Dataset Information

Distribution and Risk of Mycolactone-Producing Mycobacteria Transmission within Buruli Ulcer Endemic Communities in Cote d'Ivoire.

ABSTRACT: In Buruli ulcer (BU) endemic communities, most mycolactone-producing mycobacteria (MPM), including Mycobacterium ulcerans, the causative agent, are present in water bodies used by inhabitants; yet, their mode of transmission is still unclear. This study aimed to assess the distribution of MPM strains, both from human suspected cases and aquatic environments, for identifying possible transmission modes within two BU endemic districts, Daloa and Tiassalé (Taabo), in Côte d'Ivoire. Collected samples were processed using conventional polymerase chain reaction and screened for the presence of non-tuberculous mycobacteria (NTM) and MPMs using 16S rRNA, IS2404 and enoyl reductase (ER) primers. MPM-positive samples were further discriminated using variable number tandem repeat (VNTR) typing and sequencing. 16S rRNA and IS2404 sequences confirmed that 94% of the clinical samples contained MPMs. For environmental samples, 53% were contaminated with NTMs, of which 17% contained MPMs particularly M. ulcerans, suggesting that water-related activities could predispose inhabitants to BU transmission. MPM discrimination by VNTR at four M. ulcerans Agy99 loci identified genotype C, previously reported in Côte d'Ivoire as the most dominant profile. Phylogenetic clustering on the basis of genetic diversity in the MIRU 1 locus showed two main M. ulcerans lineages in Côte d'Ivoire.

SUBMITTER: Dassi C

PROVIDER: S-EPMC6082052 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

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Distribution and Risk of Mycolactone-Producing Mycobacteria Transmission within Buruli Ulcer Endemic Communities in Côte d'Ivoire.

Dassi Christelle C Mosi Lydia L Narh Charles A CA Quaye Charles C Konan Danièle O DO Djaman Joseph A JA Bonfoh Bassirou B

Tropical medicine and infectious disease 20170226 1

In Buruli ulcer (BU) endemic communities, most mycolactone-producing mycobacteria (MPM), including <i>Mycobacterium ulcerans</i>, the causative agent, are present in water bodies used by inhabitants; yet, their mode of transmission is still unclear. This study aimed to assess the distribution of MPM strains, both from human suspected cases and aquatic environments, for identifying possible transmission modes within two BU endemic districts, Daloa and Tiassalé (Taabo), in Côte d'Ivoire. Collected ...[more]

PMID: 30270862

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Project description:The identification of an emerging pathogen in humans can remain difficult by conventional methods such as enrichment culture assays that remain highly selective, require appropriate medium and cannot avoid misidentifications, or serological tests that use surrogate antigens and are often hampered by the level of detectable antibodies. Although not originally designed for this purpose, the implementation of polymerase-chain-reaction (PCR) has resulted in an increasing number of diagnostic tests for many diseases. However, the design of specific molecular assays relies on the availability and reliability of published genetic sequences for the target pathogens as well as enough knowledge on the genetic diversity of species and/or variants giving rise to the same disease symptoms. Usually designed for clinical isolates, molecular tests are often not suitable for environmental samples in which the target DNA is mixed with a mixture of environmental DNA. A key challenge of such molecular assays is thus to ensure high specificity of the target genetic markers when focusing on clinical and environmental samples in order to follow the dynamics of disease transmission and emergence in humans. Here we focus on the Buruli ulcer (BU), a human necrotizing skin disease mainly affecting tropical and subtropical areas, commonly admitted to be caused by Mycobacterium ulcerans worldwide although other mycolactone-producing mycobacteria and even mycobacterium species were found associated with BU or BU-like cases. By revisiting the literature, we show that many studies have used non-specific molecular markers (IS2404, IS2606, KR-B) to identify M. ulcerans from clinical and environmental samples and propose that all mycolactone-producing mycobacteria should be definitively considered as variants from the same group rather than different species. Importantly, we provide evidence that the diversity of mycolactone-producing mycobacteria variants as well as mycobacterium species potentially involved in BU or BU-like skin ulcerations might have been underestimated. We also suggest that the specific variants/species involved in each BU or BU-like case should be carefully identified during the diagnosis phase, either via the key to genetic identification proposed here or by broader metabarcoding approaches, in order to guide the medical community in the choice for the most appropriate antibiotic therapy.

Project description:Mycobacterium ulcerans, an environmental opportunistic pathogen, causes necrotic cutaneous and subcutaneous lesions, named Buruli ulcers, in tropical countries. PCR-derived tests used to detect M. ulcerans in environmental and clinical samples do not allow one-shot detection, identification, and typing of M. ulcerans among closely related Mycobacterium marinum complex mycobacteria. We established a 385-member M. marinum/M. ulcerans complex whole-genome sequence database by assembling and annotating 341 M. marinum/M. ulcerans complex genomes and added 44 M. marinum/M. ulcerans complex whole-genome sequences already deposited in the NCBI database. Pangenome, core genome, and single-nucleotide polymorphism (SNP) distance-based comparisons sorted the 385 strains into 10 M. ulcerans taxa and 13 M. marinum taxa, correlating with the geographic origin of strains. Aligning conserved genes identified one PPE (proline-proline-glutamate) gene sequence to be species and intraspecies specific, thereby genotyping the 23 M. marinum/M. ulcerans complex taxa. PCR sequencing of the PPE gene correctly genotyped nine M. marinum/M. ulcerans complex isolates among one M. marinum taxon and three M. ulcerans taxa in the African taxon (T2.4). Further, successful PPE gene PCR sequencing in 15/21 (71.4%) swabs collected from suspected Buruli ulcer lesions in Côte d'Ivoire exhibited positive M. ulcerans IS2404 real-time PCR and identified the M. ulcerans T2.4.1 genotype in eight swabs and M. ulcerans T2.4.1/T2.4.2 mixed genotypes in seven swabs. PPE gene sequencing could be used as a proxy for whole-genome sequencing for the one-shot detection, identification, and typing of clinical M. ulcerans strains, offering an unprecedented tool for identifying M. ulcerans mixed infections. IMPORTANCE We describe a new targeted sequencing approach that characterizes the PPE gene to disclose the simultaneous presence of different variants of a single pathogenic microorganism. This approach has direct implications on the understanding of pathogen diversity and natural history and potential therapeutic implications when dealing with obligate and opportunistic pathogens, such as Mycobacterium ulcerans presented here as a prototype.

Project description:BackgroundBuruli ulcer, caused by Mycobacterium ulcerans, is a neglected tropical skin disease that is primarily endemic in West and Central Africa, including Côte d'Ivoire. Studies indicate that M. ulcerans infections are caused by contact with an environmental reservoir of the bacteria, governed by specific human biological conditions. Yet, the nature of this reservoir and the exact mode of transmission remain unknown.MethodologyTo identify ecologic risk factors of Buruli ulcer in south-central Côte d'Ivoire, we pursued a qualitative study matched with geo-referencing inquiry. Embedded in a broader integrated wound management research project, we (i) mapped households and water sources of laboratory confirmed Buruli ulcer cases and (ii) interviewed 12 patients and four health care workers to assess exposure to surface water and to deepen the understanding of perceived transmission pathways.Principal findingsWater availability, accessibility, and affordability were reported as key determinants for choosing water resources. Furthermore, perceived risks were related to environmental, structural, and individual factors. Despite the presence of improved water sources (e.g., drilled wells), communities heavily relied on unprotected surface water for a multitude of activities. The nearby Bandama River and seasonal waterbodies were frequently used for washing, bathing, and collection of water for drinking and cooking. Many residents also reported to cross the river on a daily basis for agricultural chores, and hence, are exposed to stagnant water during farming activities.Conclusions/significanceOur study in two Buruli ulcer endemic villages in south-central Côte d'Ivoire revealed a wide range of water-related domestic activities that might expose people to an increased risk of contracting the disease. Environmental, biological, social, and cultural risk factors are closely interlinked and should be considered in future investigations of Buruli ulcer transmission. Active participation of the communities is key to better understand their circumstances to advance research and fight against Buruli ulcer and other neglected tropical diseases.

Dataset Information

Distribution and Risk of Mycolactone-Producing Mycobacteria Transmission within Buruli Ulcer Endemic Communities in Cote d'Ivoire.

Publications

Distribution and Risk of Mycolactone-Producing Mycobacteria Transmission within Buruli Ulcer Endemic Communities in Côte d'Ivoire.

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