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Extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) have been classified in the group of resistant bacteria of highest priority. We determined the prevalence of ESBL-E collected in feces from household and shelter pets in Guadeloupe (French West Indies). A single rectal swab was taken from 125 dogs and 60 cats between June and September 2019. The prevalence of fecal carriage of ESBL-E was 7.6% (14/185, 95% CI: 4.2-12.4), within the range observed worldwide. The only risk factor associated with a higher prevalence of ESBL-E rectal carriage was a stay in a shelter, suggesting that refuges could be hotspots for their acquisition. All but one (

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%.

Serine protease autotransporters of Enterobacteriaceae (SPATEs) are secreted proteins that contribute to virulence and function as proteases, toxins, adhesins, and/or immunomodulators. An extra-intestinal pathogenic E. coli (ExPEC) O1:K1 strain, QT598, isolated from a turkey, was shown to contain vat, tsh, and three uncharacterized SPATE-encoding genes. Uncharacterized SPATEs: Sha (Serine-protease hemagglutinin autotransporter), TagB and TagC (tandem autotransporter genes B and C) were tested for activities including hemagglutination, autoaggregation, and cytotoxicity when expressed in E. coli K-12. Sha and TagB conferred autoaggregation and hemagglutination activities. TagB, TagC, and Sha all exhibited cytopathic effects on a bladder epithelial cell line. In QT598, tagB and tagC are tandemly encoded on a genomic island, and were present in 10% of UTI isolates and 4.7% of avian E. coli. Sha is encoded on a virulence plasmid and was present in 1% of UTI isolates and 20% of avian E. coli. To specifically examine the role of SPATEs for infection, the 5 SPATE genes were deleted from strain QT598 and tested for cytotoxicity. Loss of all five SPATEs abrogated the cytopathic effect on bladder epithelial cells, although derivatives producing any of the 5 SPATEs retained cytopathic activity. In mouse infections, sha gene-expression was up-regulated a mean of sixfold in the bladder compared to growth in vitro. Loss of either tagBC or sha did not reduce urinary tract colonization. Deletion of all 5 SPATEs, however, significantly reduced competitive colonization of the kidney supporting a cumulative role of SPATEs for QT598 in the mouse UTI model.

BackgroundSelection pressure exerted by use of antibiotics in both human and veterinary medicine is responsible for increasing antimicrobial resistance (AMR). The objectives of this study were to better understand antimicrobial use in pigs, beef cattle, and poultry on farms on Guadeloupe, French West Indies, and to acquire data on AMR inEscherichia coliin these food-producing animals. A cross-sectional survey was conducted at 45 farms on Guadeloupe, and practical use of antimicrobials was documented in declarative interviews between March and July 2018. A total of 216 fecal samples were collected between January 2018 and May 2019, comprising 124 from pigs, 75 from beef cattle, and 17 from poultry litter.E. coliisolates were obtained for further testing by isolation and identification from field samples. Antimicrobial susceptibility testing and screening forblaCTX-M,blaTEM,tetA, andtetB resistance genes by polymerase chain reaction on extracted genomic DNA were performed.ResultsThe study showed rational use of antimicrobials, consisting of occasional use for curative treatment by veterinary prescription. Tetracycline was the most commonly used antimicrobial, but its use was not correlated toE. coliresistance. Extended-spectrum β-lactamase (ESBL)E. coliisolates were detected in 7.3% of pigs, 14.7% of beef cattle, and 35.3% of poultry.blaCTX-M-1was the predominant gene found in ESBL-E. coliisolates (68.8%), followed byblaCTX-M-15(31.3%).ConclusionDespite rational use of antimicrobials, the rate of ESBL-E. coliin food-producing animals in Guadeloupe, although moderate, is a concern. Further studies are in progress to better define the genetic background of the ESBL-E. coliisolates.Graphical abstract

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.

Detection and quantification of pathogenic free-living amoebae (FLA) in water samples is critical for assessing water quality and for disease management issues. The most probable number (MPN) is commonly used to account for FLA in water. Nevertheless, this requires a high number of water replicates and working volumes, and a consequent number of non-nutrient agar (NNA)-plates seeded with Escherichia coli. Herein, we aimed at optimizing this difficult method, taking also into account key factors such as (i) the counting method, (ii) the delay between sample collection and sample processing, and (iii) the temperature during water sample transportation. To simplify the MPN method, we filtrated 1 × 1000 and 1 × 100 mL water samples, and cellulose acetate filters were cut in 10 parts and inverted on NNA-plates overlaid with E. coli. The comparison between the classical and our optimized MPN method showed that the final counts were similar, therefore validating the use of the optimized method. Our results also showed that for thermophilic FLA (such as Naegleria fowleri), water samples can be kept at around +30°C and processed within 24 h. This improved MPN method is now routinely used in our laboratory to control Naegleria sp. in the water samples in Guadeloupe.

International audience; K. pneumoniae (Kp) represents nearly 10% of nosocomial infections in the Western world, including Guadeloupe, and is one of the most problematic multidrug resistant bacteria. Hypervirulent clonal groups of Kp are also responsible for invasive community-acquired infections, such as pyogenic liver abscess and meningitis. As the reservoir of clinically and epidemiologically prominent clonal groups of Kp is currently unknown, we expect to define the sources of human infections through an “One Health” approach.

International audience; Species of Enterobacter cloacae complex (ECC) are widely distributed in the environment and are part of the gut microbiota of both animals and humans. Anolis marmoratus, an endemic and anthropophilic lizard of Guadeloupe, is a natural carrier of ECC and in particular of strains resistant to third generation cephalosporins (3GCR). Bacteria belonging to this complex are also major opportunistic pathogens which draw attention by their involvement in nosocomial infections and the emergence of multidrug resistant clones. Using hsp60 genotyping, ECC is divided into 12 genetic clusters (cluster I-XII) and a loosely knit group (cluster XIII). Strains belonging to clusters III, VI and VIII are prominent in human infections.We collected ECC strains (n=155) isolated from different sources (anoles, environment and clinical samples). We characterized these strains according to their antibiotic resistance phenotypes. Nearly 28% of strains sampled from environment and anoles were resistant to 3GC. This specific resistance was related to the overexpression of the AmpC cephalosporinase. We identified the cluster for each strain by using hsp60 genotyping. Cluster VIII was predominant in A. marmoratus feces (n=37, 38.9%) and in water catchment (n=10, 45.6%). We sequenced 58 strains of C-VIII (33 from clinical isolates: 26 from France and 7 from Guadeloupe, and 25 from anoles). A Maximum likelihood analysis based on core genome alignment revealed a clear split of isolates into two main clusters depending on sources: the first one clustering most of clinical samples together (from France and Guadeloupe, n=29) but also 5 strains isolated from anoles. The second cluster was mainly constituted by strains from anoles (n=20) and by 4 clinical samples. These results suggest first that bacteria resistant to third generation cephalosporins exist in environmental reservoirs in Guadeloupe. Moreover, exchange of ECC C-VIII strains may happen between human and anoles within a One Health appro

International audience; An environment affected by anthropogenic activities can play an important role in the spread and concentration of antibiotic resistant bacteria. Under antibiotic selection pressure, exchanges of antibiotic resistance genes (ARG) can occur between bacteria from human, animal and environmental reservoirs. However, these processes are still poorly understood. This work aimed at focussing on the wastewater treatment process, which is a hotspot for selection and emergence of antibiotic resistant bacteria and constitutes a favourable interface for circulation of ARG between bacteria from the different reservoirs.Between 2018 and 2019, extended spectrum β-lactamase producing Enterobacteriaceae (ESBLE) (Enterobacter cloacae complex, Escherichia coli and Klebsiella pneumoniae) were isolated from 266 patients at University Hospital of Pointe-à-Pitre (Guadeloupe). Simultaneously, urban and hospital wastewaters were analysed at different steps of waste treatment and 120 stool samples were collected from animals living near the hospital sewers and the wastewater treatment plant. Enterobacteriaceae were characterised according to their antibiotic resistance profile. Strains with the same antibiotic resistance profile belonging to a particular human-environment-animals continuum were analysed using whole genome sequencing.We isolated a total of 905 ESBLE (292 from humans, 348 from wastewater and 265 from animals). The frequency of ESBLE carriers was important in wildlife and domestic animals living nearby polluted environments (70%). A first genome-based comparison was carried out on 30 ESBL producing E. coli isolated during this study and compared to ESBL producing E. coli collected in farms (n=12). Antibiotic resistance gene blaCTX-M-15 was frequent (17/42). IncFII type was the most common plasmid (15/42). E. coli ST131 was found on the clinical samples only (n=4). One cluster with wastewater and animal isolates differed by ≤ 5 single nucleotide polymorphisms.These first results suggest that animals living near sewers and wastewater treatment plants are subjected to antibiotic resistance pressure and some bacteria and antibiotic resistance genes could be closely related.

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.

International audience; The epidemiology of human Salmonella enterica infections in Guadeloupe (French West Indies) appears to be specific, with a higher prevalence of the subspecies enterica serovars Panama and Arechavaleta (Panama and Arechavaleta) than in other regions. A study was performed in Guadeloupe to identify the reservoir of Salmonella serovars by comparing their distribution in warm- and cold-blooded animals and in humans living in Guadeloupe and mainland France. Furthermore, a case-control study was conducted in 2012-2013 to identify the main epidemiologic risk factors for S. enterica infection among children under 15 years of age. Between June 2011 and December 2014, feces from 426 reptiles (322 anoles, 69 iguanas and 35 geckos) and 50 frogs distributed throughout Guadeloupe and nearby islands were investigated. The frequency of S. enterica carriage was 15.0% (n = 64) in reptiles but varied by species. The only significant risk factor for S. enterica infection was a more frequent presence of frogs in the houses of cases than in those of controls (P = 0.042); however, isolates were not collected. Panama and Arechavaleta were the two serovars most often recovered between 2005 and 2014 from humans living in Guadeloupe (24.5% (n = 174) and 11.5% (n = 82), respectively), which is in contrast to the low prevalence in mainland France (0.4%). Their presence at low frequencies in wild reptiles (4.6% (n = 3) and 3.1% (n = 2), respectively) and pigs (7.5% (n = 5) and 1.5% (n = 1), respectively) suggests a broad host range, and humans may be infected by indirect or direct contact with animals. These serovars are probably poorly adapted to humans and therefore cause more severe infections. The unusual subspecies houtenae serovar 43:z4,z32:- was a major subspecies in wild reptiles (24.6%, n = 16) and humans (9.4%, n = 67) but was not recovered from warm-blooded animals, suggesting that reptiles plays a key role in human infection.

International audience

Free-living amoebae (FLA) are protists frequently found in water and soils. Some FLA such as Naegleria fowleri, Acanthamoeba and Balamuthia can cause rare but fatal encephalitis. They can also harbor human pathogens (bacteria, viruses and fungi). In 2008, Nf was responsible for the death of a 9-year-old boy who swam in a geothermal bath in Guadeloupe. Our group showed that (i) Nf can be found in most of these hot baths and (ii) soil is the origin of this contamination. Herein, we aimed at analyzing the diversity and distribution of FLA in soil using a targeted metagenomic analysis. A total of 107 soil samples were collected from 27 sites, nearby water ponds, hot springs and rivers. DNA was extracted directly from soil samples or from FLA cultivated at different temperatures (30, 37 and 44ºC). Metabarcoding studies were then conducted through FLA 18S amplicons sequencing; am-plicon 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 amoe-bal enrichment step is necessary. The cultures were mainly enriched with Naegleria, Acanthamoeba and Vermamoeba, at 37ºC and 44ºC. High differences in FLA diversity were observed between the 27 sites. V. vermiformis was by far the most represented species of FLA, being detected throughout the islands while the genus Naegleria was mainly found in Basse-Terre. Nevertheless, putative pathogenic Nf was also sampled in Grande Terre and in Les Saintes Islands. Acanthamoeba were mainly found in areas where temperature is approx. 30ºC. FLA are an unrecognized risk and our results clearly show that FLA are widespread in Guadeloupe, posing a potential threat (direct or indirect) on human health in Guadeloupe. Similar studies could be performed in neighbouring Caribbean islands.

International audience; A retrospective study was conducted to identify the risk factors associated with Salmonella enterica bac-teremia in infants and children in Guadeloupe, French West Indies. The 171 patients with S. enterica infection seen between 2010 and 2014 included 155 (90.6%) with acute gastroenteritis, of whom 42 (27.1%) had concomitant bac-teremia, and 16 (9.4%) with primary bacteremia. Most cases (97.7%) were in infants and children with no underlying health condition. Two subspecies were recovered: enterica (N = 161, 94.2%) and houtenae (N = 10, 5.8%). All but one (serovar Typhi) were non-typhoidal Salmonella. The most common serovars were Panama (N = 57, 33.3% of isolates) and Arechavaleta (N = 28, 16.4%). Univariate analysis showed a strong association only between age > 6 months and infection with the Panama or Arechavaleta serovar (P = 0.002). The rate of resistance to all classes of antibiotics during the study period was low (< 15%); however, the detection of one extended-spectrum beta-lactamase-producing S. enterica strain highlights the need for continued monitoring of antimicrobial drug susceptibility. Infection with Panama (P < 0.001) or Arechavaleta (P < 0.001) serovar was significantly associated with bacteremia in a multivariate analysis. These serovars are probably poorly adapted to humans or are more virulent. A delay between onset of symptoms and hospital admission > 5 days (P = 0.01), vomiting (P = 0.001), and increased respiratory rate (P = 0.001) contributed independently to bacteremia in the multivariate analysis. Thus, if non-typhoidal infection is suspected, blood should be cultured and antibiotic treatment initiated in all patients who meet these criteria.

International audience; Objective: The first aim of this study was to evaluate the antimicrobial resistance of Enterobacteriaceae in different water environments of Guadeloupe and especially those impacted by waste water treatment plants (WWTP) effluents. The second objective was to characterize the genetic basis for antibiotic resistance of extended-spectrum beta-lactamase (ESBL) and AmpC beta-lactamase producing Enterobacteriaceae isolates (ESBLE and AmpCE).Methods: We have collected 70 surface waters (river and sea samples) impacted or not by WWTP and 18 waste waters from 2 WWTPs in 2012 and 2013. We i) determined the total and resistant bacterial counts and ii) tested isolated Enterobacteriaceae for their antimicrobial susceptibility. We also studied the genetic basis for antibiotic resistance of ESBLE and AmpCE, and the genetic background of Escherichia coli.Results: In rivers, contamination with Escherichia coli and antibiotic resistant coliforms (ARC) increased from the source to the mouth. Highest levels of river contamination with E. coli (9.26 x 105 MPN/100mL) and ARC (2.26 x 108 CFU/mL) were observed in surface water sampled near the discharge. A total of 246 Enterobacteriaceae strains resistant to antibiotics were isolated, mostly from waste waters and from river water collected near the discharge. Among these strains, 33 were Extended Spectrum Beta Lactamase (ESBLE) and 8 E. coli were AmpC beta-lactamase producers. All the ESBLE were isolated from waste waters or from river water collected near the discharge. The blaCTX-M gene was present in 29 of the 33 ESBLE strains, with 24 belonging to group 1. Numerous strains (68.7%) showed more than one acquired antibiotic resistance mechanism. E. coli strains belonged to different phylogenetic groups; among the B2 group, most strains belonged to the ST131 clone.Conclusion: Our results demonstrated that many human activities can supply antibiotic-resistant bacteria in surface water. Nevertheless, WWTPs were the most important supplier of ESBLE in water environment of Guadeloupe.

Background Urinary tract infections are bacterial infections most commonly encountered in the community. The resistance rate of uropathogens to commonly prescribed antibiotics has increased worldwide but there are no published data concerning the resistance of strains isolated from community-acquired UTI in Guadeloupe. To assess the susceptibility patterns of Enterobacteriaceae strains isolated from outpatients in Guadeloupe we conducted a prospective study from December 2012 to May 2014 among outpatients consulting at private and public laboratories for urine analysis. Risk factors for E. coli resistance to amoxicillin, third-generation cephalosporin, and ciprofloxacin were also determined. To study the trends of E. coli resistance rates over the past 10 years, data on the susceptibility patterns of E. coli from 2003 to 2014 were also collected from three major laboratories for a retrospective study. Results During the prospective study, we isolated 1293 bacterial strains from the urine of outpatients presenting for urine analysis. The most commonly isolated bacteria were E. coli (57 %) and Klebsiella pneumoniae (15.5 %). Thirty seven per cent of the E. coli strains were resistant to amoxicillin. Resistance rates to third generation cephalosporin were low for E. coli and other Enterobacteriaceae (3.1 and 12.2 % respectively) and mostly due to the presence of an Extended Spectrum Beta-lactamase. Resistance to cotrimoxazole and ciprofloxacin was moderate (17.8 and 15.6 % respectively). However, the resistance rate of E. coli to ciprofloxacin has significantly increased during the last 10 years. Risk factors were consistent with previously reported data, especially for the increasing ciprofloxacin resistance with age. Conclusion General practitioners in Guadeloupe need to be better informed to favor the prescription of fosfomycin-trometamol to reduce the risk of resistance to fluoroquinolones.

Among the many species of free-living amoebae infecting humans, only Naegleria fowleri, a few species of Acanthamoeba, Balamuthia mandrillaris recently Sappinia diploïdea and Paravahlkampfia Francina are responsible for human diseases especially deadly encephalitis outside of Acanthamoeba keratitis related . In the Central African Republic (CAR), no studies have previously been conducted about free amoebae and no suspicious cases of encephalitis or amoebic keratitis was reported even though the ecosystem supported the proliferation of these microorganisms. The objective of this study was to identify free-living amoebae present in CAR and to define the molecular characteristic. Bathing sites and cerebrospinal fluid from patients died of bacterial meningitis untagged were explored by culture and PCR and the amplicons were sequenced which allowed to characterize the species found. Only species of the genus Tetramitus, namely T. Entericus, T. waccamawensis and T.sp similar to those already described in the world and not pathogenic for humans were found in bathing sites, the cerebrospinal fluid meanwhile remained negative. Although no pathogen species such as Naegleria fowleri or species of Acanthamoeba have been isolated, this study worth pursuing because this investigation was very limited in space because of the insecurity in the country.

Objective The tuberculin skin test (TST) is an important tool in the diagnosis of tuberculosis infection in children. However, the interpretation of TST may be complicated by prior Bacillus Calmette-Guerin (BCG) vaccination. We evaluated the effect of vaccination with BCG on TST reactivity in first-year pupils attending state schools in Antananarivo. Methods STs were performed on 376 first-year schoolchildren, aged 6 and 7, attending two state primary schools. The relationships between epidemiological information, BCG status (vaccination, BCG scars) and TST reactivity were assessed to compare TST sensitivity between children with and without BCG vaccination and between those with and without a BCG scar. Result The prevalence of positive TST results of ≥5, ≥10 and ≥ 15 mm was 20.2% (76/376), 18.3% (69/376) and 11.4% (43/376), respectively. BCG vaccination was not associated with TST reactivity, whatever the threshold used: ≥5 mm (odds ratio (OR, 1.2; 95% confidence interval (CI), 0.7–2.0); ≥10 mm (OR, 0.9; 95% CI, 0.6–1.7); ≥15 mm (OR, 0.6; 95% CI, 0.3–1.2). Conclusion These results suggest that in Madagascar, a positive TST result indicates TB infection (active or latent) rather than past BCG vaccination. Therefore, high BCG vaccination coverage does not appear to impair the usefulness of the TST as a tool for diagnosing tuberculosis. Objectif: Le test cutane tuberculinique (TCT) est un outil important dans le diagnostic de l’infection tuberculeuse chez les enfants. Cependant, l’interpretation du TCT peut etre compliquee par une vaccination prealable avec le bacille de Calmette-Guerin avant (BCG). Nous avons evalue l’effet de la vaccination par le BCG sur la reactivite au TCT chez les eleves de premiere annee frequentant les ecoles publiques a Antananarivo. Methodes: Le TCT a ete effectue sur 376 ecoliers de premiere annee, âges de 6 et 7 ans, frequentant deux ecoles primaires publiques. Les relations entre l’information epidemiologique, le statut BCG (vaccination, cicatrices du BCG) et la reactivite au TCT ont eteevaluees afin de comparer la sensibilite du TCT entre les enfants avec et ceux sans vaccination BCG et entre ceux avec et ceux sans cicatrice du BCG. Resultat: La prevalence des resultats positifs au TCT de ≥5 mm, ≥10 mm et ≥15 mm etait de 20,2% (76/376), 18,3% (69/376) et 11,4% (43/376), respectivement. La vaccination par le BCG n’a pas ete associee a la reactivite au TCT, quelque soit le seuil utilise: ≥5 mm (odds ratio (OR: 1,2; IC95%: 0,7 - 2,0); ≥10 mm (OR: 0,9; IC95%: 0,6 - 1,7); ≥15 mm (OR: 0,6; IC95%: 0,3 – 1,2). Conclusion: Ces resultats suggerent qu’a Madagascar, un resultat positif au TCT indique une infection tuberculeuse (active ou latente) plutot qu’une vaccination passee par le BCG. Par consequent, une couverture elevee de vaccination par le BCG ne semble pas nuire a l’utilite du TCT comme un outil de diagnostic de la tuberculose. Objetivo: La prueba cutanea de tuberculina (PCT) es una herramienta importante para el diagnostico de la infeccion por tuberculosis en ninos. Sin embargo, la interpretacion de la PCT puede complicarse por una vacunacion previa con el bacilo Calmette-Guerin (BCG). Hemos evaluado el efecto de la vacunacion con BCG sobre la reactividad de la PCT en escolares de primer ano de escuelas publicas en Antananarivo. Metodos: Se realizaron pruebas cutaneas a 376 escolares de primer ano, con edades entre 6 y 7 anos, pertenecientes a dos escuelas primarias publicas. Se evaluo la relacion entre la informacion epidemiologica, el estatus de BCG (vacunacion, cicatrices de BCG) y la reactividad del PCT para comparar la sensibilidad del PCT entre ninos con y sin previa vacunacion con BCG y entre aquellos con y sin cicatrices. Resultados: La prevalencia de resultados de PCT positivos de ≥5 mm, ≥10 mm y ≥ 15 mm fue del 20.2% (76/376), 18.3% (69/376) y 11.4% (43/376), respectivamente. La vacunacion con BCG no estaba asociada con la reactividad del PCT, independientemente del umbral utilizado: ≥5 mm (odds ratio (OR 1.2, IC 95% 0.7–2.0); ≥10 mm (OR 0.9, 95% IC 0.6–1.7); ≥15 mm (OR 0.6, IC 95% 0.3–1.2). Conclusion: Estos resultados sugieren que, en Madagascar, un resultado positivo de PCT indica una infeccion de TB (activa o latente) mas que una vacunacion previa con BCG. Por lo tanto, una alta cobertura de vacunacion con BCG no parece afectar la utilidad de la PCT como herramienta para el diagnostico de la tuberculosis.

Background: In developing countries, knowledge of antimicrobial resistance patterns is essential to define empirical therapy. Methodology: All the surgery and intensive care wards of two hospitals in Antananarivo were included to study the antimicrobial susceptibility of the pathogenic bacteria causing nosocomial infections. A repeated cross-sectional survey was conducted between September 2006 and March 2008, one day per week. Isolates were identified using classical methods, and resistance to antibiotics was assessed according to the recommendations of the Antibiogram Committee of the French Microbiology Society. Results: Clinical specimens from 706 from 651 patients were collected. Of the 533 bacterial pathogens, 46.7% were Enterobacteriaceae , 19.3% were Staphylococcus aureus , and 19.1% were pathogens from the hospital environment ( Pseudomonas aeruginosa and Acinetobacter baumannii ).Frequencies of resistance were high, particularly in Enterobacteriaceae ; however, the rate of Staphylococcus aureus isolates resistant to oxacillin (13.6 %) was moderate and all these isolates were susceptible to glycopeptids. The percentages of isolates susceptible to ceftazidim were 81.8% for E. coli , 60.9% for Klebsiella , and 52.5% for Enterobacter spp. Resistance to third-generation cephalosporins was due to extended spectrum betalactamases (ESBL). Multivariate analysis showed that diabetes (adjusted OR: 3.9) and use of an invasive procedures (adjusted OR: 3.5) were independent risk factors for resistance to third-generation cephalosporins. Conclusion: A nationwide surveillance programme is needed to monitor the microbial trends and antimicrobial resistance in Madagascar.

The goal of the study was to investigate the in vitro antimicrobial susceptibility of Neisseria gonorrhoeae strains isolated in 2004 and 2005 in Bangui Central African Republic; Yaounde Cameroon; Antananarivo Madagascar; and Ho Chi Minh Ville and Nha Trang Vietnam. Antimicrobial susceptibility testing was performed by both disk diffusion and agar dilution methods according to Clinical and Laboratory Standards Institute (CLSI) recommendations. Minimum inhibitory concentrations (MICs) to 5 antimicrobials (penicillin G ceftriaxone ciprofloxacin spectinomycin and tetracycline) were determined when feasible. Penicillinase-producing N. gonorrhoeae (PPNG) was analyzed by the paper acidometric method (nitrocefin test). Thirty N. gonorrhoeae isolates from Bangui could be studied 79 from Yaounde 126 from Antananarivo 56 from Nha Trang and 126 from Ho Chi Minh Ville in 2004 and 2005. Unfortunately because of problems of electricity supply no strains could be recovered for the determination of MICs in Yaounde and only 68 strains could be tested in Antananarivo and 121 in Ho Chi Minh Ville. Patterns of resistance were similar in Antananarivo Bangui and Yaounde but different from those observed in Vietnam. Ciprofloxacin was highly effective in Africa but nearly all strains in Vietnam were resistant to this drug. Overall ceftriaxon and spectinomycin were the best antibiotics with one strain resistant to spectinomycin in Antananrivo and one strain resistant to ceftriaxon in Ho Chi Minh Ville. Ciprofloxacin remains highly efficient in Madagascar and Central Africa ceftriaxone and spectinomycin should be used as the first-line antimicrobial agents in treating gonorrhea in Vietnam.

Four rapid immunochromatographic assays – Determine™ HBsAg, Virucheck ® HBsAg, Cypress HBsAg Dipstick ® and Hexagon ® HBsAg – for human hepatitis B surface antigen (HBsAg) detection in human serum were evaluated. A collection of reference serum samples (91 HBsAg positive and 109 HBsAg negative) stored at −80 °C was used. Sensitivity and positive predictive value (PPV) exceeded 95%, and specificity and negative predictive value (NPV) exceeded 96% for all tests. The Determine™ HBsAg test performed best in this study, with a sensitivity of 97.8%, a specificity and PPV of 100%, a NPV of 98.2% and an accuracy rate of 99.0%. However, the differences between the tests were not significant. Other factors should therefore also be taken into account by the Ministry of Health in its decision to recommend a particular test: price, availability, delivery time and feasibility of whole-blood testing. The Determine test appears to be the most suitable for Madagascar, based on all these criteria. The use of this test, despite its lower sensitivity, could prevent blood-borne transmission of hepatitis B virus (HBV) in areas with limited resources.

Resume But de l’etude Genotypage des souches du virus de l’hepatite B (VHB) de patients centrafricains et comparaison avec les resultats obtenus dans les autres pays africains. Patients et methodes Serums collectes de patients consentants, atteints d’hepatite aigue ou chronique, hospitalises dans le service de gastroenterologie de l’hopital de l’Amitie de Bangui, Republique centrafricaine (RCA). La sequence complete du gene pre S2/S a ete definie pour la determination des genotypes. Resultats Cent quatre-vingt-seize serums ont ete collectes de 112 hommes et 84 femmes. Quatre-vingt-douze pour cent de patients ont ete infectes par le VHB (anticorps anti-HBc positifs) et la prevalence de l’Ag-HBs etait de 62 %. L’ADN viral a ete detecte dans 66 % des serums Ag-HBs positifs. Aucun ADN quantifiable n’a ete mis en evidence chez les patients Ag-HBs negatifs. Quatre-vingt-treize pour cent des souches appartiennent au genotype E ; un (3,4 %) au genotype A1 et un (3,4 %) au genotype D. Conclusion La forte prevalence de l’infection par le VHB dans la population etudiee est liee a leur recrutement. Le genotype E est predominant en RCA et la variabilite de sequences entre les souches au sein du genotype E est faible (1,8 %). Il existe egalement en RCA des souches de VHB de genotype A1 et de genotype D. Ces souches, tres minoritaires, pourraient etre d’importation.