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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">Dear Katherine<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">If the trypanosomatids circulate in the blood of infected wildlife, then it should be possible to detect and obtain enough
DNA for direct sequencing using sensitive PCR methods, such as nested-PCR, and multicopy targets, such as kDNA and ribosomal ITS/subunits.
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">Otherwise, it would be best to obtain samples from the organs where the most parasites are expected, such as lymph nodes,
bone marrow, skin, etc. <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">Best regards<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">Isabel<o:p></o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">De:</span></b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"> Sayler,Katherine A [mailto:saylerk@ufl.edu]
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<b>Enviada:</b> 5 de março de 2015 23:30<br>
<b>Para:</b> leish-l@lineu.icb.usp.br<br>
<b>Assunto:</b> [Leish-l] Confirm hits to Leishmania spp. in whole blood of mammals in the US?<o:p></o:p></span></p>
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<p><span style="font-family:"Calibri",sans-serif;color:black">Dear readers, <o:p>
</o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black">Thank you sincerely for your time. I am a tick-borne disease biologist by trade, so this is outside of my realm.
<o:p></o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black;background:white">I am trying to confirm some preliminary data that suggests the
<em><span style="font-family:"Calibri",sans-serif">Leishmania</span></em> spp. are circulating in limited geographic locales in southern Florida. Is it at all reasonable to expect to detect the DNA of the trypanosomatids in the blood of infected wildlife? My
suspicion is that this is completely improbable, as it appears that in order to detect DNA in enough quantity or quality for sequencing, you need cultured organisms. From sandflies it appears that it is possible to PCR amplify and sequence directly from the
vector, but never from the reservoir (or potential reservoir) host. Is this true? Any advice or insight would be greatly appreciated and I am tremendously naïve and this is clearly not my area of expertise</span><span style="font-family:"Calibri",sans-serif;color:black">.<o:p></o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black">Thank you again for reading my message,<o:p></o:p></span></p>
<p><span style="font-family:"Calibri",sans-serif;color:black">Katherine<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial",sans-serif;color:black">Katherine Sayler, M.Ed., Ph.D.</span><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial",sans-serif;color:black">Postdoctoral Associate | Department of Wildlife Ecology and Conservation</span><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial",sans-serif;color:black">Molecular Ecology Laboratory | 2322 Mowry Rd. Gainesville, FL 32611</span><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial",sans-serif;color:black">Phone: 352-871-3259 </span><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
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