Prevalence and characterization of extended-spectrum ß-lactamase-producing clinical Salmonella enterica isolates in Dakar, Senegal, from 1999 to 2009.

A total of 1623 clinical isolates of Salmonella belonging to 229 serotypes were received by the Senegalese Reference Center for Enterobacteria from January 1999 to December 2009. The most common serotypes were Enteritidis (19% of the isolates), Typhi (8%), Typhimurium (7%) and Kentucky (4%). A significant increase in the prevalence of resistance to amoxicillin (0.9% in 1999 to 11.1% in 2009) and nalidixic acid (0.9% in 1999 to 26.7% in 2009) was observed in non-typhoidal Salmonella serotypes. For critically important antibiotics, notably ciprofloxacin and extended-spectrum cephalosporins (ESCs), the rates of resistance were low: 0.3% and 0.5%, respectively. Seven ESC-resistant Salmonella strains and three additional ESC-resistant strains from Senegal (1990) and Mali (2007) were studied to identify the genetic basis of their antibiotic resistance. All ESC-resistant strains produced an extended-spectrum ß-lactamase (ESBL). These were CTX-M-15 (n = 6; 2000-2008), SHV-12 (n = 3; 2000-2001) and SHV-2 (n = 1; 1990). A large IncHI2 ST1 pK29-like plasmid was found in six strains (three producing SHV-12 and three CTX-M-15), whereas IncN and IncF plasmids were found in three strains and one strain, respectively. The association of plasmid-mediated quinolone resistance (PMQR) genes qnrB1 and aac(6')-Ib-cr was found in four ESBL-producing strains, leading to decreased susceptibility and even full resistance to ciprofloxacin (MIC range 0.75-2 mg/L) despite the absence of mutations in the quinolone resistance-determining region (QRDR) of gyrA, gyrB, parC and parE. This association of ESBL and multiple PMQR mechanisms within the same strains is therefore a serious concern as it hampers the use of both ESCs and fluoroquinolones for severe Salmonella infections.

Klebsiella pneumoniae resistant to third-generation cephalosporins in five African and two Vietnamese major towns: multiclonal population structure with two major international clonal groups, CG15 and CG258.

The molecular epidemiology of third-generation cephalosporin-resistant (3GC-R) Klebsiella pneumoniae in developing countries is poorly documented. From February 2007 to March 2008, we collected 135 3GC-R K. pneumoniae isolates from seven major towns in Maghreb (Morocco), West Africa (Senegal, Côte d'Ivoire), Central Africa (Cameroon), East Africa (Madagascar) and Southeast Asia (Vietnam). Their genetic diversity, assessed by multilocus sequence typing, was high (60 sequence types), reflecting multiclonality. However, two major clonal groups, CG15 (n = 23, 17% of isolates) and CG258 (n = 18, 13%), were detected in almost all participating centres. The two major clonal groups have previously been described in other parts of the world, indicating their global spread. The high diversity of enterobacterial repetitive intergenic consensus sequence-PCR banding patterns at the local level indicates that most isolates were epidemiologically unrelated. The isolates were characterized by the presence of multiple resistance determinants, most notably the concomitant presence of the aac(6')-Ib-cr, qnr and blaCTX-M-15 genes in 61 isolates (45%) belonging to 31 sequence types. These isolates were detected across a large geographical area including Cameroon (n = 1), Vietnam (n = 4), Madagascar (n = 10), Côte d'Ivoire (n = 12), Morocco (n = 13) and Senegal (n = 21). These results have major implications for patient management and highlight a potential reservoir for resistance determinants.

[Qnr and aac (6')-Ib-cr types quinolone resistance among Enterobacteriaceae isolated in Annaba, Algeria]. / Résistance aux quinolones de types qnr, aac (6')-Ib-cr chez les entérobactéries isolées à Annaba en Algérie.

AIM OF THE STUDY: To show the emergence of the qnr and aac (6')-Ib-cr genes in nalidixic acid resistant enterobacteria isolated at Annaba city in Algeria. MATERIALS AND METHODS: Enterobacterial strains (n=25) resistant to nalidixic acid have been isolated at the microbiology laboratory of the Annaba city hospital in Algeria Antibiotic susceptibility (disc diffusion method and MIC) and screening for Extended Spectrum Beta-Lactamase (ESBL) were performed according to the French Society for Microbiology guidelines. Characterization of quinolone resistance genes (qnrA, qnrB, qnrS, aac(6')-Ib-cr) was investigated by PCR. Identification of ESBL (TEM, SHV, CTX-M1, CTX-M2, CTX-M8 and CTX-M9 groups) was performed by PCR. Identification of plasmid AmpC beta-lactamases was performed by multiplex PCR. All PCR products were sequenced on both strands. Conjugation experiments were performed using azide-resistant Escherichia coli K12J5 as a recipient strain. RESULTS: Among the 25 strains selected, 24 were resistant to at least four antibiotics. Six strains showed an ESBL phenotype. The qnr gene (B1 type) was found in two ESBL producing strains with at least two types of bla gene. The aac (6')-Ib gene was detected in three strains, one with the aac (6') Ib-cr variant. With specific primers, we have shown that qnrB1, CTX-M-28, TEM1, aac (6')-Ib-cr was cotransferred together and that these genes are carried by conjugative plasmids of high molecular weight. CONCLUSION: The emergence of combination of resistance genes may pose a public health problem. Thus, a policy of surveillance of resistance seems necessary.

Epidemiology of methicillin-resistant Staphylococcus aureus lineages in five major African towns: emergence and spread of atypical clones.

The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) in Africa is poorly documented. From January 2007 to March 2008, we collected 86 MRSA isolates from five African towns, one each in Cameroon, Madagascar, Morocco, Niger and Senegal. Although one or two major clones, defined by the sequence type and staphylococcal cassette chromosome mec type, predominated at each site, genetic diversity (ten clones) was relatively limited in view of the large geographical area studied. Most of the isolates (n = 76, 88%) belonged to three major clones, namely ST239/241-III, a well-known pandemic clone (n = 34, 40%), ST88-IV (n = 24, 28%) and ST5-IV (n = 18, 21%). The latter two clones have only been sporadically described in other parts of the world. The spread of community-associated MRSA carrying the Panton-Valentine leukocidin genes is a cause for concern, especially in Dakar and possibly throughout Africa.

Epidemiology of methicillin-susceptible Staphylococcus aureus lineages in five major African towns: high prevalence of Panton-Valentine leukocidin genes.

The epidemiology of methicillin-susceptible Staphylococcus aureus (MSSA) in Africa is poorly documented. From January 2007 to March 2008, 555 S. aureus isolates were collected from five African towns in Cameroon, Madagascar, Morocco, Niger, and Senegal; among these, 456 unique isolates were susceptible to methicillin. Approximately 50% of the MSSA isolates from each different participating centre were randomly selected for further molecular analysis. Of the 228 isolates investigated, 132 (58%) belonged to five major multilocus sequence typing (MLST) clonal complexes (CCs) (CC1, CC15, CC30, CC121 and CC152) that were not related to any successful methicillin-resistant S. aureus (MRSA) clones previously identified in the same study population. The luk-PV genes encoding Panton-Valentine leukocidin (PVL), present in 130 isolates overall (57%), were highly prevalent in isolates from Cameroon, Niger, and Senegal (West and Central Africa). This finding is of major concern, with regard to both a source of severe infections and a potential reservoir for PVL genes. This overrepresentation of PVL in MSSA could lead to the emergence and spread of successful, highly virulent PVL-positive MRSA clones, a phenomenon that has already started in Africa.

[Epidemiology of the resistance of Mycobacterium tuberculosis to antituberculosis drugs at the main hospital in Dakar, Senegal. A 4-year retrospective study (2000-2003)]. / Épidémiologie de la résistance de Mycobacterium tuberculosis aux antituberculeux à l'hôpital principal de Dakar. Étude rétrospective sur quatre ans (2000-2003).

A retrospective study was carried out from January 2000 to December 2003 to assess the resistance of Mycobacterium tuberculosis to antituberculosis drugs and the impact of this on the treatment result. Two hundred and two patients' files were studied (average age: 36 years; sex-ratio: 1.7). Pulmonary localisation (85.7%) or extrapulmonary localisation (14.3%). HIV status is negative (71.3%), positive (10.8%) or unknown (17.9%). The overall recovery rate is 60.7% (61.4% in HIV-; 46.1% in HIV+), the rate of treatment failure is 2.7% (1.1% in HIV-; 15.4% in HIV+), the death rate due to tuberculosis is 6.3% (2.3% in HIV-; 23.1% in HIV+), and the rate of patients who disappeared from the system is 30.3% (35.2% in HIV-; 14.2% in HIV+). Hepatotoxicity that occurred during treatment is observed in 14.3% of cases (recovery: 56.2%; failure: 6.2%; lost from the system: 18.8%). Eighty-four percent of patients never received antituberculosis treatment (group A) versus 15.8% of patients who had already received one or more antituberculosis drugs (group B). The rates of resistance to isoniazid are 6.4% (A) and 12.5% (B), to rifampicin 1.7% (A) and 12.5% (B), to ethambutol 0.5% (A) and 0% (B), to streptomycin 24.1% (A) and 46.8% (B). The percentage of multiresistant strains is 1% in patients not treated previously and 11% in those who had already received antituberculosis treatment. When the patients are carriers of a strain that is responsive to the treatment administered, the recovery rate is 64.2% versus 46.7% in patients whose strain is resistant to at least one of the treatments administered.

Genetic characterization and antibiotic resistance of Campylobacter spp. isolated from poultry and humans in Senegal.

AIMS: The main objectives of this study were to investigate the diversity of Campylobacter genotypes circulating in Senegal and to determine the frequency of antibiotic resistance. METHODS AND RESULTS: Strains of Campylobacter jejuni isolated from poultry (n = 99) and from patients (n = 10) and Campylobacter coli isolated from poultry (n = 72) were subtyped by pulsed-field gel electrophoresis (PFGE). The pulsotypes obtained after digestion by SmaI and KpnI revealed a significant genetic diversity in both species, but without any predominant pulsotypes. However, farm-specific clones were identified in the majority of poultry houses (76.5%). Human and poultry isolates of C. jejuni had common PFGE patterns. High quinolone-resistance rates were observed for C. jejuni (43.4%) and C. coli (48.6%) isolates obtained from poultry. CONCLUSIONS: The results showed a genetic diversity of Campylobacter between farms indicating multiple sources of infection; but specific clones had the ability to colonize the broiler farms. The antimicrobial resistance patterns were not related to any specific PFGE pattern suggesting that resistance was due to the selective pressure of antibiotic usage. Campylobacter with similar genotypes were circulating in both human and poultry. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is important for the understanding of the epidemiology of Campylobacter in broiler farms in Senegal. It also emphasizes the need for a more stringent policy in the use of antimicrobial agents in food animals.

Epidemiological analysis of Salmonella enterica ssp. enterica serovars Hadar, Brancaster and Enteritidis from humans and broiler chickens in Senegal using pulsed-field gel electrophoresis and antibiotic susceptibility.

AIMS: Salmonella Hadar, Salmonella Brancaster and Salmonella Enteritidis are the main Salmonella enterica ssp. enterica serovars isolated from poultry in Senegal. Our objective was to analyse the pulsed-field gel electrophoresis (PFGE) and antibioresistance patterns of strains belonging to these serovars and to assess the significance of broiler-chicken meat as a source of human infection. METHODS AND RESULTS: A total of 142 Salmonella isolates were analysed: 79 were isolated from Senegalese patients with sporadic diarrhoea (11 S. Hadar, nine S. Brancaster and 59 S. Enteritidis) and 63 from poultry (30 S. Hadar, 17 S. Brancaster and 16 S. Enteritidis). The PFGE of XbaI- and SpeI-digested chromosomal DNA gave 20 distinct profiles for S. Hadar, nine for S. Brancaster and 22 for S. Enteritidis. Each serovar was characterized by a major pulsotype which was X3S1 in 42% of S. Hadar, X8S1 in 53.8% of S. Brancaster and X1S2 in 43% of S. Enteritidis isolates. Human and poultry isolates of Salmonella had common PFGE patterns. Antibiosensitivity tests showed multiresistance (more than two drugs) was encountered in 14.5% of S. Hadar and in 5% of S. Enteritidis isolates. Resistance to quinolones was considered to be of particular importance and 14.5% of S. Hadar isolates were found to be resistant to nalidixic acid. CONLCUSIONS: The sharing of similar PFGE profiles among isolates from humans and poultry provided indirect evidence of Salmonella transmission from contaminated broiler meat. But most of the Salmonella isolates remained drug sensitive. SIGNIFICANCE AND IMPACT OF THE STUDY: Efforts are needed to eliminate Salmonella from poultry meat intended for human consumption. This study has also highlighted the importance of continuous surveillance to monitor antimicrobial resistance in bacteria associated with animals and humans.

Risk factors for contamination of ready-to-eat street-vended poultry dishes in Dakar, Senegal.

Our objective was to investigate the Salmonella and Campylobacter contamination of traditional ready-to-eat street-vended poultry dishes and to assess the association of some restaurant characteristics and cooking practices with the contamination of these meals. One hundred and forty-eight street-restaurants were studied from January 2003 to April 2004 in Dakar. A questionnaire was submitted to the managers, and samples of ready-to-eat poultry dishes were taken. Salmonella spp. was isolated in 20.1% of the 148 street-restaurants studied and in 10.1% samples of poultry dishes. The most prevalent serovars isolated were Salmonella hadar, Salmonella enteritidis and Salmonella brancaster. Campylobacter jejuni was detected in only 3 restaurants and 3 poultry dishes. Not peeling and not cleaning vegetables and other ingredients during meal preparation (OR=3.58), dirty clothing for restaurant employees (OR=4.65), reheating previously cooked foods (OR=5.2), and no kitchen and utensils disinfection (OR=3.47) were associated with an increasing risk of Salmonella contamination. Adequate cooking procedures decreased the risk of Salmonella contamination (OR=0.15).

Evaluation of the seroprevalence of rubella in the region of Dakar (Senegal).

Data on the seroepidemiology of rubella in tropical African countries are still scarce. To determine the seroprevalence in Senegal, we conducted a 6 year retrospective survey among women of child bearing age in the urban region of Dakar, Senegal. The global seroprevalence determined with a commercial enzyme immunoassay among 3471 serological results was 90.1%. The distribution of this prevalence appeared stable with no significant difference between the years of study, age groups, and the socio-economic level of the districts of residence of the patients. Compared to seroepidemiological surveys performed in other western African countries, our data suggest an important and stable circulation of the virus in the region of Dakar. The lack of data on rubella and congenital rubella syndrome (CRS) in Senegal should encourage medical authorities to establish a national rubella surveillance network in order to develop a strategy to survey and control CRS in the country.

[Rhône-Alpes observatory of Streptococcus pneumoniae in 1999: 35 cases of meningitis]. / Observatoire Rhône-Alpes du pneumocoque en 1999: 35 cas de méningites.

In 1999, in Rhône-Alpes region, in a survey of resistance to antibiotics of Streptococcus pneumoniae, 35 cases of meningitis were observed. A retrospectic questionnary was sent to each participant. MICs to Penicillin, Amoxicillin and Cefotaxime were determined with ATB-PNEUMO gallery or E-test and by disk diffusion for the other antibiotics. The results were interpreted according to the recommendations of the CA-SFM. Mean age was 38.1 years (range : 1 month -78 years) and sex-ratio 2/5. Eight patients had previously received antibiotics, 22 patients had risk factors and 23 were transferred in intensive care unit. The patients received C3G + glycopeptide in 15 of 16 children and in 13/19 adults according to the consensus recommendations. Diagnostic was made on the direct examination of CSF in 83%, and blood cultures was positive in 74.3% of cases. The percentage of PRP was 48.6% with 17.1% of intermediate-amoxicilline and 14.3% intermediate-cefotaxime strains. Resistance to trimethoprim-sulfamethoxazole was 45.7%, to chloramphenicol 30% and to fosfomycin 6.9%. All the strains were susceptible to rifampicin and vancomycin. Among the 17 PRP strains, 7 were belonging to serotype 6 (6 in children). The clinical outcome was fatal in 7 male cases (20%), without risk factors in 3 children and 6 of 7 strains were susceptible to penicillin. Six patients (17%) had auditive and/or neurologic sequellaes. This study shows that nearly 50% of strains isolated in meningitis, in Rhône-Alpes region, were not susceptible to penicillin, and confirms the frequency of sequellaes while the mortality is not related with the resistance of strains to the antibiotics.

Distribution and susceptibility of bacterial urinary tract infections in Dakar, Senegal.

To assess the current distribution and antibiotic susceptibility patterns of uropathogens isolated from community-acquired urinary tract infections (UTIs) in Dakar, a retrospective study was carried out at the biomedical laboratory of Institut Pasteur de Dakar between January 1999 and December 2000. The most important finding, among Enterobacteriaceae, was the emergence of multidrug resistant strains (extended-spectrum beta-lactamase and both chromosomally derepressed and plasmid mediated cephalosporinase producing strains) affecting 2.9% of all Enterobacteriaceae. Of Gram-positive cocci, 15% of Staphylococcus aureus were methicillin-resistant. Our data show the original distribution of uropathogens from UTIs in ambulant patients and the emergence of multidrug resistant strains in Dakar.

Comparative in vitro activity of penicillin G, levofloxacin, moxifloxacin, telithromycin, pristinamycin, quinupristin-dalfopristin and linezolid against ofloxacin-intermediate and -resistant Streptococcus pneumoniae.

Screening by ofloxacin disk was carried out on 1158 strains of Streptococcus pneumoniae in order to investigate the in vitro bacteriostatic activity of penicillin G, levofloxacin, moxifloxacin, telithromycin, linezolid, pristinamycin and quinupristin-dalfopristin against ofloxacin-intermediate and -resistant S. pneumoniae strains. It was concluded that these new antimicrobial agents could be useful for the treatment of pneumococcal infections caused by penicillin-sensitive and -resistant S. pneumoniae, and would represent a valid therapeutic option for patients allergic to beta-lactams, should they prove to be potent in vivo.

[Antibiotic sensitivity of Salmonella isolated in army hospitals between 1998-1999]. / Sensibilité aux antibiotiques des salmonelles isolées dans les hôpitaux des armées en 1998-1999.

OBJECTIVES: Monitor the evolution in France of antibiotic sensitivity of non-typhoid salmonella isolated in fecal cultures conducted in army hospital laboratories. METHODS: A prospective study was performed from January 1998 to December 1999 in all the biology departments of the 11 army hospitals in France. All the non-repetitive strains were sent to an official center for serotyping and determination of the minimum inhibiting concentrations, by dilution in Mueller Hinton's gelose. The antibiotics currently used in treatment were tested and interpretation endpoints followed the recent recommendations of the Antibiogram committee of the French society of microbiology. Identification of beta-lactamase was conducted by iso-electric focalization and polymerization by chain reaction (PCR). For Salmonella Typhimurium, research for the specific resistance locus of the DT104 clone was made using PCR. RESULTS: Two hundred and twenty-two non-repetitive salmonella strains were isolated. The principle serotypes found were: Salmonella Enterididis (23.9%), S. Typhimurium (21.2%) and S. Hadar (10.8%). All the strains were sensitive to ciprofloxacin and cefotaxim, whereas one third exhibited reduced sensitivity to aminopenicillin. Depending on the serotype, Typhimurium and Hadar serotypes exhibited significantly lesser sensitivity to aminopenicillin, nalidixic acid and tetracycline. For S. Typhimurium, resistance is related to the diffusion of the multiresistant DT104 clone, which involves half of the strains of this serotype. For S. Hadar, 18 out of 24 strains (75%) were resistance to nalidixic acid, and 5 of them exhibited reduced resistance to ciprofloxacin. There was no difference in sensitivity to antibiotics between the strains responsible for diarrhea and those isolated in systematic examinations for capacity to work in the food trade (respectively 54 and 46% of strains). CONCLUSION: Non-typhoid salmonella are frequently isolated in diarrhea of infectious origin. The increasing resistance to antibiotics is primarily related to the diffusion of the DT104 clone, regarding S. Typhimurium and to the increase in resistance to quinolone, regarding S. Hadar.

Chromobacterium violaceum: a case of diarrhea in Senegal.

A 5-year-old infant with diarrhea had heavy growth of Chromobacterium violaceum cultured from stool. This organism is restricted geographically between latitudes 35 degrees N and 35 degrees S. It can cause sepsis and various focal infections but is not a well-known cause of diarrhea.

[Evaluation of the E-test and the ATB-PNEUMo battery for determining the beta-lactam MIC for Streptococcus pneumoniae in daily practice]. / Evaluation du E-test et de la galerie ATB-PNEUMo dans la détermination des CMI aux bêtalactamines de Streptococcus pneumoniae en pratique quotidienne.

In 1999, during the survey of resistance of Streptococcus pneumoniae to antibiotics by 31 clinical laboratories of Rhône-Alpes area, MIC to penicillin (P), amoxicillin (AMX) and cefotaxime (CTX) of 877 PRP strains or with a diameter of inhibition to oxacillin inferior to 26 mm, were determined by each institution by E-test (n = 220 strains) or ATB-PNEUMO (n = 657 strains). MICs of these three antibiotics were determined by dilution in agar medium by the coordinating center. The essential agreement was respectively for ATB-PNEUMO and E-test 89% versus 84% for P (p > 0.05), of 86% vs 79% for AMX (p < 0.01), and of 91% vs 86% for CTX (p = 0.03). When the strains were classified in clinical category, the differences were significant (p < 0.001) for AMX (85% vs 71%) and for CTX (82% vs 75%) but not for P (73% vs 78%). ATB-PNEUMO method was more sensitive than E-test for the detection of strains susceptible to P (90 vs 73%), to AMX (83 vs 78%) and to CTX (80 vs 72%) and for the strains intermediate to AMX (90 vs 78%). On the contrary, E-test is more specific than ATB-PNEUMO for the detection of P-resistant strains (94 vs 86%). Finally, the specificity of both methods is the same for detection of P-S, AMX-R and CTX-I strains.