Thanks Freida,

What better source to validate the steps I am taking to eliminate the problem 
of "AFB Contaminate".  

I would also like to mention in my haste to compose my e-mail, I may have 
stated something incorrectly.  I did not mean that a positive (+) control be 
cut at the same time as the test sample and negative (-) control.  I usually 
place the test tissue on the same pre-cut positive (+) control slide.

Thanks Rene,

For your input on recycling formalin.  This is good ammunition for my defense 
in stopping the use of recycling formalin.  I still would welcome any and all 
replies.

Akemi


Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-ta...@apmglab.com
P: E-Mail: akemiat3...@yahoo.com



--- On Thu, 5/28/09, Freida Carson <frei...@sbcglobal.net> wrote:

From: Freida Carson <frei...@sbcglobal.net>
Subject: Re: Fw: Re: AW: [Histonet] AFB & Recycling Formalin for Fresh Tissue
To: "Histonet" <Histonet@lists.utsouthwestern.edu>, "Akemi Allison-Tacha" 
<akemiat3...@yahoo.com>
Date: Thursday, May 28, 2009, 12:09 PM

Wang reported on the contamination of tissue sections with AFB by the use of 
fluorescence microscopy, and if I remember correctly, when he tested the water 
in water fountains, he found something like 33% contained AFB.  Non-pathogenic, 
but we can't tell that on our stains. The paper is in Am J Clin Pathol 51:71, 
1969.  We (Carson, Kingsley, Haberman, and Race) also reported the 
contamination in the 1964 issue of the same journal.  We stopped using tap 
water for our water baths and began using millipore filtered water that had 
already been through a deionizing column and charcoal filter.  We always cut a 
negative control from the same days workload as the patient case - uterus in 
our case.  You don't need a positive control cut the same day, just a positive 
control with only a medium number of organisms.  We
 also did not use any tap water in the deparaffinization prior to the carbol 
fuchsin.  AFB organisms have also been reported growing in 40 gal formalin 
tanks believe it or not and in the old Fisher Paraffin wafers.
 
We centrifuged tap water in 50-ml tubes, poured off the supernatent, and 
refilled and recentrifuged until we could see some sediment in the bottom.  We 
took that to Microbioloby and had it cultured to definited prove the presence 
of AFB in the tap water.
I would recommend this approach to anyone suspecting tissue contamination. 
 
Freida Carson
--- On Thu, 5/28/09, Akemi Allison-Tacha <akemiat3...@yahoo.com> wrote:


From: Akemi Allison-Tacha <akemiat3...@yahoo.com>
Subject: Fw: Re: AW: [Histonet] AFB & Recycling Formalin for Fresh Tissue
To: "Histonet" <Histonet@lists.utsouthwestern.edu>
Date: Thursday, May 28, 2009, 1:21 PM


Hello,
I responded to Gudrun personally, but I thought that the information I am 
supplying him below might be a good educational source for other histologists 
who are having similar scenario's for AFB contamination.  

I would also like to say that I have an extremely eager histology team that 
have expressed an interest in learning theory and practice.  Most of my team 
are OJT.  They have had several pathologists approach them with these concerns 
regarding AFB, and because of their knowledge, or lack of it, couldn't fix the 
problem.  We are starting with the basics and I am giving in-services along the 
way.

Hi  Gudrun,

They are shipping the recycled formalin directly to the outside hospital 
surgery to put their fresh tissues into.....

As far as the AFB, we have had intermitant (+) AFB on the edges of tissues, 
that has been extremely questionable.  We currently are not
 cutting a negative (-) & positive(+) control on the same water bath as the 
test sample. That will be remedied beginning Monday. The water here is also in 
question.  I will be sending it out for microbacteria analysis.

We are going through a thorough housekeeping and in-service process.  We are 
starting with the tissue processors, which are currently only changed weekly 
(not rotated daily), embedding centers, which have to my knowledge, never been 
drained, and the hopper cleaned-out.  The water baths are not scrubbed out with 
soap and HOT water daily and
covered at night to prevent contamination.  The water bath is not skimmed of 
excess tissue debris, each and every time another specimen is placed on it.  
The Carbol Fuchsin is not being filtered prior to use.  The working solution is 
currently being poured back into the stock bottle.  Need I go on................

Akemi Allison-Tacha BS, HT (ASCP)
 HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-ta...@apmglab.com
P: E-Mail: akemiat3...@yahoo.com



--- On Thu, 5/28/09, Gudrun Lang <gu.l...@gmx.at> wrote:

From: Gudrun Lang <gu.l...@gmx.at>
Subject: AW: [Histonet] Recycling Formalin for Fresh Tissue
To: akemiat3...@yahoo.com
Cc:
histonet@lists.utsouthwestern.edu
Date: Thursday, May 28, 2009, 10:34 AM

Just for interest. Your lab takes the recycled formalin to make 4% neutral
buffered formalin (NBF)? Or do they ship the recycled formalin directly in
bottles to the surgery?
AFB = acid fast bacili?

Do you have concerns, that they produce false-positive AFB stains? I cannot
imagine, that a (hopefully) small number of dead bacili on the surface of
tissue will pretend this. An other question: Isn't there a filter in the
recycler to prevent such contamination?
Gudrun

-----Ursprüngliche Nachricht-----
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von
akemiat3...@yahoo.com
Gesendet: Donnerstag, 28. Mai 2009 18:14
An: Histonet
Betreff: [Histonet] Recycling Formalin for Fresh Tissue

Good Morning Histo Land,

I am asking all you out there to give me your input on recycIing formalin. 
I realize this has been discussed in the not too distant past, but this may
be a little different situation.  I realize some labs are recycling formalin
to put on their tissue processors, but this is a totally different
 scenario.

I have been a manager at my lab for only a month.  Much to my surprise, I
just found out yesterday that our lab is recycling formalin and shipping it
out to one of our clients to put their FRESH SPECIMENS into. 


I just had a lengthy conversation with Robert Lott the day before yesterday,
regarding pre-analytical protocol's effecting the test results of AFB.  By
the way, we were in full agreement.  This formalin issue was unknown to me
then.  I realize that the pre-analytical, pre-analytical, pre-analytical,
process effects the final out come.  I need to present my case to our
Director of Pathology why this is compromising patient care.

Any and all responses are gladly welcome.

Thanks,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell:
 425.941.4287 
W: E-Mail: aallison-ta...@apmglab.com
P: E-Mail: akemiat3...@yahoo.com

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