I am trying to isolate some halophilic streptomyces, but it's not working very well.
I've tried to use Starch Casein Agar with different amounts of NaCl up to 2%. Different types of microorganisms including some bacillus growth on my plates.
Is there any way to make a selective media for halophilic streptomyces?
If I add 1g of CaCo3 to 10 g of some soil samples today in order to reduce the number of other microorganisms, will it work? Please help me find the best media for the isolation.
Thanks.
Hii, I would suggest you to use Actinomycetes Isolation Agar (AIA) for the isolation of actinomycetes from soil. during isolation increase the salt concentration according to ur need and use nystatin to avoid fungal contamination. Once you get powdery and pigmented colonies try to subculture them in ISP medium from 1 to 7. I hope it will work and let me know whether it is working or not.
Good luck.
Hi, I am also agree with surjit singh answer...But in that you should make some changes...AIA medium for isolation of actinomycetes from soil with different Nacl concentration and supplement with Antibiotics (to inhibit the growth of fungal and bacterial dominance)
Actidione 20mg/l
Nalidixic acid 100mg/l
in the medium..you will get good number of streptomyces colonies..I am sure you will get good results in this protocol...
All d best
Salam. Ba tavajoh be in ke man chandin kar darmorede streptomyces dashtam.ebtedaan chand soal.1. Khakhayei ke be onvane sample estefade mikoni az che manateghi mibashand? 2. Az che raveshi braye joda sazeie sterptomyces estefade mikoni? Manzoram culture methods?
Beast regard
Hello, I would like to suggest, Enrich the soil in Actinomycete isolation broth along with concentration of salt (5-20%) depending on your requirement (addition of Nalidixic acid and Clotrimazole can avoid contaminants) for few days, then isolate on Actinomycete isolation agar for Plate isolaion. Hope it helps...
use one of the International streptomyces media (e.g. ISP-4) and add salts (like those used in a salt water fish tank) You can obtain them at any good pet store with a fish section. I am using these to isolate microbes that grow in the ocean water. They would likely work with halophilic microbes as well but you will need to titrate the concentration.
I do agree with the professionals above on the isolation medium & the antibiotic
supplement(s),,, however as for the amendment with sodium chloride
you could use a gradient concentration(s) of the salt starting from 2%-20%
I managed to work with halophiles bacteria isolated from the Dead Sea/Jordan.
Wish You Good Luck.
DR.AL-DOORI-MAY
I am not experts like above. Is it not possible to increase salt concentration little bit high and avoid utilization of antibiotics. This will selectively enrich halophiles. Looking for response
Hi, thanks to giving a opportunity, I would suggest you to use Actinomycetes Isolation Agar (AIA) and Starch Casein Agar for the isolation of actinomycetes from soil. during isolation, when you have prepared the solid media used as source water or full strength seawater / 50% seawater , its very useful to growing the halotolerant actinomycetes in culture media also you have used as specific anti bacterial and antifungal drugs due to the avoiding the other contaminants. If you dought plz download and following method this paper links
http://nopr.niscair.res.in/bitstream/123456789/24828/4/IJMS%2042%287%29%20888-897.pdf
best of luck.
With Regards
Dr. S. Prakash
Changing the culture medium and rising the concentration of NaCl maybe help.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC239490/pdf/aem00167-0161.pdf
Please see the ref. above. What is the salt concentration at the source of isolation. Make it similar to that and use the selective medium-it sure will work. Good Luck. Swaranjit Singh Cameotra
Hi! you will try in Kuster’s agar medium with 50% sea water (from sample native). Before that u will do pretreatment (kept at 55 °C for 5 min) in sediment sample. Then start isolation. all the best..
It will be interesting to see the production of Biosurfactants by these interesting animal cules! Lets work together....ssc
You can use the medium Chitin Vitamins B + 80µg/mL of cycloheximide to eliminate the fungi+ 5% that will serve you to isolate Halotolerant streptomyces and Bacillus (which resist also the the salt and the pretreatment of your samples)
As already mentioned above it is a good way to mimic the salt concentration the organisms were taken from. To do that, you can use a salt mixture called "instant ocean" that contains almost the same amount of minerals as they are present in sea water. Hope it helps.
i think you can do that using high Nacl concentration medium
Hi Farzaneh
The isolation of slow growing MOs is not a simple problem.
Maybe your media is not the problem, many streptomyces are very slow in growth anyway, but under more harsh conditions, it might take even longer. Control the plates after f.e. two weeks, if there is still no growth.
In your case, it might be helpful to increase the salt level and vary the pH. Streptomyxes griseus or others are alcalophilic and are able to grow in a window from 5 to 11. Lower the pH under 6 will reduce the growth of bacteria, the same like increase it at 10. You can make a preselection in a liquid submer culture, make several transfers until you get a specificly enriched culture, adapted to this pH and salinity and free from the complex sources of the original soil. Don't add antibiotics or antifungals in the liquid culture, because you disturb the natural mostly symbiotic mixed culture. Finally you plate on the same media that you used for screening, maybe on antibiotic or anti-fungal plates too. Best isolation method for this screenings is a FACS-Cellsorter, but mostly it is not available. You can add 1g/LCarbopol 965 to the liquid media, it avoids the formation of the strong streptomyces mycel and gives you a better chance to catch an isolated cell.
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