Phylogenetic tree of COI gene sequences among sand fly species.

<p>The COI (<i>S. ingrami</i> HLE-9, <i>S. ingrami</i> HLE-22, <i>S. ingrami</i> KLE-2, <i>S. ingrami</i> TAV-51, <i>S. hamoni</i> KLE-18 and S. <i>africana africana</i> HLE-82) gene was amplified from infected sand flies and sequenced. Analysis of the sequences together with those from <i>Sergentomyia</i> species, <i>Phlebotomus</i> species and <i>Lutzomyia</i> species registered in GeneBank were performed. The bar scale represents 0.02% divergences. Bootstrap values are shown above or below branches. HLE, Hlefi community; KLE, Klefe community; TAV, Taviefe community.</p

Phylogenetic tree of 18S rRNA gene sequences among sand fly species.

<p>The 18S rRNA (<i>S. ingrami</i> HLE-9, <i>S. ingrami</i> HLE-22, <i>S. ingrami</i> KLE-2, <i>S. ingrami</i> TAV-51, <i>S. hamoni</i> KLE-18 and S. <i>africana africana</i> HLE-82) gene was amplified from infected sand flies and sequenced. Analysis of the sequences together with those from <i>Sergentomyia</i> species, <i>Phlebotomus</i> species and <i>Lutzomyia</i> species registered in GeneBank were performed. The bar scale represents 0.005% divergences. Bootstrap values are shown above or below branches.</p

Detection of <i>Leishmania</i> DNA in sand flies.

<p><b>A.</b> PCR of <i>Leishmania</i> internal transcribed spacer 1 (ITS1) region amplified within pools of female <i>Sergentomyia</i> sand flies captured indoors and <i>Leishmania</i> spp. controls. M: 100 bp size marker; Lanes 1 and 2 (<i>L. major</i> and <i>L. tropica</i> reference strains, respectively); Lanes 3 to 6, <i>S. ingrami</i> pools, Lane 7, <i>S. hamoni</i> pool; Lane 8, <i>S. africana africana</i> pool (∼500 bp) and Lane 9, negative control. <b>B. </b><i>Hae</i>III digestion of restriction fragment length polymorphisms of ITS1 PCR products shown in A. M: 100 bp size marker; Lane 1, <i>L. major</i> (IPAP/EG/1989/S1-177); Lane 2, <i>L. tropica</i> (MHOM/SU/1974/K27); Lanes 3 to 6, <i>S. ingrami</i> pools; Lane 7, <i>S. hamoni</i> pool and Lane 8, <i>S. africana africana</i> pool.</p

Summary of <i>Sergentomyia</i> sand flies screened by molecular biological method in this study.

*<p>Ten female specimens in each pool.</p>¶<p>Identified human-blood fed specimens (Boakye et al, unpublished results).</p>a<p><i>L. tropica</i> DNA detection according to sequence analysis.</p>b<p><i>Trypanosoma</i> sp. DNA detection according to sequence analysis.</p>c<p><i>L. major</i> DNA detection according to sequence analysis.</p

Phylogenetic tree of ITS1 gene sequences among species.

<p><i>Leishmania</i> ITS1 gene was amplified within <i>Leishmania</i> DNA positive <i>S. ingrami</i> pools (IING/GH/2007/HLE-9, IING/GH/2007/KLE-2, IING/GH/2007/HLE-22 and IING/GH/2007/TAV-51) and <i>S. hamoni</i> pool (IHAM/GH/2007/KLE-18) and sequenced. Analysis was performed by the neighbor-joining method on the sequences together with those from 10 <i>Leishmania</i> species including the 2 human isolates from Ghana, <i>L. major</i> (MHOM/GH/2004/HO-004) and an uncharacterized <i>Leishmania</i> sp. (MHOM/GH/2006/TAVE). The scale bar represents 0.05% divergence. Bootstrap values are reported at nodes.</p

Phylogenetic tree of SSU rRNA gene sequences among species.

<p>The SSU rRNA (IAFR/GH/2007/HLE-82) gene was amplified within <i>S. africana africana</i> pool and sequenced. Phylogenetic analysis of the SSU rRNA gene sequences was performed by the neighbor-joining method on the sequence together with those from 25 <i>Trypanosoma</i> species. The sequences from the database are represented by the name of the species, isolates and GeneBank (accession number). The scale bar represents 0.02% divergence. Bootstrap values are reported at nodes.</p

A sample gel image.

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Accession numbers of influenza B sequences in NCBI influenza Virus Resource and GISAID used for the phylogenetic reconstruction of HA genes.

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Lineage specific markers of influenza B HA gene.

Multiple sequence alignment was carried out using ClustalW in BioEdit with a boostrap replicates of 1000 in line with Edgar [16]. Influenza B/Brisbane/60/2008 was used as the reference sequence for B/Victoria lineages while B/Wisconsin/1/2010, Clade 3 and B/Massachusetts/2/2012, Clade 2 were used as the reference sequence for B Yamagata lineages. Influenza B virus lineage-specific markers (nts 522, 540–542, 548, 549, 555, 558 and 568) are shown in yellow, whereas the clade specific markers (nts 538, 562 and 589) have been highlighted as green.</p

HA amino acid alignment for influenza B Yamagata-lineage.

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