[Leish-l] inquiry
Patrick Bastien
patrick.bastien at univ-montp1.fr
Sat Jun 4 05:44:40 BRT 2011
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 écrit :
> 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 écrit :
>
>> 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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>>
>> ________________________________
>>
>>
>>
>>
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