Toxin Production and Resistance of Staphylococcus Species Isolated from Fermented Artisanal Dairy Products in Benin.

Staphylococcus species are considered as one of the major pathogens causing outbreaks of food poisoning. The aim of this work was to assess the toxinogenic and antibiotic susceptibility profiles of the strains of Staphylococcus spp isolated from three types of fermented dairy products (yoghourt, millet dêguê, and couscous dêguê). The isolation of the Staphylococcus strains was performed on selective media, and their identification was done using biochemical and molecular methods. The susceptibility at 15 antibiotics tested was assessed using the disc diffusion method. The immunodiffusion method was used to evaluate the toxin (luk-E/D, luk-S/F, ETA, and ETB) production. Biofilm formation was qualitatively researched on microplates. Less than half (42.77%) of the collected samples were contaminated with Staphylococcus spp. The yoghourt and millet dêguê samples collected in the afternoon were more contaminated than those collected in the morning. The S. aureus, S. capitis, and S. xylosus strains, respectively, were the most present. S. aureus was the only coagulase-positive species identified in our samples. The highest resistance to antibiotics was observed with penicillin (100%) irrespective of the nature of the sample. S. aureus strains were highly (71.4%) resistant to methicillin. The S. aureus strains were the most biofilm-forming (27.6%), followed by S. capitis strains. Panton and Valentine's leukocidin (luk-S/F) was produced by only S. aureus strains at a rate of 8.33%. Only coagulase-negative Staphylococcus (CNS) produced Luk-E/D. The high rates of Staphylococci contamination indicate bad hygiene quality during the production and distribution of dairy products. It is, therefore, necessary to improve the quality of fermented milk products.

Extended spectrum beta-lactamase and fluoroquinolone resistance genes among Escherichia coli and Salmonella isolates from children with diarrhea, Burkina Faso.

BACKGROUND: The emergence and spread of multidrug-resistant gram-negative bacteria (MDR) has become a major public health concern worldwide. This resistance is caused by enzymes-mediated genes (i.e., extended spectrum beta-lactamases) that are common in certain Enterobacterioceae species. However, the distribution of these genes is poorly documented in Burkina Faso. This study aims to determine the prevalence and distribution of the resistant genes coding for broad spectrum beta-lactamases and quinolones in rural Burkina Faso. METHODS: Multiplex PCR assays were carried out to detect ESBL-encoding genes, including blaOXA, blaTEM, blaCTX-M, blaSHV. The assays also assessed the presence of quinolone resistance gene namely qnrA, qnrB and qnrS in the quinolone-resistance DEC and Salmonella strains. RESULTS: The Extended-Spectrum Beta-Lactamases (ESBL) resistance phenotype was reported in all the E. coli isolates (5/5). Cross-resistance phenotype to quinolones (CRQ) was shown by one Salmonella strain (1/9) and three E. coli (3/5). Cross-resistance phenotypes to fluoroquinolones (CRFQ) were harboured by one Salmonella (1/9) and carbapenemase phenotypes were detected in two E. coli strains (2/5). Whilst the blaOXA genes were detected in 100% (5/5) of E. coli isolates and in 33.33% (3/9) Salmonella isolates. One strain of E. coli (1/5) harbored the blaCTX-M gene and the qnrB gene simultaneously. CONCLUSIONS: This study identified ß-lactam (bla) and quinolone resistance (qnr) genes in multidrug-resistant E. coli and Salmonella spp. in rural Burkina Faso. Our finding which highlighted the enterobacteriaceae strains resistance to ß-lactams and quinolones are of high interest for adequate management of antimicrobial resistant genes outbreak in Burkina Faso.

Distribution of resistance genes encoding ESBLs in Enterobacteriaceae isolated from biological samples in health centers in Ouagadougou, Burkina Faso.

OBJECTIVE: Resistance to antibiotics most especially third generation cephalosporins has assumed a worrisome dimension globally. Genes conferring these resistance which are mediated by enzymes known as extended spectrum beta-lactamases (ESBLs) are now wide spread among several Enterobacteriaceae species. However there is paucity of data regarding the distribution of these genes in Burkina Faso. Hence this prospective study aims to determine the prevalence and distribution of ESBL encoding genes in ESBL producing Enterobacteriaceae strains isolated from clinical samples of patients attending the three major hospitals in Ouagadougou Burkina Faso. RESULTS: ESBL-encoding genes were assayed in 187 ESBL producing Enterobacteriaceae strains. Among these isolates, the prevalence of ESBL-producing strains with blaTEM, blaSHV and blaCTX-M genes were 26.2% (49/187), 5.9% (11/187) and 40.1% (75/187) respectively. The association of ESBL encoding genes with health centers was statistically significant (p = 0.0209). Approximately 39.6% of E. coli harbored CTX-M and Klebsiella spp. 5.9%. This study demonstrates the dissemination of TEM, SHV and CTX-M genes in ESBL producing Enterobacteriaceae strains in Ouagadougou. Continuous spread of these bacteria poses great public health risk, thus increased surveillance and regulation of antibiotics use is imperative in Burkina Faso.

Comparative study of the impact of the administration of Amoxicillin and Algo-Bio® alternative substance to antibiotics, on the level of selection of resistant Enterobacteriaceae in the digestive flora of piglets.

OBJECTIVES: The aim of study was to evaluate by comparative study the level of selection of antibiotic-resistant Enterobacteriaceae in the digestive microbiota of piglets when using amoxicillin and Algo-Bio®. METHODS: Amoxicillin and Algo-Bio® administration was carried out over a period of 5 days (D0-D4) at a dose of 1mL/10kg body weight. A phenotypic study was carried out with enumeration of resistant Enterobacteriaceae on MacConkey agar plates in the presence and absence of amoxicillin. Escherichia coli isolates were identified and were subjected to antimicrobial susceptibility testing. RESULTS: The percentages of amoxicillin-resistant Enterobacteriaceae before treatment ranged from 10-15% for the four groups of piglets. Following treatment initiation, on the second day (D1) to the fifth day (D4) of treatment, the percentages increased to 54-87% for the groups treated with amoxicillin. In the group treated with Algo-Bio® and the controls, the percentages were <50%. The percentage of amoxicillin-resistant E. coli strains to the associated antibiotics increased during days of amoxicillin treatment, whereas in the control and Algo-Bio® groups the percentages of E. coli resistant to antibiotics did not increase. CONCLUSION: The results indicated that Algo-Bio® constitutes a good alternative prophylactic to antibiotics to reduce bacterial growth in the digestive tract of animals.

Epidemiology and Antibiotic Resistance Phenotypes of Diarrheagenic Escherichia Coli Responsible for Infantile Gastroenteritis in Ouagadougou, Burkina Faso.

The emergence and persistence of multidrug-resistant (MDR) diarrheagenic Escherichia coli (DEC) causing acute diarrhea is a major public health challenge in developing countries. The aim of this study was to evaluate the resistance phenotypes of DEC isolated from stool samples collected from children less than 5 years of age with acute diarrhea living in Ouagadougou/Burkina Faso. From August 2013 to October 2015, this study was carried out on 31 DEC strains of our study conducted in "Centre Médical avec Antenne Chirurgicale (CMA)" Paul VI and CMA of Schiphra. DEC were isolated and identified by standard microbiological methods and polymerase chain reaction (PCR) method was used to further characterize them. Antimicrobial susceptibility testing was done based on the disk diffusion method. DEC isolates were high resistant to tetracycline (83.9%), amoxicillin (77.4%), amoxicillin clavulanic acid (77.4%), piperacillin (64.5%), and colistin sulfate (61.3%). The most resistant phenotype represented was the extended spectrum ß-lactamase (ESBL) phenotype (67.7%). Aminoglycosides were 100% active on enteroinvasive E. coli (EIEC) and enterohemorrhagic E. coli (EHEC). All the DEC isolates exhibited absolute (100%) sensitivity to ciprofloxacin. Monitoring and studying the resistance profile of DEC to antibiotics are necessary to guide probabilistic antibiotic therapy, especially in pediatric patients.

Molecular Characterization of Diarrheagenic Escherichia Coli in Children Less Than 5 Years of Age with Diarrhea in Ouagadougou, Burkina Faso.

Diarrheagenic Escherichia coli (DEC) is important bacteria of children's endemic and epidemic diarrhea worldwide. The aim of this study was to determine the prevalence of DEC isolated from stool samples collected from children with acute diarrhea living in Ouagadougou, Burkina Faso. From August 2013 to October 2015, stool samples were collected from 315 children under 5 years of age suffering from diarrhea in the "Centre Médical avec Antenne Chirurgicale (CMA)" Paul VI and the CMA of Schiphra. E. coli were isolated and identified by standard microbiological methods, and the 16-plex PCR method was used to further characterize them. Four hundred and nineteen (419) E. coli strains were characterized, of which 31 (7.4%) DEC pathotypes were identified and classified in five E. coli pathotypes: 15 enteroaggregative E. coli (EAEC) (48.4%), 8 enteropathogenic E. coli (EPEC) (25.8%) with 4 typical EPEC and 4 atypical EPEC, 4 enteroinvasive E. coli (EIEC) (12.9%), 3 enterohemorrhagic E. coli (EHEC) 9.67%, and 1 enterotoxigenic E. coli (ETEC) 3.2%. The use of multiplex PCR as a routine in clinical laboratory for the detection of DEC would be a useful mean for a rapid management of an acute diarrhea in children.

Characterization of Virulence Potential of Pseudomonas Aeruginosa Isolated from Bovine Meat, Fresh Fish, and Smoked Fish.

Pseudomonas aeruginosa owns a variability of virulence factors. These factors can increase bacterial pathogenicity and infection severity. Despite the importance of knowledge about them, these factors are not more characterized at level of strains derived from local food products. This study aimed to characterize the virulence potential of P. aeruginosa isolated from various animal products. Several structural and virulence genes of P. aeruginosa including lasB, exoS, algD, plcH, pilB, exoU, and nan1 were detected by polymerase chain reaction (PCR) on 204 strains of P. aeruginosa. They were isolated from bovine meat (122), fresh fish (49), and smoked fish (33). The 16S rRNA gene was detected on 91.1% of the presumptive strains as Pseudomonas. The rpoB gene showed that 99.5% of the strains were P. aeruginosa. The lasB gene (89.2%) was the most frequently detected (p < 0.05). In decreasing importance order, exoS (86.8%), algD (72.1%), plcH (72.1%), pilB (40.2%), and exoU (2.5%) were detected. The lasB gene was detected in all strains of P. aeruginosa serogroups O11 and O16. The prevalence of algD, exoS, and exoU genes in these strains varied from 51.2% to 87.4%. The simultaneous determination of serogroups and virulence factors is of interest for the efficacy of surveillance of infections associated with P. aeruginosa.

Population structure of clinical Pseudomonas aeruginosa from West and Central African countries.

BACKGROUND: Pseudomonas aeruginosa (PA) has a non-clonal, epidemic population with a few widely distributed and frequently encountered sequence types (STs) called 'high-risk clusters'. Clinical P. aeruginosa (clinPA) has been studied in all inhabited continents excepted in Africa, where a very few isolates have been analyzed. Here, we characterized a collection of clinPA isolates from four countries of West and Central Africa. METHODOLOGY: 184 non-redundant isolates of clinPA from hospitals of Senegal, Ivory Coast, Nigeria, and Central African Republic were genotyped by MLST. We assessed their resistance level to antibiotics by agar diffusion and identified the extended-spectrum ß-lactamases (ESBLs) and metallo-ß-lactamases (MBLs) by sequencing. The population structure of the species was determined by a nucleotide-based analysis of the entire PA MLST database and further localized on the phylogenetic tree (i) the sequence types (STs) of the present collection, (ii) the STs by continents, (iii) ESBL- and MBL-producing STs from the MLST database. PRINCIPAL FINDINGS: We found 80 distinct STs, of which 24 had no relationship with any known STs. 'High-risk' international clonal complexes (CC155, CC244, CC235) were frequently found in West and Central Africa. The five VIM-2-producing isolates belonged to CC233 and CC244. GES-1 and GES-9 enzymes were produced by one CC235 and one ST1469 isolate, respectively. We showed the spread of 'high-risk' international clonal complexes, often described as multidrug-resistant on other continents, with a fully susceptible phenotype. The MBL- and ESBL-producing STs were scattered throughout the phylogenetic tree and our data suggest a poor association between a continent and a specific phylogroup. CONCLUSIONS: ESBL- and MBL-encoding genes are borne by both successful international clonal complexes and distinct local STs in clinPA of West and Central Africa. Furthermore, our data suggest that the spread of a ST could be either due to its antibiotic resistance or to features independent from the resistance to antibiotics.