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Aims: This study aims to characterize of toxigenic Vibrio cholerae strains for improved cholera surveillance in Benin. Methodology: 304 diarrheal stool samples were collected from people with watery diarrhea of unknown etiology and vomiting during epidemics from 2016 to 2018 in Benin. Toxigenic Vibrio cholerae strains were isolated and then biochemical tests, serogrouping and serotyping were performed. Antibiotic susceptibility tests were performed using the disc diffusion method and E-tests. Multiplex and real-time PCR were used to identify and detect virulence genes (CtxA, OmpW and TcpA). Results: The results showed a 21.71% prevalence of toxigenic Vibrio cholera in Benin. All strains were Vibrio cholerae O1 serotype Inaba (100%) and showed a high sensitivity to doxycycline (96.97 %), chloramphenicol (95.45 %) and ciprofloxacin (90.91 %). However, antibiotic resistance was observed, especially for erythromycin (74.24 %), Trimethoprim-sulfamethoxazole (71.21 %) and ampicillin (43.94 %). The CtxA and TcpA virulence genes were respectively detected in 100% and 96.97% of the toxigenic strains of V. cholerae isolated. While the OmpW gene was identified on all the toxigenic strains of Vibrio cholerae isolated. Conclusion: Vibrio cholerae strains isolated from patients suspected of cholera were highly virulent and resistant to antimicrobials.

The study aims to document the level of contamination of the aquatic ecosystem of the Cotonou-Lake Nokoué canal hydrographic complex by multidrug-resistant bacteria and their resistance genes. For this purpose, water samples were taken from several points of the complex and from the sediments at the depth of the lake. Samples of several species of freshly caught fish products from the lake were also collected. Bacteriological analyses were carried out according to the AFNOR standard (NF U: 47–100). The identification of the different bacterial species isolated was then carried out using the API 20E gallery and specific biochemical tests. The antibiogram of the strains was performed according to the recommendations of the EUCAST. Molecular characterization of the identified strains was carried out by searching for resistance and virulence genes. The results obtained revealed the presence of several bacterial species in water samples and in sediment and intestine samples of fishery products with a predominance of Gram-negative bacilli. The resistance profile of Gram-negative bacilli showed a total resistance to metronidazole (100%). 23% of the strains were also resistant to ciprofloxacin, 41% to amoxicillin, and 60% to aztreonam. Of the Gram-positive cocci identified, 66% was resistant to vancomycin, 7.5% to ciprofloxacin, 71% to erythromycin, and 22% to tetracycline. Regarding the genes sought, blaTEM (46%), blaSHV (24%), and blaCTX-M-15 (31%) were present in the genome of Gram-negative bacilli as resistance genes and fimH (41%) as virulence gene. As for Gram-positive cocci, the van B gene was completely absent. The van A was present at 6.25% in Staphylococcus aureus and mecA at 21.88 and 33.33%, respectively, in Staphylococcus aureus and coagulase-negative staphylococci strains. The high resistance of isolated bacterial strains is a matter of concern and calls for a rational use of antibiotics in order to avoid the transmission of antibiotic resistance from the environment to humans.

This study aims to carry out a bacteriological characterization and determine the resistance profile of Vibrio cholerae O1 strains isolated during the epidemiological season of 2020 in Benin. To achieve this goal, 43 diarrheal stool samples were analyzed. The samples were taken during the epidemic period of 2020. Bacteriological analyses consisted of enrichment of the samples in buffered peptone water followed by culture on SBCT agar. Then the characteristic colonies were subjected to microscopy, biochemical identification (oxidase, seeding and reading of TSI agar and API 20 E gallery), serotyping, and antibiotic sensitivity tests using the diffusion technique in agar medium according Kirby-Bauer method. The median age of the patients included in this study was 25 years (IQR: 15-40) with predominantly female patients. Individuals aged 11 to 25 were the most represented. Of the 43 stool samples analyzed, 22 were culture positive for V. cholerae and belonged to serogroup O1. The clinical manifestations observed in patients with cholera were watery diarrhea, vomiting and severe dehydration before admission to hospital. It should be noted that all of V. cholerae O1 strains isolated were multidrug resistant with a strong resistance to erythromycin (81.13%), ampicillin (79.96%), chloramphenicol (79.06%), and cotrimoxazole (78.12%). Key words: Bacteriological analyses, Vibrio cholerae O1, antimicrobial resistance, Benin.

Background: The environment plays an important role in the dissemination of multidrug resistant bacteria, especially through the aquatic ecosystem, including hospital effluents, rivers, but also spring water and drinking water. This study aims to determine selected antimicrobial resistance genes in some aquatic matrices in southern Benin. Methods: Collected water samples were filtered through a membrane 0.22 µm thick. After filtration, the membrane was deposited on Muëller Hinton agar. Then the colonies resulting from this subculture were subjected to a microbiological examination by the conventional method. The antibiotic sensitivity test was carried out by the Kirby Bauer method according to the recommendations of the French Society of Microbiology. Resistance genes were looked for by PCR. Results: Of the 222 water samples collected, 265 bacterial strains were isolated, the majority of which were strains of Coagulase Negative Staphylococcus (CNS) with 37.74% (n = 100), followed by strains of Klebsiella pneumoniae (21.89%; n = 58), Escherichia coli (10.57%; n = 28). All isolated gram-negative bacilli strains are multidrug resistant with resistance of all strains to amoxicillin, ampicillin and amoxicillin + clavulanic acid. Of the 15 resistance genes searched in the genome of Gram-negative bacilli strains, 8 were detected, namely the TEM, SHV, CTX-M15, VIM, NDM, SUL1, SUL2 and AADA genes. Resistance of CNS strains to amoxicillin, oxacillin and cefoxitin was observed. The meca gene was detected in all CNS strains. The vanA and VanB genes were only detected in strains isolated from drinking water in sachets collected from producers and street sellers. Conclusion: These results show the dissemination of resistance genes in Benin and once again confirms the urgency of a global fight against antimicrobial resistance.

This study aimed to provide data on the subacute toxicity and fetotoxicity of aqueous and hydroethanolic extracts of Daniellia oliveri (Rolfe) Hutch. & Dalziel, Anacardium occidentale L., Diospyros mespiliformis Hochst. ex A. DC., Khaya senegalensis (Desv.) A. Juss., Ocimum gratissimum L., Vernonia amygdalina Delile, Pterocarpus erinaceus Poir., and Manihot esculenta Crantz used in the treatment of diarrheal diseases in Benin. For subacute toxicity, each male and female Wistar rat received by esophageal gavage a maximum of 1000 mg/kg over 28 days following the Organization for Economic Co-operation and Development (OECD) Guideline No. 407. Bodyweight changes, biochemical and hematological parameters were assessed. The fetotoxicity of the extracts was evaluated on Dutch Blue hen eggs at 100 mg/kg and 300 mg/kg. The hatching rate, biochemical and hematological constants were subsequently determined. The subacute toxicity data did not indicate any mortality or signs of toxicity. Also, no significant difference in the bodyweight of the rats and the hematological parameters was noted. Concerning fetotoxicity, a low hatching rate (varying from 20% to 50%) was observed in the batches treated with the extracts at 300mg/kg compared to the rate obtained in the negative control batch (80%). The batches that received the hydroethanolic extracts of Vernonia amygdalina and Manihot esculenta at the tested doses had a hatching rate of 0%. When looking at hematological parameters, no significant difference was noted for all batches. However, an increase in the Aspartate Amino-Transferase (ASAT) values was noted in the batches that received the hydroethanolic extract of Khaya senegalensis and the aqueous extract of Diospyros mespiliformis at 100 and 300 mg/kg. These results explicitly show that the hydroethanolic extracts of Manihot esculenta and Vernonia amygdalina are fetotoxic at the tested doses.

Cholera is an epidemic disease and a real public health problem throughout the world, particularly in West Africa. This study provides a comprehensive overview of the pathogenicity, epidemiology and Vibrio cholerae strains’s antibiotics resistance in West Africa. A literature review was conducted online in English using the keywords “Cholera”, “Vibrio cholerae” “West Africa”, “Epidemiology”, “Antibiotic resistance”. These keywords were entered into using electronic databases such as PubMed, Google Scholar, Scopus and Elsevier and articles were used according to the reliability of their sources, study areas, and subjects. This review was based on the collected data from different databases. One hundred and twenty-three articles were identified. After the initial and final sorting of the collected data in order to eliminate duplicate copies, eighty-three were retained while seventy articles were selected, respectively, for this review. Though some studies had recommended for a system of monitoring cholera in West African countries, nevertheless, there is the need to create more awareness. Furthermore, hygienic practices and environmental wastes management in these countries need to be improved. Key words: Cholera, epidemiology, Vibrio cholerae, antibiotic resistance, West Africa.

Cholera is a disease that Benin has been fighting against for years but which continues to create economic losses for households. The aim of this study is to assess the level of knowledge of the population about this disease and to assess hygiene practices in order to identify the factors responsible for its sustainability. The data were collected through a survey of 720 households in the 12 departments of the country. These data were analyzed with SAS software and three groups of people were identified on the basis of hygiene measures. The majority of respondents know cholera. The main symptoms they mentioned were vomiting (84.83%) and diarrhea (81.04%). Diarrhea was more reported in group 3 (88.96%) than in groups 1 (86.58%) and 2 (83.19%). Low levels of hygiene were the main cause of this disease. In the absence of toilets, the interviewees pass stool in the bush and at the edge of water sources used in households. Some of them have bins (in particular group 1) for household waste but do not subscribe to refuse collection structures. Therefore, they throw garbage in sometimes unofficial landfills. Others, on the other hand, have not garbage cans (mainly group 2) and they empty garbage everywhere (street, gutters and bushes). In addition, hand washing is hardly ever done after using the toilet. Hygiene practices in some households are insufficient, which does not protect the population from cholera.

Objectives. This study aims to study the epidemiological and geographic characteristics of the meningococcal serogroups four years after the introduction of serogroup A meningococcal conjugate vaccine. Methods. This is a prospective, descriptive, analytical study, and it took place from 2016 to 2018. Cerebrospinal fluid (CSF) samples were taken after the identification of meningitis cases. The samples, thus, taken were sent to the laboratory for culture and identification of Neisseria meningitidis in accordance with WHO standards. Results. Eight hundred and ninety-nine bacterial strains were identified, of which 219 were strains of Neisseria meningitidis. The majority of N. meningitidis-positive samples were from male patients (59.8%) with a median age of 4 (IQR: 1–13). Four of N. meningitidis serogroups were identified, namely, serogroups C (6.8%), W (19.6%), X (1.8%), and A (0.5%). Geographically, 92.7% of the identified N. meningitidis serogroups came from patients who lived in the northern region of the country. The departments most concerned were Alibori (N. meningitidis C (66.7%) and N. meningitidis W (20.9%)); Atacora (N. meningitidis W (41.9%), N. meningitidis X (75.0%), and N. meningitidis C (13.3%)); and Borgou (N. meningitidis W (23.3%)). Conclusion. The results of this study showed that there is an emergence of cases of meningococcal of serogroup C four years after the introduction of MenAfricVac in Benin. These results demonstrated the effectiveness of case-by-case surveillance in detecting small changes in the distribution of serogroups that could have important implications for public health strategies in the coming seasons.

Objective Poultry is commonly considered to be the primary vehicle for Campylobacter infection in humans. The aim of this study is to assess the risk of Campylobacteriosis in chicken meat consumers in southern Benin by assessing the prevalence and resistance profile of Campylobacter coli and Campylobacter jejuni isolated from chicken thigh in Southern Benin. Results The contamination rate of Campylobacter in the samples was 32.8%. From this percentage, 59.5% were local chicken thighs and 40.5% of imported chicken thighs (p = 0.045). After molecular identification, on the 256 samples analyzed, the prevalence of C. jejuni was 23.4% and 7.8% for C. coli, with a concordance of 0.693 (Kappa coefficient of concordance) with the results from phenotypic identification. Seventy-two-point seven percent of Campylobacter strains were resistant to Ciprofloxacin, 71.4% were resistant to Ampicillin and Tetracycline. 55.8% of the strains were multi-drug resistant.

Background: Bacterial meningitis is an inflammation of the meninges caused mainly by three bacterial species Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae that are transmitted by nasopharyngeal secretions emitted by carriers. Meningitis is a public health problem in Benin, like all countries in the African meningitis belt. This study aims to analyze the epidemiological surveillance data of meningitis in Benin from 2016 to 2018. Methods: Each suspect case of meningitis was recorded and Cerebrospinal Fluid (CSF) samples were collected. CSF collection was accompanied by the Integrated Disease Surveillance and Response form. This sheet provides information on the patient’s social-demographic and epidemiological data. CSF specimens were sent to the laboratory for analysis and identification (Gram stain, biochemical parameters, and latex agglutination test) of pathogens according to the WHO standards. Results: Of the 2992 patients with suspected meningitis, 2893 were hospitalized with a death rate of 9.4% (281/2992). The sex ratio of registered patients was 1.29 in favor of men. The median age was 4 years (min: 0; max: 90). Patients younger than five years were the most represented (44.8%). During the study period, there was a decrease in the incidence of meningitis per 100,000 inhabitants (6.3 to 3.2 from 2016 to 2018). Of 2928 CSF samples collected we were able to identify 899 pathogenic bacterial species. The most represented species are S. pneumoniae (63.4%), N. meningitidis (24.4%) and H. influenzae (12.2%). Conclusion: The burden of disease is disproportionate in the northern departments as in others. The frequency of bacterial meningitis in the northern region increased during the study period. However, deaths have been recorded in the departments of the South (“Atlantic”, “Plateau”). This suggests an improvement in epidemiological surveillance and case management throughout the national territory.

Foodborne infections, mainly those attributable to Campylobacter, are one of the most common causes of intestinal diseases, of bacterial origin in humans. Although the vehicle of transmission is not always identified, the most common vehicles are poultry, poultry products, and contaminated water. In Southern Benin, an excessive use of poultry manure as fertilizer in vegetable farms was noted. This survey aimed to determine the prevalence and concentration of Campylobacter spp., especially Campylobacter jejuni and Campylobacter coli, in selected environmental samples (poultry manure, and irrigation water) and freshly harvested leafy vegetables in two (Houeyiho and Sèmè-Kpodji) vegetable farms in southern Benin. To achieve this objective, we analyzed 280 samples, including 224 samples of leafy vegetables (Solanum macrocarpon and Lactuca sativa capita), 28 samples of irrigation water, and 28 samples of poultry manure. The analysis of the samples taken was carried out according to the modified NF EN ISO 10272-1 standard. Of the 280 samples analyzed in this survey, 63 were positive for Campylobacter contamination. For leafy vegetable samples analyzed in this survey, the contamination rate was of 15.63%. 60.71% of poultry manure samples analyzed were contaminated with Campylobacter spp. and 39.29% of irrigation water samples were contaminated. The statistical analysis of these results showed that there is a correlation between the contamination of leafy vegetables, poultry manure, and irrigations (p<0.01). Campylobacter jejuni (53.97%) was more involved in contaminations than Campylobacter coli (36.57%). This study has shown that there is a real risk of food poisoning by Campylobacter jejuni and Campylobacter coli among consumers of leafy vegetables in southern Benin. The origin of contamination of these leafy vegetables is poultry manure used as fertilizer in vegetable gardens and irrigation water used.

Background and Aim: Salmonella spp. are among the world's leading foodborne pathogens, found naturally in the intestines of many animals. Lactic acid bacteria, mainly Lactobacillus, are a promising alternative to antibiotics for animal and human health. This study aimed to assess the in vitro antibacterial activity of Lactobacillus spp. strains against virulent Salmonella spp. isolated from slaughter animals in Benin. Materials and Methods: Eleven samples of raw cow's milk, five samples of breast milk, and six infant stool samples were taken. From these samples, strains of Lactobacillus were isolated and identified. The probiotic potential of each of the identified strains was characterized, and finally in vitro antibacterial activity of these strains was evaluated against three virulent strains of Salmonella spp. and a reference strain of Salmonella Typhimurium ATCC 14028. Results: Out of the 22 samples collected, 20 strains of Lactobacillus spp. were isolated and identified. These strains included Lactobacillus plantarum (30%), Lactobacillus delbrueckii (25%), Lactobacillus casei (25%), Lactobacillus salivarius (15%), and Lactobacillus acidophilus (05%). Characterization of the probiotic potential of these strains showed that only 16 strains were resistant to pH=1.5. Fourteen of them were able to withstand the simulated gastric juice (pH 1.5+pepsin). The 14 probiotic strains showed very good antibacterial activity against virulent strains of Salmonella spp. with inhibition zone diameters ranging from 12.36±0.03 mm to 35.33±0.05 mm (R values>6 mm). Conclusion: From this study, Lactobacillus strains isolated from raw cow milk, breast milk, and infantile stool might be used as some valid candidates for probiotics. It also represents good alternatives for antibiotics in the fight against animal and human salmonellosis.

Background Healthcare-associated infections have become a public health problem, creating a new burden on medical care in hospitals. The emergence of multidrug-resistant bacteria poses a difficult task for physicians, who have limited therapeutic options. The dissemination of pathogens depends on “reservoirs”, the different transmission pathways of the infectious agents and the factors favouring them. Contaminated environmental surfaces are an important potential reservoir for the transmission of many healthcare-associated pathogens. Pathogens can survive or persist in the environment for months and be a source of infection transmission when appropriate hygiene and disinfection procedures are inefficient. The aim of this study was to identify bacterial species from hospital surfaces in order to effectively prevent healthcare-associated infections. Methods Samples were taken from surfaces at the University Hospital of Abomey-Calavi/So-Ava in South Benin (West Africa). To achieve the objective of this study, 160 swab samples of hospital surfaces were taken as recommended by the International Organization for Standardization (ISO 14698-1). These samples were analysed in the bacteriology section of the National Laboratory for Biomedical Analysis. All statistical analyses were performed using SPSS Statistics 21 software. A Chi Square Test was used to test the association between the Results of culture samples and different care units. Results Of the 160 surface samples, 65% were positive for bacteria. The frequency of isolation was predominant in Paediatrics (87.5%). The positive samples were 64.2% Gram-positive bacteria and 35.8% of Gram-negative bacteria. Staphylococcus aureus predominated (27.3%), followed by Bacillus spp. (23.3%). The proportion of other microorganisms was negligible. S. aureus and Staphylococcus spp. were present in all care units. There was a statistically significant association between the Results of culture samples and different care units (χ2 = 12.732; p = 0.048). Conclusion The bacteria found on the surfaces of the University Hospital of Abomey-Calavi/So-Ava’s care environment suggest a risk of healthcare-associated infections. Adequate hospital hygiene measures are required. Patient safety in this environment must become a training priority for all caregivers.

Additional file 1: Table S1. Distribution of the study population by age and gender. Figure S1a. Sample collection sites. (1) Abomey Calavi in the Atlantic Region (a = Central Clinic of Abomey Calavi) and (2) Cotonou in the Central Region (b = Anastasis Hospital, c = Polyclinic St Vincent de Paul and d = Mênontin Hospital). Figure S1b. Distribution of genotypes by age in Cotonou. Figure S1c. Distribution of genotypes from 90 stool samples in Abomey-Calavi. Figure S1d. Distribution of genotypes according to age in Abomey-Calavi.

With the introduction of the new meningitis A conjugate vaccine in 2012, Benin has opted for case-by-case surveillance for bacterial meningitis. The study aims to assess the case-by-case surveillance system for the meningitis epidemic in Benin during the period 2016 to 2018. A retrospective and evaluative study with a mixed approach (qualitative and quantitative) was conducted, on the three sites identified for sentinel surveillance of meningitis in Benign. The evaluation of the performance of the surveillance system was based on the updated guidelines of the Center for Disease Prevention and Control to evaluate a public health surveillance system. All criteria except sensitivity, specificity and positive predictive value were measured. Semi-structured individual interviews were conducted with the agents of the surveillance system surveyed and the normative documents as well as the notice forms were examined. This study reveals that all the centers had the case definition and notification forms. The ratio of notified cases to registered cases was 0.77. We found that the monitoring system in place had five levels. The system performance was recognized with simplicity at 80.8%, good acceptability (completeness: 98.5%; Promptitude: 88.9%) and responsiveness of 85%. The completeness of the health facilities was 45%. This study shows that it is imperative to strengthen the knowledge of the actors involved in epidemiological surveillance through periodic training to improve the performance of the surveillance system.

Rotavirus remains the main causative agent of gastroenteritis in young children, in countries that have not yet introduced the vaccine. Benin, in order to implement the WHO recommendations, projects to introduce the rotavirus vaccine in 2018 as part of its Expanded Program on Immunization. But before the introduction of this vaccine, epidemiological data on rotavirus infections and rotavirus genotypes circulating in Benin should be available. The aim of this study is to generate epidemiological data on infantile rotavirus diarrhea in Benin. In order to determine the epidemiological characteristics and electrophoretypes of rotavirus responsible for gastroenteritis in diarrheic children aged 0 to 5 years, 186 stool samples were collected according to the WHO Rotavirus Laboratory Manual from March 2014 to February 2015 at Suru-Lere University Hospital Center. Detection of rotavirus antigen was performed by the ELISA test, followed by molecular characterization using polyacrylamide gel electrophoresis. 186 stool samples were analyzed for rotavirus, and seventy-three (39.2%) were found to be positive for rotavirus antigen by ELISA. Children aged 3 to 24 months were the most affected by rotavirus diarrhea in this study. Of the seventy-three children affected with rotavirus diarrhea, 27 (37%) had vomiting accompanied by dehydration and fever. Results based on electrophoresis showed that, among the 73 samples tested, 38 yielded typical rotavirus electrophoretic migration profiles.

Les legumes-feuilles constituent d’excellents apports pour la diversification de l’alimentation humaine. La presente etude a permis d’identifier les legumes-feuilles a proprietes anti-diarrheiques consommes a Cotonou et Abomey-Calavi (Sud-Benin). A l’aide d’enquetes basees sur la methode d’Interview Semi-Structuree, il a ete recense vingt-sept (27) especes de legumes feuilles regroupees en vingt-trois genres (23) et appartenant a dix-sept (17) familles botaniques. Les familles les plus representees, en termes d’espece, sont les Amaranthaceae et les Lamiaceae. Les especes les plus consommees pour le traitement des infections diarrheiques sont : Ocimum gratissimum L . (29,05%), Vernonia amygdalina Delile. (16,80%), Crateva adansonii DC. ssp adansonii (13,49%) et Sesamum radiatum L . (11,41%). Ces legumes-feuilles sont prepares a l’etat frais ou seches (poudres) puis consommes comme legume d’accompagnement (sauce) ou utilises comme tisane. Ces resultats constituent la base d’etudes ulterieures visant a evaluer experimentalement les potentialites antibacteriennes et anti-diarrheiques de ces legumes-feuilles. Cela permettra de mettre a la disposition des populations, des Medicaments Traditionnels Ameliores. Mots cles : Ethnobotanique, diarrhees, legumes feuilles, medecine traditionnelle.