Diana Goodwin,  HT in Trenton  NJ asked on Histonet a couple of days ago - 
and I cross-posted this query to PATHO-L - here's a digest of the replies -

>>Well, the inevitable has happened. Here we are in historic Trenton NJ, 
where Washington crossed the Delaware in the famous surprise attack on the 
Hessian army. Now we have a new claim to fame:  Anthrax.

My histology department has received a skin biopsy from a patient with 
suspected
cutaneous anthrax.  Can anyone give me a reference as to the safe processing 
of such specimens, and, specifically, one that addresses if formalin kills 
Bacillus anthracis?<<
********
Vann Schaffner, MD
Lovelace Medical Center-Pathology
5400 Gibson SE
Albuquerque, NM   87108
505-262-7429
vann.schaffner@lovelace.com
replied:
>>There should not be significant numbers of spores in a biopsy of a skin
lesion, and formalin should kill the vegetative forms. Universal precautions
would protect against the low likelihood of  transmitting the vegetative
forms in an unfixed biopsy. Skin biopsy might recover organisms if submitted
fresh and cultured, or they might be seen on a gram stain of sections.  The
recommendations I have seen ask for a routine culture of a vesicular lesion,
or the edge, and under the roof of an eschar.<<
***************
Tim Morken EMT(MSA), HTL(ASCP)
Centers for Disease Control and Prevention
Infectious Disease Pathology
MS-G32 
1600 Clifton Road
Atlanta, GA 30333
PH 404-639-3964
FAX 404-639-3043
email tim9@cdc.gov 
replied:
>>These must be reported to the State Health Department, who will contact CDC.
They will apply a "case definition" to see if it fits. Any biopsy should be
handled normally with sterile technique and universal precautions. Fix in
formalin. 

The State Health Department and your clinicians will decide what to do with
it. If it is highly suspicious (as determined by the State) they may want to
send the biopsy to CDC right away.  If there is not a very high suspicion,
they will probably have you process the biopsy as a diagnostic case. If,
after testing it is still suspicious the State Health Department will send
it on to CDC for more testing. 

There is a very specific channel of communication for these cases because
they are treated as criminal cases. Therefore chain-of-custody documentation
for the specimens must be met. Work with the State Health Department and
they will give the guidence you need. - Hopefully no one else will need this 
information!<<
******************
Sate Hamza, M.D. 

http://peir.net
at the U of AL at Birmingham, provided some amplification of some citations:

John < RHD101@AOL.COM > provided some citations. 


The first article referred to, < J Clin  Microbiol 1990;28:223 >

(Identification of Bacillus anthracis by using monoclonal antibody to cell

wall galactose-N-acetylglucoseamine polysaccharide ) is not about

identifying the organism in paraffin-embedded tissue sections. 


And the following reference, < Am J Surg Pathol 1986;10:595 >  , is about

"Histology, immunohistochemistry, and ultrastructure of the verruga in

Carrion's disease". The other refernce noted, < Mol Microbiol 1992;6:1801 >,

also is not about B. anthracis. It is entitled : "Phylogenetic relationships

among the agent of bacillary angiomatosis, Bartonella bacilliformis, and

other alpha-proteobacteria". 


I did a medline search looking for answers. Not much is in the literature.

The findings in the skin biopsy in cutaneous anthrax don't seem to be that

specific. On H&E alone, I think the most that can be done is see changes

"consistent with" anthrax, if the clinical history, exam, and cultures all

go along with the diagnosis.  Lever shows 3 black and white pictures of the

changes. I did not find any citations regarding immunohistochemistry in this

setting. I did find the following though. The first is a case of

inhalational anthrax, and the second reports on using PCR to make the

diagnosis.


1) Fritz et al in 1995 < Lab Invest 1995 Nov;73(5):691-702 > reported using

"Immunolabeling techniques".."to identify the anthrax bacillus by light and

electron microscopy" in an experimental case of inhalation anthrax in a

rhesus monkey. 


2) Jackson et al < Proc Natl Acad Sci U S A 1998 Feb 3;95(3):1224-9 > used

PCR to identify the bacillus in formalin-fixed tissue sections from the 1979

Sverdlovsk anthrax victims. One of the samples was from a "vaccination site"

(the rest were mostly from spleen or meninges)

(You can see the article through the following link: 

http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=944

8313 )
************
Hope this information helps.

Bob Richmond
Samurai Pathologist
Knoxville TN