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Isolation and characterization of new Leptospira genotypes from patients in Mayotte (Indian Ocean).

ABSTRACT: BACKGROUND: Leptospirosis has been implicated as a severe and fatal form of disease in Mayotte, a French-administrated territory located in the Comoros archipelago (southwestern Indian Ocean). To date, Leptospira isolates have never been isolated in this endemic region. METHODS AND FINDINGS: Leptospires were isolated from blood samples from 22 patients with febrile illness during a 17-month period after a PCR-based screening test was positive. Strains were typed using hyper-immune antisera raised against the major Leptospira serogroups: 20 of 22 clinical isolates were assigned to serogroup Mini; the other two strains belonged to serogroups Grippotyphosa and Pyrogenes, respectively. These isolates were further characterized using partial sequencing of 16S rRNA and ligB gene, Multi Locus VNTR Analysis (MLVA), and pulsed field gel electrophoresis (PFGE). Of the 22 isolates, 14 were L. borgpetersenii strains, 7 L. kirschneri strains, and 1, belonging to serogoup Pyrogenes, was L. interrogans. Results of the genotyping methods were consistent. MLVA defined five genotypes, whereas PFGE allowed the recognition of additional subgroups within the genotypes. PFGE fingerprint patterns of clinical strains did not match any of the patterns in the reference strains belonging to the same serogroup, suggesting that the strains were novel serovars. CONCLUSIONS: Preliminary PCR screening of blood specimen allowed a high isolation frequency of leptospires among patients with febrile illness. Typing of leptospiral isolates showed that causative agents of leptospirosis in Mayotte have unique molecular features.

SUBMITTER: Bourhy P

PROVIDER: S-EPMC2889827 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature

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Isolation and characterization of new Leptospira genotypes from patients in Mayotte (Indian Ocean).

Bourhy Pascale P Collet Louis L Clément Sabine S Huerre Michel M Ave Patrick P Giry Claude C Pettinelli François F Picardeau Mathieu M

PLoS neglected tropical diseases 20100622 6

<h4>Background</h4>Leptospirosis has been implicated as a severe and fatal form of disease in Mayotte, a French-administrated territory located in the Comoros archipelago (southwestern Indian Ocean). To date, Leptospira isolates have never been isolated in this endemic region.<h4>Methods and findings</h4>Leptospires were isolated from blood samples from 22 patients with febrile illness during a 17-month period after a PCR-based screening test was positive. Strains were typed using hyper-immune a ...[more]

PMID: 20582311

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Project description:IntroductionHuman angiostrongyliasis (HA) is a neurological helminthic disease caused by the lung worm Angiostrongylus cantonensis. It is suspected in the combination of travel or a residence in an endemic area and eosinophilic meningitis. In Mayotte, an island in the Indian Ocean, cases are rare but regular. The main objective of our study was to describe the epidemiological and diagnosis clues of HA in Mayotte. The secondary objectives were to evaluate the contribution of Real-Time Polymerase Chain Reaction (RT- PCR) for the diagnosis of HA, delineate the characteristics of the local transmission and ascertain the presence of A. cantonensis in Achatina fulica, the potential vector of the disease.Materials and methodsBetween 2007 and 2012, all cases of eosinophilic meningitis were retrospectively included and investigated by RT- PCR in the CSF. Descriptive analysis was conducted for clinical, biological and radiological features, and were analyzed for all patients together with the search for prognostic factors for mortality. Concurrently, geolocalization and temporal parameters were studied to correlate the occurrence of the cases with rainfall seasons and snails were collected to enhance a parasitic carriage with real time PCR.ResultsDuring the 6-year period of the study, 14 cases were identified (2.3 cases/year) and 9 among 10 remaining CSF were positive in PCR. Among 14 cases of EM, 13 were less than 2 year-old children. The 1 year mortality rate was 5/14 (35.7%). Among survivors, 3/7 (42.8%) presented neurological sequelae. Factors associated with mortality were dysfunction of cranial nerves, abnormal brain imaging, and CSF glucose level inferior to 2 mmol/l. Occurrence of cases was temporarily and spatially correlated to the rainy season. Among the 64 collected giant snails, 6 (9.4%) were positive with A. cantonensis PCR. The likely main route of transmission was the children licking snails, carriers of the parasite.ConclusionIn Mayotte, HA was mainly found in paediatric cases under 2 years old, and evidenced a life-threatening disease. PCR seems to be a promising tool in the definitive diagnosis of HA. Population should be aware of the role of A. fulica, and not let the children have direct contact with the snails.

Project description:BackgroundThe south-west insular territories of the Indian Ocean have recently received attention concerning the diversity of arthropods of medical or veterinary interest. While a recent study highlighted the circulation of Culicoides-borne viruses, namely bluetongue and epizootic hemorrhagic disease, with clinical cases in Mayotte (comprising two islands, Petite-Terre and Grand-Terre), Comoros Archipelago, no data have been published concerning the species diversity of Culicoides present on the two islands.ResultsA total of 194,734 biting midges were collected in 18 sites, covering two collection sessions (April and June) in Mayotte. Our study reports for the first time livestock-associated Culicoides species and recorded at least 17 described Afrotropical species and one undescribed species. The most abundant species during the April collection session were C. trifasciellus (84.1%), C. bolitinos (5.4%), C. enderleini (3.9%), C. leucostictus (3.3%) and C. rhizophorensis (2.1%). All other species including C. imicola represented less than 1% of the total collection. Abundance ranged between 126-78,842 females with a mean and median abundance of 14,338 and 5111 individuals/night/site, respectively. During the June collection, the abundance per night was low, ranging between 6-475 individuals. Despite low abundance, C. trifasciellus and C. bolitinos were still the most abundant species. Culicoides sp. #50 is recorded for the first time outside South Africa.ConclusionsOur study reports for the first time the Culicoides species list for Mayotte, Comoros Archipelago, Indian Ocean. The low abundance and rare occurrence of C. imicola, which is usually considered the most abundant species in the Afrotropical region, is unexpected. The most abundant and frequent species is C. trifasciellus, which is not considered as a vector species so far, but its role needs further investigation. Further work is needed to describe Culicoides sp. #50 and to carry on faunistic investigations on the other islands of the archipelago as well as in neighboring countries.

Project description:Lake Dziani Dzaha is a thalassohaline tropical crater lake located on the "Petite Terre" Island of Mayotte (Comoros archipelago, Western Indian Ocean). Stromatolites are actively growing in the shallow waters of the lake shores. These stromatolites are mainly composed of aragonite with lesser proportions of hydromagnesite, calcite, dolomite, and phyllosilicates. They are morphologically and texturally diverse ranging from tabular covered by a cauliflower-like crust to columnar ones with a smooth surface. High-throughput sequencing of bacterial and archaeal 16S rRNA genes combined with confocal laser scanning microscopy (CLSM) analysis revealed that the microbial composition of the mats associated with the stromatolites was clearly distinct from that of the Arthrospira-dominated lake water. Unicellular-colonial Cyanobacteria belonging to the Xenococcus genus of the Pleurocapsales order were detected in the cauliflower crust mats, whereas filamentous Cyanobacteria belonging to the Leptolyngbya genus were found in the smooth surface mats. Observations using CLSM, scanning electron microscopy (SEM) and Raman spectroscopy indicated that the cauliflower texture consists of laminations of aragonite, magnesium-silicate phase and hydromagnesite. The associated microbial mat, as confirmed by laser microdissection and whole-genome amplification (WGA), is composed of Pleurocapsales coated by abundant filamentous and coccoid Alphaproteobacteria. These phototrophic Alphaproteobacteria promote the precipitation of aragonite in which they become incrusted. In contrast, the Pleurocapsales are not calcifying but instead accumulate silicon and magnesium in their sheaths, which may be responsible for the formation of the Mg-silicate phase found in the cauliflower crust. We therefore propose that Pleurocapsales and Alphaproteobacteria are involved in the formation of two distinct mineral phases present in the cauliflower texture: Mg-silicate and aragonite, respectively. These results point out the role of phototrophic Alphaproteobacteria in the formation of stromatolites, which may open new perspective for the analysis of the fossil record.

Project description:Extensive studies on the taxonomic resolution required for bioassessment purposes have determined that resolution above species level (genus, family) is sufficient for their use as indicators of relevant environmental pressures. The high-throughput sequencing (HTS) and meta-barcoding methods now used for bioassessment traditionally employ an arbitrary sequence similarity threshold (SST) around 95% or 97% to cluster sequences into operational taxonomic units, which is considered descriptive of species-level resolution. In this study, we analyzed the effect of the SST on the resulting diatom-based ecological quality index, which is based on OTU abundance distribution along a defined environmental gradient, ideally avoiding taxonomic assignments that could result in high rates of unclassified OTUs and biased final values. A total of 90 biofilm samples were collected in 2014 and 2015 from 51 stream sites on Mayotte Island in parallel with measures of relevant physical and chemical parameters. HTS sequencing was performed on the biofilms using the rbcL region as the genetic marker and diatom-specific primers. Hierarchical clustering was used to group sequences into OTUs using 20 experimental SST levels (80%-99%). An OTU-based quality index (IdxOTU) was developed based on a weighted average equation using the abundance profiles of the OTUs. The developed IdxOTU revealed significant correlations between the IdxOTU values and the reference pressure gradient, which reached maximal performance using an SST of 90% (well above species level delimitation). We observed an interesting and important trade-off with the power to discriminate between sampling sites and index stability that will greatly inform future applications of the index. Taken together, the results from this study detail a thoroughly optimized and validated approach to generating robust, reproducible, and complete indexes that will greatly facilitate effective and efficient environmental monitoring.

Dataset Information

Isolation and characterization of new Leptospira genotypes from patients in Mayotte (Indian Ocean).

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Isolation and characterization of new Leptospira genotypes from patients in Mayotte (Indian Ocean).

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