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how to do Aspergillus Niger count?


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#1 dingdong

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Posted 04 April 2014 - 05:44 PM

Hi, I'm facing problem in counting my Aspergillus Niger. 

I grew the Aspergillus Niger on Sabouraud dextrose agar with chloramphenicol(SDAC), incubated for 5 days. Then I washed the plate with 9ml of 0.001% Tween 80, this I considered as 108 suspension.

Then I diluted to 102 suspension and took 0.1ml to do spread plate on SDAC, incubated for 5 days.

The problem is after 5 days incubation, my Aspergillus grows all over the agar surface, I cannot get separate colonies.

 

May I know what is the proper way to count the Aspergillus? I need to know the viable count for my 102 suspension.

Thank you.



#2 Phil Geis

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Posted 05 April 2014 - 02:57 AM

Sounds like USP <51> based protocol.  If you're using ATCC 16404 - the correct species name is Aspergillus brasiliensis,  Folks typically count colonies earlier than 5 days - look every day.



#3 dingdong

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Posted 14 April 2014 - 07:14 PM

I'm using ATCC 6275. I tried to count on the 3rd days, but I only got 4 colonies this time.

Any proper protocol on how to collect the spores and prepare exactly 102 suspension?

Thanks.



#4 Phil Geis

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Posted 15 April 2014 - 03:06 AM

That's a matter of your consistent execution of protocol and appropriate dilution.  Looks like your assumption of 10E8 is not correct.






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