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December 2015


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Tim Cser <[log in to unmask]>
Reply To:
The Pharmaceutical Microbiology Forum Email List <[log in to unmask]>
Mon, 30 Nov 2015 20:27:21 +0000
text/plain (161 lines)
ISO 29981:2010 is a good start for TAC of Bifidobacterium.  It's for milk and starter cultures so it should be a good reference for you even if it isn't exactly what you're looking for.  The formula does require a supplement (MUP) to inhibit Lactic acid bacteria.

Here's a link to the TOS-MUP media:,MDA_CHEM-100043#documentation,nav

Tim Cser
[log in to unmask]

-----Original Message-----
From: The Pharmaceutical Microbiology Forum Email List [mailto:[log in to unmask]] On Behalf Of Don English/SUF/US
Sent: Monday, November 30, 2015 7:51 AM
To: [log in to unmask]
Subject: Re: [PMFLIST] TAC of probiotics.

Dear Lim,

Enumeration of probiotic products can be somewhat difficult because they will generally contain more than one probiotic species such as a Lactobacillus species and a Bifidobacterium species.  To get counts for species of these 2 genera, you will generally need to use 2 different types of microbial growth agar for enumeration.  For conducting Lactobacilli counts of a probiotic product formulation, M17 or MRS Agars can be used.  However, there is no standard method that has been described for the selective enumeration of bifidobacteria in probiotic supplements.  In February 2015, there was an article that had been published in the Journal of Micro. Methods which indicated that both BSM Mupirocin and WSP Mupirocin Agars were found to be superior in the enumeration of Bifidobacteria species in probiotic supplements.  The reference for this article is as follows:

Bunesova, V, Musilva, S., Geigerova, M., Pechar, R., and Rada, V.
Comparison of Mupirocin-based media for selective enumeration of bifidobacteria in probiotic supplements.
J. Microbiol. Methods
2015, Feb., 109:106-0109.

From my experience, it is a waste of time to conduct Total Aerobic Plate count enumeration of probiotic supplements by using the methodology described in the USP due to the interfering recovery of Lactobacilli, but I have conducted pathogen screening of these products by using the enrichment techniques that are described in the USP with no issues.  The biggest problem that you are going to face is if you are going make a label claim on the product that it contains x amount of these species when it comes to stability of the counts for these species with the passage of time.  To get around this, you will need to add more bacteria in the product than what is indicated on the label.  In this case, the label should read contains a minimum of x amount of these species.


Donald J. English
Senior Manager - Microbiology, Research and Development One Avon Place, Suffern, New York 10901 Phone 1-845-369-2805 Tieline 3435 [log in to unmask]

Please consider the environment before printing

-----Original Message-----
From: The Pharmaceutical Microbiology Forum Email List [mailto:[log in to unmask]] On Behalf Of Tony Cundell
Sent: Friday, November 27, 2015 10:53 AM
To: [log in to unmask]
Subject: Re: [PMFLIST] TAC of probiotics.

Modified Mann-Rogosa-Sharpe with anaerobic incubation at 37 degree C for 72 hours by Champagne et al, 2011 Intern, J Food Microbiol. 149: 185-193 is recommended for Bifidobacterium spp.

On Thu, Nov 26, 2015 at 8:01 PM, Lim Foong Choo <[log in to unmask]>

> How about Bifidobacterium sp.  ? shall we perform TAMC  using method
> described in USP <61> ?
> Bifidobacterium is  anaerobic bacteria, but after incubate more than 2
> days, result showed TNTC on Tryptic soy agar plate at lower dilution .
> How to interpret  Microbial Contamination of probiotics ? shall we use
> bile tolerant Gram negative bacteria count or using modified method ?
> Thank you ..
> fc LIm
> --------------------------------------------------
> From: "Tony Cundell" <[log in to unmask]>
> Sent: Friday, November 27, 2015 12:28 AM
> To: <[log in to unmask]>
> Subject: Re: [PMFLIST] TAC of probiotics.
> I wrote a book chapter on this topic. USP <61> testing for TAMC would
> not
>> be appropriate for a probiotic with billions of lactobacilli per g.
>> the chapter was entitled " The Application of Modern Microbial
>> Methods to the QC testing of Probiotics" which appeared in volume 4
>> of the Encyclopedia of Rapid Microbiological Methods edited by
>> Michael Miller
>> On Thu, Nov 26, 2015 at 5:03 AM, Conor Shambrook
>> <[log in to unmask]>
>> wrote:
>> Yes we do.
>>> We find that we dont get any LAB growing on our TVC or Y&M plates.
>>> Oddly though we've found that if tested in the same run without
>>> being careful to segregate the samples (normal non-probiotic sample
>>> plates poured before bring the product into the laminar flow
>>> cabinet) that you can get LAB growth on the other samples plates.
>>> This is typically quite obvious due to the distinctive LAB colony
>>> morphology when this occurs.
>>> Cheers,
>>> Conor
>>> On 23/11/2015 9:30 PM, "Lim Foong Choo" <[log in to unmask]> wrote:
>>> > Dear PMF member :
>>> >
>>> >
>>> > Any one perform Total aerobic count of Probiotics , eg : >
>>> Bifidobacterium
>>> ,
>>> > lactobacilli ?
>>> > Should we test for TAC for those sample containing probiotics ?
>>> > what is the specification
>>> >
>>> >
>>> >
>>> >

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