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From:
derek freeman <[log in to unmask]>
Reply To:
The Pharmaceutical Microbiology Forum Email List <[log in to unmask]>
Date:
Tue, 3 Dec 2019 15:12:05 +0000
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Hi Jovan,

Mea Culpa - Sorry  for previously asking for information that you had already provided in your original post.

Moving on, I assume you have checked for changes associated indicator (lot or type) , gas-pak (lot or type)  jar ( type or gasket).

On review I'm wondering what prompted you cross check the jar conditions with aerobes if the indicators were  working and when you were normally getting suitable growth recovery of Clostridia spp. during GP testing?

Like others I would suggest that your indicator system may not be very sensitive and probably you have  sufficient anaerobic conditions to support growth of more aerotolerant anaerobes such as Clostridia spp. particularly towards the latter stages of incubation.  In contrast during the early stages there is insufficient anaerobiosis to prevent aerobic growth.

Reminds me of Clin Micro demonstrations for students where in the absence of sufficient gas-paks we would incubate Ps. aeruginosa plates alongside Clostridia perfringens in the same jar to use up excess O2 present.

Anyway, tolerance to Oxygen by anaerobes is dependent on immediate metabolic status, as well as being strain, species and genera related, i.e. is a spectrum. Despite being referred to as obligate anaerobes, strains of Clostridia spp. can  grow adequately albeit in microaerophilic environments. In addition being spore formers, spores may survive in aerobic environments and grow out when anaerobic conditions are eventually established. It could well be that despite recovering Clostridia during your GP tests that you may have always had a relatively poor anaerobic environment or more likely a  delay in establishing one in your test system, but an insensitive indicator system along with test outcome (suitable recovery of indicator organism on test completion) was masking this.

I would concur with other responders that you should perhaps look at your indicator system sensitivity but might suggest also conducting trials using obligate anaerobes including Clostridia spp. or Bacteroides spp. to  establish suitability of the test system.

By the way regarding aerotolerant anaerobes- I am led to believe that Propionibacterium acnes is now  reclassified as Cutibacterium acnes  -  (got to love the new name).

Best Regards & Good Luck

Derek

________________________________
From: The Pharmaceutical Microbiology Forum Email List <[log in to unmask]> on behalf of Jovan Rankov <[log in to unmask]>
Sent: 29 November 2019 09:40
To: [log in to unmask] <[log in to unmask]>
Subject: Re: [PMFLIST] Anaerobic jars - aerobic strains

Hi Derek, we had A. lwoffii and M. luteus

Jovan Rankov
Specijalista procesa obezbeđenja sterilnosti
Služba za mikrobiološka ispitivanja
Kontrola Kvaliteta EEC
Hemofarm AD
tel: + 381 (13) 803 368
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-----Original Message-----
From: The Pharmaceutical Microbiology Forum Email List <[log in to unmask]> On Behalf Of derek freeman
Sent: Tuesday, November 26, 2019 9:59 PM
To: [log in to unmask]
Subject: Re: [PMFLIST] Anaerobic jars - aerobic strains

Hi Jovan,
Can you confirm what  genera or species of "obligate aerobes" were used in the  trial test?
Regards

Derek
________________________________
From: The Pharmaceutical Microbiology Forum Email List <[log in to unmask]> on behalf of Jovan Rankov <[log in to unmask]>
Sent: 26 November 2019 15:04
To: [log in to unmask] <[log in to unmask]>
Subject: [PMFLIST] Anaerobic jars - aerobic strains

Dear forum,

This a correction of my query, to avoid confusion.
Recently in our microlab we obrained growth of strictly aerobic species in our anaerobic jars. We then tested our anaerobic jars (with oxygen indicators) by incubating strictly aerobic species from our library of isolates, and we obtained full growth on all petri dishes. Indicators suggested that the atmosphere was anaerobic after a few hours. We even sent our strains to genetic ID and got the same results - species are strictly aerobic (Micrococcus luteus, Acinetobacter lwoffii).

In GP test we use these jars for anaerobic strains and we do obtain growth of strictly anaerobic strains regularly (Clostridium spp.).

We tested old and new jars, lids were tight, indicators were within shelf life, everything seemed allright.

Does anyone else think this is weird? How would you proceed?

Jovan
QC Microbiologist

________________________________


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