[Leish-l] inquiry

Thu Jun 9 14:15:43 BRT 2011

Different colleagues have different experience in different places.

KP

-----Original Message-----
From: Arezki Izri [mailto:arezki.izri at avc.aphp.fr] 
Sent: Thursday, June 09, 2011 1:43 AM
To: Patrick Bastien; Chang, Kwang-Poo
Cc: vishwamohan_katoch at yahoo.co.in; lalit Kant ICMR; leish-l at lineu.icb.usp.br
Subject: Re: [Leish-l] inquiry

Dear all,
I agree with Pr Bastien. All L. infantum do not have difficulties to grow in 
vitro. In my knowledge, the NNN medium with rabbit blood is the best to 
isolate strains. Some strains need 4 to 5 weeks before growing.
Dr Arezki IZRI

----- Original Message ----- 
From: "Patrick Bastien" <patrick.bastien at univ-montp1.fr>
To: "Chang, Kwang-Poo" <KwangPoo.Chang at rosalindfranklin.edu>
Cc: <vishwamohan_katoch at yahoo.co.in>; "lalit Kant ICMR" 
<lalitkant at icmr.org.in>; <leish-l at lineu.icb.usp.br>
Sent: Saturday, June 04, 2011 10:44 AM
Subject: Re: [Leish-l] inquiry

Dear K.P.,
I did not know you even worked in Tukey !...

Just to correct one of your last messages: I did not say we have no
problem in isolating L infantum. I wrote "that all L. infantum do NOT
have difficulties in growing in vitro: (a) it depends upon the culture
medium you are using; (b) it seems to depend upon the "taxon" (here
zymodeme), dermotropic zymodemes being in general (but not always)
more difficult to grow.". So: we indeed noted that some isolates are
more difficult to grow; and we do experience failures in isolating L
infantum.

As regards the medium used for isolating from sandflies, it was the
simple and classical NNN medium (for 1 liter: NaCl 6 gr , BactoAgar 10
gr; then blood 12%). One important point is the type of blood used.
Horse blood works less well than rabbit blood, it seems. Human blood
does not work. But I believe evrybody knows that.
Best wishes
P. Bastien

Professeur Patrick Bastien
Laboratoire de Parasitologie-Mycologie, Faculté de Médecine
UMR MIVEGEC (CNRS 5290 - IRD 224 - Université Montpellier 1)
National Reference Centre for Leishmania
CHU de Montpellier
39 Avenue Charles Flahault
34295 Montpellier Cedex 5, France

"Chang, Kwang-Poo" <KwangPoo.Chang at rosalindfranklin.edu> a Ã©critÂ :

> Dr. Bastien,
>
> Could you kindly share with us the magic medium you used ?
>
> Anything is worth a try, although we have tried a variety of   different 
> formulations of 3N.
>
> KP
>
> -----Original Message-----
> From: Patrick Bastien [mailto:patrick.bastien at univ-montp1.fr]
> Sent: Wednesday, June 01, 2011 7:39 AM
> To: Chang, Kwang-Poo
> Cc: Sharman; Petr Volf; Hiro Goto; elfadil abass; 
> vishwamohan_katoch at yahoo.co.in; lalit Kant ICMR; 
> leish-l at lineu.icb.usp.br
> Subject: Re: [Leish-l] inquiry
>
> Hi everybody !
> Just to let you know that all L. infantum do NOT have difficulties in
> growing in vitro: (a) it depends upon the culture medium you are
> using; (b) it seems to depend upon the "taxon" (here zymodeme),
> dermotropic zymodemes being in general (but not always) more difficult
> to grow.
> Once they are adapted to in vitro culture, there is no more problems.
> Re: isolation from sandflies, Pr Rioux (from whom I learnt in the
> field) used to soak the fly in successive baths of saline with
> antibiotics, before dissecting it upon a sterilized glass slide using
> alcohol-soaked instruments ! A bit awckward but try it !..
> Best wishes
> P. Bastien
>
> Professeur Patrick Bastien
> Laboratoire de Parasitologie-Mycologie, Faculté de Médecine
> UMR MIVEGEC (CNRS 5290 - IRD 224 - Université Montpellier 1)
> Centre National de Référence des Leishmania
> CHU de Montpellier
> 39 Avenue Charles Flahault
> 34295 Montpellier Cedex 5, France
>
>
>
>
>
> "Chang, Kwang-Poo" <KwangPoo.Chang at rosalindfranklin.edu> a Ã©critÂ :
>
>> Dr. Sharma's cultures produce promastigotes, which cannot be
>> passaged to build up the population, if I remember correctly.
>>
>> Petr will be great to help Dr. Sharma out with the sandfly studies.
>>
>> KP
>>
>> ________________________________
>>
>> From: Sharman [mailto:nandlals at hotmail.com]
>> Sent: Tue 5/31/2011 10:13 AM
>> To: Petr Volf; Chang, Kwang-Poo; Hiro Goto; elfadil abass;
>> vishwamohan_katoch at yahoo.co.in; lalit Kant ICMR
>> Cc: leish-l at lineu.icb.usp.br
>> Subject: Re: [Leish-l] inquiry
>>
>>
>> Hi Petr,
>> Thanks for enclosing the pdf. In fact the strain which we are
>> dealing with is not very difficult to grow but difficult to
>> maintain, It is rK 39 positive and as I mentioned in the
>> conversation below that even the serous exudate from the ulcer is
>> also positive for rK 39 dipstick. In between because of my
>> relocation to a different institute I was little away from this work
>>  but I intend to resume this work in couple of months. Although in
>> past we tried to isolate the parasite from sand flies but did not
>> succeed. We will surely attempt again with the techniques described
>> by you and others. Contamination is the problem.
>> This particular focus seems to be a complex one.
>> NL Sharma
>>
>> From: Petr Volf <mailto:volf at cesnet.cz>
>> Sent: Tuesday, May 31, 2011 12:36 PM
>> To: Chang, Kwang-Poo <mailto:KwangPoo.Chang at rosalindfranklin.edu>  ;
>>  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
>> 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
>> >> _______________________________________________
>> >> Leish-l mailing list
>> >> Leish-l at lineu.icb.usp.br
>> >> http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l
>> >> _______________________________________________
>> >> Leish-l mailing list
>> >> Leish-l at lineu.icb.usp.br
>> >> http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l
>> >>
>> >
>> >
>> >
>> > 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
>> >
>> > _______________________________________________
>> > Leish-l mailing list
>> > Leish-l at lineu.icb.usp.br
>> > http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l
>> >
>> _______________________________________________
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>>
>>
>> ________________________________
>>
>>
>>
>>
>> _______________________________________________
>> Leish-l mailing list
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>>
>>
>>
>
>
>
>
>

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