Investigation of a new focus of Cutaneous Leishmaniasis in Ghana

Leishmaniasis is a disease of significant public health importance, which burdens a number of countries around the world, particularly in the tropics and subtropics. An outbreak of suspected cutaneous leishmaniasis (CL) has been witnessed in the Ho district of the Volta region in the south-eastern part of Ghana since 1999, where chronic ulcers typical of CL are being diagnosed. In this part of Ghana leishmaniasis has remained endemic to date. To add to the improvement of the level of understanding of the diseases in Ghana; the identity of the parasite, vector incrimination, non-invasive and field friendly diagnosis, and compound susceptibility tests were investigated. Patients presenting with cutaneous lesions suggestive of CL were selected where skin aspirates were collected from the sites of active lesion(s). Portions of the aspirates were cultured in M199 medium and DNA extracted from the promastigotes generated, while portions of the aspirates were inoculated onto FTA cards. PCR and PCR-RFLP were directly performed on the isolated DNA and the FTA cards. The pattern of bands produced from the patient samples were a complete deviation from DNAs of all the positive controls of Leishmania species. The sequenced PCR products and the further phylogenetic analysis revealed close relatedness to Leishmania enriettii species. The Leishmania species (GH5) responsible for the CL cases in that part of Ghana were successfully isolated into culture for the first time and proved to be distinct from the known species but closely related to non-pathogenic Leishmania enriettii. The transmission and the scanning electron micrograph evidence of the parasite confirmed their Leishmania identity. A peroxidoxin-based simple field friendly antigen detection test device was found diagnostically sensitive to Ghana species (GH5) and the other species of Leishmania used as controls in the diagnostic investigation. In the compound susceptibility test, the species isolated from Ghana (GH5) was found to be relatively resistant to cryptolepine, at concentrations to which the control species Leishmania mexicana was susceptible

First isolation of Leishmania from Northern Thailand:case report, identification as Leishmania martiniquensis and phylogenetic position within the Leishmania enriettii complex

Since 1996, there have been several case reports of autochthonous visceral leishmaniasis in Thailand. Here we report a case in a 52-year-old Thai male from northern Thailand, who presented with subacute fever, huge splenomegaly and pancytopenia. Bone marrow aspiration revealed numerous amastigotes within macrophages. Isolation of Leishmania LSCM1 into culture and DNA sequence analysis (ribosomal RNA ITS-1 and large subunit of RNA polymerase II) revealed the parasites to be members of the Leishmania enriettii complex, and apparently identical to L. martiniquensis previously reported from the Caribbean island of Martinique. This is the first report of visceral leishmaniasis caused by L. martiniquensis from the region. Moreover, the majority of parasites previously identified as "L. siamensis" also appear to be L. martiniquensis

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

The therapeutic landscape of HIV-1 via genome editing

Abstract Current treatment for HIV-1 largely relies on chemotherapy through the administration of antiretroviral drugs. While the search for anti-HIV-1 vaccine remain elusive, the use of highly active antiretroviral therapies (HAART) have been far-reaching and has changed HIV-1 into a manageable chronic infection. There is compelling evidence, including several side-effects of ARTs, suggesting that eradication of HIV-1 cannot depend solely on antiretrovirals. Gene therapy, an expanding treatment strategy, using RNA interference (RNAi) and programmable nucleases such as meganuclease, zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), and clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins (CRISPR–Cas9) are transforming the therapeutic landscape of HIV-1. TALENS and ZFNS are structurally similar modular systems, which consist of a FokI endonuclease fused to custom-designed effector proteins but have been largely limited, particularly ZFNs, due to their complexity and cost of protein engineering. However, the newly developed CRISPR–Cas9 system, consists of a single guide RNA (sgRNA), which directs a Cas9 endonuclease to complementary target sites, and serves as a superior alternative to the previous protein-based systems. The techniques have been successfully applied to the development of better HIV-1 models, generation of protective mutations in endogenous/host cells, disruption of HIV-1 genomes and even reactivating latent viruses for better detection and clearance by host immune response. Here, we focus on gene editing-based HIV-1 treatment and research in addition to providing  perspectives for refining these techniques

Identification of Sand Flies (Diptera: Psychodidae) Collected from Cutaneous Leishmaniasis Endemic Focus in the Ho Municipality, Ghana

Leishmaniasis, is a vector-borne disease transmitted to humans through the bite of infected female sand flies. Active and continuous monitoring of the sand fly is an important aspect of disease control. Thus, the correct identification of its vectors is paramount in this regard. Objective: The study was conducted to morphologically and molecularly identify female sand fly species in a cutaneous leishmaniasis endemic focus collected in three villages in the Ho Municipality of the Volta region. CDC light traps and sticky paper traps was used for the collection of the sand flies. The morphologically identified sand flies was molecularly confirmed by running PCR with the mitochondrial cytochrome c oxidase gene subunit I (COI) primers and DNA sequenced. A total of 537 sand flies was collected, made up of 363 females and 174 males.  Eleven different species of sand flies was morphologically identified – one Phlebotomus species and ten Sergentomyia species. The PCR amplified products showed bands of molecular weights 658 base pairs for the primers. The molecular identification using the 658-bp fragment of the (COI) gene was congruent with the morphological identification

Immunologic and virological response to ART among HIV infected individuals at a tertiary hospital in Ghana

Abstract Background The need to study the outcome of Antiretroviral Therapy (ART) among Human Immunodeficiency Virus (HIV) infected individuals in Ghana, a sub-Saharan African country crucial in the era of the “Treat All” policy. The aim of this study was to analyze selected determinants of immunological and virological response to ART among HIV infected individuals in a tertiary facility in Cape Coast, Ghana. Methods An analytical cross sectional study with a retrospective component was conducted in the Cape Coast Teaching Hospital (CCTH), Central Region. Clients aged 18 years and above attending the HIV Clinics for ART and who were on ART for 6 months or more were recruited. The viral loads, CD4 count and other socio-demographic data were analyzed using STATA version 13 (STATA Corp, Texas USA). Descriptive analysis was done and presented with appropriate measures of central tendencies. In addition, bivariate and multivariate analysis was carried out with p value of 0.05 interpreted as evidence of association between variables. Results A total of 440 participants were included in this study with a mean age of 45.5 (±11.6) years. The mean CD4 count at baseline, 6 months on ART and currently at study recruitment were 215.1 cells/mm3 (±152.6), 386.6 cells/mm3 (±178.5), and 579.6 cells/mm3 (±203.0) respectively. After 6 months and 12 months on ART, the number who had achieved viral copies  1000 copies/ml after 6 months on ART (aOR 2.0, 95% CI 1.2–3.2, p = 0.01). Conclusion There was good response to ART among clients, high virological suppression and immunological recovery hence low rates of change to second line ART regimen in this cohort studied. With strict adherence to the national policy on HIV testing, management of positive clients and full implementation of the “Treat All” policy, Ghana could achieve, if nothing at all, the third “90, 90, 90” target by 2020

Chromosome-Scale Assembly of the Complete Genome Sequence of Leishmania (Mundinia) sp. Ghana, Isolate GH5, Strain LV757

Leishmania (Mundinia) sp. Ghana is a kinetoplastid parasite isolated in 2015 in Ghana. We report the complete genome sequence of L. (M.) sp. Ghana, sequenced using combined short-read and long-read technologies. This will facilitate greater understanding of this novel pathogen and its relationships within the subgenus Mundinia

Chromosome-Scale Assembly of the Complete Genome Sequence of Leishmania (Mundinia) procaviensis Isolate 253, Strain LV425

Leishmania (Mundinia) procaviensis is a parasitic kinetoplastid that was first isolated from a rock hyrax in Namibia in 1975. We present the complete genome sequence of Leishmania (Mundinia) procaviensis isolate 253, strain LV425, sequenced using combined short- and long-read technologies. This genome will contribute to our understanding of hyraxes as a Leishmania reservoir

Chromosome-scale genome sequencing, assembly and annotation of six genomes from subfamily Leishmaniinae

We provide the raw and processed data produced during the genome sequencing of isolates from six species of parasites from the sub-family Leishmaniinae: Leishmania martiniquensis (Thailand), Leishmania orientalis (Thailand), Leishmania enriettii (Brazil), Leishmania sp. Ghana, Leishmania sp. Namibia and Porcisia hertigi (Panama). De novo assembly was performed using Nanopore long reads to construct chromosome backbone scaffolds. We then corrected erroneous base calling by mapping short Illumina paired-end reads onto the initial assembly. Data has been deposited at NCBI as follows: raw sequencing output in the Sequence Read Archive, finished genomes in GenBank, and ancillary data in BioSample and BioProject. Derived data such as quality scoring, SAM files, genome annotations and repeat sequence lists have been deposited in Lancaster University's electronic data archive with DOIs provided for each item. Our coding workflow has been deposited in GitHub and Zenodo repositories. This data constitutes a resource for the comparative genomics of parasites and for further applications in general and clinical parasitology