<div dir="ltr"><div class="gmail_default" style="font-family:verdana,sans-serif;font-size:small;color:#000000">A very important and challenging study, indeed, Joaquina! Congratulations to the Spaniards! Carlos.</div></div><div class="gmail_extra"><br><div class="gmail_quote">2015-03-06 9:32 GMT-03:00 joaquina <span dir="ltr"><<a href="mailto:joaquina@ugr.es" target="_blank">joaquina@ugr.es</a>></span>:<br><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex">Dear Katherine,<br>
Your approach is wrong. Leishmania DNA can be amplified by PCR and directly sequenced from host or vector biological samples. However, you must carefully choose a convenient PCR technique as not every of them give the same results. Also, you should take into account the presence of other trypanosomatid species apart from Leishmania, both in vector and in the reservoir and it is necessary to differentiate them properly. L. infantum is endemic in southwestern Europe but it is becoming clear that the classic epidemiological cycle dog-sandfly-human is turning into a network of animal species that collaborate with the dog in the maintenance of the parasite under natural conditions. Our research in southeastern Spain involved as reservoir, apart from the dog, the wild rabbit (Oryctolagus cuniculus) (please find attached the paper about it). Our studies also support the importance of rodents such as Rattus rattus, Apodemus sylvaticus and Mus musculus in the life cycle of L. infantum (article in revision in Parasitology Research)<br>
Joaquina Martin-Sanchez<br>
Dpto Parasitologia<br>
Fac Farmacia<br>
Granada University<br>
Granada, Spain<span class=""><br>
<br>
El 2015-03-06 00:30, Sayler,Katherine A escribió:<br>
</span><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex"><span class="">
Dear readers,<br>
<br>
Thank you sincerely for your time. I am a tick-borne disease biologist<br>
by trade, so this is outside of my realm.<br>
<br>
I am trying to confirm some preliminary data that suggests the<br></span>
_Leishmania_ spp. are circulating in limited geographic locales in<span class=""><br>
southern Florida. Is it at all reasonable to expect to detect the DNA<br>
of the trypanosomatids in the blood of infected wildlife? My suspicion<br>
is that this is completely improbable, as it appears that in order to<br>
detect DNA in enough quantity or quality for sequencing, you need<br>
cultured organisms. From sandflies it appears that it is possible to<br>
PCR amplify and sequence directly from the vector, but never from the<br>
reservoir (or potential reservoir) host. Is this true? Any advice or<br>
insight would be greatly appreciated and I am tremendously naïve and<br>
this is clearly not my area of expertise.<br>
<br>
Thank you again for reading my message,<br>
<br>
Katherine<br>
<br>
Katherine Sayler, M.Ed., Ph.D.<br>
Postdoctoral Associate | Department of Wildlife Ecology and<br>
Conservation<br>
Molecular Ecology Laboratory | 2322 Mowry Rd. Gainesville, FL 32611<br>
Phone: <a href="tel:352-871-3259" value="+13528713259" target="_blank">352-871-3259</a><br>
<br>
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<br></blockquote></div><br><br clear="all"><div><br></div>-- <br><div class="gmail_signature"><div dir="ltr"><div style="text-align:left"><span style="font-family:verdana,sans-serif">Carlos H. N. Costa, MD, DSc.</span></div><div style="text-align:left"><span style="font-family:verdana,sans-serif">Coordenador</span></div><div style="text-align:left"><span style="font-family:verdana,sans-serif">Laboratório de Leishmanioses - LabLeish</span></div><div style="text-align:left"><span style="font-family:verdana,sans-serif">Universidade Federal do Piauí</span><br></div><div><div style="text-align:left"><span style="font-family:verdana,sans-serif">Instituto de Doenças Tropicais Natan Portella</span></div><div style="text-align:left"><font face="verdana, sans-serif">Rua Artur de Vasconcelos 151-Sul</font></div><div><div style="text-align:left"><span style="font-family:verdana,sans-serif">64001-450 Teresina-PI</span></div><font face="verdana, sans-serif"><div style="text-align:left">Brazil</div><div style="text-align:left">Telephones: +55 86 3222-4377 (W), <div style="text-align:center;display:inline!important"><div style="text-align:center;display:inline!important">+55 86 3221-3062 (W), </div></div><div style="text-align:center;display:inline!important">+55 86 3237-1075 (R). </div></div><div style="text-align:left"><br></div><div style="text-align:left"><br></div><span style="font-size:x-small"><div style="text-align:left">Aviso: As informações contidas nesta mensagem são CONFIDENCIAIS, protegidas pelo sigilo legal, por direitos autorais e destinadas exclusivamente à pessoa ou organização para a qual a mensagem foi destinada.</div><div style="text-align:left">Warning: This message is meant only for the intended recipient of the transmission. It is forbidden any unauthorized use, alteration, reproduction and distribution. If you are not the correct recipient, please notify us immediately by return e-mail and delete this message from your system.</div></span></font></div></div></div></div>
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