Prevalence and characterization of ESBL-producing Escherichia coli in healthy pregnant women and hospital environments in Benin: an approach based on Tricycle

IntroductionExtended-Spectrum Beta-Lactamase (ESBL)-producing Enterobacterales are recognized as significant pathogens due to their resistance to multiple antibiotics. This study aimed to determine the prevalence of ESBL-producing Escherichia coli (E. coli) in different settings, including healthy pregnant women, the food chain, and the environment of tertiary hospitals in Benin.MethodsSamples were collected from various sources, including fecal samples from healthy pregnant women, food samples from hospital canteens, and hospital effluents from four tertiary hospitals in southern Benin. Fecal samples were plated on MacConkey agar supplemented with cefotaxime (4 μg/mL), while food and water samples were plated on Tryptone Bile X agar supplemented with cefotaxime (4 μg/mL). Urea indole tests were used for preliminary identification of E. coli colonies, followed by confirmation of ESBL production using the double disk synergy technique. Antibiotic susceptibility testing of ESBL-producing E. coli strains was conducted using the disk diffusion method on MH agar. Polymerase Chain Reaction (PCR) was used to investigate the presence of ESBL-encoding genes.ResultsAmong the 296 fecal samples collected from four tertiary hospitals, ESBL-producing E. coli was isolated from 22.30% (66) of the samples. All E. coli isolates from hospital effluents exhibited ESBL production, while ESBL-producing E. coli was not detected in food and drinking water samples. The analysis of variable associations showed no significant associations (p > 0.05) for the studied factors. Antibiotic susceptibility testing revealed high resistance rates among the ESBL-Ec isolates against several tested antibiotics, including amoxicillin, aztreonam, ceftriaxone, ciprofloxacin, and trimethoprim-sulfamethoxazole. However, most isolates remained susceptible to ertapenem, amoxicillin-clavulanate, and imipenem. The most prevalent ESBL-encoding genes were blaTEM (37.50%), blaOXA-1 (19.44%), and blaSHV (11.11%), while a smaller proportion of isolates carried blaCTXM-1/blaCTXM-15 (5.55%) and blaCTXM-9.DiscussionThis study provides insights into the prevalence of ESBL-producing E. coli carriage in the feces of healthy pregnant women in southern Benin. Additionally, it highlights hospital wastewater as a potential reservoir of ESBL-producing bacteria in the environment. The detection of ESBL-producing E. coli in hospital effluents raises concerns about the dissemination of antibiotic resistance genes into the environment. The high resistance rates observed among ESBL-Ec isolates against commonly used antibiotics emphasize the urgent need for antimicrobial stewardship and infection control measures. The identification of prevalent ESBL-encoding genes contributes to understanding the genetic basis of ESBL resistance in the studied population. Further research is warranted to explore the mechanisms of transmission and potential interventions to mitigate the spread of ESBL-producing Enterobacterales

Molecular characterization of Enterobacteriaceae producing β-lactamase and methicillin-resistant staphylococci isolated from the hospital environment and catheters in two public hospitals in Benin, Republic of Benin

Antimicrobial resistance is a real public health problem. All over the world, it has a considerable impact in hospitals. The present study was conducted to ascertain the bacterial ecology in two hospitals in Benin as well as the resistance genes present in the recovered isolates. A total of 146 environmental and catheter samples were collected at the University Hospital Center of Abomey-Calavi / So-Ava and at the Beninese Army Hospital of Cotonou. These samples were inoculated on Mannitol Salt and Eosin Methylene Blue agars. The colonies obtained were identified and their sensitivity to antibiotics were tested, using the Kirby Bauer technique. Four resistance genes encoding the production of extended-spectrum beta-lactamases (blaCTX-M1, blaCTX-M2, blaCTX-M9, blaCTX-M15) and the gene coding for methicillin resistance (mecA) were screened.  The gene coding for methicillin resistance (mecA) was sought in staphylococci. A total of 69 (53,49%) and 60 (46,51%) strains belonging to Enterobacteriaceae family and staphylococci were identified, respectively. A predominance of Staphylococcus aureus (25.6%) followed by Enterobacter cloacae (21.0%) and coagulase negative staphylococci (21.0%) was observed. These bacterial strains showed multidrug-resistance, particularly to beta-lactams, fluoroquinolones, aminoglycosides, and macrolides. Beta-lactamases were identified in the genome of bacterial strains with a predominance of blaCTX-M15 (42.8%). The frequency of the mecA gene in staphylococci was 50%. These results show the magnitude of the antimicrobial resistance situation in the hospitals investigated. They can be used to support advocacy for urgent action at the national level, especially with regards to the management and efficient use of antimicrobials in Benin

Microbiological Characterization of Grilled Meat “Tchatchanga” in Cotonou (Southern Benin): Enumeration, Isolation and Resistance Profile of Staphylococcus aureus and Escherichia coli

Collective food-borne diseases are the reason for a considerable number of deaths in developing countries. The contamination of meat is often noticed. The purpose of the present study was to enumerate, isolate and evaluate the resistance profile of Staphylococcus aureus and Escherichia coli in grilled meat consumed in Southern Benin. To achieve this goal, 30 thirty samples of grilled meat “Tchatchanga” were collected in three districts of the city of Cotonou. After collection, the samples were identified, stored in a cooler containing cold accumulator’s and sent to the laboratory for analysis. The resistance profile of the different isolated strains was then sought: 30% of the samples were contaminated with Escherichia coli and 100% with Staphylococci. In terms of hygienic quality, 70% of the samples were of unsatisfactory. Of the 30 staphylococcal strains, 11 were identified as Staphylococcus aureus. Regarding the strain resistance profile, 88.89% of the Escherichia coli strains were multi-resistant compared to 72.72% of Staphylococcus aureus. This study revealed the necessity of urgent actions to ensure food safety in Benin

Isolation and characterization of three novel Acinetobacter baumannii phages from Beninese hospital wastewater

Acinetobacter baumannii is an opportunistic pathogen that is mostly associated with hospital-acquired infections. The rapid emergence of multi- and pan-drug-resistant Acinetobacter strains poses an increasing challenge in hospitals. Phage therapy offers one treatment option for infections caused by A. baumannii. We isolated three phages from Beninese hospital wastewater – fBenAci001, fBenAci002, and fBenAci003 – that infected clinical A. baumannii strains from Finnish patients. Phylogenetic analysis showed that these phages resemble phages of the genus Friunavirus, family Autographiviridae. The isolated phages meet the requirements set for phages used for phage therapy. However, they were found to have a narrow host range, which may limit their therapeutic use.Peer reviewe

Data_Sheet_1_Prevalence and characterization of ESBL-producing Escherichia coli in healthy pregnant women and hospital environments in Benin: an approach based on Tricycle.pdf

IntroductionExtended-Spectrum Beta-Lactamase (ESBL)-producing Enterobacterales are recognized as significant pathogens due to their resistance to multiple antibiotics. This study aimed to determine the prevalence of ESBL-producing Escherichia coli (E. coli) in different settings, including healthy pregnant women, the food chain, and the environment of tertiary hospitals in Benin.MethodsSamples were collected from various sources, including fecal samples from healthy pregnant women, food samples from hospital canteens, and hospital effluents from four tertiary hospitals in southern Benin. Fecal samples were plated on MacConkey agar supplemented with cefotaxime (4 μg/mL), while food and water samples were plated on Tryptone Bile X agar supplemented with cefotaxime (4 μg/mL). Urea indole tests were used for preliminary identification of E. coli colonies, followed by confirmation of ESBL production using the double disk synergy technique. Antibiotic susceptibility testing of ESBL-producing E. coli strains was conducted using the disk diffusion method on MH agar. Polymerase Chain Reaction (PCR) was used to investigate the presence of ESBL-encoding genes.ResultsAmong the 296 fecal samples collected from four tertiary hospitals, ESBL-producing E. coli was isolated from 22.30% (66) of the samples. All E. coli isolates from hospital effluents exhibited ESBL production, while ESBL-producing E. coli was not detected in food and drinking water samples. The analysis of variable associations showed no significant associations (p > 0.05) for the studied factors. Antibiotic susceptibility testing revealed high resistance rates among the ESBL-Ec isolates against several tested antibiotics, including amoxicillin, aztreonam, ceftriaxone, ciprofloxacin, and trimethoprim-sulfamethoxazole. However, most isolates remained susceptible to ertapenem, amoxicillin-clavulanate, and imipenem. The most prevalent ESBL-encoding genes were blaTEM (37.50%), blaOXA-1 (19.44%), and blaSHV (11.11%), while a smaller proportion of isolates carried blaCTXM-1/blaCTXM-15 (5.55%) and blaCTXM-9.DiscussionThis study provides insights into the prevalence of ESBL-producing E. coli carriage in the feces of healthy pregnant women in southern Benin. Additionally, it highlights hospital wastewater as a potential reservoir of ESBL-producing bacteria in the environment. The detection of ESBL-producing E. coli in hospital effluents raises concerns about the dissemination of antibiotic resistance genes into the environment. The high resistance rates observed among ESBL-Ec isolates against commonly used antibiotics emphasize the urgent need for antimicrobial stewardship and infection control measures. The identification of prevalent ESBL-encoding genes contributes to understanding the genetic basis of ESBL resistance in the studied population. Further research is warranted to explore the mechanisms of transmission and potential interventions to mitigate the spread of ESBL-producing Enterobacterales.</p

Exploration of the antibacterial and chemical potential of some Beninese pharmacopoiea traditional plants

Objectives. This study aims to evaluate the antibacterial and chemical properties of some medicinal plants used in the fight against enteropathogens in Benin. Methods. Aqueous and ethanolic extracts of Senna siamea, Uvaria chamae, Lantana camara and Phyllantus amarus were tested on 10 bacterial strains. Well diffusion technique, coupled with the microdilution determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (CMB) was used for antibacterial testing. The larval cytotoxicity was evaluated by using Artemia salina crustacean larvae. flavonoids and polyphenols were also assayed by the method using aluminum trichloride (AlCl3) and the method using the folin-Ciocalteu reagent, respectively. Results. The results of the study revealed that extracts had an effective antibacterial activity at 100 mg/mL, with MIC between 100 and 25 mg/mL and CMB between 100 and 50 mg/mL. The inhibition diameters of the extracts varied between 7.5 and 21 mm. The ethanolic extract of Phyllantus amarus leaves showed the best antibacterial activity. None of the extracts tested was found to be cytotoxic at the dose of 20 mg/mL. The aqueous Uvaria chamae root extract has the highest polyphenol content (231.896552±0.27586207 in μg EAG/100 mg extract), whereas the aqueous leaf extract of Uvaria chamae is the richest in flavonoids (41.061082 0.43180737 in μg ER/100 mg of extract). Conclusions. These interesting results can be used in the development of improved traditional medicines against enteropathogens

Toxicological Characterization of Ten Medicinal Plants of the Beninese Flora Used in the Traditional Treatment of Diarrheal Diseases

The use of medicinal plants in traditional medicine is a common practice in developing countries. However, this unregulated or poorly rational use may present a dose-dependent risk of toxicity to humans. This study aimed to explore the phytochemical and toxicological characteristics of ten (10) plant species used in the traditional treatment of infectious diarrhea in Benin. The acute toxicity of aqueous and hydroethanolic extracts of Khaya senegalensis, Daniellia oliveri, Rauvolfia vomitoria, Vernonia amygdalina, Manihot esculenta, Ocimum gratissimum, Senna italica, Diospyros mespiliformis, Pterocarpus erinaceus, and Anacardium occidentale was evaluated following the OECD 423 protocol at a single dose of 2000 mg/kg. This safety test was complemented by a larval cytotoxicity test. Hematological and biochemical examinations, as well as a histological study of the liver and kidneys, were performed. Larval cytotoxicity was assessed by the sensitivity of Artemia salina larvae to different concentrations of the plant extracts studied. Testing for chemical compounds was performed on the basis of differential staining and precipitation reactions. The mean lethal concentration (LC50) was determined by the probit method. The qualitative phytochemical screening of the plants studied revealed the presence of catechic tannins, gallic tannins, flavonoids, anthocyanins and sterol-terpenes, alkaloids, saponosides, and reducing compounds. This composition varied according to the plants studied. Acute toxicity data indicated that there was no mortality and no structural and functional alterations of the liver and kidneys of treated animals. Larval cytotoxicity data suggest that the plants studied are not cytotoxic (LC50 ≥ 0.1 mg/mL). These observations reflect the safety of these plants and justify their use in traditional medicine in the treatment of many diseases including diarrheal diseases

Pathogenicity, Epidemiology and Virulence Factors of Salmonella species: A Review

Salmonella infections are major public health problems worldwide. The hereby review aimed to establish an overview on the pathogenicity, epidemiology and virulence factors of Salmonella spp. in the world. A systematic search was conducted online using the keywords ‘Salmonella’, ‘Salmonella spp.’, ‘Salmonella spp. Epidemiology’, ‘virulence factors of Salmonella spp. in the world’, ‘bacteria responsible for the contamination of meat products’, ‘non-typhoid salmonella’. These keywords were entered into databases such as PubMed and Google Scholar using mainly French language. The obtained articles were included based on the reliability of their source, the study area (usually Benin and Africa) and the subject. The review revealed that Salmonella spp. is motile Gram-negative rod-shaped bacteria, of the family Enterobacteriaceae, currently counting more than 2,600 serovars. Human contamination occurs through the ingestion of contaminated water and food and can cause gastroenteritis or typhoid fever, which are two serious public health problems. A gene set constituting the pathogenicity islands determines the pathogenesis of Salmonella spp. The diagnosis is based on bacteriological, serological and molecular techniques. Salmonella infections are usually treated using antibiotics; however, emergence of antibiotic resistance in these microorganisms suggests that the anti-salmonella control should explore new sources such as medicinal plant

Antibiotic profiling of multidrug resistant pathogens in one-day-old chicks imported from Belgium to benin

Abstract Background Little data exist on the presence of resistant pathogens in day-old chicks imported into Benin. The occurrence of pathogenic bacteria was assessed in 180 one-day-old chicks imported from Belgium and received at the Cardinal Bernardin Gantin International Airport in Cotonou (Benin). The samples included swabbing the blisters of 180 chicks, followed by 18 pools of 10 swabs for bacterial isolation. Classic bacteriological methods based on Gram staining, culture on specific media and biochemical characterization were used. Antibacterial susceptibility screening to antibiotics was conducted using the Kirby–Bauer disc diffusion method, and the results were interpreted according to guidelines from the European Committee on Antimicrobial Susceptibility Testing (EUCAST). DNA extraction was performed by the heat treatment method. Resistance genes were screened by real-time PCR. Results We isolated 32 bacteria, including Escherichia coli (50%), Enterococcus spp. (28%), and coagulase-negative staphylococci (10%). The isolates were investigated for antibiotic resistance against antibiotics using the disk diffusion method and showed that in the Escherichia coli strains isolated, the highest rate of resistance was obtained against ciprofloxacin (81%), followed by trimethoprim + sulfamethoxazole (62%). Enterobacter cloacae was sensitive to all the antibiotics tested. Pseudomonas spp. resistant to amoxicillin and trimethoprim + sulfamethoxazole was noted. The SulII gene was found in all cloacal samples, while the SulI and bla TEM genes were present at 44.44% and 16.67%, respectively. Conclusion This study confirms that imported day-old chicks can be a potential source of dissemination of resistant bacteria in poultry production. A system for immediate detection of resistant bacteria in chicks upon arrival in the country is thus needed

Whole genomes from bacteria collected at diagnostic units around the world 2020

Abstract The Two Weeks in the World research project has resulted in a dataset of 3087 clinically relevant bacterial genomes with pertaining metadata, collected from 59 diagnostic units in 35 countries around the world during 2020. A relational database is available with metadata and summary data from selected bioinformatic analysis, such as species prediction and identification of acquired resistance genes