[Leish-l] inquiry

Tue May 31 13:21:41 BRT 2011

Petr,

Thanks a lot for the info. I wish to mention several preliminary studies of samples from Turkey as follows:

1.	Most, if not all infantum CL patients in the Trop Med Center of Cukurova University, Adana are actually from the hilly area of Duzici, Kadirli, Osmaniye, etc in the north, but not from the Cukurova plain. Only 4 positive cultures were obtained by us out of ~100 lesion aspirates. All 4 gave us sequences, indicative of L infantum. The question is really the identity of the remaining ~96 samples, which produced no positive cultures and thus cannot be studied, except by direct examination of the clinical samples. 

2.	In Urfa, my record shows that cultivation of dozens of lesion aspirates has a successful rate of ~50%. Analyses of several dozens gave us the identical nagt sequences, indicative of L tropica, all being alleles I and II heterogeneity with 7 nt substitutions. Yet, 7% of the patient sera turned out rK39 positive. The question is whether L infantum accounts for some of the CL whose lesion aspirates produced no positive cultures ?

3.	In the coastal villages of Hatay, more than 20 lesion aspirates gave us no growth on cultivation and were examined directly by PCR/RFLP and sequencing, producing a mixture of L infantum or L tropica or both in a single sample - a picture very similar to the samples from Dr. NL Sharma in our hand. Enough controls were included to exclude the possibility of sample cross contamination. Two successful cultures were subsequently reported from dogs (or patients ?), both being L infantum according to our Turkish colleagues.

These preliminary studies raised several issues, which have been already noted by others in the literature:

1.	Have we missed something by studying cultured parasites ? 
2.	Can we make adequate predictions about the diseases based on the data from one, two or several isolates in culture from patients or sand flies or reservoirs ?

KP

________________________________

From: Petr Volf [mailto:volf at cesnet.cz] 
Sent: Tuesday, May 31, 2011 2:06 AM
To: Chang, Kwang-Poo; Sharman; Hiro Goto; elfadil abass
Cc: leish-l at lineu.icb.usp.br
Subject: Re: [Leish-l] inquiry

Hi K.P.,

in Cukurova region, Turkey, cutaneous L. infantum (now it seems that we are dealing with L.donovani/L. infantum hybrid) grew very poorely if we isolated them from patients. Only 1 of 25 isolations was succesfull. However, the same strain (confirmed by molecular methods) grew repeatedly and very well if we isolated them from sandflies. It might be useful for Dr. Sharma try to get isolates from sand flies. It is very laborious but very useful, in Cukurova we got about dozen of isolates by this method (all identical).

Patients are rK39 negative, see attached paper.

Best wishes

Petr

	----- Original Message ----- 

	From: Chang, Kwang-Poo <mailto:KwangPoo.Chang at rosalindfranklin.edu>  

	To: Sharman <mailto:nandlals at hotmail.com>  ; Hiro Goto <mailto:hgoto at usp.br>  ; elfadil abass <mailto:elfadil_abass at yahoo.com>  

	Cc: leish-l at lineu.icb.usp.br 

	Sent: Sunday, May 22, 2011 1:39 AM

	Subject: Re: [Leish-l] inquiry

	Dr. NL Sharma has been working on an important CL endemic area along the Satluj River valley to the south of Himalaya in India (Please correct me should I be wrong for anything I said here and below). I had the good fortune of visiting the site several years back courtesy of Dr. Sharma. 

	I believe Dr. Sharma's finding is important, since the parasites there are different from the familiar Indian L donovani in Bihar. The parasites are refratory to in vitro cultivation. They do differentiate into promastigotes and may grow a little, but  can't really be subcultured to establish passageable lines. This is very much reminescent of L infantum in the Mediterranean area. I recall Dr. Sharma has also found rK39+ dogs, if I remember correctly. If so,  Satluj river valley is endemic to the infantile CL.

	Analyses of several batches of basically clinical CL samples from Dr. Sharma showed evidence of L infantum, but also L tropica as well as a mixture of the two in one sample. This is the same picture we have noted for samples from Hatay, Turkey.

	These observations make me wonder a lot about our current knowledge on the clinico-epidemiology based on data collected previously by analyses of cultured promastigotes from one or few 'representative samples'. Nowaday, technology makes it very doable to work with biological samples for Leish DNAs directly from sand flies, patients and reservoir animals.

	KP

________________________________

	From: leish-l-bounces at lineu.icb.usp.br on behalf of Sharman
	Sent: Fri 5/20/2011 11:31 PM
	To: Hiro Goto; elfadil abass
	Cc: leish-l at lineu.icb.usp.br
	Subject: Re: [Leish-l] inquiry

	Dear all
	I agree with Hiro gito, We are working on a focus where the CL is
	predominantly caused by L. donovani, and the rK 39 STRIPS GIVE POSITIVE
	RESULTS WITH SERA as well as serous exudate from the lesion. The results are
	dependent upon species.
	NL Sharma

	--------------------------------------------------
	From: "Hiro Goto" <hgoto at usp.br>
	Sent: Wednesday, May 18, 2011 12:09 AM
	To: "elfadil abass" <elfadil_abass at yahoo.com>
	Cc: <leish-l at lineu.icb.usp.br>
	Subject: Re: [Leish-l] inquiry

	> Dear all,
	> In our oppinion, DAT and rK39 for those samples are not indicated  since
	> these tests are produced for the diagnosis of visceral  leishmaniasis.
	> In case of tegumentary leishmaniasis, it is very appropriate the
	> observation of J.J. Shaw appointing species specificity of antibody
	> response in these cases. Titers of antibodies are in general low in  these
	> cases therefore depending on the species, it may result  negative. We have
	> published a review recently in Expert Rev. Anti  Infect. Ther. 8(4),
	> 419?433 (2010), Current diagnosis and treatment of  cutaneous and
	> mucocutaneous leishmaniasis, where we raise this point.
	> Hiro Goto
	>
	> Citando elfadil abass <elfadil_abass at yahoo.com>:
	>
	>> Dear all I would recommend using DAT and rK39 strip test to measure
	>> antibody
	>> responses and to evaluate the diagnostic efficiency for both tests  in
	>> such group
	>> of patients.
	>>
	>>  Elfadil Abass
	>> ________________________________________________________________________________
	>> Institute of Medical Microbiology and Hospital Hygiene
	>> Philipps University Marburg
	>> BMFZ / Hans-Meerwein Straße 2
	>> D-35033 Marburg, Germany
	>> ________________________________________________________________________________
	>>
	>>
	>>
	>>
	>> ________________________________
	>> From: Nuha Nuwayri-Salti <racha at aub.edu.lb>
	>> To: saad saad <saad1426 at gmail.com>; "leish-l at lineu.icb.usp.br"
	>> <leish-l at lineu.icb.usp.br>
	>> Sent: Fri, May 13, 2011 9:39:33 AM
	>> Subject: Re: [Leish-l] inquiry
	>>
	>> Dear Saad first precaution to take is to separate your samples into
	>> several
	>> portions each (at least 5 each being no more than a few hundred(200-400)
	>> microliters). This is a necessary precaution to avoid freezing and
	>> thawing
	>> several times the same sample which will be the case should you do
	>> different
	>> studies at different times which is unavoidable.
	>>
	>> What you can do is correlate the type(ulcerated, abscess, furuncle
	>>  etc.. ) the
	>> locale, the number and age of lesions with the levels of antibody in  the
	>> sera of
	>> these patgients and also monitor cell mediated immunity with leishmanin
	>> skin
	>> test!
	>> I have just published (in print) an article about having circulating
	>> parasites
	>> in some of these patients with apparently pure cutaneous disease. It is
	>> the
	>> first paper that revealed this fact. It would be interesting to confirm
	>> or
	>> de-confirm this fact repeating what we did.
	>> Best wishes
	>>
	>> Nuha Nuwayri-Salti MD
	>>
	>>
	>>
	>> AOA
	>> Medical Honor Society
	>>
	>>
	>>
	>>
	>> -----Original Message-----
	>> From: leish-l-bounces at lineu.icb.usp.br
	>> [mailto:leish-l-bounces at lineu.icb.usp.br]
	>> On Behalf Of saad saad
	>> Sent: Friday, April 29, 2011 11:23 PM
	>> To: leish-l at lineu.icb.usp.br
	>> Subject: [Leish-l] inquiry
	>>
	>> Dear all
	>> Hi. i am Saad from Saudi Arabia. i have 60 sera from positive case of
	>> CL from south west of the country. I would like to have your
	>> recommendations to start a good research line in CL using these sera.
	>> Thanks in advance for your help
	>> _______________________________________________
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	>>
	>
	>
	>
	> Profa. Dra. Hiro Goto
	> Laboratório de Soroepidemiologia e Imunobiologia
	> Instituto de Medicina Tropical de São Paulo, USP
	> Av. Dr. Enéas de Carvalho Aguiar, 470, prédio II, quarto andar
	> 05403-000 - São Paulo, SP
	> Tel. +55-11-3061 7023, 3061 7056 ou 3061 7027
	> Fax. +55-11-3061-8270
	>
	> _______________________________________________
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