trapping the gravid fruitfly and the pupa is fully developed we can see the red eyes of the fly.Hoping to see flies tomorrow morning. Homelab Kolkata BatulP 7/1/24
My three bottles with TRSV medium developed by Cubists ready for fruitflies from single mother.I can make replicates of the first generation flies.Homelab Kolkata BatulP 7/1/24
[07/01, 11:06 am] Batul Ma’am: PREPARATION OF MEDIA FOR CULTURE OTHER THAN CORNMEAL AGAR MEDIUM
1.BSRV MEDIUM FOR CULTURE
U can use BSRV medium so it contains banana, sugar, rava and vinegar.
take 1 banana 1/4 teaspoon of sugar, 5 teaspoon of rava, smashed and mixed them well and then added 100ml of water using sanitizer bottle and then cooked it on low flame for around 5 minutes.
Switch off the flame.
After 2 minutes of turning off the flame At the end add 1ml of vinegar.
Transfer using funnel into the bottle which was dried well in sunlight…
Close ur bottle with the cotton to avoid any contamination
- TRSV MEDIA CULTURE
TRSV stands for
Tomato Rava Sugar Vinegar media …
Ingredients required
• 1/2 tomato
• 1tbsp Rava
• 1/2 tbsp sugar
• 75 ml water
• 7.5 ml vinegar…
Add rava tomato sugar in a pan …
Then add water to it …
Cook on the flame for 10 mins.
.{When you see the ingredients getting mixture n it’s thick enough to pour in the bottle}
Switch off the flame and let it cool.
Later add vinegar to it and mix well…
Then transfer into ur bottle which u had dried in the sun…
Close ur bottle with the cotton to avoid any contamination…
@Batul Ma’am
[07/01, 11:07 am] Batul Ma’am: TRSV medium was prepared as above👆🏼
BREAKING NEWS My first baby fruitfly is a girl ecloded at 8am today 8/1/24 The gravid mother fruitfly was trapped on 30th Dec so after 9days the adult fruitfly has ecloded.
Homelab Kolkata BatulP
This second baby fruitfly is a male
This fly has only one wing and it cannot fly
This fly has both wings damaged.
Fly4 no wings
Context to Curriculum WhatsApp group discussions:
[08/01, 1:01 pm] Rahul CUBE: Why do you think the wings are damaged? The flies are born with reduced and shrunken wings. Soon after the enclosion they inflate the wings. It takes sometime.
I guess the wall of your culture bottle are very sticky and wet. The flies are dying stuck to it before they could completely inflate their wings.
This is my guess! What do you think? @Batul Ma’am ??!
[08/01, 1:02 pm] Rahul CUBE: If you can just wipe the inside of the bottle surface with some tissue before the others eclose. That might help.
[08/01, 1:03 pm] Rahul Master CUBE: And if the food at the bottom if very goopy and slurry. You can put a tissue paper in it to soak moisture. It will also help with pupation by providing surface for the larvae to pupate on.
[08/01, 1:04 pm] Rahul CUBE: They aren’t damaged wings. They have just not been inflated yet. The flies are dying before the wings could inflate. The flies fill their early wings with a king of body fluid to expand them after their enclosion from the pupae. That process needs them to be on a drier surface.
[08/01, 1:40 pm] Batul Ma’am: Ok,will do the needful and I hope the other flies survive
[08/01, 1:54 pm] Batul Ma’am: The inside of the plastic cup is dry as you can see in my pics sent earlier.The base does have water but I remove it everyday with a pipette.Now I have kept a tissue paper inside.I can see atleast 8-10 dead flies with shrunken wings.
[08/01, 1:55 pm] Batul Ma’am: Meanwhile the fly that had one proper wing I could transfer to another bottle.It is still trying to inflate its other wing
[08/01, 1:58 pm] Rahul CUBE: All the best @Batul Ma’am ! Enjoy your Seymour Benzer moments. 
The fruitfly with one inflated wing is trying to inflate the other wing and is also feeding on the TRSV medium
[09/01, 3:59 pm] Batul Ma’am: In TRSV bottle 1 of 8/1/24 which had 1 female fruitfly with only one wing I have now transferred a male fruitfly today on 9/1/24.Cube homelab Kolkata BatulP
[09/01, 4:00 pm] Batul Ma’am: @Rahul CUBE the flies which ecloded today 4 of them are flying.Thanks for your idea of putting a tissue paper inside the container.
Rahul CUBE:Small but ingenious modification that fly biologists do in the lab to keep the culture healthy!! You’re most welcome. Thanks for joining the CUBE home lab with the mainstream laboratories around the country. Kudos!!
Morphological observations of the fruitfly whose single line culture is at my homelab in Kolkata. 11/1/24
[16/01, 8:27 am] Batul Ma’am: BREAKING NEWS Today is 6th day since the single line fruitflies were kept in A1 and A2 for breeding. There is NO LARVA seen.The temperature has dropped to 13°C minm and maxm at 22°C.The fruitfly mother had given adults in 8 to 9 days on 8thjan when temperatures were higher.Homelab Kolkata BatulP 16/1/24
[16/01, 8:53 am] Batul Ma’am: The maximum temperature during the development of the mother fruitfly from Dec31st to 8thjan was 26°C and minimum 15°C
[16/01, 8:56 am] Batul Ma’am: As temperature falls development of the egg and larva slows down according to the above observation.On 4thjan 3rd instar larvae were seen in 4days,this time with lower temperatures the larvae have not formed even after 6days
https://depts.washington.edu/cberglab/wordpress/outreach/an-introduction-to-fruit-flies/
According to this reference the lifecycle of the fruitfly doubles at lower temperatures of 22°C.
I have focussed on the white spots inside the bottle which looked like eggs.Are these eggs of the fruitfly @Arunan sir @Rahul CUBE and others
Four fruitflies transferred to A2 10/1Homelab Kolkata BatulP. please help me to know the males and females
Six flies transferred to A1 today 10/1/24 Homelab Kolkata BatulP
Mating behaviour seen in my fruitflies today at 11am.The fruitflies have been transferred to A1 and A2 on 10/1/24.No larvae seen yet.Temperatures both minm and maxm have become low in Kolkata since a week.Homelab Kolkata BatulP 18/1/24
Both males with dark tip to abdomen and females with white abdomen are present in A2 single line culture of fruitfly at homelab Kolkata BatulP 21/1/24 @Himanshu Joshi @Arunan sir @shalini CUBE and others
Males are smaller than females and have a darker abdomen last segment.In A1 also we can see both males and females.Homelab Kolkata BatulP 21/1/24@@Himanshu Joshi and others
Breaking News!! 7larvae seen today in single line culture A1 at Homelab Kolkata BatulP 22/1/24.The male and female flies were kept in the TRSV medium bottle A1 on 10/1/24 however some eggs were noticed only on 20th January on bottle walls.The reason for late mating and egg laying was probably the drop in temperature in Kolkata to 12°C minm and 22°C max since the last week.
The lifecycle of mother fruitfly from gravid to next generation virgin flies was 9days.At that time temperatures in Kolkata were higher with minm17°C and max27°C
All the flies in my cultures A1 and A2 were born from same mother.A single gravid fly had been trapped on 30th Dec 23.Within 9 days on 8thjan 24,I had the next generation fruitflies. On 10thjan 24 these fruitflies were kept in A1 andbA2 TRSV medium.The eggs were first noticed by Dr Arunan on 20thjan on my bottle wall.On 22nd Jan 24,I can see larvae 4mm in size.Within a day I expect them to form pupae and pupae take four days to become adult fly so by Independence day 26th I should get the next generation.From 10thjan to 26th Jan is 16days.The life cycle will be double the number of days.I am attributing this to late oogenesis in fruitfly due to drop in temperatures in Kolkata.
Discussions: Context to curriculum WhatsApp group
[30/01, 8:05 pm] Batul Ma’am: To study the effect
of
Temperature on
the length of the
life cycle of fruitflies
of same mother fly.
A single line culture
was established
using a single gravid
mother fly.
The first generation
took 9 days.from 30th Dec to 8th of January.
The temperature average in kolkata during this period was 14.6°C min and 25°C max
The second generation of fruit flies was started on 10thjan when a male and virgin female fruitfly from first generation was kept in Tomato rava Sugar Vinegar medium.The second generation flies ecloded on 24thJan after 14 days.
The average temperature during this period in Kolkata was 13°Cmin and 21°6°C max
The third generation was started on 25thjan with a virgin female and male put in a TRSV medium bottle B1.On 30th of Jan 3rd instar larvae of 4mm size are seen in the bottle.The expectation is they will develop into pupae which will eclode to 3rd Generation fruitflies on 3rd Feb that is after 8days.The temperature average during this period is 14.5°Cmin and 25°Cmax.in Kolkata
[30/01, 8:13 pm] Rahul Master CUBE: What was the temperature at the place where the bottles were kept during the first generation that took 9 days??
The temperature of Kolkata is of no relevance of you’ve kept it in your house with a regulated temperature due to the enclosed nature of the house. The temperature inside our houses are mostly warmer than the cold outside. @Batul Ma’am
How can we ensure that the temperature that might be affecting the development of flies is recorded properly with precision and reliability??? @Batul Ma’am ?
[30/01, 8:34 pm] Batul Ma’am: Well the effect of being indoors for all the three generations is same as the bottles were in the same place throughout.
[30/01, 10:11 pm] Rahul Master CUBE: But how do we know that? Isn’t there any objective method to collect reliable data of the temperature around the bottle at your place? How are we going to pose a hypothesis with just the word of mouth???
[30/01, 10:16 pm] Batul Ma’am: -the temperature in kolkata is from reliable reference not word of the mouth.would you like to suggest a method to measure temperature inside the bottle as even that will be different from temperature inside my room.
[30/01, 10:18 pm] Rahul CUBE: No. But I can certainly suggest how to measure the temperature around the bottle in the room which might be different from the Kolkata’s temperature. Simply put a thermometer adjacent to the bottle.
That’s what scientists like Benzer and we in our lab do to reliably tell the ambient temperature of the fly.
Hope it helps!!
[30/01, 10:21 pm] Rahul CUBE: Delhi is 7-8 degrees today. But my lab is 25 (maintained by precision ACs, and my apartment room is 16-18, without any conditioning of the air.)
Now, if I want to hypothesise something, I need to be very clear of the ambient temperature in which I am keeping the flies.
[30/01, 10:22 pm] Rahul CUBE: Now, coming to your question of how temperature affects development of flies. What’s your hypothesis @Batul Ma’am ??
[30/01, 10:22 pm] Batul Ma’am: True
[30/01, 10:23 pm] Rahul CUBE: If I keep flies in my room or lab and report the temperature of Delhi as a reference temperature. I am surely going to get wrong correlation!! What do you think?? @Batul Ma’am ??
[30/01, 10:23 pm] Batul Ma’am: Temperature affects the Circadian clock of the fruitfly and hence the number of days required to complete the lifecycle.
[30/01, 10:25 pm] Batul Ma’am: One question how do we find the max and min temperature inside the room?Do we measure around dawn,then noon and at sunset,at midnight?
[30/01, 10:26 pm] Rahul CUBE: Does circadian rhythm regulates development/embryogenesis??
[30/01, 10:27 pm] Batul Ma’am: Yes I think so.let me look for references
[30/01, 10:27 pm] Rahul CUBE: Yes. Exactly like that. We keep a thermometer and take the readings at/between dusk and dawn. Do you think enclosed rooms will fluctuate as much as Kolkata itself??
[30/01, 10:28 pm] Rahul Master CUBE: Do fertilised eggs of fruitfly have circadian rhythm?? @Batul Ma’am ?!
[30/01, 10:35 pm] Batul Ma’am: Yes every cell has a Circadian rhythm. The enzymes/hormones are released/activated at different point of time in the Circadian cycle,so any disruption will affect their actions,and hence metabolism and therefore growth.
[30/01, 10:36 pm] Rahul CUBE: Any reference for this?
[30/01, 10:39 pm] Rahul CUBE: We can perturb circadian rhythm by perturbing light and dark cycles for an organism. Does it mean, if I keep the embryo in total darkness, or total light. The fruitfly development will be perturbed too? @Batul Ma’am ?!
[30/01, 10:44 pm] Rahul CUBE: Also, human babies develop in complete darkness inside the womb. Do they have Circadian rhythms? If yes, How do they get light-dark, temperature cues inside to set a rhythm of the day in the first place???
And if no, why their development is not affected?? @Batul Ma’am?
Four pupae seen in the second generation single line culture B1 31/1/24 Homelab Kolkata BatulP
Two breaks in costal vein,an incomplete subcostal vein and presence of anal cell show that my fruitfly single line culture belongs to Drosophilidae family.Homelab kolkata BatulP 30/1/24
Graph of temperature vs duration of lifecycle of fruitfly at Homelab Kolkata, BatulP Jan’24
[31/01, 1:52 pm] Rahul CUBE: What can be inferred from this above graph? How does temperature affects the development??? @Batul Ma’am ?!
[31/01, 1:53 pm] Batul Ma’am: From the above graph we can see that increase in temperature shortens the lifecycle of fruitfly .
[31/01, 1:55 pm] Rahul CUBE: Great hypothesis!! But how do you suggest that might be happening? What role can temperature play in development of an organism?? You mentioned something about circadian rhythms yesterday. Let’s take it forward from there to understand it better.
[31/01, 1:56 pm] Rahul CUBE: Also, how’s the temperature measured in this data??
Why you have reported average of the maximum temperature and not the minimum??
[31/01, 1:58 pm] Batul Ma’am: We can also plot the average minimum temperature but interestingly the average max temp for both first and 3rd generation is 25°C so I chose the max
[31/01, 2:00 pm] Rahul CUBE: What does this paper concludes? A short layman summary will help us all. Does it relate circadian rhythm to early development of an organism??? @Batul Ma’am ??!
[31/01, 2:01 pm] Batul Ma’am: The Cicadian rhythms is regulated by period protein.This cycle of production of period protein is linked with production of growth hormones .So if this cycle is altered it will affect the development of the organism.
[31/01, 2:01 pm] Rahul CUBE: But don’t you think minimum temperature will be greater determinant for the development as the maximum is closer to the optimal temperature of baseline normal behaviour of fruitflies. But going higher or lower than the baseline is what determines greater changes.
[31/01, 2:06 pm] Batul Ma’am: It could also be the difference in max and min temperature which is likely to affect the Circadian rhythm. Just as long days (light)followed by a short night (dark)will affect Circadian cycle.
[31/01, 2:09 pm] Rahul CUBE: It is the same thing. Isn’t it? The difference between the max temp. and min. temp tell us about the extent of deviation from the max. temp. (which in this case is around 21 to 25.)
[31/01, 2:11 pm] Rahul CUBE: Also, you did not report the light and dark duration in your experiment. The circadian rhythm is also gated by the lIght-Dark ques. Isn’t it? @Batul Ma’am ??1
[31/01, 2:17 pm] Batul Ma’am: The light dark periods should be similar in the three generations as all of them in Jan’24.
[31/01, 2:23 pm] Rahul CUBE: But what was the periods? 12hrL-12hrD? 14hrL-10hrD? 16hrL-8hrD ?? When you are trying to make connections with Circadian Rhythm don’t you think that will be important for making any hypothesis?? @Batul Ma’am ?
Also, you are talking about Circadian dependent developmental changes but reporting the average temperature of the starting and and day of the cycle. Instead you should report the average daily temperature. (Circadian rhythm is a 24hr rhythm in organisms)
[31/01, 2:24 pm] Rahul CUBE: were your flies kept outside and were subjected to the natural light of the sun? or were they kept inside a room and were subjected to artificial light?? @Batul Ma’am ?
[31/01, 2:26 pm] Rahul Master CUBE: The same question holds for temperature too!! @Batul Ma’am !!
No no I have taken average temperature from daily temperature readings and not first and last.
[31/01, 2:30 pm] Rahul CUBE: Then we can’t say that the temperature and light conditions outside are the direct causal factors for the development. Can we??? Our home interiors have a different temperature range than the exposed outside. Also different light dark regimes depending on when we switch on/off the electrical light source. Isn’t it??? @Batul Ma’am !!
[31/01, 2:32 pm] Rahul CUBE: Then don’t you think reporting Kolkata’s weather report and correlating it with the fly development which is not directly exposed to it is not the ideal way.
[31/01, 2:33 pm] Batul Ma’am: How do they regulate the light dark periods in an experiment in the lab? Because the sunlight is way more powerful than our led lights?
[31/01, 2:35 pm] Batul Ma’am: Technically you are right but I am still excited to observe the change in length of lifecycle when temperatures dipped and their switch back to normal when temperatures gained.
[31/01, 2:36 pm] Rahul CUBE: sun sets around 6 in the evening but out LEDs are on. So it’s not aligned to the sunlight cycle if the fly is in our homes.
Good question about the lab conditions. We maintain a 12hr-Light and 12hr-Dark cycle strictly by having a time gated switch for the lights. It is proven than anything around a tubelight is enough to give them the light signals.
[31/01, 2:41 pm] Rahul CUBE: So are you not excited enough to figure out the technically correct reason for that observation? I am sure you are. We often mistake correlation as causation in Science.
let me give you an example: There is a common notion prevalent in society that if someone is fatigued then they sleep for longer and deeper. Turns out this correlation is not true when you carefully examine the quality of sleep in human volunteers (fatigued vs well-rested).
[31/01, 2:46 pm] Rahul CUBE: You are correlating two things (development of the flies and the outside environmental conditions) that are not directly interacting with each other! The ambient temperature to which the drosophila is exposed is more important to give you a better result. It might still hold the same trend but with more accurate controls on the factors.
[31/01, 2:54 pm] Batul Ma’am: There would be more accuracy in the factors but it would still be a corelation and not a cause.But the corelation is equally important. @Rahul CUBE
[31/01, 3:09 pm] Rahul CUBE: Not at all; I strongly disagree with this. Correlation studies cannot be as important as the causation.
Correlation has led to many non-specific drugs coming into the market and then later retracted upon non-targetted adverse effects.
Correlation studies also tell us that when the springs comes in the Scandinavian village, the number of migratory storks increases and so does the birth rate in the country. People used to correlate in earlier times and think that it’s the storks that deliver the babies to the mothers from their gods.
Correlation tells about a phenomenon, not that it’s true and holds direct relationship!
[31/01, 3:09 pm] Rahul Master CUBE: If the method is scientific, you’ll not just correlate. You can find direct-interaction.
In your case, the method has a lot of loopholes. It’s not scientifically rigorous. It’s brilliant to begin with. But will be much appreciated if you take positive from peer suggestions and make it more rigorous in data collection and reporting.
[31/01, 3:16 pm] Batul Ma’am: Well that’s absurd the storks bringing the babies.I don’t think my work merits such comparisions.
What I meant to say is that it’s not always possible to find one cause there are many interrelated causes.
[31/01, 3:25 pm] Rahul CUBE: You missed the drug research example and chose to be offended by the least scientific example.
The fact that correlation studies are not comparable is established by many real lab work that’s published.
You’re work is brilliant. But not scientifically rigorous as you’re not even comparing two directly interacting factors. But instead correlating two factors that are not directly interacting. All I am suggesting are the following points: (which are very easily doable)
-
Keep a thermometer besides the bottle to keep a track of direct ambient temperature fluctuations during the 24hr cycle.
-
Keep a record of when the light is switched on and off in the room where the flies are kept.
-
Draw a graph correlating these factors.
How impossible can this be? You are correct that you’ll still be plotting correlation as you are not keeping other internal factors constant. But at least you’ll be reporting directly interacting factors.
And if you want to prove that temperature is causal. The experimental for that is not impossible too.
Simply do this:
- Keep a control bottle at a constant temperature (hot or cold) you can use a space heater/water in a bowl to maintain constant (more or less, not exactly) room temperature. And then compare the development .
- If the ones that are exposed to daily fluctuations show a different developmental plot than the control. It tells you that temperature is causal!
[31/01, 3:27 pm] Rahul CUBE: Science is more fun if one is not offended by the critics. That’s how scientific peer reviews work. No one can prove/claim/publish without going through reviewers. What do you think? We are friends in life but critics in science!
[31/01, 3:28 pm] Rahul CUBE: @Batul Ma’am ! Cheers to your work. And Cheers to much more that we are eager to see from you.
[31/01, 3:30 pm] Batul Ma’am: Thanks these are simple,doable changes.
More flies of third generation of single line culture C1 ecloded on 4/2/24 at Homelab Kolkata BatulP
Activity in the fruitfly culture C1 at 10am on 5/2/24Homelab Kolkata BatulP
Gravid fruitfly with a fat white belly in C1 Homelab Kolkata BatulP
Identification of male & female fruitfly: described by @Rahul
Males are distinctly differentiated by their genitals. Their lower abdomen is black on the dorsal side. Also, they have sex combs (fine black hair bristles on their first pair of appendage) that helps them during courtship.
Smaller than females in general.
Arrow pointing to the sex combs
How to Distinguish Between Male and Female Fruit Flies Steps https://web.as.uky.edu/biology/faculty/cooper/Population%20dynamics%20examples%20with%20fruit%20flies/Sex-Determining%20Guide.pdf
. Fungi taking all over the spices… 2/2/24. Kalyan west.
