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Grow Acetobacter

Acetobacter

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7 replies to this topic

#1 Adriana Reis

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Posted 30 July 2014 - 05:37 AM

Hello Everyone

This is my first post.

 

I am having quite the problematic bug here.

 

I have been trying with no success to grow Acetobacter pasteurianus.

 

First I have tried with this broth 

  • 500ml 

7.5g of malt extract

2.5g of yeast extract

add 470ml of water

and autoclave

 

then add 30ml of ethanol 50% filtered

when it left the autoclave broth was filled with a precipitate

 

grow at 26C

but had no growth

 

  • tried this one instead 1L

peptone 3g

yeast extract 5g

mannitol 25g

grow at 28C with no luck

 

tried after to grow it at 26C but also with no luck

 

  • I am trying to get CaCO3 so I could try a different one

1L

glucose 100g

yeast 10g

CaCO3 20g

pH 6.8

 

Anyone can suggest something to help me with this?

 

I am probably doing something wrong!!!

 

 

Thank you

Adriana Reis

 

 



#2 Phil Geis

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Posted 30 July 2014 - 09:52 AM

Try lowering p H to < 5

#3 El Crazy Xabi

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Posted 30 July 2014 - 05:38 PM

All of them are listed by the DSMZ for different strains of the species. How long have you incubated the cultures? In Acetobacter pasteurianus DSM 25273 , it states 3-7 days of incubation in medium 105 (the 3rd one). If you had it delivered as freeze-dried or frozen you should expect an even longer incubation time at the beginning. They don't state the incubation time in the other strains and media though. Better ask about it to the culture collection. You should have received some documentation with the vial.

However, if you already had the bug in the lab... are you sure the inoculum is still viable?



#4 Adriana Reis

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Posted 31 July 2014 - 01:50 AM

Thank you for your replies!!

I did bought it from DSMZ. 

Incredibly there is no pH mention or days of incubation so I thought it would have been the average.

I will definitely leave it for longer this time.

My Acetobacter pasteurianus is DSM 2006 and it is a new one bought recently.

 

THank you!

Im enjoying this new forum already!



#5 pito

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Posted 01 August 2014 - 04:54 AM

http://www.jcm.riken...umber?JCM=21215

 

The JCM sometimes offers more information.

 

But the pH should have not been the problem, you should not adjust it to grow it if you use the medium mentioned by DSMZ


If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#6 Adriana Reis

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Posted 04 August 2014 - 12:39 AM

Thank you

Funny enough this is a mix of two different broths from different websites. Will attempt this one as well.!



#7 El Crazy Xabi

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Posted 05 August 2014 - 08:10 PM

Sometimes is good to go and check the LPSN for the ofiginal description.

http://www.bacterio....ml#pasteurianus

For Acetobacter pasteurianus it can be tricky to find as it was initially described in 1879 but there is a link for a paper about reassesment of the genus that includes info about culturing:

http://www.sciencedi...723202083800204

 

Th e majority of the strains were grown on GYC-medium, containing per liter : 10 g yeast
extract (Oxoid), 50 g D-glucose, 30 g CaC0 3 and 25 g agar. Th e A. perox ydans, A. pasteurianus
subsp. paradoxus and subsp. ascendens strains did not develop on this medium ;
they were grown on a modified Carr-medium (1968) containing per liter : 30 g yeast
extract (Oxoid), 20 g ethanol and 20 g agar. On this medium, however, these strains died
off quickly. MYP-medium, cont ain ing per liter: 5 g yeast extract (Oxoid), 3 g peptone,
25 g D-mannitol and 25 g agar was used for maintenance. The temperature for growth
was always 28°C.



#8 Adriana Reis

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Posted 06 August 2014 - 12:45 AM

Thanks

I sticked with MYP broth and Im starting to see some cloudiness. Just repeated the process on another culture and will wait and see.

Thanks for all the advice!!!






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