GMU:Bioelectronics, aesthetics and other interesting things/Trina Ukmata: Difference between revisions
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=Spot the antibiotics producer: STREPTOMYCES= | =Spot the antibiotics producer: STREPTOMYCES= | ||
[[File:Micro.png| | [[File:Micro.png|800px]] | ||
---- | ---- | ||
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Think no further than ''the soil''. | Think no further than ''the soil''. | ||
''Streptomyces''. | |||
---- | |||
===ISOLATION OF STREPTOMYCES=== | ===ISOLATION OF STREPTOMYCES=== | ||
====1st step: Air dry the soil samples for 3-4 h at 45° for the selection of Actinomyces spores, considering that many other bacteria die during this pre-treatment==== | ====1st step: Air dry the soil samples for 3-4 h at 45° for the selection of Actinomyces spores, considering that many other bacteria die during this pre-treatment==== | ||
<gallery> | |||
File:Soil1.png | |||
File:Soil.png | |||
</gallery> | |||
====2nd step:Dilute 1-3g of soil in 10 ml saline solution (NaCl 0,9%) and let the solution stand for 15 min==== | ====2nd step:Dilute 1-3g of soil in 10 ml saline solution (NaCl 0,9%) and let the solution stand for 15 min==== | ||
<gallery> | |||
File:NACL.png | |||
File:Solution .png | |||
</gallery> | |||
====3rd step: Prepare 3 dilutions (1:10, 1:100, 1:1000) out of it==== | ====3rd step: Prepare 3 dilutions (1:10, 1:100, 1:1000) out of it==== | ||
<gallery> | |||
File:Diluting.png | |||
File:Dilutions&Solution.png | |||
</gallery> | |||
====4th step: Prepare the Mannitol Soy Agar (MSA) petri dishes==== | ====4th step: Prepare the Mannitol Soy Agar (MSA) petri dishes==== | ||
< | <gallery> | ||
MSA RECIPE: | File:Mannitol.png | ||
File:Plates.png | |||
</gallery> | |||
''MSA RECIPE:'' | |||
In a 250 mL conical flask mix: | In a 250 mL conical flask mix: | ||
2g Agar | *2g Agar | ||
2g Mannitol | *2g Mannitol | ||
2g Soya flour (Holland and Barrett) | *2g Soya flour (Holland and Barrett) | ||
10 mM CaCl2 | *10 mM CaCl2 | ||
100 mL Deionised water | *100 mL Deionised water | ||
<tt> | |||
Medium alternative: Inorganic Salt Starch Agar (ISSA) | Medium alternative: Inorganic Salt Starch Agar (ISSA) | ||
</ | </tt> | ||
====5th step: Plate an aliquot of 0.1ml of each dilution on the medium (MSA)==== | ====5th step: Plate an aliquot of 0.1ml of each dilution on the medium (MSA)==== | ||
<gallery> | |||
File:Put.png | |||
File:Spread.png | |||
</gallery> | |||
<tt> | |||
The loop used for spreading the solution on the medium (shown on the 2nd photo) is a DIY loop, created from paper clips | |||
</tt> | |||
====6th step: Incubate at 30° for 5-7 days==== | ====6th step: Incubate at 30° for 5-7 days==== | ||
::Optional: Supplementation with cyclohexamide (50 μg/ml) and nystatin (50 μg/ml) to further remove competing bacteria | '''Important:''' Use parafilm to fortify the covers of each petri dish, in order to avoid evaporation, as it can lead to contamination. | ||
[[File:Incubation.png|800px]] | |||
Optional: Supplementation with cyclohexamide (50 μg/ml) and nystatin (50 μg/ml) to further remove competing bacteria | |||
---- | |||
==3-DAY OUTCOMES== | |||
<gallery> | |||
File:After3days.png | |||
File:After3dayss.png | |||
</gallery> | |||
'''Observations:''' | |||
The no-dilution petri dish shows a tendency towards contamination. Other petri dishes seem on the right track by far. The small white powdery looks like a good sign. | |||
==6-DAY OUTCOMES== | |||
<gallery> | |||
File:Sample1.png|1:10 | |||
File:Sample2.png|1:100 | |||
File:Sample3.png|1:1000 | |||
File:Sample4.png|1:1000 (2) | |||
</gallery> | |||
'''Observations:''' | |||
There has been overgrowing a hybrid of organisms, but assumingly the pinky/white powdery dots are streptomyces colonies.Such colonies are mostly to be found on the 1:100 and 1:1000 dilution plates. In this case, creating subcultures is the best solution, in order to get a unified-type of biotope in the petri dish! | |||
Only one sample has been contaminated, as it was the first one to get done during the isolation process. | |||
==FINAL DAY MICROSCOPICAL VIEWS== | |||
[[File:Micro2.png|800px]] | |||
==Insight== | |||
'''Streptomyces''' species are the most abundant source of antibiotics. More specifically, they produce the two-thirds of all existent antibiotics. Except that, this genus produces a rife scope of bioactive compounds. It usually inhabits the soil and plays a significant role in decomposing. | |||
So, next time when you get ill, decide to take another pill, in order to re-feel the thrill... | |||
'''...take a minute to think and ''praise the genus of good welfare.''''' | |||
---- | ---- | ||
'''Inspiration''': | '''Inspiration''': | ||
http://annadumitriu.tumblr.com/ - Anna Dumitriu | http://annadumitriu.tumblr.com/ - Anna Dumitriu | ||
https://exploringtheinvisible.com/ - Dr. Simon Park | https://exploringtheinvisible.com/ - Dr. Simon Park | ||
Latest revision as of 16:03, 14 July 2016
Spot the antibiotics producer: STREPTOMYCES
Antibiotics.
The human gets ill, goes downhill, forgets the dollar bill, takes a pill, regains will, re-feels the thrill.
It’s simply the recuperation cycle.
But, do you ever think about the fundamentals of having the chance of recuperation from infections?
Sure.
It’s thanks to the pill - the antibiotic.
Good job!
Now, zoom in deeper…
Do you actually know where this savior comes from?
Think no further than the soil.
Streptomyces.
ISOLATION OF STREPTOMYCES
1st step: Air dry the soil samples for 3-4 h at 45° for the selection of Actinomyces spores, considering that many other bacteria die during this pre-treatment
2nd step:Dilute 1-3g of soil in 10 ml saline solution (NaCl 0,9%) and let the solution stand for 15 min
3rd step: Prepare 3 dilutions (1:10, 1:100, 1:1000) out of it
4th step: Prepare the Mannitol Soy Agar (MSA) petri dishes
MSA RECIPE:
In a 250 mL conical flask mix:
- 2g Agar
- 2g Mannitol
- 2g Soya flour (Holland and Barrett)
- 10 mM CaCl2
- 100 mL Deionised water
Medium alternative: Inorganic Salt Starch Agar (ISSA)
5th step: Plate an aliquot of 0.1ml of each dilution on the medium (MSA)
The loop used for spreading the solution on the medium (shown on the 2nd photo) is a DIY loop, created from paper clips
6th step: Incubate at 30° for 5-7 days
Important: Use parafilm to fortify the covers of each petri dish, in order to avoid evaporation, as it can lead to contamination.
Optional: Supplementation with cyclohexamide (50 μg/ml) and nystatin (50 μg/ml) to further remove competing bacteria
3-DAY OUTCOMES
Observations: The no-dilution petri dish shows a tendency towards contamination. Other petri dishes seem on the right track by far. The small white powdery looks like a good sign.
6-DAY OUTCOMES
1:10
1:100
1:1000
1:1000 (2)
Observations: There has been overgrowing a hybrid of organisms, but assumingly the pinky/white powdery dots are streptomyces colonies.Such colonies are mostly to be found on the 1:100 and 1:1000 dilution plates. In this case, creating subcultures is the best solution, in order to get a unified-type of biotope in the petri dish!
Only one sample has been contaminated, as it was the first one to get done during the isolation process.
FINAL DAY MICROSCOPICAL VIEWS
Insight
Streptomyces species are the most abundant source of antibiotics. More specifically, they produce the two-thirds of all existent antibiotics. Except that, this genus produces a rife scope of bioactive compounds. It usually inhabits the soil and plays a significant role in decomposing.
So, next time when you get ill, decide to take another pill, in order to re-feel the thrill...
...take a minute to think and praise the genus of good welfare.
Inspiration:
http://annadumitriu.tumblr.com/ - Anna Dumitriu
https://exploringtheinvisible.com/ - Dr. Simon Park