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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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===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==== | |||
<gallery> | <gallery> | ||
File:Soil1.png | File:Soil1.png | ||
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File:Plates.png | File:Plates.png | ||
</gallery> | </gallery> | ||
''MSA RECIPE:'' | ''MSA RECIPE:'' | ||
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*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==== | ||
:[[File:Incubation.png| | '''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 | Optional: Supplementation with cyclohexamide (50 μg/ml) and nystatin (50 μg/ml) to further remove competing bacteria | ||
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==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.''''' | |||
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'''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 |
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