Yaoundé-like virus in resident wild bird, Ghana

Tissue and swab samples from 551 wild birds collected in Ghana (October-November 2007) were assayed for alphaviruses, flaviviruses, and influenza A viruses using polymerase chain (PCR) techniques. One pool sample tested positive for Flavivirus RNA; further testing revealed that the amplified sequence was Yaoundé virus (YAOV), or closely related to it. YAOV is an apparently rare Flavivirus closely related to medically important human pathogens Japanese Encephalitis virus and West Nile virus. It is known only from West Africa. This is the first detection from Ghana, and only the second detection from a bird. Samples were negative for alphaviruses and Influenza A virus.We thank field companions including E. Bonaccorso, M. Robbins, and M. Thompson; laboratory companions, including A. Negredo and N. Reyes; Dr. F. Pozo and I. Casas from ISCIII for their collaboration; and L. Benitez for introduction to ISCIII. This study was supported by the US National Biological Information Infrastructure, the GAINS program of the Wildlife Conservation Society, and the Centers for Disease Control and Prevention, grants FIS PI07/1308 of the Red de Investigacion de Centros de Enfermedaes Tropicales RD06/0021, and the agreement signed between the Institute of Health Carlos III and the Spanish Ministry of Health and Social Policy for the surveillance of imported viral hemorrhagic fevers. Influenza work was supported by grant GR09/0040 (MPY-1440/09) ISCIII. RW is supported by grant CGL2010-15734/BOS, Ministerio de Ciencia e Innovación, Spain

First isolation of a new species of Leishmania responsible for human cutaneous leishmaniasis in Ghana and classification in the Leishmania enriettii complex

An active case detection approach with PCR diagnosis was used in the Ho District of the Volta Region, Ghana that identified individuals with active cutaneous leishmaniasis. Three isolates were successfully cultured and DNA sequences from these were analysed (ribosomal RNA internal transcribed spacer 1; ribosomal protein L23a intergenic spacer; RNA polymerase II large subunit), showing them to be Leishmania, identical to each other but different from all other known Leishmania spp. Phylogenetic analysis showed the parasites to be new members of the Leishmania enriettii complex, which is emerging as a possible new subgenus of Leishmania parasites containing human pathogens

An outbreak of suspected cutaneous leishmaniasis in Ghana: lessons learnt and preparation for future outbreaks

Human cutaneous leishmaniasis (CL) has previously been reported in West Africa, but more recently, sporadic reports of CL have increased. Leishmania major has been identified from Mauritania, Senegal, Mali, and Burkina Faso. Three zymodemes (MON-26, MON-117, and MON-74, the most frequent) have been found. The geographic range of leishmaniasis is limited by the sand fly vector, its feeding preferences, and its capacity to support internal development of specific species of Leishmania. The risk of acquiring CL has been reported to increase considerably with human activity and epidemics of CL have been associated with deforestation, road construction, wars, or other activities where humans intrude the habitat of the vector. In the Ho Municipality in the Volta Region of Ghana, a localised outbreak of skin ulcers, possibly CL, was noted in 2003 without any such documented activity. This outbreak was consistent with CL as evidenced using various methods including parasite identification, albeit, in a small number of patients with ulcers

First detection of Leishmania tropica DNA and Trypanosoma species in Sergentomyia sand flies (Diptera: Psychodidae) from an outbreak area of cutaneous leishmaniasis in Ghana.

BackgroundLeishmania major and an uncharacterized species have been reported from human patients in a cutaneous leishmaniasis (CL) outbreak area in Ghana. Reports from the area indicate the presence of anthropophilic Sergentomyia species that were found with Leishmania DNA.Methodology/principal findingsIn this study, we analyzed the Leishmania DNA positive sand fly pools by PCR-RFLP and ITS1 gene sequencing. The trypanosome was determined using the SSU rRNA gene sequence. We observed DNA of L. major, L. tropica and Trypanosoma species to be associated with the sand fly infections. This study provides the first detection of L. tropica DNA and Trypanosoma species as well as the confirmation of L. major DNA within Sergentomyia sand flies in Ghana and suggests that S. ingrami and S. hamoni are possible vectors of CL in the study area.Conclusions/significanceThe detection of L. tropica DNA in this CL focus is a novel finding in Ghana as well as West Africa. In addition, the unexpected infection of Trypanosoma DNA within S. africana africana indicates that more attention is necessary when identifying parasitic organisms by PCR within sand fly vectors in Ghana and other areas where leishmaniasis is endemic

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