Our team in Europe has developed the routine clinical laboratory identification of microorganisms by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). To evaluate the utility of MALDI-TOF MS in tropical Africa in collaboration with local teams, we installed an apparatus in the Hôpital Principal de Dakar (Senegal), performed routine identification of isolates, and confirmed or completed their identification in France. In the case of discordance or a lack of identification, molecular biology was performed. Overall, 153/191 (80.1%) and 174/191 (91.1%) isolates yielded an accurate and concordant identification for the species and genus, respectively, with the 2 different MALDI-TOF MSs in Dakar and Marseille. The 10 most common bacteria, representing 94.2% of all bacteria routinely identified in the laboratory in Dakar (Escherichia coli, Klebsiella pneumoniae, Streptococcus agalactiae, Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus haemolyticus, Enterobacter cloacae, Enterococcus faecalis, and Staphylococcus epidermidis) were accurately identified with the MALDI-TOF MS in Dakar. The most frequent misidentification in Dakar was at the species level for Achromobacter xylosoxidans, which was inaccurately identified as Achromobacter denitrificans, and the bacteria absent from the database, such as Exiguobacterium aurientacum or Kytococcus schroeteri, could not be identified. A few difficulties were observed with MALDI-TOF MS for Bacillus sp. or oral streptococci. 16S rRNA sequencing identified a novel bacterium, “Necropsobacter massiliensis.” The robust identification of microorganisms by MALDI-TOF MS in Dakar and Marseille demonstrates that MALDI-TOF MS can be used as a first-line tool in clinical microbiology laboratories in tropical countries.

BackgroundOnly limited data are available relating to the etiology of diarrhea in children and adults in Senegal. The aim of this prospective study was to describe the epidemiology and etiology of community-acquired diarrheal infections in children and adults living in urban settings.MethodsA prospective study was carried out from March 2009 to December 2010, in the urban region of Dakar, Senegal. Patients with acute diarrhea were enrolled, interviewed to collect their clinical history, and their stools were tested for bacteria, virus and parasites.ResultsA total of 223 patients (including 112 children younger than five years old) with diarrhea were included. At least one enteropathogen was detected in 81% (180/223) of the patients: 29% (64/223) had bacterial infections (mainly diarrheagenic E. coli and Shigella spp), 21% (39/185) viral infections (mainly rotavirus) and 14% (31/223) parasitic infections. Co-infection was identified in 17.8% (32/180) of the patients. Viral infection was significantly more frequent in children under five years old during the dry season. Bacteria and parasites were equally frequent in all age groups. There was a seasonal variation of bacterial infections during the study period, with a higher proportion of infections being bacterial, and due to Salmonella spp. in particular, during the rainy season.ConclusionOur study suggests that in urban settings in Senegal, rotavirus is the principal cause of pediatric diarrhea during the dry season and that the proportion of bacterial infections seems to be higher during the rainy season. Further work is needed to document the burden of diarrheal diseases in sub-Saharan urban communities and to identify risk factors, including those linked to the rapid and unplanned urbanization in Africa.

Strain FF7T was isolated from the peritoneal fluid of a 44-year-old woman who suffered from pelvic peritonitis. This strain exhibited a 16S rRNA sequence similarity of 94.8 % 16S rRNA sequence identity with Haemophilus parasuis, the phylogenetically closest species with a name with standing in nomenclature and a poor MALDI-TOF MS score (1.32 to 1.56) that does not allow any reliable identification. Using a polyphasic study made of phenotypic and genomic analyses, strain FF7T was a Gram-negative, facultatively anaerobic rod and member of the family Pasteurellaceae. It exhibited a genome of 2,442,548 bp long genome (one chromosome but no plasmid) contains 2,319 protein-coding and 67 RNA genes, including 6 rRNA operons. On the basis of these data, we propose the creation of Haemophilus massiliensis sp. nov. with strain FF7T (= CSUR P859 = DSM 28247) as the type strain.Electronic supplementary materialThe online version of this article (doi:10.1186/s40793-016-0150-1) contains supplementary material, which is available to authorized users.

Biting midges of the genus Culicoides are implicated as vectors for a wide variety of pathogens. The morphological identification of these arthropods may be difficult because of a lack of detailed investigation of taxonomy for this species in Africa. However, matrix-assisted laser desorption ionization؊time of flight mass spectrometry (MALDI-TOF MS) profiling is efficient for arthropod identification at the species level. This study established a spectrum database of Culicoides spp. from Senegal using MALDI-TOF. Identification of Culicoides insects to the species level before mass spectrometry was performed on the basis of morphological characters. MALDI-TOF MS reference spectra were determined for 437 field-caught Culicoides of 10 species. The protein profiles of all tested Culicoides revealed several peaks with mass ranges of 2 to 20 kDa. In a validation study, 72 Culicoides specimens in the target species were correctly identified at the species level with a similarity of 95 to 99.9%. Four Culicoides protein profiles were misidentified. Nevertheless, six SuperSpectra (C. imicola, C. enderleini, C. oxystoma, C. kingi, C. magnus, and C. fulvithorax) were created. Abdomens of midges were used to amplify and sequence a portion of the mitochondrial cytochrome oxidase I gene (COI). The results obtained using the MALDI-TOF MS method were consistent with the morphological identification and similar to the genetic identification. Protein profiling using MALDI-TOF is an efficient approach for the identification of Culicoides spp., and it is economically advantageous for approaches that require detailed and quantitative information of vector species that are collected in field. The database of African Culicoides MS spectra created is the first database in Africa. The COI sequences of five Culicoides species that were previously noncharacterized using molecular methods were deposited in GenBank. Biting midges of the genus Culicoides are among the smallest hematophagous insects, and they can be a nuisance to humans and animals (1). These insects are implicated as vectors for a wide variety of viruses, such as bluetongue virus (2), African horse sickness virus (3), epizootic hemorrhagic disease virus (4), the Schmallenberg virus that recently emerged in Europe (5-7), Toggenburg orbivirus (8), and the Oropouche virus (9), which is the only human virus transmitted by Culicoides. Additionally, biting midges are involved in the transmission of various human filarial parasites, such as Mansonella perstans (10-13), Mansonella streptocerca, Mansonella ozzardi, and Mansonella rodhaini (14). Recently, Leishmania spp. and Leishmania infantum were detected in Culicoides imicola and Culicoides spp. (15).Vector identification is an important step in the epidemiology of vector-borne diseases. Information on the major vector species may provide a clearer indication of the geographic distribution of the disease or its potential distribution, the location of danger points for high risks of contact with the vector, and access to alternative...

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Nosocomial infections are very common in African hospitals, particularly in neonatal units. These infections are most often caused by bacteria such as Escherichia coli, Klebsiella spp and Staphylococcus spp. Salmonella strains are rarely involved in nosocomial infections. Here, we report the first description of S. Grumpensis in neonatal infections in Senegal. Seventeen Salmonella strains were isolated from hospitalized infants’ stool samples. The following resistance phenotype was described in strains: AMXRTICRCFR FOXRCFXRCTXRCAZRIMPSATMRNARNORRCIPRTMRGMRTERSXTR. All isolates were susceptible to imipenem, 15 out of 17 produced an extended spectrum ß-lactamase (ESBL). blaOXA-1, blaSHV-1, blaTEM-1, blaCTX-M1 genes were detected in strains 8, 13, 5 and 8, respectively. blaCTX-M1 sequencing revealed the presence of blaCTX-M-109. Thirteen of the 17 Salmonella Grumpensis strains were analyzed by PFGE. These 13 isolates belonged to a single pulsotype and were genotypically identical. This is the first report of neonatal S. Grumpensis infections in Senegal, and the first report of blaCTX-M-109 in the genus Salmonella.

Diarrheal diseases remain a leading cause of death with 14.7 million deaths in 2001 and 26% of global mortality worldwide according to WHO. Shigella species are prevalent in tropical areas; they are present all the year, with epidemic outbreaks in rainy season. Between 2001 and 2010 one hundred ninety (190) strains of Shigella flexneri isolated from National Senegalese Enterobacteriaceae Center located at the Pasteur Institute in Dakar were studied. Susceptibility was performed by antibiogram following the CASFM recommendations. Detection and characterization of integrons and resistance genes was done by PCR using specific primers and sequencing. Antibiotic susceptibility showed high percentage resistance to tetracycline: 95%, cotrimoxazole 60%, ampicillin 55%. Nineteen strains were cephalosporin resistant (10%). Two isolates were resistant to quinolones and one was imipenem resistant. Genes tet, dfr, cat, bla tem1 , bla oxa30 , bla shv , bla CTX-M , blakpc,bla IMP , gyrA, gyrB, parC and parE were detected on isolates. Integrons harbored genes resistance. The class 1 integron predominated followed by class 2 integron. Genes bla oxa30 , aadA1/aadA2 dfrA1, dfrA7 were found on class 1 integron. Class 2 integron showed three different types cassettes. No class 3 integron was detected. Genes dfrA1, dfrA7, sat, and aadA1 were harbouring by integrons. Antibiotic susceptibility showed that Shigella flexneri strains are resistant to the first line drugs used to treat shigellosis in Senegal. Resistance to 3rd generation of cephalosporins and fluoroquinolones emerged and is of great concern. These molecules must be used with caution in the treatment of shigellosis.

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