Assessment of the pathogen genomics landscape highlights disparities and challenges for effective AMR Surveillance and outbreak response in the East African community.

The East African Community (EAC) grapples with many challenges in tackling infectious disease threats and antimicrobial resistance (AMR), underscoring the importance of regional and robust pathogen genomics capacities. However, a significant disparity exists among EAC Partner States in harnessing bacterial pathogen sequencing and data analysis capabilities for effective AMR surveillance and outbreak response. This study assesses the current landscape and challenges associated with pathogen next-generation sequencing (NGS) within EAC, explicitly focusing on World Health Organization (WHO) AMR-priority pathogens. The assessment adopts a comprehensive approach, integrating a questionnaire-based survey amongst National Public Health Laboratories (NPHLs) with an analysis of publicly available metadata on bacterial pathogens isolated in the EAC countries. In addition to the heavy reliance on third-party organizations for bacterial NGS, the findings reveal a significant disparity among EAC member States in leveraging bacterial pathogen sequencing and data analysis. Approximately 97% (n = 4,462) of publicly available high-quality bacterial genome assemblies of samples collected in the EAC were processed and analyzed by external organizations, mainly in Europe and North America. Tanzania led in-country sequencing efforts, followed by Kenya and Uganda. The other EAC countries had no publicly available samples or had all their samples sequenced and analyzed outside the region. Insufficient local NGS sequencing facilities, limited bioinformatics expertise, lack of adequate computing resources, and inadequate data-sharing mechanisms are among the most pressing challenges that hinder the EAC's NPHLs from effectively leveraging pathogen genomics data. These insights emphasized the need to strengthen microbial pathogen sequencing and data analysis capabilities within the EAC to empower these laboratories to conduct pathogen sequencing and data analysis independently. Substantial investments in equipment, technology, and capacity-building initiatives are crucial for supporting regional preparedness against infectious disease outbreaks and mitigating the impact of AMR burden. In addition, collaborative efforts should be developed to narrow the gap, remedy regional imbalances, and harmonize NGS data standards. Supporting regional collaboration, strengthening in-country genomics capabilities, and investing in long-term training programs will ultimately improve pathogen data generation and foster a robust NGS-driven AMR surveillance and outbreak response in the EAC, thereby supporting global health initiatives.

Genomic surveillance of severe acute respiratory syndrome coronavirus 2 in Burundi, from May 2021 to January 2022.

BACKGROUND: The emergence and rapid spread of new severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) variants have challenged the control of the COVID-19 pandemic globally. Burundi was not spared by that pandemic, but the genetic diversity, evolution, and epidemiology of those variants in the country remained poorly understood. The present study sought to investigate the role of different SARS-COV-2 variants in the successive COVID-19 waves experienced in Burundi and the impact of their evolution on the course of that pandemic. We conducted a cross-sectional descriptive study using positive SARS-COV-2 samples for genomic sequencing. Subsequently, we performed statistical and bioinformatics analyses of the genome sequences in light of available metadata. RESULTS: In total, we documented 27 PANGO lineages of which BA.1, B.1.617.2, AY.46, AY.122, and BA.1.1, all VOCs, accounted for 83.15% of all the genomes isolated in Burundi from May 2021 to January 2022. Delta (B.1.617.2) and its descendants predominated the peak observed in July-October 2021. It replaced the previously predominant B.1.351 lineage. It was itself subsequently replaced by Omicron (B.1.1.529, BA.1, and BA.1.1). Furthermore, we identified amino acid mutations including E484K, D614G, and L452R known to increase infectivity and immune escape in the spike proteins of Delta and Omicron variants isolated in Burundi. The SARS-COV-2 genomes from imported and community-detected cases were genetically closely related. CONCLUSION: The global emergence of SARS-COV-2 VOCs and their subsequent introductions in Burundi was accompanied by new peaks (waves) of COVID-19. The relaxation of travel restrictions and the mutations occurring in the virus genome played an important role in the introduction and the spread of new SARS-COV-2 variants in the country. It is of utmost importance to strengthen the genomic surveillance of SARS-COV-2, enhance the protection by increasing the SARS-COV-2 vaccine coverage, and adjust the public health and social measures ahead of the emergence or introduction of new SARS-COV-2 VOCs in the country.

Serotype distribution and antimicrobial susceptibility of invasive Streptococcus pneumoniae isolates among adult and elderly population before and after introduction of pneumococcal conjugate vaccine in Casablanca, Morocco.

BACKGROUND: Streptococcus pneumoniae (S. pneumoniae), remains a major cause of mortality and morbidity worldwide. The objective of this study was to determine the trends of invasive pneumococcal diseases (IPD) in adult and elderly population in Casablanca (Morocco) before and after introduction of pneumococcal conjugate vaccine (PCV) by determining the distribution of pneumococcal serotypes and antibiotic resistance profile of isolated strains. METHOD: The proposed study is a retrospective laboratory-based surveillance of IPD in hospitalized adult (15-59 years old) and elderly (≥ 60 years old) patients in Ibn Rochd University Hospital Centre from 2007 to 2019 (13 years). All the 250 non-duplicate clinical invasive isolates from adult and elderly patients, confirmed as S. pneumoniae according to the laboratory standard identification procedures, are included in this study. RESULTS: A significant decrease of the overall incidence in IPD was observed only in adults from 0.71 to 0.54/100000 populations (P = 0.02) and to 0.47/100000 populations (P = 0.0137) in the early and mature post-vaccine period respectively compared to the pre-vaccine period. Our results also showed a significant reduction in the overall prevalence of vaccine serotypes from 28.17 to 6.90% (P = 0.0021) for the PCV-10 serotypes, and from 46.48 to 25.86% (P = 0.0164) for the PCV-13 serotypes only in the mature post-vaccine period (2015-2019). In parallel, the rate of non-vaccine serotypes did not significantly change in the early post-vaccine period (2011-2014) while it increased considerably from 54 to 74.14% (P = 0.0189) during the mature post-vaccine period. The rate of penicillin non-susceptible pneumococcal isolates decreased significantly from 23.94 to 8.77% (P = 0.02) in adult patients, and the rate of cotrimoxazole non-susceptible pneumococcal isolates significantly decreased from 29.58 to 8.77% in the early post-vaccine period (P = 0.003) and to 7.24% in the mature post-vaccine period (P = 0.0007). CONCLUSION: Although childhood vaccination has considerably reduced the incidence of IPD in adult population through the herd effect, IPD remain a real public health problem due to the alarming increase in non-vaccine serotypes (NVS) and the lack of herd effect among elderly population. The rate of antibiotic resistance was relatively low. Nevertheless, resistance constitutes a serious problem to the therapeutic arsenal due to the known capacity for genetic dissemination in the pneumococcus.

Whole genomic comparative analysis of Streptococcus pneumoniae serotype 1 isolates causing invasive and non-invasive infections among children under 5 years in Casablanca, Morocco.

BACKGROUND: Streptococcus pneumoniae serotype 1 remains a leading cause of invasive pneumococcal diseases, even in countries with PCV-10/PCV-13 vaccine implementation. The main objective of this study, which is part of the Pneumococcal African Genome project (PAGe), was to determine the phylogenetic relationships of serotype 1 isolates recovered from children patients in Casablanca (Morocco), compared to these from other African countries; and to investigate the contribution of accessory genes and recombination events to the genetic diversity of this serotype. RESULTS: The genome average size of the six-pneumococcus serotype 1 from Casablanca was 2,227,119 bp, and the average content of coding sequences was 2113, ranging from 2041 to 2161. Pangenome analysis of the 80 genomes used in this study revealed 1685 core genes and 1805 accessory genes. The phylogenetic tree based on core genes and the hierarchical bayesian clustering analysis revealed five sublineages with a phylogeographic structure by country. The Moroccan strains cluster in two different lineages, the five invasive strains clusters altogether in a divergent clade distantly related to the non-invasive strain, that cluster with all the serotype 1 genomes from Africa. CONCLUSIONS: The whole genome sequencing provides increased resolution analysis of the highly virulent serotype 1 in Casablanca, Morocco. Our results are concordant with previous works, showing that the phylogeography of S. pneumoniae serotype 1 is structured by country, and despite the small size (six isolates) of the Moroccan sample, our analysis shows the genetic cohesion of the Moroccan invasive isolates.

Impact and prevalence of comorbidities and complications on the severity of COVID-19 in association with age, gender, obesity, and pre-existing smoking: A meta-analysis.

Background: COVID-19 patients usually present multiple comorbidities and complications associated with severe forms of SARS-CoV-2 infection. This study aimed to assess the risk factors and prevalence of comorbidities and complications contributing to the severity of COVID-19. Methods: This meta-analysis was performed according to PRISMA guidelines. We searched various databases, including PubMed, Google Scholar, and Scopus (between 2020 and 2023), for eligible studies for this meta-analysis. Results: Thirty-three studies were eligible, including 85,812 patients, of which 36 % (30,634/85,812) had severe disease, whereas 64 % (55,178/85,812) had non-severe disease. Severe cases were potentially correlated with the following factors: gender (male) (odd ratio (OR) = 1.52, 95 % CI: 1.34-1.73), advanced age (OR = 3.06, 95 % CI: 2.18-4.40) pre-existing smoking (OR = 1.33, 95 % CI: 1.01-1.75), obesity (OR = 2.11, 95 % CI: 1.47-3.04), diabetes (OR = 1.81, 95 % CI: 1.35-2.43), hypertension (OR = 2.22, 95 % CI: 1.72-2.87), coronary heart disease (OR = 2.17, 95 % CI: 1.42-3.31), CKD (OR = 2.27, 95 % CI: 1.26-4.06), COPD (OR = 1.95, 95 % CI: 1.22-3.09), malignancy (OR = 1.63, 95 % CI: 1.07-2.49) and cerebrovascular disease (OR = 2.76, 95 % CI: 1.63-4.62). All these comorbidities were significantly higher in the severe COVID-19 group compared with the non-severe COVID-19 group. In addition, the most severe complications were associated with shock (OR = 28.08, 95 % CI: 3.49-226.03), ARDS (OR = 13.09, 95 % CI: 5.87-29.18), AKI (OR = 16.91, 95 % CI: 1.87-152.45) and arrhythmia (OR = 7.47, 95 % CI: 2.96-18.83). However, these complications were the most likely to prevent recovery in patients with severe affections compared with non-severe affection groups. Conclusion: All the comorbidities and complications listed above are more likely to cause severe forms of COVID-19 in some patients and hinder recovery. They are therefore risk factors to be controlled to minimize the undesirable effects of the disease.

Neisseria meningitidis with decreased susceptibility to penicillin G and molecular characterization of the penA gene in strains isolated at University Hospital Centers of Casablanca and Marrakech (Morocco).

Introduction: the laboratory diagnosis of meningococcal meningitis relies on conventional techniques. This study aims to evaluate the correlation between the reduced sensitivity to penicillin G of Neisseria meningitidis (N.m) strains and the expression of the altered PBP 2 gene. Methods: out of 190 strains of N.m isolated between 2010 and 2021 at the bacteriology laboratories of Ibn Rochd University Hospital Centre (IR-UHC) in Casablanca and the UHC Mohammed VI in Marrakech, 23 isolates were part of our study. We first determined their state of sensitivity to penicillin G by E-Test strips and searched for the expression of the penA gene by PCR followed by Sanger sequencing. Results: of all the confirmed cases of N.m, 93.15% (n=177) are of serogroup B, 75.2% (n = 143) are sensitive to penicillin G and 24.73% (n = 47) are of intermediate sensitivity. No resistance to penicillin G was observed. Reduced sensitivity to penicillin G in N.m is characterized by mutations namely F504 L, A510 V, I515 V, G541 N and I566 V located in the C-terminal region of the penA gene encoding the penicillin-binding protein 2 (PBP2) (mosaic gene). Conclusion: our study presents useful data for the phenotypic and genotypic monitoring of resistance to penicillin G in N.m and can contribute to the analysis of genetic exchanges between different Neisseria species.

Evaluation of SARS-CoV-2 infection risks after primary vaccination with BNT162b2, BBIBP-CorV, or ChAdOx1-nCOV-19 and after homologous and heterologous booster vaccinations with these vaccines and evaluation of SARS-CoV-2 reinfection profiles.

Background: The emergence of SARS-CoV-2 variants has significantly increased the number of cases of COVID-19 among vaccinated individuals, raising concerns about the effectiveness of current vaccines. The aim of this study was to analyze the SARS-CoV-2 infection risks after primary vaccination with BNT162b2, BBIBP-CorV, or ChAdOx1-nCOV-19 and after homologues and heterologous booster vaccinations with these vaccines, as well as the profiles of reinfected patients. Methods: We analyzed retrospectively 1082 patients vaccinated or unvaccinated with BNT162b2, BBIBP-CorV, and/or ChAdOx1nCoV-19 vaccines to determine their SARS-CoV2 infection statuses using the reverse transcription-polymerase chain reaction (RT-PCR) in addition to their clinical features. The infection risks of patients receiving the different vaccine regimens were compared using multivariate logistic regression analysis, comparing the adjusted OR of a positive COVID-19 test result. Results: Among 596 vaccinated patients, 53%(n = 286) tested positive for SARS-CoV-2 and 57%(n = 310) tested negative. Among positive cases, 10 were reinfection cases. The risk of SARS-CoV-2 infection was 1.6 (adj. OR) for patients who received one dose compared with those who received two doses (95% CI = 1.3-1.8; p < 0.01).The risk was 2.6 (adj. OR) for patients who received one dose compared with those who received three doses (95%CI = 2.1-3.3; p < 0.01), and 1.6 (adj. OR) for patients who received two doses compared with those who received three doses (95% CI = 1.3-2; p < 0.01). The patients who received two doses that were heterologous to that of the primary vaccine had the lowest risk of infection. Booster vaccinations (third dose) significantly reduced the number of positive cases with an acceptable safety profile. Higher cycle-threshold (Ct) values (indicative of viral load) were observed in vaccinated patients, whereas low Ct values were observed in unvaccinated patients. Conclusion: A complete cycle of vaccination with homologous vaccines or heterologous vaccines resulted in an acceptable reduction in SARS-CoV-2 infection. Further, vaccination was associated with a reduction in viral load.

Epidemiology, clinical characteristics and risk factors of coronavirus disease 2019 (COVID-19) in Casablanca.

This is an analytical cross-sectional study of coronavirus disease 2019 (COVID-19) based on data collected between 1 November 2020 and 31 March 2021 in Casablanca focusing on the disease's epidemiological status and risk factors. A total of 4569 samples were collected and analysed by reverse-transcription polymerase chain reaction (RT-PCR); 967 patients were positive, representing a prevalence of 21.2 % for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The mean age was 47.5±18 years, and infection was more common in young adults (<60 years). However, all age groups were at risk of COVID-19, and in terms of disease severity, the elderly were at greater risk because of potential underlying health problems. Among the clinical signs reported in this study, loss of taste and/or smell, fever, cough and fatigue were highly significant predictors of a positive COVID-19 test result (P<0.001). An assessment of the reported symptoms revealed that 27 % of COVID-19-positive patients (n=261) experienced loss of taste and/or smell, whereas only 2 % (n=72) of COVID-19-negative patients did (P<0.001). This result was consistent between univariate (OR=18.125) and multivariate (adjusted OR=10.484) logistic regression analyses, indicating that loss of taste and/or smell is associated with a more than 10-fold higher multivariate adjusted probability of a positive COVID-19 test (adjusted OR=10.48; P<0.001). Binary logistic regression model analysis based on clinical signs revealed that loss of taste and/or smell had a performance index of 0.846 with a P<0.001, confirming the diagnostic utility of this symptom for the prediction of COVID-19-positive status. In conclusion, symptom evaluation and a RT-PCR [taking into account cycle threshold (C t) values of the PCR proxy] test remain the most useful screening tools for diagnosing COVID-19. However, loss of taste/smell, fatigue, fever and cough remain the strongest independent predictors of a positive COVID-19 result.

Burden of invasive pneumococcal disease in children in Casablanca, Morocco four years after the introduction of pneumococcal vaccination.

Streptococcus pneumonia is a common bacterium that can cause several types of infections, including invasive infections especially in children aged <5 years. The aim of this work is to report the different aspects of invasive pneumococcal disease (IPD) in a pediatric hospital in Casablanca, Morocco 4 years after the implementation of pneumococcal vaccination. We conducted a descriptive, retrospective study over a 4-year period from January 2015 to December 2018 in A. Harouchi Pediatric Hospital in Casablanca. This study included hospitalized children aged 0 to 14 years´ old who had an IPD. The data was collected using a data collection sheet from archived patient records and computerized laboratory database; organization of data was done using Microsoft Excel 2016 and analysis was done using SPSS-20. A total of 68 patients were included in this series over the 4-year period. Meningitis was the most common IPD (54.41%) followed by bacteremia (19.17%) and then pneumonia (16.17%). Of the 35 serogrouped strains, 7 were included in the pneumococcal conjugate vaccine (PCV) 10 (20%), 6 were PCV13-nonPCV10 serotypes (17.14%) and 6 were non-vaccine serotypes (17.14%). Among the strains tested for their antibiotic resistance profile, 32.70% were resistant to penicillin, tetracycline (29.78%), erythromycin (20.75%) and cotrimoxazole (17.31%). One strain was intermediate to ceftriaxone. The evolution was unfavorable for 18 patients (26.47%). This study reported high resistance rates to penicillin, tetracyclin and erythromycin. The mortality essentially concerned meningitis patients. Ongoing surveillance of antibiotic susceptibility and serotype distribution is needed by a national surveillance network.

A Rare Case of Pneumococcal Appendicitis in a Child.

Appendicitis is the most common cause for abdominal surgery in children. It is usually caused by Escherichia coli and Streptococcus species and is generally polymicrobial. However, Streptococcus pneumoniae is a rare cause of appendicitis. We report a rare case of pneumococcal appendicitis in a 7-year-old child with no underlying conditions, in association with E. coli and group F ß-hemolytic Streptococcus. The isolated pneumococcal strain was sensible to all tested antibiotics. The patient had a full recovery after surgery and antibiotics. This case emphasizes that S. pneumoniae can cause a variety of unusual infections like appendicitis, in patients with or without underlying conditions. Thus, even though being a rare entity, physicians should always be aware of S. pneumoniae as a possible causative agent.

Evaluation of the Performances of the Rapid Test RESIST-5 O.O.K.N.V Used for the Detection of Carbapenemases-Producing Enterobacterales.

BACKGROUND: The emergence of carbapenemase-producing Enterobacterales (CPE) is a public health problem, requiring rapid and reliable diagnostic methods. The aim is to compare the new rapid immunochromatographic (IC) test: RESIST-5 O.O.K.N.V with PCR and the predictive model of EUCAST algorithm for the detection of CPE. METHODS: A longitudinal cross-sectional study was carried out in the bacteriology-virology laboratory of the Ibn Rochd-Casablanca University Hospital, from 1 February 2019 to 28 February 2020, concerning strains with reduced sensitivity to Ertapenem. The identification of bacterial species was carried out according to the standard criteria of microbiology and antibiogram according to CASFM-EUCAST 2019 recommendations. The sensitivity and specificity of the rapid IC test were calculated. RESULTS: The results of the new IC test showed a sensitivity and specificity of 100% for the detection of OXA-48 and NDM. These carbapenemases were detected simultaneously with a sensitivity and specificity of 100%. OXA-48 was the most common carbapenemas found (36%), followed by NDM (24%) and (13.4%) cases of OXA-48 and NDM coexistence. CONCLUSION: The rapid IC test could be a rapid and effective diagnostic tool for detecting the most common carbapenemases in our context, and to accelerate the implementation of adequate antibiotic therapy and infection control measures in patients with CPE infections.

Early neonatal respiratory distress revealing meningitis caused by Streptococcus pneumoniae serotype 17F: a case report.

Background: Streptococcus pneumoniae (S. pneumoniae) is the first leading cause of invasive diseases such as meningitis, bacteremia and pneumoniae in children. In this case we report an early neonatal respiratory distress revealing meningitis caused by S. pneumoniae Serotype 17F through vertical transmission, in the newborn of 3 hours of live. Case description: A male late preterm newborn was born by vaginal delivery at a gestational age of 34 weeks. At 3 hours of life, he was admitted for early moderate neonatal respiratory distress in the Neonatal Medicine and Resuscitation Service. Cerebrospinal fluid culture yielded S. pneumoniae belonging to serotype 17F while the blood culture was negative. The same pneumococcal serotype was recovered from the high vaginal swab of the mother. Both isolates were found susceptible to all tested antibiotics except tetracycline and chloramphenicol to which the strain was resistant. Antibiotherapy management of the child included ceftriaxone at 150mg/kg/day for 21 days, in combination with gentamycin at 5 mg/kg/day for 5 days. ciprofloxacin was added at 40mg/kg/day in two doses for a period of three weeks as the baby presented a hydrocephalus. Conclusion: This finding shows that clinical manifestations of neonatal pneumococcal meningitis may be atypical and/or misleading.

Global Landscape Review of Serotype-Specific Invasive Pneumococcal Disease Surveillance among Countries Using PCV10/13: The Pneumococcal Serotype Replacement and Distribution Estimation (PSERENADE) Project.

Serotype-specific surveillance for invasive pneumococcal disease (IPD) is essential for assessing the impact of 10- and 13-valent pneumococcal conjugate vaccines (PCV10/13). The Pneumococcal Serotype Replacement and Distribution Estimation (PSERENADE) project aimed to evaluate the global evidence to estimate the impact of PCV10/13 by age, product, schedule, and syndrome. Here we systematically characterize and summarize the global landscape of routine serotype-specific IPD surveillance in PCV10/13-using countries and describe the subset that are included in PSERENADE. Of 138 countries using PCV10/13 as of 2018, we identified 109 with IPD surveillance systems, 76 of which met PSERENADE data collection eligibility criteria. PSERENADE received data from most (n = 63, 82.9%), yielding 240,639 post-PCV10/13 introduction IPD cases. Pediatric and adult surveillance was represented from all geographic regions but was limited from lower income and high-burden countries. In PSERENADE, 18 sites evaluated PCV10, 42 PCV13, and 17 both; 17 sites used a 3 + 0 schedule, 38 used 2 + 1, 13 used 3 + 1, and 9 used mixed schedules. With such a sizeable and generally representative dataset, PSERENADE will be able to conduct robust analyses to estimate PCV impact and inform policy at national and global levels regarding adult immunization, schedule, and product choice, including for higher valency PCVs on the horizon.

Epidemiological profile of Neisseria meningitidis in Casablanca, Morocco: 2010-2019.

Surveillance of invasive meningococcal diseases (IMD) must be carried out regularly and continuously in order to detect the emergence of strains of reduced susceptibility to antibiotics for therapeutic and prophylactic use and the appearance of new invasive clones. Molecular-typing approaches allow reliable traceability and powerful epidemiological analysis. This is an epidemiological study of Neisseria meningitidis causing meningitis in Casablanca, Morocco. The grouping was confirmed by PCR mainly on the isolates from cerebrospinal fluid (CSF). A total of 245 confirmed isolates of N .meningitidis were obtained between 2010 and 2019 of which 93 % are of group B. Overall, 24 % of all the isolates have a reduced susceptibility to penicillin G, but no resistance to penicillin G has been reported. All the isolated strains are susceptible to third-generation cephalosporins (3GCs). Genotyping by multilocus sequence typing (MLST) of a selection of 18 strains showed that the majority of isolates belong to the invasive clonal complex CC 32(9/18) followed by the CC 41/44(3/18).