Analysis of chlorhexidine, antibiotics and bacterial community composition in water environments from Brazil, Cameroon and Madagascar during the COVID-19 pandemic.

The widespread of chlorhexidine and antibiotics in the water bodies, which grew during the global COVID-19 pandemic, can increase the dispersion of antibiotic resistance. We assessed the occurrence of these pharmaceutical compounds as well as SARS-CoV-2 and analysed the bacterial community structure of hospital and urban wastewaters from Brazil, Cameroon, and Madagascar. Water and wastewater samples (n = 59) were collected between January-June 2022. Chlorhexidine, azithromycin, levofloxacin, ceftriaxone, gentamicin and meropenem were screened by Ultra-High-Performance Liquid Chromatography coupled with mass spectrometer. SARS-CoV-2 was detected based on the nucleocapsid gene (in Cameroon and Madagascar), and envelope and spike protein-encoding genes (in Brazil). The total community-DNA was extracted and used for bacterial community analysis based on the 16S rRNA gene. To unravel likely interaction between pharmaceutical compounds and/or SARS-CoV-2 with the water bacterial community, multivariate statistics were performed. Chlorhexidine was found in hospital wastewater effluent from Brazil with a maximum concentration value of 89.28 µg/L. Additionally, antibiotic residues such as azithromycin and levofloxacin were also present at concentrations between 0.32-7.37 µg/L and 0.11-118.91 µg/L, respectively. In Cameroon, azithromycin was the most found antibiotic present at concentrations from 1.14 to 1.21 µg/L. In Madagascar instead, ceftriaxone (0.68-11.53 µg/L) and levofloxacin (0.15-0.30 µg/L) were commonly found. The bacterial phyla statistically significant different (P < 0,05) among participating countries were Proteobacteria, Patescibacteria and Dependentiae which were mainly abundant in waters sampled in Africa and, other phyla such as Firmicutes, Campylobacterota and Fusobacteriota were more abundant in Brazil. The phylum Caldisericota was only found in raw hospital wastewater samples from Madagascar. The canonical correspondence analysis results suggest significant correlation of azithromycin, meropenem and levofloxacin with bacteria families such as Enterococcaceae, Flavobacteriaceae, Deinococcaceae, Thermacetogeniaceae and Desulfomonilaceae, Spirochaetaceae, Methanosaetaceae, Synergistaceae, respectively. Water samples were also positive for SARS-CoV-2 with the lowest number of hospitalized COVID-19 patients in Madagascar (n = 7) and Brazil (n = 30). Our work provides new data about the bacterial community profile and the presence of pharmaceutical compounds in the hospital effluents from Brazil, Cameroon, and Madagascar, whose limited information is available. These compounds can exacerbate the spreading of antibiotic resistance and therefore pose a risk to public health.

Viral etiology of lower respiratory tract infections in adults in the pre-COVID-19 pandemic era: A cross-sectional study in a single center experience from Cameroon.

Background and Aims: Respiratory viruses are responsible for the majority of lower respiratory tract infections (LRTIs) worldwide. However, there is a gap on the epidemiology of viral LRTIs in adults in sub-Saharan African countries. In Cameroon, like in other countries, the role of viral respiratory pathogens in the etiology of LRTIs in adults is helpful for clinical management. This study aimed to determine the viral aetiologies of LRTIs among hospitalized adults in a reference center for respiratory diseases in the town of Yaounde in Cameroon and its surroundings. Methods: A cross-sectional study was conducted from January 2017 to January 2018 at Jamot Hospital in Yaounde (Cameroon). Clinical and demographic information; BAL and sputa were collected from hospitalized patients meeting LRTI case definitions. The clinical samples were investigated for respiratory pathogens with a commercial Reverse Transcriptase Real-Time Polymerase Chain Reaction (RT-PCR) targeting 21 viruses, cultures for bacterial and fungal infections. Results: The 77 included adult patients with LRTIs had an appropriate clinical sample for microbial investigations. A viral agent was detected in 22.1% (17/77) samples. The main viruses detected included rhinovirus (10/77), coronavirus (hCoV-OC43 and hCoV-229E), and influenza A virus (3/77 each). A concomitant viral and bacterial co-infection occurred in 7.8% of patients (6/77) while viral co-infection occurred in one patient (1.3%). No Severe acute respiratory syndrome coronavirus 2 (SARS­CoV­2) was detected in clinical samples. Most patients were under antimicrobials before getting diagnosed. Conclusions: Respiratory viruses account for 22.1% of LRTIs in hospitalized patients in this study. Despite prior antimicrobial therapy and delay, rhinovirus, coronavirus and influenza A virus were the most detected in patients in the pre-COVID-19 pandemic era in a single center experience from Cameroon.

Regional sequencing collaboration reveals persistence of the T12 Vibrio cholerae O1 lineage in West Africa.

Background: Despite recent insights into cholera transmission patterns in Africa, regional and local dynamics in West Africa-where cholera outbreaks occur every few years-are still poorly understood. Coordinated genomic surveillance of Vibrio cholerae in the areas most affected may reveal transmission patterns important for cholera control. Methods: During a regional sequencing workshop in Nigeria, we sequenced 46 recent V. cholerae isolates from Cameroon, Niger, and Nigeria (37 from 2018 to 2019) to better understand the relationship between the V. cholerae bacterium circulating in these three countries. Results: From these isolates, we generated 44 whole Vibrio cholerae O1 sequences and analyzed them in the context of 1280 published V. cholerae O1 genomes. All sequences belonged to the T12 V. cholerae seventh pandemic lineage. Conclusions: Phylogenetic analysis of newly generated and previously published V. cholerae genomes suggested that the T12 lineage has been continuously transmitted within West Africa since it was first observed in the region in 2009, despite lack of reported cholera in the intervening years. The results from this regional sequencing effort provide a model for future regionally coordinated surveillance efforts. Funding: Funding for this project was provided by Bill and Melinda Gates Foundation OPP1195157.

Trends of Antibacterial Resistance at the National Reference Laboratory in Cameroon: Comparison of the Situation between 2010 and 2017.

INTRODUCTION: Antimicrobial resistance represents a growing public health threat. One of the World Health Organization's strategic objectives is "strengthening knowledge through surveillance and research." Sub-Saharan African countries are still far from achieving this objective. We aimed to estimate and compare the prevalence of antibacterial resistance in 2010 and 2017 in Cameroon. METHODS: We conducted a retrospective study on all clinical specimens cultured in Centre Pasteur du Cameroun (CPC) in 2010 and 2017. Data were extracted from the CPC's laboratory data information system software and then managed and analyzed using R. Bacterial resistance rates were calculated in each year and compared using chi-square or Fisher's tests, and relative changes were calculated. Outcomes included acquired resistance (AR), WHO priority resistant pathogens, some specific resistances of clinical interest, and resistance patterns (multi, extensively, and pan drug resistances) for five selected pathogens. RESULTS: A total of 10,218 isolates were analyzed. The overall AR rate was 96.0% (95% CI: 95.4-96.6). Most of WHO priority bacterial resistance rates increased from 2010 to 2017. The most marked increases expressed as relative changes concerned imipenem-resistant Acinetobacter (6.2% vs. 21.6%, +248.4%, p = 0.02), imipenem-resistant Pseudomonas aeruginosa (13.5% vs. 23.5%, +74.1%, p < 0.01), 3rd generation-resistant Enterobacteriaceae (23.8% vs. 40.4%, +65.8%, p < 10-15), methicillin-resistant Staphylococcus aureus (27.3% vs. 46.0%, +68.6%, p < 0.002), fluoroquinolone-resistant Salmonella (3.9% vs. 9.5%, +142.9%, p = 0.03), and fluoroquinolone-resistant Enterobacteriaceae (32.6% vs. 54.0%, +65.8%, p < 10-15). For selected pathogens, global multidrug resistance was high in 2010 and 2017 (74.9% vs. 78.0% +4.1%, p = 0.01), intensively drug resistance rate was 5.8% (7.0% vs. 4.7%; p = 0.07), and no pan drug resistance has been identified. CONCLUSION: Bacterial resistance to antibiotics of clinical relevance in Cameroon was high and appeared to increase between 2010 and 2017. There is a need for regular surveillance of antibacterial resistance to inform public health strategies and empirically inform prescription practices.

Analysis of Haemophilus species in patients with respiratory tract infections in Yaoundé, Cameroon.

OBJECTIVES: To identifyHaemophilus species and characterize antimicrobial susceptibility of isolates from patients with respiratory tract infections (RTIs) in Cameroon. METHODS: Isolates (n = 95) were from patients with RTIs obtained from two Hospitals in Yaoundé, Cameroon. Isolates were identified by biochemical assay, PCR-based method, MALDI-TOF and whole genome sequencing. Antibiotic minimum inhibitory concentrations were determined by E-test. RESULTS: H. influenzae was the most prevalent species varying from 76.8% to 84.2% according to different methods. The isolates were mainly nontypable (n = 70, 96%). Three isolates of H. influenzae were capsulated (b, e and f). The isolates were genetically diverse and 40 unique sequence types were identified including 11 new ones. Resistance to ampicillin was observed among 55.3% (52/94) and 9% (14/52) produced TEM-1 ß-lactamase. PBP3 mutations occurred in 57.7% of ampicillin resistant isolates (30/52). Eleven isolates were chloramphenicol resistant with 80% producing chloramphenicol acetyltransferase (8/10). Four Haemophilus isolates were rifampicin resistant with two mutations in rpoB gene. Five isolates were ciprofloxacin resistant and harbored mutations in the quinolone resistance determining regions of gyrA and parC genes. CONCLUSION: H. influenzae isolates are highly diverse and show high levels of antibiotic resistance. H. influenzae serotype b is still circulating in the post-vaccination era.

Hospital-based Surveillance Provides Insights Into the Etiology of Pediatric Bacterial Meningitis in Yaoundé, Cameroon, in the Post-Vaccine Era.

BACKGROUND: Meningitis is endemic to regions of Cameroon outside the meningitis belt including the capital city, Yaoundé. Through surveillance, we studied the etiology and molecular epidemiology of pediatric bacterial meningitis in Yaoundé from 2010 to 2016. METHODS: Lumbar puncture was performed on 5958 suspected meningitis cases; 765 specimens were further tested by culture, latex agglutination, and/or polymerase chain reaction (PCR). Serotyping/grouping, antimicrobial susceptibility testing, and/or whole genome sequencing were performed where applicable. RESULTS: The leading pathogens detected among the 126 confirmed cases were Streptococcus pneumoniae (93 [73.8%]), Haemophilus influenzae (18 [14.3%]), and Neisseria meningitidis (15 [11.9%]). We identified more vaccine serotypes (19 [61%]) than nonvaccine serotypes (12 [39%]); however, in the latter years non-pneumococcal conjugate vaccine serotypes were more common. Whole genome data on 29 S. pneumoniae isolates identified related strains (<30 single-nucleotide polymorphism difference). All but 1 of the genomes harbored a resistance genotype to at least 1 antibiotic, and vaccine serotypes harbored more resistance genes than nonvaccine serotypes (P < .05). Of 9 cases of H. influenzae, 8 were type b (Hib) and 1 was type f. However, the cases of Hib were either in unvaccinated individuals or children who had not yet received all 3 doses. We were unable to serogroup the N. meningitidis cases by PCR. CONCLUSIONS: Streptococcus pneumoniae remains a leading cause of pediatric bacterial meningitis, and nonvaccine serotypes may play a bigger role in disease etiology in the postvaccine era. There is evidence of Hib disease among children in Cameroon, which warrants further investigation.

Bacterial Aetiologies of Lower Respiratory Tract Infections among Adults in Yaoundé, Cameroon.

Lower respiratory tract infections (LRTIs) remain a challenge in African healthcare settings and only few data are available on their aetiology in Cameroon. The purpose of this study was to access the bacterial cause of LRTIs in patients in Cameroon by two methods. Methods. Participants with LRTIs were enrolled in the referral centre for respiratory diseases in Yaoundé city and its surroundings. To detect bacteria, specimens were tested by conventional bacterial culture and a commercial reverse-transcriptase real-time polymerase chain reaction (RT-PCR) assay. One hundred forty-one adult patients with LRTIs were enrolled in the study. Among the participants, 46.8% were positive for at least one bacterium. Streptococcus pneumoniae and Haemophilus influenzae were the most detected bacteria with 14.2% (20/141) followed by Klebsiella pneumoniae, 9.2% (13/141), Staphylococcus aureus, 7.1% (10/141), and Moraxella catarrhalis, 4.3% (6/141). Bacterial coinfection accounted for 23% (14/61) with Haemophilus influenzae being implicated in 19.7% (12/61). The diagnostic performance of RT-PCR for bacteria detection (43.3%) was significantly different from that of culture (17.7%) (p< 0.001). Only Streptococcus pneumoniae detection was associated with empyema by RT-PCR (p<0.001). These findings enhance understanding of bacterial aetiologies in order to improve respiratory infection management and treatment. It also highlights the need to implement molecular tools as part of the diagnosis of LRTIs.

Prevalence of respiratory bacterial infections in people with lower respiratory tract infections in Africa: the BARIAFRICA systematic review and meta-analysis protocol.

INTRODUCTION: The burden of lower respiratory tract infections (LRTIs) is a substantial public health concern. However, the epidemiology of LRTI and its bacterial aetiologies are poorly characterised, particularly in the African continent. Providing accurate data can help design cost-effective interventions to curb the burden of respiratory infections in Africa. Therefore, the aim of this systematic review and meta-analysis will be to determine the prevalence of respiratory Bacterial Aetiologies in people with low Respiratory tract Infections in Africa (BARIAFRICA) and associated factors. METHODS AND ANALYSIS: We will search PubMed, EMBASE, Web of Science, African Journals Online, Cumulative Index to Nursing and Allied Health Literature, and Global Index Medicus to identify studies that reported the prevalence (of enough data to compute this estimate) of respiratory bacterial infections in people with LRTIs in Africa from 1 January 2000 to 31 March 2018, without any linguistic restrictions. Study selection, data extraction and risk of bias assessment will be conducted independently by two investigators. Heterogeneity will be evaluated using the χ² test on Cochran's Q statistic and quantified with H and I² statistics. Prevalence will be pooled using a random-effect meta-analysis model. Subgroup and meta-regression analyses will be used to identify sources of heterogeneity of prevalence estimates. This study will be reported according to the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses. ETHICS AND DISSEMINATION: Since this study will be based on published data, it does not require ethical approval. This systematic review and meta-analysis is intended to serve as a basis for determining the burden of LRTIs, for identifying data gaps and for guiding future investigations in Africa. The final report will be published in peer-reviewed journals, presented in conferences and submitted to relevant health policy makers. PROSPERO REGISTRATION NUMBER: CRD42018092359.

The Lake Chad Basin, an Isolated and Persistent Reservoir of Vibrio cholerae O1: A Genomic Insight into the Outbreak in Cameroon, 2010.

The prevalence of reported cholera was relatively low around the Lake Chad basin until 1991. Since then, cholera outbreaks have been reported every couple of years. The objective of this study was to investigate the 2010/2011 Vibrio cholerae outbreak in Cameroon to gain insight into the genomic make-up of the V. cholerae strains responsible for the outbreak. Twenty-four strains were isolated and whole genome sequenced. Known virulence genes, resistance genes and integrating conjugative element (ICE) elements were identified and annotated. A global phylogeny (378 genomes) was inferred using a single nucleotide polymorphism (SNP) analysis. The Cameroon outbreak was found to be clonal and clustered distant from the other African strains. In addition, a subset of the strains contained a deletion that was found in the ICE element causing less resistance. These results suggest that V. cholerae is endemic in the Lake Chad basin and different from other African strains.

Multidrug-resistant Salmonella enterica serotype Typhi, Gulf of Guinea Region, Africa.

We identified 3 lineages among multidrug-resistant (MDR) Salmonella enterica serotype Typhi isolates in the Gulf of Guinea region in Africa during the 2000s. However, the MDR H58 haplotype, which predominates in southern Asia and Kenya, was not identified. MDR quinolone-susceptible isolates contained a 190-kb incHI1 pST2 plasmid or a 50-kb incN pST3 plasmid.

Multiple contaminations of chickens with Campylobacter, Escherichia coli and Salmonella in Yaounde (Cameroon).

INTRODUCTION: Food-borne diseases associated with Campylobacter, Escherichia coli, and Salmonella are mainly caused by the consumption of raw or undercooked poultry meat. The objective of this study was to evaluate the prevalence of Campylobacter, Escherichia coli, and Salmonella in chickens. METHODOLOGY: One hundred and fifty chickens collected from eight retail markets in Yaounde were examined for the presence of Campylobacter, Escherichia coli, and Salmonella using standard bacteriological procedures. RESULTS: Of the 150 chickens collected, 135 (90%) were contaminated with Campylobacter (68.9% C. coli and 31.1% C. jejuni). All the chickens were positive for E. coli. Among the 150 isolates, 17 (11.3%) were enteropathogenic E. coli (EPEC). Additionally, 103 Salmonella strains were recovered from 90 chickens. Salmonella Enteritidis (45.6%) and Salmonella Hadar (28.1%) were the most frequent serotypes. Multiple contamination was found in 142 chickens (94.6%), of which 83 (55.3%) were concurrently contaminated with Campylobacter, Escherichia coli, and Salmonella. CONCLUSION: These results show that chickens in Cameroon are highly contaminated with Campylobacter, Escherichia coli, and Salmonella. The multiple contaminations of chickens is a potential risk of infection for consumers and highlights the necessity of public awareness for food safety.

Prevalence and antimicrobial resistance of Salmonella serotypes in chickens from retail markets in Yaounde (Cameroon).

From February 2006 to January 2007, 150 chickens were collected from eight retail markets in Yaounde, and 90 (60%) tested positive for Salmonella. Seventy-nine chickens were contaminated with only one Salmonella serotype, 10 with two different serotypes, and 1 with four serotypes. The most prevalent serotypes were Enteritidis (47 strains) and Hadar (29 strains). The isolates were tested for their susceptibilities to amoxicillin, amoxicillin/clavulanic acid, cefoxitin, cefotaxime, gentamicin, streptomycin, tetracycline, chloramphenicol, sulfonamides, nalidixic acid, ciprofloxacin, and trimethoprim/sulfamethazole by disk diffusion assay. Minimum inhibitory concentrations of ampicillin, streptomycin, tetracycline, sulfonamides, and nalidixic acid were determined for the resistant strains by agar dilution method. Eleven isolates (10.7%) of the 103 tested were susceptible to all antimicrobials. Resistance was most observed to tetracycline (84.5%), streptomycin (44.7%), and nalidixic acid (34%). Forty-one isolates (39.8%) were multidrug resistant (resistant to three or more antimicrobials from different classes), of which 68.3% were Hadar and 21.9% Enteritidis. The most frequent resistant pattern in Hadar was streptomycin-tetracycline-nalidixic acid. These results highlight once more the need for surveillance of Salmonella contamination in poultry.