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Here, we describe the genome sequence of ECC486. This Enterobacter oligotrophicus strain was isolated from a wild Anolis marmoratus speciosus specimen, a lizard endemic to the territory of Guadeloupe (French West Indies). Its draft genome sequence consists of 40 contigs and contains a total of 4,504,233 bp, with a G+C content of 54.1%.

14 pages; International audience; Irregular urchins exclusively live in marine soft bottom habitats, dwelling either upon or inside sediments and selectively picking up sediment grains and organic particles, or swallowing bulk sediment to feed on the associated organic matter. The exact food source and dietary requirements of most irregular echinoids, however, remain incompletely understood. The schizasterid species Abatus cordatus (Verrill, 1876) is a sub-Antarctic spatangoid that is endemic to the Kerguelen. The feeding behaviour of A. cordatus was investigated using simultaneously metabarcoding and stable isotope approaches. Comparison of ingested and surrounding sediments by metabarcoding revealed a limited selective ingestion of prokaryotes and eukaryotes by the urchin. Compared to surrounding sediments, the gut content had (i) higher carbon and nitrogen concentrations potentially due to selective ingestion of organic matter and/or the sea urchin mucus secretion and (ii) δ15N enrichment due to the selective assimilation of lighter isotope in the gut. Feeding experiments were performed using 13C and 15 N-enriched sediments in aquariums. The progression of stable isotope enrichment in proximal and distal parts of the digestive track of A. cordatus revealed that all particles are not similarly transported likely due to siphon functioning. Ingestion of water with associated dissolved and particulate organic matter should play an important role in urchin nutrition. A. cordatus had a gut resident time fluctuating between 76 and 101 h and an ingestion rate of 36 mg dry sediment h−1 suggesting that dense populations of the species may play a key ecological role through bioturbation in soft bottom shallow-water habitats of the Kerguelen Islands.

International audience; Wastewater treatment plants are considered hot spots for antibiotic resistance. Most studies have addressed the impact on the aquatic environment, as water is an important source of anthropogenic pollutants. Few investigations have been conducted on terrestrial animals living near treatment ponds. We isolated extended-spectrum-β-lactamase Enterobacter cloacae complex-producing strains from 35 clinical isolates, 29 samples of wastewater, 19 wild animals, and 10 domestic animals living in the hospital sewers and at or near a wastewater treatment plant to study the dissemination of clinically relevant resistance through hospital and urban effluents. After comparison of the antibiotic-resistant profiles of E. cloacae complex strains, a more detailed analysis of 41 whole-genome-sequenced strains demonstrated that the most common sequence type, ST114 (n = 20), was present in human (n = 9) and nonhuman (n = 11) samples, with a close genetic relatedness. Whole-genome sequencing confirmed local circulation of this pathogenic lineage in diverse animal species. In addition, nanopore sequencing and specific synteny of an IncHI2/ST1/blaCTX-M-15 plasmid recovered on the majority of these ST114 clones (n = 18) indicated successful worldwide diffusion of this mobile genetic element.

Wastewater treatment plants are considered hot spots for antibiotic resistance. Most studies have addressed the impact on the aquatic environment, as water is an important source of anthropogenic pollutants., Wastewater treatment plants are considered hot spots for antibiotic resistance. Most studies have addressed the impact on the aquatic environment, as water is an important source of anthropogenic pollutants. Few investigations have been conducted on terrestrial animals living near treatment ponds. We isolated extended-spectrum-β-lactamase Enterobacter cloacae complex-producing strains from 35 clinical isolates, 29 samples of wastewater, 19 wild animals, and 10 domestic animals living in the hospital sewers and at or near a wastewater treatment plant to study the dissemination of clinically relevant resistance through hospital and urban effluents. After comparison of the antibiotic-resistant profiles of E. cloacae complex strains, a more detailed analysis of 41 whole-genome-sequenced strains demonstrated that the most common sequence type, ST114 (n = 20), was present in human (n = 9) and nonhuman (n = 11) samples, with a close genetic relatedness. Whole-genome sequencing confirmed local circulation of this pathogenic lineage in diverse animal species. In addition, nanopore sequencing and specific synteny of an IncHI2/ST1/blaCTX-M-15 plasmid recovered on the majority of these ST114 clones (n = 18) indicated successful worldwide diffusion of this mobile genetic element.

Limited data are available on the contribution of wildlife to the spread of antibacterial resistance. We determined the prevalence of resistance to antibiotics in Escherichia coli isolates collected from wild animals in 2013 and 2014 and the genetic basis for resistance to third-generation cephalosporin in Guadeloupe. We recovered 52 antibiotic-resistant (AR) E. coli strains from 48 of the 884 (5.4%) wild animals tested (46 iguanas, 181 birds, 289 anoles, and 368 rodents at 163 sampling sites). Rodents had higher rates of carriage (n = 38, 10.3%) than reptiles and birds (2.4% and 1.1%, respectively, p < 0.001). A significant association (p < 0.001) was found between the degree of anthropization and the frequency of AR E. coli carriage for all species. The carriage rate of ciprofloxacin- and cefotaxime-resistant isolates was 0.7% (6/884) and 1.5% (13/884), respectively. Most (65.4%) AR E. coli were multi-drug resistant, and the prevalence of extended-spectrum beta-lactamase (ESBL)-producing E. coli was low (n = 7, 0.8%) in all species. Eight ESBL-producing E. coli were recovered, two genetically unrelated isolates being found in one bird. These isolates and 20 human invasive ESBL E. coli isolates collected in Guadeloupe during the same period were investigated by whole genome sequencing. blaCTX–M–1 was the only ESBL gene shared by three animal classes (humans, n = 2; birds, n = 2; rodents, n = 2). The blaCTX–M–1 gene and most of the antimicrobial resistance genes were present in a large conjugative IncI1 plasmid that was highly similar (>99% nucleotide identity) to ESBL-carrying plasmids found in several countries in Europe and in Australia. Although the prevalence of ESBL-producing E. coli isolates was very low in wild animals, it is of concern that the well-conserved IncI1 plasmid-carrying blaCTX–M–1 is widespread and occurs in various E. coli strains from animals and humans.

Free-living amoebae (FLA) are ubiquitous protists. Pathogenic FLA such as N. fowleri can be found in hot springs in Guadeloupe, soil being the origin of this contamination. Herein, we analyzed the diversity and distribution of FLA in soil using a targeted metataxonomic analysis. Soil samples (n = 107) were collected from 40 sites. DNA was extracted directly from soil samples or from FLA cultivated at different temperatures (30, 37 and 44 &deg, C). Metabarcoding studies were then conducted through FLA 18SrDNA amplicons sequencing, amplicon sequence variants (ASV) were extracted from each sample and taxonomy assigned against SILVA database using QIIME2 and SHAMAN pipelines. Vermamoeba were detected in DNA extracted directly from the soil, but to detect other FLA an amoebal enrichment step was necessary. V. vermiformis was by far the most represented species of FLA, being detected throughout the islands. Although Naegleria were mainly found in Basse-Terre region, N. fowleri was also detected in Grand Terre and Les Saintes Islands. Acanthamoeba were mainly found in areas where temperature is approx. 30 &deg, C. Vannella and Vahlkampfia were randomly found in Guadeloupe islands. FLA detected in Guadeloupe include both pathogenic genera and genera that can putatively harbor microbial pathogens, therefore posing a potential threat to human health.

International audience; Extended-spectrum beta-lactamases (ESBLs), especially those of the CTX-M type, represent a major public health problem throughout the world. Although the carriage of ESBL-producing Enterobacteriaceae (EPE) in feces of horses is now well recognized, little is known about the diversity of EPE after treatment of horses with antibiotics. We undertook this study to assess and follow the diversity of EP Escherichia coli isolated from horses after antibiotic treatment for an infection. Fecal samples from two horses treated and two that were untreated were tested for the presence of EPE on different days. All isolated E. coli strains were evaluated for antimicrobial resistance (AMR) and by whole-genome sequencing. Multi locus sequence typing, phylogrouping, resistance genes and plasmid content were extracted from genomic data. A phylogenetic analysis based on single nucleotide polymorphism (SNP) divergence was also performed on the core genome. We isolated 35 strains belonging to the A, B1 and C phylo-groups. All but one expressed SHV-12 enzymes and one expressed CTX-M-1. Intra- and inter-horse genetic diversity of E. coli strains was identified in the genome analysis and 10 AMR profiles. Two distinct EP E. coli-resistant populations (phylo-group B1: ST4164-AMR3 and ST155-AMR2) were found in one horse, and five other resistant populations were found in the second horse (phylo-group A: ST1250-AMR1; phylo-group B1: ST1250-AMR1, ST6981-AMR1 and phylo-group C: ST10-AMR4). Some persistent EP E. coli strains were detected at least 1 month after treatment. These results indicate that EP E. coli strains isolated from horse feces show intra- and inter-host genetic diversity, even in a region with low ESBL prevalence and in horses that are rarely treated with third-generation cephalosporins. These results also suggest that horizontal gene transfer and/or selection of resistance genes probably occurs in vivo within the horse gut microbiome. Follow-up of EP E. coli resistance profiles for at least 1 month after treatment is warranted to prevent persistence of EP E. coli.

Background: Immature stages of Aedes aegypti develop in many man-made aquatic habitats in which mosquito larvae are exposed to physicochemical elements and microorganisms that may influence their life cycle and their ability to transmit human arboviruses. Despite the omnipresence of Ae. aegypti in tropical and subtropical regions, little is known about the natural bacterial communities associated with this mosquito or their relation to the biotic and abiotic characteristics of their aquatic habitats. We comprehensively characterized the physicochemical properties and bacterial microbiome of Ae. aegypti breeding sites and larvae on Guadeloupe and in French Guiana. In addition, we explored whether geographical location, the type of breeding site and physicochemical parameters of the water influenced the microbiota associated with this mosquito species.Method: We used large-scale 16S rRNA gene sequencing of 160 breeding sites and 147 pools of Ae. aegypti larvae from sites widely distributed across Guadeloupe and French Guiana and recorded 13 physicochemical parameters at the sampled breeding sites. Ordination plots and multiple linear regression were used to assess the influence of environmental factors on the bacterial microbiome of water and larvae.Results: We found territory-specific differences in physicochemical properties (dissolved oxygen, conductivity and metal content) and the composition of bacterial communities in Ae. aegypti breeding sites that influenced the relative abundance of several bacteria genera (Methylobacterium, Roseoccocus) on the corresponding larvae. A significant fraction of the bacterial communities identified on Ae. aegypti larvae, dominated by Herbiconiux and Microvirga genera, were consistently enriched in mosquitoes regardless the location. Conclusion: Territory-specific differences observed in the biotic and abiotic properties of Ae. aegypti breeding sites influenced a fraction of the microbial communities of the corresponding larvae, raising concern about the impact of these changes on pathogen transmission by different Ae. aegypti populations.

International audience; The East African Indian (EAI) and Central Asian (CAS) lineages of Mycobacterium tuberculosis complex (MTBC) mainly infect tuberculosis (TB) patients in the eastern hemisphere which contains many of the 22 high TB burden countries including China and India. We investigated if phylogeographical, epidemiological and demographical characteristics for these 2 lineages differed in SITVIT2 database. Genotyping results and associated data (age, sex, HIV serology, drug resistance) on EAI and CAS lineages (n = 10,974 strains) were extracted. Phylogenetic and Bayesian, and other statistical analyses were used to compare isolates. The male/female sex ratio was 907/433 (2.09) for the EAI group vs. 881/544 (1.62) for CAS (p-value

Background Limited epidemiological and antimicrobial resistance data are available on Salmonella enterica from sub-Saharan Africa. We determine the prevalence of resistance to antibiotics in isolates in the Central African Republic (CAR) between 2004 and 2013 and the genetic basis for resistance to third-generation cephalosporin (C3G). Methodology/Principal findings A total of 582 non-duplicate human clinical isolates were collected. The most common serotype was Typhimurium (n = 180, 31% of the isolates). A randomly selected subset of S. Typhimurium isolates were subtyped by clustered regularly interspaced short palindromic repeat polymorphism (CRISPOL) typing. All but one invasive isolate tested (66/68, 96%) were associated with sequence type 313. Overall, the rates of resistance were high to traditional first-line drugs (18–40%) but low to many other antimicrobials, including fluoroquinolones (one resistant isolate) and C3G (only one ESBL-producing isolate). The extended-spectrum beta-lactamase (ESBL)-producing isolate and three additional ESBL isolates from West Africa were studied by whole genome sequencing. The blaCTX-M-15 gene and the majority of antimicrobial resistance genes found in the ESBL isolate were present in a large conjugative IncHI2 plasmid highly similar (> 99% nucleotide identity) to ESBL-carrying plasmids found in Kenya (S. Typhimurium ST313) and also in West Africa (serotypes Grumpensis, Havana, Telelkebir and Typhimurium). Conclusions/Significance Although the prevalence of ESBL-producing Salmonella isolates was low in CAR, we found that a single IncHI2 plasmid-carrying blaCTX-M-15 was widespread among Salmonella serotypes from sub-Saharan Africa, which is of concern., Author summary Salmonella enterica infections are common causes of bloodstream infection in sub-Saharan Africa and associated with a high mortality rate. Levels of multidrug resistance have become alarmingly high. Then, third-generation cephalosporin (C3G) and fluoroquinolones have become standard for first-line empirical treatment. Recently, C3G-resistant Salmonella populations have emerged and spread over all continents. This resistance is mainly mediated by acquired extended-spectrum beta-lactamase (ESBL) genes carried by mobile genetic elements such as plasmids. We report here the prevalence of resistance to antibiotics in isolates in the Central African Republic (CAR) between 2004 and 2013 and the genetic basis for resistance to C3G. Overall, resistance rates to antimicrobials were low during the study period, for all classes other than conventional antimicrobials, confirming recommendations for first-line treatment based on C3G and fluoroquinolones. Only one ESBL-producing isolate was recovered. The ESBL gene and the majority of antimicrobial resistance genes found were present in a large plasmid highly similar to ESBL-carrying plasmids found in East and West Africa, highlighting its significant role in the spread of ESBL genes in Salmonella isolates in sub-Saharan Africa. These finding have implications for treatment of salmonellosis and support the growing necessity for increased microbiological surveillance based on networks of clinical laboratories in order to control dissemination of antibiotic resistance among Salmonella isolates.

Aims and objectives: Bioinformatics and computational tools are increasingly used in a wide variety of scientific studies. Genotyping, genomics and whole genome sequencing (WGS) methods allowed the generation of high amounts of data that are difficult to analyze manually. The purpose of this review is to establish a potential list of available databases and bioinformatics tools/resources related to Mycobacterium tuberculosis complex (MTBC), the causative agent of tuberculosis (TB). Methods: At present, a total of 36 tools with freely-accessible links were recovered from publications indexed in PubMed or Google Scholar. Following keywords were searched: “Mycobacterium tuberculosis AND database”, “Mycobacterium tuberculosis AND web tool”, “Mycobacterium tuberculosis AND tool”, “Mycobacterium tuberculosis AND bioinformatics”. Resources found were classified into several thematic sections/categories such as biological databases, drug resistance, or prediction/classification tools. Among recently available resources (published in 2019), one can mention COMBAT-TB-NeoDB ( https://neodb.sanbi.ac.za/browser/ ), CPLP-TB ( http://cplp-tb.ff.ulisboa.pt/ ), DeepAMR ( http://www.robots.ox.ac.uk/~davidc/code.php ), SecProMTB ( http://server.malab.cn/SecProMTB/index.jsp ), SITVIT2 ( http://www.pasteur-guadeloupe.fr:8081/SITVIT2 ), SNP-IT ( https://github.com/samlipworth/snpit ), StackTB ( https://stacktb.cs.sfu.ca/ ), and TBProfiler ( http://tbdr.lshtm.ac.uk/ ). Results: Although several tools/resources have been developed over time, many are either not regularly updated, or not made available permanently. We argue in favor of sustainable development of resources in order to assist the scientific community and policy makers to take timely concerted action for TB control. At “Institut Pasteur de la Guadeloupe”, we intend to build a newer generation of SITVIT databases including classical genotyping, epidemiologic and demographic data, information on drug resistance, as well as WGS data. Several thousands of complete genome sequences are today available for MTBC strains on public sequence repositories (such as GenBank, DDBJ, or ENA). Thanks to recently developed tools, data extracted from available complete sequences would be juxtaposed and stored for comparative and molecular epidemiological studies. Conclusion: Although, the present non-exhaustive list is certainly not “revolutionary”, it would nonetheless foster the construction of integrated pipelines/tools related to specific biological questions concerning MTBC. This list will be made available soon through Wikipedia; and in the meantime it is already available for consultation via link: http://www.pasteur-guadeloupe.fr:8081/SpolSimilaritySearch/info.jsp .

International audience; Mycobacterium tuberculosis genetic structure, and evolutionary history have been studied for years by several genotyping approaches, but delineation of a few sublineages remains controversial and needs better characterization. This is particularly the case of T group within lineage 4 (L4) which was first described using spoligotyping to pool together a number of strains with ill-defined signatures. Although T strains were not traditionally considered as a real phylogenetic group, they did contain a few phylogenetically meaningful sublineages as shown using SNPs. We therefore decided to investigate if this observation could be corroborated using other robust genetic markers. We consequently made a first assessment of genetic structure using 24-loci MIRU-VNTRs data extracted from the SITVIT2 database (n = 607 clinical isolates collected in Russia, Albania, Turkey, Iraq, Brazil and China). Combining Minimum Spanning Trees and Bayesian population structure analyses (using STRUCTURE and TESS softwares), we distinctly identified eight tentative phylogenetic groups (T1-T8) with a remarkable correlation with geographical origin. We further compared the present structure observed with other L4 sublineages (n = 416 clinical isolates belonging to LAM, Haarlem, X, S sublineages), and showed that 5 out of 8 T groups seemed phylogeographi-cally well-defined as opposed to the remaining 3 groups that partially mixed with other L4 isolates. These results provide with novel evidence about phylogeographically specificity of a proportion of ill-defined T group of M. tuberculosis. The genetic structure observed will now be further validated on an enlarged worldwide dataset using Whole Genome Sequenc-ing (WGS).

International audience; A Q fever epidemic occurred in 2013 in a small military residential area in Cayenne, French Guiana. A retrospective cohort study was conducted to identify Q fever risk factors. Confirmed acute Q fever case was defined as positive serology (IgM >= 50 and phase II IgG >= 200) and/or positive qPCR on serum or blood. In addition, wild mammals were captured at the study site and tested by serology and real-time PCR performed on blood, vaginal swabs and ticks. The attack rate was 20 percent (11/54). All the cases were symptomatic with fever > 38.5 degrees C and community-acquired pneumonia for four cases. Log binomial multivariate models identified two independent risk factors associated with Q fever: to clean the house (RRa = 7.5 CI95% [1.03-55.3]) and to carry a three-toed sloth in arms (RRa = 2.6 CI95% [1.1-5.8]). Eighteen marsupial individuals were captured, all PCRs were negative but 17% (3/18) had a positive serology. Another study conducted after the epidemic found only one (1/4) three-tooth sloth (Bradypus tridactylus) with feces highly infectious for C. burnetii MST17. The same strain C. burnetii genotype 17 has been laboratory-confirmed in this mammal and in human cases. These results support the implication of three-toed-sloth in this epidemic. Human contamination mainly occurs through inhalation of infectious aerosols as suggested by high relative risk associated with house cleaning activities and pulmonary forms of the disease, and through direct contact with three- toed-sloth. Positive serological results among marsupials confirm wildlife exposure and suggest a more complex sylvatic transmission cycle among wild mammals.

International audience; Leptospirosis is a cosmopolitan zoonosis caused by bacteria of the genus Leptospira. Whether the distribution is worldwide, the hot and humid climate of the tropics is particularly conducive to its expansion. In most French overseas departments and territories, leptospirosis is considered as a public health problem. In French Guiana, a French department located in the northeastern part of the Amazon rainforest, it is supposed to be rare. The objective of this review was to make an inventory of the knowledge on human and animal leptospirosis in French Guiana and neighboring countries. A comprehensive search was conducted through the indexed and informal medical literature in English, French, Spanish and Portuguese. Thus, respectively ten and four publications were identified on human and animal leptospirosis in French Guiana, published between 1940 and 1995 in the form of case reports or case series. The publications concerning this disease in the other countries of the Guiana Shield, eastern Venezuela, Guyana, Suriname, and Brazilian state of Amapá, also scarce or nonexistent. However recent data from the French National Centre of leptospirosis showed a recent and sudden increase in the number of cases in the department, probably partly due to the development of diagnostic tools such as Elisa IgM serology. It is likely that leptospirosis is a neglected disease in the region, due to the lack of diagnostic tools readily available, the lack of knowledge of the local clinicians on this disease and the existence of many other pathogens with similar clinical presentation such as malaria, arboviruses and Q fever and Amazonian toxoplasmosis. The establishment of more large-scale studies on animal and human leptospirosis is necessary and urgent to know the true burden of this disease in our region.

Summary Background Mycobacterium ulcerans infection is the third most common mycobacterial disease in the world after tuberculosis and leprosy. To date, transmission pathways from its environmental reservoir to humans are still unknown. In South America, French Guiana has the highest reported number of M ulcerans infections across the continent. This empirical study aimed to characterise the epidemiology of M ulcerans infection in French Guiana between 1969 and 2013. Methods Data were collected prospectively mainly by two dermatologists at Cayenne Hospital's dermatology department between Jan 1, 1969, and Dec 31, 2013, for age, date of diagnosis, sex, residence, location of the lesion, type of lesion, associated symptoms, and diagnostic method (smear, culture, PCR, or histology) for all confirmed and suspected cases of M ulcerans . We obtained population data from censuses. We calculated mean M ulcerans infection incidences, presented as the number of cases per 100 000 person-years. Findings 245 patients with M ulcerans infections were reported at Cayenne Hospital's dermatology department during the study period. M ulcerans infection incidence decreased over time, from 6·07 infections per 100 000 person-years (95% CI 4·46–7·67) in 1969–83 to 4·77 infections per 100 000 person-years (3·75–5·79) in 1984–98 and to 3·49 infections per 100 000 person-years (2·83–4·16) in 1999–2013. The proportion of children with infections also declined with time, from 42 (76%) of 55 patients in 1969–83 to 26 (31%) of 84 in 1984–98 and to 22 (21%) of 106 in 1999–2013. Most cases occurred in coastal areas surrounded by marshy savannah (incidence of 21·08 per 100 000 person-years in Sinnamary and 21·18 per 100 000 person-years in Mana). Lesions mainly affected limbs (lower limbs 161 [66%] patients; upper limbs 60 [24%] patients). We diagnosed no bone infections. Interpretation The decrease of M ulcerans infection incidence and the proportion of children with infections over a 45 year period in this ultra-peripheral French territory might have been mostly driven by improving living conditions, prophylactic recommendations, and access to health care. Funding Agence Nationale de la Recherche.

BACKGROUND:In recent years, first-line therapy for Mycobacterium ulcerans infection in French Guiana has consisted of antibiotics active against this organism. Two regimens are used comprising rifampicin associated with clarithromycin or amikacin.PATIENTS AND METHODS:We describe four patients presenting apparent worsening of their lesions during treatment: ulceration of a nodular lesion in a 32-year-old woman and worsening of an ulcerated lesion in three patients aged 16, 27 and 79 years.DISCUSSION:In these 4 patients, we concluded that the symptoms were caused by a paradoxical response or a reaction, a phenomenon already described in tuberculosis and leprosy. Such worsening is transient and must not be misinterpreted as failure to respond to treatment. The most plausible pathophysiological hypothesis involves the re-emergence of potentially necrotizing cellular immunity secondary to the loss of mycolactone, a necrotizing and immunosuppressive toxin produced by M. ulcerans, resulting from the action of the antibiotics., Introduction. — Depuis quelques années, en Guyane, l’infection à Mycobacterium ulcerans (ulcère de Buruli) est traitée en première intention par des antibiotiques actifs sur cette mycobactérie. Deux schémas thérapeutiques sont utilisés, comprenant de la rifampicine associée àde la clarithromycine ou à de l’amikacine.Observation. — Nous décrivons quatre cas de patients présentant une apparente aggravation des lésions durant leur traitement, soit à type d’ulcération de lésions initialement nodulaires chez une patiente de 32 ans, soit à type d’aggravation d’une ulcération préexistante chez des patients de respectivement 16, 27 et 79 ans.Discussion. — Chez ces quatre patients, nous concluons à des réponses ou des réactions paradoxales, phénomène déjà décrit au cours de la tuberculose et de la lèpre. Ces aggravations sont transitoires et ne doivent pas être interprétées comme des échecs thérapeutiques. L’hypothèse physiopathologique la plus probable implique la restauration d’une immunitécellulaire antérieurement inhibée par la mycolactone, une toxine nécrosante et immunosuppressive produite par M. ulcerans. La disparition de cette toxine, sous l’effet des antibiotiques, serait responsable de la réaction immunitaire à l’origine d’une nécrose des tissus infectés.

International audience; Tuberculosis (TB) remains broadly present in the Americas despite intense global efforts for its control and elimination. Starting from a large dataset comprising spoligotyping (n = 21183 isolates) and 12-loci MIRU-VNTRs data (n = 4022 isolates) from a total of 31 countries of the Americas (data extracted from the SITVIT2 database), this study aimed to get an overview of lineages circulating in the Americas. A total of 17119 (80.8%) strains belonged to the Euro-American lineage 4, among which the most predominant genotypic family belonged to the Latin American and Mediterranean (LAM) lineage (n = 6386, 30.1% of strains). By combining classical phylogenetic analyses and Bayesian approaches, this study revealed for the first time a clear genetic structuration of LAM9 sublineage into two subpopulations named LAM9C1 and LAM9C2, with distinct genetic characteristics. LAM9C1 was predominant in Chile, Colombia and USA, while LAM9C2 was predominant in Brazil, Dominican Republic, Guadeloupe and French Guiana. Globally, LAM9C2 was characterized by higher allelic richness as compared to LAM9C1 isolates. Moreover, LAM9C2 sublineage appeared to expand close to twenty times more than LAM9C1 and showed older traces of expansion. Interestingly, a significant proportion of LAM9C2 isolates presented typical signature of ancestral LAM-RDRio MIRU-VNTR type (224226153321). Further studies based on Whole Genome Sequencing of LAM strains will provide the needed resolution to decipher the biogeographical structure and evolutionary history of this successful family.

Beijing lineage of Mycobacterium tuberculosis constitutes the most predominant lineage in East Asia. Beijing epidemiology, evolutionary history, genetics are studied in details for years revealing probable origin from China followed by worldwide expansion, partially linked to higher mutation rate, hypervirulence, drug-resistance, and association with cases of mixed infections. Considering huge amount of data available for 24-loci Mycobacterial Interspersed Repetitive Units-Variable Number of Tandem Repeats, we performed detailed phylogenetic and Bayesian population structure analyses of Beijing lineage strains in mainland China and Taiwan using available 24-loci MIRU-VNTR data extracted from publications or the SITVIT2 database (n = 1490). Results on genetic structuration were compared to previously published data. A total of three new Beijing clonal complexes tentatively named BSP1, BPS2 and BSP3 were revealed with surprising phylogeographical specificities to previously unstudied regions in Sichuan, Chongqing and Taiwan, proving the need for continued investigations with extended datasets. Such geographical restriction could correspond to local adaptation of these “ecological specialist” Beijing isolates to local human host populations in contrast with “generalist pathogens” able to adapt to several human populations and to spread worldwide.

Introduction L’infection a Mycobacterium ulcerans (IMU) est la troisieme maladie mycobacterienne la plus repandue apres la tuberculose et la lepre. A ce jour, les voies de transmission de son reservoir environnemental aux humains sont encore inconnues. En Amerique du sud, la Guyane francaise a le plus grand nombre d’IMU sur le continent. Cette etude visait a caracteriser l’epidemiologie de l’IMU en Guyane francaise entre 1969 et 2013. Materiel et methodes Les donnees ont ete recueillies prospectivement par les medecins du seul service de dermatologie de Guyane a l’hopital de Cayenne entre 1969 et 2013. Elles portaient sur l’âge, la date du diagnostic, le sexe, la residence, la localisation de la lesion, le type de lesion, les symptomes associes et les elements diagnostiques (frottis, culture, PCR, histopathologie) pour tous les cas d’IMU. Nous avons calcule les incidences moyennes d’IMU, presentees comme le nombre de cas pour 100 000 personnes–annees, a partir des donnees demographiques de la population generale provenant des recensements (INSEE). Resultats Au total, 245 patients atteints d’IMU ont ete diagnostiques au Service de dermatologie de l’hopital de Cayenne au cours de la periode d’etude. L’incidence de l’IMU en nombre de cas pour 100 000 personnes–annees a diminue au fil du temps passant de 6,07 en 1969–1983 a 4,77 en 1984–1998 et a 3,49 en 1999–2013, cette baisse etant statistiquement significative (β–0,14 ; p = 0,02). La proportion d’enfants atteints a egalement diminue significativement avec le temps, passant de 76 % en 1969–1983 a 31 % en 1984–1998 et a 21 % en 1999–2013 (p = 0,01). La plupart des cas se sont produits dans les zones cotieres entourees de savanes marecageuses (incidence de 21,8 pour 100 000 personnes–annees a Sinnamary et de 21,8 ans pour 100 000 personnes–annees a Mana). Les lesions ont principalement affecte les membres (membres inferieurs : 66 % ; membres superieurs : 24 %). Nous n’avons diagnostique aucune infection osseuse. Discussion Cette etude, la plus longue serie mondiale sur l’IMU, met en evidence une modification de l’epidemiologie de cette infection en Guyane ces 45 dernieres annees : d’une part, une baisse d’incidence globale, d’autre part, une augmentation de la moyenne d’âge des patients avec passage d’une epidemiologie de type africain (surtout des enfants) a une epidemiologie de type australien (surtout des adultes). L’amelioration des conditions de vie et de l’acces aux soins sont les probables principaux facteurs explicatifs de cette evolution.

Summary We recently showed that the Mycobacterium tuberculosis sublineage LAM9 could be subdivided as two distinct subpopulations – each reflecting its unique biogeographical structure and evolutionary history. We subsequently attempted to verify if this genetic structuration could be traced in an enlarged global sample. For this purpose, we analyzed global evolutionary relationships of LAM strains in a large dataset (n = 1923 isolates from 35 countries worldwide) with concomitant spoligotyping and MIRU-VNTR data, followed by a deeper analysis of LAM9 sublineage (n = 851 isolates). Based on a combination of phylogenetical analysis and Bayesian statistics, a total of three different clusters, tentatively named LAM9C1, C2 and C3 were described in this dataset. Closer inspection of the phylogenetic tree with concomitant data on origin of isolates with genetic clusterization revealed LAM9C3 being the most tightly knit group exclusively found in the Old World as opposed to LAM9C2 being a loosely-knit group without any phylogeographical specificity; while LAM9C1 appeared with a majority of strains being well-clustered despite some isolates that intermixed with unrelated LAM clusters. Subsequently, we hereby describe a new M . tuberculosis LAM sublineage named LAM9C3 with phylogeographical specificity for the Old World. These findings open new perspectives to study respective migration histories and adaptation to human hosts of specific M . tuberculosis clones during the exploration and conquest of the New World. We therefore plan to reevaluate the nomenclature and evolutionary history of various LAM sublineages using Whole Genome Sequencing (WGS).

International audience; Buruli ulcer is an emerging and neglected tropical disease caused by Mycobacterium ulcerans. Few cases have been reported so far in the Americas. With 250 cases reported since 1969, French Guiana is the only Buruli ulcer endemic area in the continent. Thus far, no genetic diversity studies of strains of M. ulcerans from French Guiana have been reported. Our goal in the present study was to examine the genetic diversity of M. ulcerans strains in this region by using the Multilocus Variable Number Tandem Repeat Analysis (MLVA) approach. A total of 23 DNA samples were purified from ulcer biopsies or derived from pure cultures. MVLA was used in the study of six previously-described Variable Number of Tandem Repeat (VNTR) markers. A total of three allelic combinations were characterized in our study: genotype I which has been described previously, genotype III which is very similar to genotype I, and genotype II which has distinctly different characteristics in comparison with the other two genotypes. This high degree of genetic diversity appears to be uncommon for M. ulcerans. Further research based on complete genome sequencing of strains belonging to genotypes I and II is in progress and should lead soon to a better understanding of genetic specificities of M. ulcerans strains from French Guiana.

Vibrio vulnificus is a ubiquitous marine bacterium that is responsible for infections and some seafood-related illnesses and deaths in the United States, mainly in individuals with compromised health status in the Gulf of Mexico region. Most phylogenetic studies focus on V. vulnificus strains isolated in the southern United States, but almost no genetic data are available on northeastern bacterial isolates of clinical or environmental origin. Our goal in this study was to examine the genetic diversity of environmental strains isolated from commercially-produced oysters and in clinical strains of known pathogenicity in northeastern United States. We conducted analyses of a total of eighty-three strains of V. vulnificus, including 18 clinical strains known to be pathogenic. A polyphasic, molecular-typing approach was carried out, based upon established biotypes, vcg, CPS, 16S rRNA types and three other genes possibly associated with virulence (arylsulfatase A, mtlABC, and nanA). An established Multi Locus Sequence Typing (MLST) method was also performed. Phylogenetic analyses of these markers and MLST results produced similar patterns of clustering of strains into two main lineages (we categorized as 'LI' and 'LII'), with clinical and environmental strains clustering together in both lineages. Lineage LII was comprised primarily but not entirely of clinical bacterial isolates. Putative virulence markers were present in both clinical and environmental strains. These results suggest that some northeastern environmental strains of V. vulnificus are phylogenetically close to clinical strains and probably are capable of virulence. Further studies are necessary to assess the risk of human illness from consuming raw oysters harvested in the northeastern US.

Extracting DNA from deep subsurface sediments is challenging given the complexity of sediments types, low bio-masses, resting structures (spores, cysts) frequently encountered in deep sediments, and the potential presence of enzymatic inhibitors. Promising results for cell lysis efficiency were recently obtained by use of a cryogenic mill (Lipp et al., 2008). These findings encouraged us to devise a DNA extraction protocol using this tool. Thirteen procedures involving a combination of grinding in liquid nitrogen (for various durations and beating rates) with different chemical solutions (phenol, chloroform, SDS, sarkosyl, proteinase, GTC), or with use of DNA recovery kits (MagExtractor (R)) were compared. Effective DNA extraction was evaluated in terms of cell lysis efficiency, DNA extraction efficiency, DNA yield and determination of prokaryotic diversity. Results were compared to those obtained by standard protocols: the FastDNA (R) SPIN kit for soil and the Zhou protocol. For most sediment types grinding in a cryogenic mill at a low beating rate in combination with direct phenol-chloroform extraction resulted in much higher DNA yields than those obtained using classical procedures. In general (except for clay-rich sediments), this procedure provided high-quality crude extracts for direct downstream nested-PCR, from cell numbers as low as 1.1 x 10(6) cells/cm(3). This procedure is simple, rapid, low-cost, and could be used with minor modifications for large-scale DNA extractions for a variety of experimental goals. (C) 2011 Elsevier B.V. All rights reserved.

Vibrio nigripulchritudo , the etiological agent of Litopenaeus stylirostris summer syndrome, is responsible for mass mortalities of shrimp in New Caledonia. Epidemiological studies led to the suggestion that this disease is caused by an emergent group of pathogenic strains. Genomic subtractive hybridization was carried out between two isolates exhibiting low and high virulence. Our subtraction library was constituted of 521 specific fragments; 55 of these were detected in all virulent isolates from our collection ( n = 32), and 13 were detected only in the isolates demonstrating the highest pathogenicity ( n = 19), suggesting that they could be used as genetic markers for high virulence capacity. Interestingly, 10 of these markers are carried by a replicon of 11.2 kbp that contains sequences highly similar to those of a plasmid detected in Vibrio shilonii , a coral pathogen. The detection of this plasmid was correlated with the highest pathogenicity status of the isolates from our collection. The origin and consequence of this plasmid acquisition are discussed.

In a previous study, we demonstrated the existence of an emerging cluster of Vibrio nigripulchritudo that proved to be associated with shrimp mortality events in New Caledonia. Using sequence polymorphisms evidenced in this previous MultiLocus Sequence Typing study, we developed two new quantitative PCR assays permitting the detection and quantification of V. nigripulchritudo at the genospecies level using SYBR Green I chemistry and at the emerging cluster level using Fluorescence Resonance Energy Transfer technology with hybridization probes. The use of this molecular diagnostic tool evidenced the colonization of the shrimp pond ecosystem by the pathogenic cluster at least at the onset of the disease. This new tool will allow better investigation of the dynamics of this bacterial pathogen in the shrimp farm ecosystem. (C) 2007 Elsevier B.V. All rights reserved.

Selon l’Office International des Epizooties (OIE) une maladie émergente désigne une maladie grave récemment reconnue, dont la cause peut, ou non, avoir déjà été établie, et qui est susceptible de se propager au sein d’une population ou entre des populations, par exemple à l’occasion d’échanges internationaux d’animaux aquatiques et/ou de produits d’animaux aquatiques. Si les maladies émergentes qui affectent la santé humaine ont été très étudiées, celles qui touchent les organismes marins et les organismes aquacoles d’intérêt économique en particulier sont en revanche peu documentées. C’est en restreignant l’émergence aux seules maladies infectieuses que seront présentés de façon non exhaustive quelques scénarios de l’émergence des maladies chez les organismes d’intérêt aquacole en les illustrant par trois exemples disponibles dans la littérature scientifique : l’un relatif à l’apparition d’un agent pathogène chez un nouvel hôte avec le cas de l’herpesvirus de la carpe Koï, l’autre à l’évolution d’un agent pathogène existant avec le cas de la vibriose à Vibrio nigripulchritudo sévissant dans les élevages de crevettes pénéides de Nouvelle-Calédonie et enfin le dernier lié à l’introduction d’un pathogène préexistant avec le cas de Bonamia ostreae infectant l’huître plate Ostrea edulis. Les causes d’émergence de maladies sont multiples et font intervenir de façon intercurrente l’agent pathogène, l’environnement, l’hôte ou les espèces hôtes et des facteurs anthropiques. Dans le milieu marin, ces causes sont bien souvent méconnues. Dans ce contexte le développement des réseaux de surveillance et des techniques de diagnostic revêtent un intérêt considérable afin d’anticiper, de prévenir et/ou d’intervenir sur l’émergence des maladies en limitant leur conséquences sanitaires, écologiques et politiques., According to the world organisation for animal health (OIE) an emerging disease is defined as a recently admitted serious illness, whose aetiology can, or not, have already been established, and which is likely to be propagated within a population or between populations, for example at the time of international exchanges of aquatic animals and/or products of aquatic animals. Even though the emerging diseases that affect human health have been much studied, those which affect marine organisms and species of aquaculture interest in particular are poorly documented. By restricting emergence to only infectious diseases, we aim at presenting in a non-exhaustive way some scenarios of the emergence of the diseases of aquacultured species by illustrating them with three examples available in the scientific literature: one relating to the appearance of a pathogenic agent in a new host with the case of the herpesvirus of the Koi carp, the other with the evolution of a pre-existing pathogenic agent with the case of shrimp vibriosis due to Vibrio nigripulchritudo in New Caledonia, and the last example relating to the introduction of one pathogenic pre-existing pathogen in an unscathed area with the case of Bonamia ostreae infecting the flat oyster Ostrea edulis. The causes of the emergence of diseases are multiple and implicate in an intercurrent way pathogenic agents, the environment, the host or host species and anthropogenic factors. In the marine environment, these causes are very often ignored. In this context, the development of zoosanitary surveillance networks and diagnostic tools present a considerable interest in order to anticipate, prevent and/or intervene on the emergence of the diseases by limiting their sanitary, ecological and political consequences.

A collection of 57 isolates of Vibrio nigripulchritudo from either diseased or healthy shrimp and from shrimp farms environment was studied in order to gain a better understanding of the epidemiology of this pathogen, notably isolated from two distinct shrimp disease complexes. Molecular typing using two different techniques, arbitrarily primed PCR (AP-PCR) and multi-locus sequence typing (MLST), studied together with experimental pathology data allowed a relevant epidemiological insight into this possibly emerging pathogen. Additionally, results obtained with the two molecular typing techniques were congruent and allowed discriminating the strains associated with the "Summer Syndrome" from strains isolated from other contexts, especially the other shrimp vibriosis "Syndrome 93". These results highlight that the "Summer Syndrome" is most probably caused by an emergent clonal pathogen that therefore deserves surveillance and that AP-PCR can satisfactorily be used for that purpose.