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Problem in bacterial cell culture

microbiology cell culture bacteria

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12 replies to this topic

#1 Celz

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Posted 28 January 2014 - 08:54 PM

Dear all, 

 

I would like to seek for your opinion. What's the condition will causing the bacterial strain to become weaker? I have 2 probiotic bacterial strains which will growth actively within 18 hours in milk, however now the strain seems like become weaker and grow very slow even I incubate for 48 hours. Did anyone know the reason?

 

Thank you. 



#2 bob1

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Posted 30 January 2014 - 11:58 AM

Maybe the strains aren't what you think they are (maybe have become contaminated with something else)? It could also be that there are antibiotic residues in the milk, or something has been altered in the milk composition with time of year (if pasture fed animals).  It could also be that there is some lab based evolution, or you have subconsciously selected for a slow growing strain, or haven't stored them properly so they take a long time to recover.



#3 phage434

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Posted 30 January 2014 - 06:47 PM

Yoghurt cultures, for example, are required to contain at least three different species of bacteria. Perhaps your probiotic cultures are also mixed cultures. If they are, then continuous culture could easily lead to overgrowth of one of the constituent species.



#4 Celz

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Posted 01 February 2014 - 04:42 PM

Dear Thelymitra pulchella,

May I know is there any way to recover back the bacteria to make it become active? Thanks



#5 Celz

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Posted 01 February 2014 - 04:46 PM

Dear Veteran, 

I am using 6 bacterial strains, but culture it separately. It was strange and I find out that, the same bacterial strain (A), using the same technique, same material to culture into same volume of milk, but it growth very well in one bottle and totally no growth at all in another bottle. 



#6 bob1

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Posted 01 February 2014 - 05:24 PM

Dear Thelymitra pulchella,

May I know is there any way to recover back the bacteria to make it become active? Thanks

You can select for fast growing strains by just doing growth curves on lots of different isolates, and repeatedly choosing only those that grow faster than others.  For incorrect storage - you should get the bacteria growing actively again and then make more stocks under appropriate conditions for your bacteria.



#7 Celz

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Posted 02 February 2014 - 04:41 PM

 

Dear Thelymitra pulchella,

May I know is there any way to recover back the bacteria to make it become active? Thanks

You can select for fast growing strains by just doing growth curves on lots of different isolates, and repeatedly choosing only those that grow faster than others.  For incorrect storage - you should get the bacteria growing actively again and then make more stocks under appropriate conditions for your bacteria.

 

 

Do you think the bacteria can be activate back if I culture it onto the agar plate and isolate the single colony? Thank you. 



#8 pito

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Posted 04 February 2014 - 01:28 AM

 

 

Dear Thelymitra pulchella,

May I know is there any way to recover back the bacteria to make it become active? Thanks

You can select for fast growing strains by just doing growth curves on lots of different isolates, and repeatedly choosing only those that grow faster than others.  For incorrect storage - you should get the bacteria growing actively again and then make more stocks under appropriate conditions for your bacteria.

 

 

Do you think the bacteria can be activate back if I culture it onto the agar plate and isolate the single colony? Thank you. 

Its not so much about "activating" them back, its more about selecting the right isolate again...

Some will be fast growers, others slow growers, its just up to you to get that fast growing one back...

(and yes, some might indeed be slow growers, but due to a mutation become fast growers..)


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


#9 Celz

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Posted 04 February 2014 - 04:08 PM

 

 

 

Dear Thelymitra pulchella,

May I know is there any way to recover back the bacteria to make it become active? Thanks

You can select for fast growing strains by just doing growth curves on lots of different isolates, and repeatedly choosing only those that grow faster than others.  For incorrect storage - you should get the bacteria growing actively again and then make more stocks under appropriate conditions for your bacteria.

 

 

Do you think the bacteria can be activate back if I culture it onto the agar plate and isolate the single colony? Thank you. 

Its not so much about "activating" them back, its more about selecting the right isolate again...

Some will be fast growers, others slow growers, its just up to you to get that fast growing one back...

(and yes, some might indeed be slow growers, but due to a mutation become fast growers..)

 

 

Do you think if I streak it on an appropriate agar plate, incubate and isolate the single colony will be a good way to get back the active colony among all the weakest one? Thank you.



#10 bob1

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Posted 04 February 2014 - 05:07 PM

No- growth on agar is very difficult to give an indication of the growth rate - you need to pick single colonies and screen them for growth rate in liquid culture.



#11 Celz

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Posted 04 February 2014 - 05:15 PM

Do you mean I need to screen for all the single colonies on the plate and find out the one which can growth faster? 



#12 bob1

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Posted 04 February 2014 - 05:16 PM

Maybe not all, but pick maybe 10-20, and work from there.



#13 perneseblue

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Posted 06 February 2014 - 06:06 PM

Could the bacteria strain simply be acidifying its media or throwing out waste products that are inhibiting its own growth (eg acetic acid. lactic acid)?

Does a change in growth media help?


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