Summary of CUBE chatshaala

[saswathy679 (Aswathy suresh)]

450th Day : CUBE Chatshaala.

Discussed about 12.06.2021 fruit flies culture conditions in Bottukadavu Thrissur Kerala.
SLBK A18…SLBK B18…SLBK C18…SLBK E18 - 28.05.2021 subcultures of fruit flies.
Important points we were discussed about how many days want transfer flies from pervious subcultures to new subculture bottle!..importance of gravid females in a new culture!.. How to identify Gravid flies!.. How distinguish between female flies and male flies…How can we transfer flies from old bottle to new media bottles!..

N.B
Criteria of Transfer
Transfer Flies Between 15th to 25th Day
At least 8 - 10 Gravid Flies to be transferred

Difference between Male and Female Fly

Pointed Abdomen - Female.
Rounded Abdomen - Male.

also we were discussed about how to find a relabeled article from google…
What are the points we can make sure it’s relabel article or not…
•Author names.
•Journals.
•Published year.
•about Authors.
• Peer reviewed
• Cross reference

What is meaning of Peer reviewing ?

The peer-review process subjects an author’s work, research, or ideas to critical observation or examination. of others who are experts in the same field (peers) and is considered necessary to ensure academic scientific quality.

450th Day : CUBE Chatshaala.
Picture is showed size of microworms present in fruit flies culture bottles less than 1mm sized. Taken photo keeping culture bottle between torch light & phone camera digital zoom it taken photo. Magnification of this photo 8x… How to take better photo graph of worm…using better setup!?.

Picture showing Panagrellus.
One of thirteen currently recognized species of Panagrellus, P. redivivus is about 50 μm in diameter and just over 1 mm in length, barely visible to the naked eye.
Panagrellus redivivus - Wikipedia.
Look like quite similar to fruit flies culture bottle worms…coming under phylum nematode.

•Kingdom: Animalia
•Phylum: Nematoda
•Class: Secernentea
•Order: Rhabditida
•Family: Panagrolaimidae
•Genus: Panagrellus
•Species: P. redivivus

What is difference between microworms & nematodes…!? Is there any difference.
cubist are searching C.elegans in Indian native soil…
Caenorhabditis elegans is a free-living transparent nematode about 1 mm in length
Caenorhabditis elegans - Wikipedia.

Why the importance of Model organisms… Why we are taking particular model system for studying behavior!!?
For example :
Cardamine plant & Phyllanthus plant… We are taking phyllanthus plant for studying sleep wake cycle… Because only this plant leaves showing opening and closing behavior.
We are choosing model systems based on what kind of research question we are addressing.
Like ;
•Fruit fly taking for study Circadian rhythm, activities patterns…
•Moina taking for study hypoxia conditions…
•Zebra fish - immunity study.

450th Chatshaala : CUBE Chatshaala.
Moina
•Easy to culture.
•Maintenance is low
•Survived in low oxygen level
•short life cycle
•Translucent to red

Addressing research questions.

•Hypoxia in Moina (The ability to adapt)
•Ephippia.
Moina have 2 Reproduction
Asexual & sexual…asexual in normal conditions…sexual in abnormal conditions (harsh conditions).

Discussed about moina video taken setup of @Drishtant… In that video background quite black colour, Moina clearly able see that video …

Dark-field microscopy (also called dark-ground microscopy) describes microscopy methods, in both light and electron microscopy, which exclude the unscattered beam from the image. As a result, the field around the specimen (i.e., where there is no specimen to scatter the beam) is generally dark.

[2020ugchsncnseethala (Seethalakshmi .C. R.)]

Does the growth of the micro worms adversely affect the fruit flies culture bottle?@aswathy

[saswathy679 (Aswathy suresh)]

In my culture I observed this worm were growing after 16 days later… But still I got approximation number of Larave,Pupae, Adult flies inside these culture bottle. So, that means this worm is not harmful to Larave, Pupae, Adult flies. Not a parasitic nematode I guesses.

[saswathy679 (Aswathy suresh)]

451th Day 2021 : CUBE Chatshaala.

1st Part

Moina
Discussed about CUBE model system moina.

Why moina taken as a model system!?
• Very short life cycle.
• Maintain very easily.
• Survive in low oxygen level.
• Easy to culture.

Cubits are maintain this water flea for major purpose to studying Hypoxia, Epiphia.

•What is Hypoxia conditions ?
Low oxygen level moin body turn to red survive in low oxygen level (ability to adaptation).

•What is Epiphia ?
Moina have 2 mode reproduction ;
Asexual and sexual
Normally moina reproduction is asexual reproduction. But some harsh conditions moina reproduce sexually.

These are the major research question Cubits were addressing…

@man masih beck was sharing his experience culturing moin in his home lab Ranchi

Moina source (he got) : Moina macrocopa JSK 1 (Lab bred).

Ranchi Group Have Moina JSK 1.

Standard method of Culturing moina in Home Lab!

250mL Dechlorinated water, add 1 drop of milk every alternative days.

@man masih beck currently facing some problems culturing moin in his home. Have very few number of moina currently now (Man can correct).

1. Mother moina give 5 - 7 number of progeny’s .
   Number of moina he transferred : 5, 3 were gravid.

What is algae - moina relationship!?
Do moina feed on spirulina?

@Zahra Moina Story starting from 15th January 2021. Decrease and Increase growth of moina every month ways. How did she culture moina using subculture.

Moina Growth Curve

Factors which are affecting growth curve of moina.
1.Nutrition.
2.Area covered by population.
3.Crowding.
4.Toxicity.

@manmasih
The shortest summary of lessons that the Moina collaborators could learn from yesterday’s discussion is that Replicates/Subculturing should never be avoided and that the daily updates encourage young researchers like the CUBists to be consistent and at times even alert the CUBists about possible dangers.

[saswathy679 (Aswathy suresh)]

452th Day : CUBE Chatshaala

14.06.2021 Discussed about what is dark microscopy…? Why get background is more dark color but object is more clear…
In our CUBE people were taking photos different way to observe their object. For example Moina, Fruit Fly, Nematode…etc.
Light source coming from down - picture background will be White. If Light source coming from Up picture background will more black.

@Abhishak Unnikrishan from Kerala joined yesterday discussion…he was shared his experience in Drosophila melanogaster culturing in lab…How to making standard cornmeal agar media…what are ingredients they are using for making this media…
Autoclave bottles are preferred to culture Drosophila melanogaster in lab.
Standard cornmeal agar media adding :-
Ingredients
•Corn powder - Source of Carbohydrates.
•Agar - Solidified agent.
•Yeast extract - Source nutrients.
•Sucrose - Carbohydrates.
•Dextrose - Carbohydrates.
•Propionic acid - Antifungal agent.
•Orthoboric acid - Antibacterial agent.

In Home we are making alternative media for culturing fruit flies.
TRSV - media contain ingredients are :
•Tomato - Source of nutrients, some kinds of acids (malic acid, citric acid, acetic acid), Vitamin C are present in tomato.
•Rava - Source of Carbohydrates, Good binding agent.
•Sugar - Contains sucrose
•Vinegar - Antifungal reagent, it contain acetic acid.

TRSV media is a alternative form of Standard cornmeal agar media.

Also @Abhishak shared olfactory experiment procedures, importance done in his college lab…

Comparing Native Drosophila melanogaster with Lab bred Drosophila melanogaster 2nd instar larvae for this experiment…

Hypothesis : Native one have better olfaction compared to Lab bred flies. Because Native fly want to find their own food but lab bred fly have much availability of food (Culturing inside bottle). Highly chances to increase mutant flies.

Experiment : Using agar petri dish added 15 larave middle of plate 2 ends added : 1 side added IAA (Banana smells) Opposite side added Paraffin oil (No smell). Keep this setup in black box (Covered with black cover). After 1min check observation. Like ways 3 times, different concentration IAA checked.

Conclusion : Native D.melanogaster larvae have better olfaction compared to Lab D.melanogaster.

How to trap fruit flies ?
2 setups : 2 bottles with tomato pieces.
1st setup keep inside kitchen
2nd setup keep outside kitchen

Time @9:00am keeping setup.
Expecting 5 to 10 flies around @10:00am
Expecting 10 flies around @12:00pm

Also expecting Life Cycle of Fruit fly inside bottles…
•Eggs - 15/06/2021
•1st instar - 16/06/2021
•2nd instar - 17/06/2021
•3rd instar - 18/06/2021
•Pupation - 19/06/2021
•Next generation - 22/06/2021

How to identify 1st instar larvae, 2nd instar larave, 3rd instar larvae…? Size difference we can able see…! Other than size how can we distinguish 1st,2nd,3rd instar.

This paper is from one article says that other than size we can identify larvae looking their hook part… 1st instar 2 dots like…2nd instar 3rd instar have much developed hook (mouth part) can see. Also 3rd instar body end can able to see 2 orange ring structure. Shape of body end also distinguish 2nd & 3rd instar larvae.

@KiranKalakotiR In the last session, we had the discussion on how to descriminate 1st instar,2nd Instar & 3rd Instar larva? Does size is the only factor ro differentiate different stages of Drosophila larva, we also found one reference which says there are some structures through which we can easily identify the stage. 1st Instar although is of 1mm in size and 2nd instar is 2mm in size but there’s a difference in their mouth hook. 1st instar larva has mouth hooks which appears like two pairs of black spots and in 2nd instar it’s fully developed and structured. What do you think about 3rd instar larva? What’s the crucial change or development has happened and also it’s to be visible so that we can identify it, any comments how do we take good photos so that it’s visible?

How to save Fruit flies from Nematode attacking… Observed nematode were growing all culture bottle… We don’t know this nematodes is harmful or not in that case immediately want to save current cultures from nematodes.

How it possible ?
Adding more amount of vinegar will help to prevent the worm growth inside culture…save them in this way.
But it will may effect Fruit flies as well…. So first need to to check which amount of (20ml,25ml,30ml) effects nematodes not effect fruit flies… This is experiment need to conform test…

@2020ugchsncnseethala @KiranKalakotiR @drishtantmkawale @manmasih @saida786110.

[saida786110 (Saida Sayyed)]

If we cut hydra in 2 part auto part will regenerate the lost body that is will get two hydra if we cut one hydra into two pieces. So how is Regeneration different from Growth? What is Regeneration? The regeneration of the lost part is done by cell division then is Cell division (Mitosis) causing Regeneration? What is cell differentiation?
@KiranKalakotiR @manmasih

[saida786110 (Saida Sayyed)]

@⁨Shravani FY⁩ lost her hydra and Moina culture. Can it be because of the Kitchen smell? Can we also get the story of losing hydra in Christmas vacation @⁨Shravani FY⁩

@shravani reply During Christmas vacation December 2019 @⁨Shruti Salunkhe FY⁩ @karan we had bought our hydra culture from Elphinstone college to HBCSE lab the hydras was fed by Moina we were transferring the hydra to a beaker at the same time fruit fly Group was putting the orthophosphoric acid and Propionic acid in The media the next day we lost our hydra culture later we brought hydra from Sophia college Mumbai

We think it was because of the smell of these acids that we lost our hydra culture and that is the reason why I think the reason why my hydra died in my home lab could be because of kitchen smell.

[saida786110 (Saida Sayyed)]

How can fruitfly locate the fruit by sense of smell? And If Fruitfly can locate the fruit by a sense of smell is Hydra getting similarly affected by the smell? @saswathy679 @manmasih @2020ugchsncnseethala

[saida786110 (Saida Sayyed)]

Celebration of Goof ups Alexander Fleming Movement Moina surviving on algae self-sustaining ecosystem -@⁨Shravani

Moina collaborators are maintaining Moina using different methods like Milk,Yeast,Coconut Water, Ro water and now celebrating goof ups we have found we can culture Moina with Algae @⁨Shravani FY⁩ what will be Moina feeding on in Natural environment? @Rechel_tirkey @drishtantmkawale @manmasih @Zahra_R17 @Man

[saswathy679 (Aswathy suresh)]

454th Day : CUBE Chatshaala

Biological Rhythm

Biological rhythm is a phrase often used interchangeably with circadian rhythm. These rhythms are a series of bodily functions regulated by your internal clock. They control cycles like sleep and wakefulness, body temperature, hormone secretion, and more.

Every organisms have their own Biological rhythm.
For example :
•Flowering fruiting every year in mango tree, Jack fruit…etc. It is annual rhythm.
•Rhythm of heartbeat.
•One of the rarest flowers in the world, the Neelakurinji blooms just once every 12 years in India’s south-western states.
•Sleep wake cycle (Plants & Animals).

We are taking particular model systems to study biological rhythms in plant taking Phyllanthus, animals taking Fruit Fly.

Phyllanthus plants leàves are opening closing before sunrise and after sunset. Also is repeated every day…Is it any factors controlling their process ? Some internal Genes are control them.

CIRCADIAN CLOCK ASSOCIATED 1
(CCA1) and LATE ELONGATED HYPOCOTYL (LHY) in plants
Indeed, most organisms do not simply respond to sunrise but, rather, anticipate the dawn and adjust their biology accordingly. When deprived of exogenous time cues, many of these diurnal rhythms persist, indicating their generation by an endogenous biological circadian clock.

Plant Circadian Rhythms

defining attribute of circadian rhythms is that they are endogenously generated and self-sustaining, so they persist under constant environmental conditions, typically constant light (or dark) and constant temperature. Under these controlled conditions, the organism is deprived of external time cues, and the free-running period of ∼24 h is observed. A third characteristic of all circadian rhythms is temperature compensation; the period remains relatively constant over a range of ambient temperatures (Pittendrigh, 1954).Plant Circadian Rhythms - PMC

The interactive regulation between clock genes is central for oscillator function. Here, we show interactions between theArabidopsis clock genes LATE ELONGATED HYPOCOTYL(LHY), CIRCADIAN CLOCK ASSOCIATED 1(CCA1), and TIMING OF CAB EXPRESSION 1(TOC1). The MYB transcription factors LHY and CCA1 negatively regulate TOC1 expression. We show that both proteins bind to a region in the TOC1 promoter that is critical for its clock regulation. Conversely, TOC1 appears to participate in the positive regulation of LHY andCCA1 expression. Our results indicate that these interactions form a loop critical for clock function inArabidopsis.

Animals eg Fruit Flies active at day time and less active at night time. Why !?

Drosophila circadian keeps time daily fluctuations of clock-related proteins which interact in what is called a transcription-translation feedback loop. The core clock mechanism consists of two interdependent feedback genes, namely the PER/TIM genes and the CLK/CYC genes.
PER/TIM Genes were producing proteins stating from morning and at the evening proteins starting bind each other inside the cytoplasm (Cell). After it will degraded inside the cell.

drosophila-lg.wmv - YouTube.
CYC & CLK gene was together & then they bind to promoter region of PER & TIM gene.

Nail Regeneration Study

In the last Nail Session on 9th June 2021, Wednesday, we had a brief discussion on the anomalies seen in three (out of ~40) observations/data of Nail. Imagine, if three observations are intriguing this much, then what will happen when we continue this and slowly analyse each observation?
A Nail data bank will be created!
Let us make use of that data to understand simple yet interesting concepts not only of Life Sciences but cousin-fields of STEM!i) There are consistent observations that are being documented, but further processing of the photos, that become data later, is important. The aspect of discussing in brief on the observations, the nail growth/week, whether it is according/not according to the hypothesis, any inconsistent nail growth, etc. should be seen.
ii) At the same time, increasing the sample size i.e. involving more people in the study will increase the reliability on the processed data and ultimately, we can call it reliable data
For this, a standard method (that is replicable by all) needs to be devised to involve everyone.
In yesterday’s nail session, @⁨Aswathy suresh @⁨Kiran @⁨Dp⁩@⁨SeethaLakshmy⁩ @⁨Arunan and I made an attempt to formulate a standard method.
Yes, the nail is pushed from the proximal end to the distal end. Pushing is nothing but the deposition of dead keratinised cells. The older cells are pushed forward and new ones start depositing there.
This active process is what we call Nail Growth.
Yes, keratinocytes are differentiated from common stem cells.
“…its surrounding interfollicular epidermis (IFE). The IFE contains its own progenitor cells to ensure tissue renewal in the absence of injury, and HFs contain multipotent SCs that are activated at the start of a new hair cycle and upon wounding to provide cells for HF regeneration and repair of the epidermis.”
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2405915/: Epidermal Stem Cells of the Skin
Though the above reference tells of the progenitor stem cells (keratinocytes) involved in the growth of hair, I am sure there must be a same/similar mechanism in the development of Nail too!
@drishtantmkawale

@KiranKalakotiR @drishtantmkawale @manmasih @Rechel_tirkey @saida786110 @2020ugchsncnseethala

[saswathy679 (Aswathy suresh)]

455th Day : CUBE Chatshaala.

Discussed about model system FRUIT FLIES
from Bottukadavu - Thrissur Kerala.

@saswathy679 has been maintaining single line cultures of native fruit fly and these are very Single lines came from one mother. Currently have 4 single line cultures SLBK A, SLBK B, SLBK C, SLBK E - (Bottukadavu - Thrissur Kerala)

Research Question : To Find Drosophila melanogaster around native area

Currently facing some problems in some of the single lines SLBK A,SLBK B which are started on 26/05/2020 around last year.

SLBK A19 - 13.06.2021, SLBK B18 - 28.06.2021 subcultures of A & B species.
SLBK A19 - 13.06.2021 = ~5 Adult flies (2 Gravid), ~2 Larvae, 1 Pupate.
SLBK B19 - 28.05.2021 = ~4 Adult flies (4 Male), 0 Larave, 0 Pupae.

Flies will loose at any particular time. These cultures are precious to me because it started since 1 year ago…
How to save culture from this critical situation.!!!?

Happy Fruit Fly Culture
What are the criteria can tell this is a happy culture. Major talking about Larvae, Pupae, Adult flies, these all were in there in side culture can conclude these culture is going good.
Happy cultures are in my home lab SLBK C19 - 13.06.2021, SLBK E14 -13.06.2021.

SLBK C19 - 13.06.2021 = ~30 Adult flies (15 Gravid), ~20 Larvae, 0 Pupate.
SLBK B19 - 28.05.2021 = ~30 Adult flies (12 Male), ~25 Larave, 0 Pupae.

Expecting pupation will happen today evening get ~10 (5th Day).

What are the difference between 1st instar, 2nd instar, 3rd instar Larvae? Why 3rd instar larvae pupate upper side of culture bottle (Cotton plugs, upper side of bottle wall.) What could be reason.

Negative geotaxis behavior showing in fruit flies. So flies were always going upward direction. So that’s why flies pupae can able to see in upper parts of sides culture.

@Ishita tried to trapped 2 flies on 12.06.2021 which were present in the electric wire around @9:30pm.she trapped inside the empty bottle, after one day 13.06.2021 put some banana pieces on this bottle (Food). Today 17.06.2021 observed around 10/12 Larvae were moving bottles (Different sizes). Ishita made media TRSV for transfer these flies, her plan to culture them in a suitable media for further experiment study. But 2 were escaped. Only 1 were left… What she can do now!? That fly is not going up direction.

What will do in order to transfer these fly.!

What kind of research questions are addressing in Drosophila.
• Finding D.melanogaster native.
• Life span study of Drosophila.

What is Life span study ? What is the difference between Life Span & Life Cycle!?

What are other research questions can study in Drosophila

@Ishita Life span refers to the time period of an individual drosophila from birth to death.

Where else life cycle referes to the reproductive cycle in which the egg goes to become a larvae to a pupa and finally to an adult

@drishtantmkawale @2020ugchsncnseethala @KiranKalakotiR @saida786110 @manmasih

[saida786110 (Saida Sayyed)]

In today’s Chatshaala we discussed the basic identifiable features of the Cardamine plant TO FAMILIARIZE WITH CARDAMINE SPECIES Cardamine - Burke Herbarium Image Collection Cardamine is a large genus of flowering plants in the mustard family, Brassicaceae, known as bittercresses and toothworts. It contains more than 200 species of annuals and perennials.[1] Species in this genus can be found worldwide, except the Antarctic,[Cardamine - Wikipedia.
Cardamine is identified based on the Leaf pattern starting from cotyledons, simple leaf and compound leaf. Is there any plant showing this same pattern?

What all features we should look for in the Cardamine plant to identify it? Four species of bittercress were detected in the nursery industry of the United States, including New Zealand bittercress, hairy bittercress, flexuous bittercress, and little bittercress.On the Identity of the Weedy Bittercresses (Cardamine : Brassicaceae) in United States Nurseries: Evidence from Molecules and Morphology | Weed Science | Cambridge Core. Can we get an update of Cardamine home lab

Answering why Cardamine… Cardamine hirsuta is related to the model plant Arabidopsis thaliana and comparisons between these two experimentally tractable species have advanced our understanding of development and diversity. The power of forwarding genetics to uncover new biology was evident in the isolation of REDUCED COMPLEXITY, a gene that is present in C. hirsuta but lost in A. thaliana and shapes crucifer leaf diversity.

Cardamine hirsuta: a comparative view - PubMed

Genetic engineering in Cardamine plant through Floral Dip method What is the difference between conventional genetic engineering and floral dip method? What is the floral dip method? Can a gene be transferred by a solution of bacteria? Why kanamycin resistance Gene?

[saida786110 (Saida Sayyed)]

Breaking news fungal growth was observed in TRSV medium @ishita’s fruitfly culture Starting from 12 June she transferred some flies which were not active in her kitchen into a plastic bottle. And then added food that usually attracts flies larvae, pupae were observed Expectation - Newly emerging flies are expected on 23rd June based on observation of fruitfly collaborators. To transfer the flies TRSV media was made In which fungal growth was observed on 4th-day @ishita please update about the current situation of your fruitfly culture Vinegar does not act as an antifungal agent?

Yet another citizen science project To spread awareness of Aedes mosquito In this pandemic there is a lot of news about epidermic caused by dengue During monsoon mosquitoes are seen even more Aedes mosquito breeding in small stagnant water bodies indoors or outdoors The way to identify them is the body of this mosquito is usually black in colour with white spots on it Great initiative by Cube Kanpur @Hina how can we identify non Aedes like culex and other mosquito? What are the most prominent breeding site?

Lifecycle of Aedes Mosquito

Aedes Larvae

None Aedes

[saswathy679 (Aswathy suresh)]

458th Day of CUBE Chatshaala
Summary of Discussion

ZEBRA FISH

Discussed about @⁨Tejasri⁩ CUBE Home Lab updates from Nellore (Andhra Pradesh).

Tejasri and her collaborators from Nellore group working on zebra fish quite long time… They were culturing Zebra fishes in Collage Lab as well as Home Lab… After the pandemic time Tejasri & Ujvala were started culturing Zebra fish using minimum requirement which are available in home.

Tejasri have 5 Zebra fish in her home lab 3 Females and 2 males. Source of zebra fishes from collage lab 19 February 2021.She still culturing them using what are available in home lab. Made a setup In order to culture Zebra fish she taken 2.5L beaker for to culturing them.

Many of the new cubist were joined today started to asking about basics Questions…
How to tell about ZEBRA FISHES in to in a New Cubists in a simple way!?

“How to Identify Zebra Fish? What are the identifiable features of Zebra Fish?” - @Shrudhika (New Cubist).

Zebra Fish are ornamental fishes.we can able to see some strips were go through their body. Blue, White, Golden colored strips can able to see their body… Color depending on different species. Size of Zebra fish around 4 - 5 cm.

“Zebra Fishes breeds in Monsoon season, Because of the temperature is quite low, it’s also favorable to zebra fish to lay eggs” - @Tejas Dhepe.

" How to identify Male & Female Zebra fishes?"

•Size of the abdomen.
Female is quite bigger abdomen compared to Male fishes.

•Male fish is longer than Female fish.

Breeding
The mating and production of offspring by zebra fishes.

Zebra Fishes are eating their own eggs. So the progeny’s will not get in the same setup which we were kept Zebra fishes…
In that cases we are transferring Female & Male Fishes in other setup which we called Breeding setup (Collecting eggs).

Male and Females are puting together for breeding setup… Here is Zebra fishes External fertilization is taking place in side the water.
Above picture is from Nellor College lab : Picture shared by Tejasri.

“How to identified Fertilized & Unfertilized eggs”?!?

•Fertilized eggs are more transparent Yolk is visible in the center of the Eggs.
•Unfertilized eggs are more white colored Yolk is present in the surface of Egg.

What kind of research Question Cubists are addressing in Zebra Fish model system.

@Sujith from Kerala, also maintain different variety of Zebra fish (December) in his home lab Chalissery - Palakad.

Research Question : Immunology
Hypothesis : Look for anti - angiogenic molecules in zebra fish.

ZEBRA FISH MODEL FOR REGENERATION RESEARCH FIN REGENERATION
The zebrafish (Danio rerio) is one of the most accessible vertebrate models to study regeneration. In this Primer, we highlight the tools available to study regeneration in the zebrafish, provide an overview of the mechanisms underlying regeneration in this system and discuss future perspectives for the field.https://www.researchgate.net/publication/335959587_Model_systems_for_regeneration_Zebrafish

@Tejas from Bajaj science, Wardha, maintain 5 fishes of different species.

Tejas addressing research Question quite interesting came ZEBRA FISH DON’T SLEEP AT NIGHT!

“Fishes do not sleep at night” according to Tejas.

We can see their eyes in not close while resting time (During night). But fishes not much active in night.

How find out answer of this interesting research question…!@Tejas

How make simple Breeding setup in Home Lab, using what are available in home…?

@manmasih @2020ugchsncnseethala @drishtantmkawale @saswathy679

[saswathy679 (Aswathy suresh)]

459th Day : CUBE Chatshaala.

Summary continue Yesterday were stopped.
Discussed about model system…
ZEBRA FISH

@⁨Tejasri⁩ were talking about how did she zebra fish eggs collecting from Breeding setup and after collected fertilized eggs what was next!? How did maintain eggs in Petri plate…!!?How to safely maintain them in fungal contamination…?

What are the stages in life cycle of zebra fish?
How long is zebrafish development?
The zebrafish lifecycle is divided into four major periods: Embryo, larva, juvenile, and adult.

Picture from @Tejasri College Lab

Giving Artemia feed for to larval Zebra fishes…

What is Artemia?
Artemia is a genus of aquatic crustaceans also known as brine shrimp.

Brine shrimp

Why Artemia?
Why can’t give Moina,Paramecium,Microorganisms…which are easily available in Natural.
Zebrafish are primarily eating zooplankton and insects, although they can eat a variety of other foods, such as worms, small crustaceans or algae.

“Moina & Artemia (Both are Crustaceans). So can we take Artemia as a model system ?” - @Shrudhika

•Culture them easy.
•Maintain them easy
•Both of them are crustaceans.

How to culture Artemia?!

Artemia a Marine water organisms.
•Dried Artemia eggs adding in to saltwater and a container.
•After hatching collect them and transfer in to dechlorinated water.

**When we buy Artemia from a shop, it is in the form of packets…if they are eggs (Artemia) how they survive in packets!?** - @Chitra ma’am.

**What is Artemia Cysts?**
Artemia in the form of Cysts when we buy markets. Cysts are more hard. Which are protecting.

Artemia Cysts - Brine Shrimp eggs with high eclosion rate I BernAqua

Life cycle of Artemia

Under optimal conditions brine shrimp can live for several months, grow from nauplius to adult in only 8 days time and reproduce at a rate of up to 300 nauplii or cysts every 4 days.

http://www.fao.org/3/w3732e/w3732e0m.htm

Moina - another crustacean

Moina can find in Natural source like pont, streams ect… It’s a fresh water organisms.
Also we can buy from pet shop.

Moina have 2 type of reproduction
•Asexual
•Sexual

Horizontal gene transfer happen in asexual organisations…?

Mutation in plants Golden apple, Navel Orange.

Moina - 1.Parthenogenic Female, 2. Sexual Female.

[saswathy679 (Aswathy suresh)]

460th Day : CUBE Chatshaala.

Seasonomics

What are the questions we are addressing in Mango Model System?

Why no sprouting and no fruiting in other plants from shrudiga’s observation?.

•Today being Summer Solstice in northern hemisphere, are we recording in the next one week how Mango trees behave in different Latitudes and Longitudes in India and SriLanka?

I-net Farms Mango – Yet another common mango of Kerala, Moovandan is deeply linked with Kerala culture too. It takes only three years for its tree to blossom. That’s why this mango variety got this name (‘Moonnu’ means three and ‘Aand’ means year). They grow as branches, and filled with leaves.Moovandan Mango fruit plants are available here

Villagers indulge in lot of myths about Mangoes as well as on fruiting pattern! Can we have authentic data?
List of mango cultivars
From Wikipedia, the free encyclopediahttps://en.wikipedia.org/wiki/List_of_mango_cultivars

Paclobutrazol (PBZ) is a plant growth retardant and triazole fungicide. It is a known antagonist of the plant hormone gibberellin. It acts by inhibiting gibberellin biosynthesis, reducing internodial growth to give stouter stems, increasing root growth, causing early fruitset and increasing seedset in plants such as tomato[1] and pepper.[2]

Paclobutrazol

Can the PBZ method used for any tree?
Paclobutrazol (PBZ) is a plant growth retardant and triazole fungicide. It is a known antagonist of the plant hormone gibberellin. It acts by inhibiting gibberellin biosynthesis, reducing internodial growth to give stouter stemshttps://en.wikipedia.org/wiki/Paclobutrazol
Gibberllin is a common plant hormone and so," A known gibberellin biosynthesis inhibitor is paclobutrazol (PBZ), which in turn inhibits growth and induces early fruitset as well as seedset."Gibberellin - Wikipedia.

•Can we understand that this growth as Vegetative growth?

•What is the role of Gibberellins as plant hormones? How do Mango plantation groups utilize this knowledge?

•Why so little studies on Mango in India? Can we try collating some salient scientific literature? Wonderful HomeLab Model through out the country!

@drishtantmkawale @SukritiM @2020ugchsncnseethala @manmasih

Fruitflies which are pests
http://www-naweb.iaea.org/nafa/ipc/tephritid-fruit-flies.html

pictures of pest fruitflieshttps://preventfruitfly.com.au/why-is-fruit-fly-a-problem/the-pest-species/

The adult female fruit fly lay eggs in the flesh of ripening and ripe fruits and vegetables… Once the eggs hatch, the larvae begin to feed within the fruit, causing it to rot and drop to the ground.
https://preventfruitfly.com.au/controlling-fruit-fly/gardeners/choosing-a-control-strategy/

•What do they taste with?
•Tasting sensory organs? What can hairs do? Any thing similar in Antenna for smelling?
•How do they taste?
Labellum…
•What does the bristles do?

How to identify Drosophila Genus!!? What are the characters looking for…!

[saswathy679 (Aswathy suresh)]

461th Day : CUBE Chatshaala
Summary

Nail Regeneration Study

Discussed about :
1)How often do we clip our nails?
2)Which fingers’ nail grows the fastest?
3)Can nails be used as an indicator of health?

•Age
•Gender
•Diet
•Co-morbidities
Effecting factors of Nail growth…

4)How does a nail grow?
5)What is it made up of?

•Epidermis
•Epithelial

6)Is it Nail growth or Nail Regeneration?

Growth: Increase in size.
Regeneration: Regrowth of damaged and missing organ.

7)What is the nail made up of?
8)Just protein or something else alongwith protein?
9)What are keratinocytes?
Will an injury affect the growth of nails?

Structure of Nail

Onychophagia, or the chronic habit of biting your nails, has actually been associatedTrusted Source with a faster growth rate. This may be because biting causes trauma to the nail, stimulating circulation in the nail bed.

This also supports the theory that frequent nail clipping makes your nails grow a little faster. Regular clipping doesn’t carry the same risks as nail biting, so if you want longer nails, clipping is the better route.

Bifunctional ectodermal stem cells around the nail display dual fate homeostasis and adaptive wounding response toward nail regeneration

https://www.researchgate.net/publication/266581292_Bifunctional_ectodermal_stem_cells_around_the_nail_display_dual_fate_homeostasis_and_adaptive_wounding_response_toward_nail_regeneration

A team of researchers has identified a new population of nail stem cells, which have the ability to either self-renew or undergo specialization or differentiation into multiple tissues.
Under normal circumstances, the stem cells contribute to the growth of both the nails and the adjacent skin. However, if the nail is injured or lost, a protein called “Bone Morphogenic Protein,” or BMP, signals to the stem cells to shift their function exclusively to nail repair.

Nail growth. Thirty-five years of observation Nail growth. Thirty-five years of observation - PubMed

People of youg age group show faster nail growth compared to elders.

A 35-year observation of the growth of my nails indicates the slowing of growth with increasing age. The average daily growth of the left thumbnail, for instance, has varied from 0.123 mm a day during the first part of the study when 1 was 32 years of age to 0.095 mm a day at the age of 67.

HYPOTHESIS Need not be correct! Our DESIGN OF EXPERIMENT WILL TEST and PROVE OR DISPROVE it! After that we can declare to the whole world what is correct!

The term Different needs to be defined as faster or slower?
For testing @2020ugchsncnseethala Hypothesis, suppose you get FIVE 18 yr old FEMALES and another FIVE 18 yr old MALES.
Answer to your hypothesis questions,We can declare to world if 1.sample sizes is large enough 2.The study is repeatable or can be reproduced.

461th Day : CUBE Chatshaala
19years old female nail data collected by Seethalakshmy. Showing growth of nail.

How easy to study Nail Growth/Regeneration…!
•Henna /Mehandi can apply for making mark in nail.
Requirements : Mehandi cone.
How to measure proper Nail growth using graph paper!?

[saswathy679 (Aswathy suresh)]

462nd Day : CUBE Chatshaala.
Summary

FRUIT FLIES

24.06.2021 - Yesterday discussed about single line cultures in (Bottukadavu - Thrissur Kerala) current situation…

Summary of Work
SLBK A, B, C, E - total 4 single line cultures SLBK A, B were not good past 4 months. These cultures started almost one year ago… Now their conditions is really bad.
Flies number, Larval,Pupae were very less.
SLBK B almost lost conditions.There is only 2 larvae were left…No flies, No Pupate observed yet.

SLBK C, E are going good condition.
Current culture bottles : SLBK A19 ( 13.06.2021), B18 ( 28.05.2021), C18 (28.05.2021), C19( 13.06.2021, E18 (28.05.2021), E19( 13.06.2021)…subculture bottles have.

Description.
1)SLBK A19 - •15 Flies, •10Larvae, 15 Pupae ( 24.06.2021 - Observation)
2)SLBK B18 - •0 Flies, •2 Larave, •0 Pupae ( 24.06.2021 - Observation)
3)SLBK C18 - •+30 Flies, •+20Larvae, •+25Pupae. ( 24.06.2021 - Observation)
4)SLBK C19 - •+40 Flies, •20Larvae, 25 Pupae.( 24.06.2021 - Observation)
5)SLBK E18 - •15 Flies, •15Larvae, +15 Pupae.( 24.06.2021 - Observation)
6)SLBK E19 - •+30 Flies, •20Larvae, •25 Pupae.( 24.06.2021 - Observation)

SLBK B18 - The last bottle of B species culture only 2 (3mm sized) Larvae were there. Then how to save B culture ? Is it possible ?

Summary
1)SLBK C18 ( 28.05.2021) - •+30flies •20Larvae •25Pupae.
Dead flies - +25
2)SLBK C19 ( 13.06.2021) - •+40flies •20Larvae •25Pupae.
Dead flies - +20

Dead flies also increasing day by day inside the culture bottles
Importance of Replicate Bottles!
Facing issues availability of bottles.
How to solve these problems!? .

•We can reuse old culture bottles and take in to new culture as well.
(But rainey times can’t wash/dry up properly)
•Can we use Seven up bottles (Green colored bottles)!? any colored bottles for culturing these flies.

Different colors of bottle (Red, Blue, Green) will effects fruit flies badly ? Flies Is there any color preference?

Reference

Study gives the green light to the fruit fly’s color preference

462nd Day : CUBE Chatshaala.
Summary

PHYLLANTHUS PLANT

Color effect on sleep wake study!? (Red & Green)

To study effect of water on sleep wake cycle!
Water them once a day.

How to check the PH of soil at home!?

Potting mixture
[Sand/Cocopeat + Soil + Crowdung]

•20% Cocopeat.
•70% Soil.
•10% Crowdung.
Using potting mixture…

Design of the experiment
(Using 5 pots;)
POT 1, POT 2, POT 3, POT 4, POT 5.
•10 seed will sowing each 5 pots.
•Water then in alternative days.

[saswathy679 (Aswathy suresh)]

463rd Day : CUBE Chatshaala.
Summary

Discussed about model system CUBE Model Systems.

@Janhavi from Wardha Mumbai joined today discussion, She was new cubist.She really qurious about :

•What is model system!?
•Why we are taking model system for studying!?
•What type of research questions we are addressing in our Cubists.?

We are taking model system based on which kind of research question we are addressing…

Cubist maintain different types of model organisms

1)Fruit Fly (Drosophila) - Development Biology.
2)Moina - Hypoxia.
3)Snail - Learning & Memory.
4)Zebra Fish - Immunology.
5)Nematodes - Olfactory.
6)Phyllanthus - Sleep wake study.
7)Cardamine - Genetic engineering.
8)Ants - Communication.
9)Earthworms - Regeneration.
10)Hydra - Regeneration.

(Nail regeneration study, Mango mapping, Mosquitoes, Koel calls…etc)
These all model systems using based on related research questions.

What is model system! importance of model system ?

Taken model organisms which ;
•Easy to maintain.
•Very short life cycle.
•Easy to culture.

Models organisms have always been important in science and continue to be used to test hypotheses and predict information.

A model organism is a species that has been widely studied, usually because it is easy to maintain and breed in a laboratory setting and has particular experimental advantages. Model organisms are non-human species that are used in the laboratory to help scientists understand biological processes.

Summary

FRUIT FLIES

What is D. melanogaster!?
Why D. melanogaster?

D.melanogaster is also Six Nobel prizes – what’s the fascination with the fruit fly? Medical research.

The 1995 laureates in physiology or medicine are developmental biologists who have discovered important genetic mechanisms which control early embryonic development. They have used the fruit fly, Drosophila melanogaster , as their experimental system. This organism is classical in genetics.

Reference from public chat box giving below ;

In 1910, Thomas Hunt Morgan performed an experiment at Columbia University, in New York City, New York, that helped identify the role chromosomes play in heredity.

For their extensive researches on Drosophila, T.H. Morgan became the first geneticist to be awarded Nobel Prize in Medicine in 1934
Morgan apparently began breeding Drosophila in 1908. In 1909 he observed a small but discrete variation known as white-eye in a single male fly in one of his culture bottles.

Aroused by curiosity, he bred the fly with normal (red-eyed) females. All of the offspring (F1) were red-eyed. Brother–sister matings among the F1 generation produced a second generation (F2) with some white-eyed flies, all of which were males. To explain this curious phenomenon, Morgan developed the hypothesis of sex-limited—today called sex-linked—characters, which he postulated were part of the X-chromosome of females.

Gregg L. Semenza
The Nobel Prize in Physiology or Medicine 2019 Prize motivation: “for their discoveries of how cells sense and adapt to oxygen availability.”

•Fruit flies can easy to maintain in Lab/Home.
•Culturing them very easy.
•maintains simple.

@Janhavi made a media for to culture fruit flies using which are available in her homelab.Rava, Sugar, Tomato, Lemon juice (she don’t had the vinegar) made the media on 20/06/2021. today she were observed some microbial growth Also she don’t transferred any flies in to this bottles.

•Why we are making Media ?
•Importance of Media!!!

Its depending on our research questions…
Example : - If you want to study life cycle (Development Biology), you don’t need to make media for to culture these flies. We can use Tomato, Banana pieces to make trap flies, as well as we can keep them inside bottle more than 1 week. In this 1 week we can observe the life cycle of fruit flies.
Life cycle study in Fruit flies (Objective). Design of the experiment based on research question. Requirements : Transparent bottles, Tomato/Banana (food for flies).

another research study done by @⁨Seethalakshmy⁩.
Fruit fly larvae gut color change!
Design of the experiment : only used •Water bottle (transparent) • Fruits/Vegetables.
Experiment was used different colored Fruits (Tomato,Pumpkin,Guava, Water melon)
Used Tomatoes she observed some of visible larvae (3mm) shown the red color…
Repeated experiment using ;
Pumpkin - Yellow color
Guava - Pale pink
Watermelon - Red.
Result was some larvae (Not all Larvae) showing gut color change (Emitting fruit color)

• If we feed fruitflies with turmeric, can we see different colors at gut region with alkality different from acidity?

• If we transfer 10 gravid females, how many eggs we can ecpect on 1st day?

• Then transfer all the flies after 24 hrs into another bottle for laying eggs.After that, transfer all flies to a third bottle for another 24 hrs, to get in each bottle all larvae of more or less same mean age + or - 12 hrs!

[saswathy679 (Aswathy suresh)]

464th Day : CUBE Chatshaala
Summary of Discussion

Nirupam Samant from Goa was joined today chatshaala discussion, He is new to CUBE and first time joined attended Chatshaala, Nirupam is from Physics background.
Also we were discussed about Birds in goa,Common Birds in goa - Crows, Cuckoo, Kingfishers.

Koel calls in Goa - according to Nirupam winter season around November - December months mostly hears about koel calls.

High wind time Crows were together seems in Goa.

Existing story of Vulchers.
•Reason of existing
•Vulchers are normally eating dead body of animals, birds etc.
• Vulchers died because of the human body started to decomposing producing there some chemical were killed these birds.

According to @⁨Batul ma’am usage of DDT is causing the existing of Vulcher.

DDT - chemical is normally fusing for pesticides, insecticides etc. And also it’s cause to mutation. Dichlorodiphenyltrichloroethane.

DDT - Using to kill ants!?
Which chemical harmful for ants!?

Discussed model system

ANTS

Objective : Study Communication of Ants.
@⁨Shrudhika⁩ observed 1week ago Weaver ants were died because of war, war was 2 different species.
Weaver ants & Red ants… After war @⁨Shrudhika⁩ observed most of the Weaver were died surrounding in her formicarium and also they produced an acid chemical calling Oleic acid. After the war, till now she couldn’t see any ants in her formicarium as well as surroundings.

What could be the reason!?

She didn’t replaced her current setup! Kept in old place! And also she didn’t cleaned her correct formicarium…

26.06.2021 Didn’t observed any dead bodies of Ants.

Hypothesis : Chemical released by dead ants are persistent for a weeks.

How to prove/disprove this hypothesis using experiment.

Design of Experiment

Make a new Formicarium setup and keep in new place and compare with old setup.

Setup A : New formicarium
Setup B : Old formicarium

Both of the keeping in different area…for to trap ants.

Old setup made up of using glass jar, stones, soil, Sugar, Water… Similar way making new setup but this one will keep in different place.

According to @2020ugchsncnseethala adding more soil and less stones will give good culture of ants.

Ant Study
@seethlakshmy focused on black crazy ants.
Antenna non clubbed, abdomen showing black color rings.

•Why can we identified Black crazy ants.?
•Common name of these ants?
•Which kind of research question you are addressing in this type ants?