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.

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).

Surgical wound infection caused by a multi drug resistant Streptococcus pneumoniae Serotype 19A after a total coloproctectomy with ileostomy.

Streptococcus pneumoniae (S. pneumoniae) colonizes asymptomatically the human nasopharynx. This pathogen is responsible for sinusitis, otitis media, pneumonia, bacteremia and meningitis. We report the case of a 35-year-old female patient who developed a surgical wound infection by a multi drug resistant S. pneumoniae serotype 19A after a total coloprotectomy. This first found in Morocco shows the implication of multidrug resistant S. pneumoniae in surgical wound infections.

Molecular Characterization of Invasive Isolates of Neisseria meningitidis in Casablanca, Morocco.

Meningococcal epidemiology may change unpredictably, and typing of Neisseria meningitidis isolates is crucial for the surveillance of invasive meningococcal disease (IMD). Few data are available regarding the meningococcal epidemiology in countries of North Africa. We aimed to explore invasive meningococcal isolates from the Casablanca region in Morocco. We used whole-genome sequencing (WGS) to characterize 105 isolates from this region during the period of 2011 to 2016. Our data showed that the majority (n = 100) of the isolates belonged to serogroup B. Genotyping indicated that most of the isolates (n = 62) belonged to sequence type 33 of clonal complex 32. The isolates also showed the same PorA and FetA markers and clustered together on the basis of WGS phylogenetic analysis; they seemed to correspond to an expansion of local isolates in the Casablanca region, as reported for similar isolates in several other countries. These data suggest that serogroup B isolates may predominate in Morocco, which may have an important impact in the design of vaccination strategies.

Invasive pneumococcal disease among children younger than 5 years of age before and after introduction of pneumococcal conjugate vaccine in Casablanca, Morocco.

OBJECTIVES: The purpose of this study was to compare the incidence rate of invasive pneumococcal disease, the rates of antibiotic resistance and serotype distribution among children ≤5 years old before and after PCVs introduction in Casablanca, Morocco. METHODS: This study was conducted at the Ibn Rochd University Hospital Centre of Casablanca during two periods encompassing pre-and post-implementation of PCVs, respectively from January 2007 to October 2010 and from January 2011 to December 2014. All the non-duplicate invasive S. pneumoniae isolates recovered during the study periods were included. RESULTS: There were 136 cases of IPD, 91 before and 45 after PCVs introduction. The greatest decrease in incidence rate of IPD occurred in children ≤ 2 years of age declining from 34.6 to 13.5 per 100,000 populations (p<0.0001) before and after vaccination, respectively. The incidence rate of PCV-7, PCV-10 non-PCV-7 and PCV-13 non-PCV-10 serotypes decrease significantly from 18.0 to 4.6, from 5.7 to 1.3 and from 5.7 to 0.8/100,000 population (p<0.001) in the same age, respectively. CONCLUSION: Shifts in the distribution of IPD serotypes and reductions in the incidence rate of disease suggest an effective reduction of the burden of IPD in children, but continued high quality surveillance is critical to assess the changes in serotype distributions.

Changing epidemiology of pediatric Streptococcus pneumoniae isolates before vaccine introduction in Casablanca (Morocco).

The objective of this study was to describe the evolution over time of serotypes and antimicrobial resistance of Streptococcus pneumoniae isolates responsible for invasive pneumococcal disease (IPD) in children ≤5 years in Casablanca (Morocco), before vaccine introduction. Isolates recovered from cases of IPD in children ≤5 years during two study periods (1994-2001 and 2006-2010) were compared in terms of serotypes and antimicrobial susceptibility. Serogrouping was done using Pneumotest Kit and serotyping was done by the Quellung capsular swelling. Antibiotic susceptibility pattern was determined by Etest method. The theoretical coverages of the 7-, 10- and 13-valent pneumococcal vaccines were determined. A total of 85 and 102 isolates were studied for the two periods, respectively. During the first period, the most prevalent serogroups/serotypes were 5, 7, 1, 14, 6, 23, 19, 2, 15 and 18, whereas during the second study period, the most frequent were 19, 14, 6, 23, 5, 1 and 18. From 2006 to 2010, serotype 19A accounted for 7.8% of the isolates, whereas serogroups 33 and 22 were rarely isolated (1% each). Assuming cross protection among serogroups, the respective coverage rates for the first vs. second periods reached 40% vs. 53.9% for PCV7, 74.1% vs. 71.6% for PCV10 and 75.3% vs. 82.4% for PCV13. The prevalence of penicillin-nonsusceptibility increased (22.4% vs. 48.5%) as well as the levels of penicillin-resistance (5.9% vs. 15.8%). Penicillin-nonsusceptibility was relatively low across both study periods in serogroups 7, 15, 1 and 5, whereas it was high in serogroups 14, 19 and 23. In conclusion, serotype fluctuations and increases in antibiotic resistance of S. pneumoniae occurred in Casablanca before vaccine introduction. Both of these variations are probably linked to the increase in antibiotic use that has occurred in Casablanca over time. There is a need for a continuous surveillance to assess the vaccine formulations, monitor vaccine impact and to guide treatment recommendations.

Eleven-year surveillance of antibiotic resistance in Streptococcus pneumoniae in Casablanca (Morocco).

OBJECTIVE: To analyze trends of antibiotic resistance rates in Streptococcus pneumoniae from 1998 to 2008 in Casablanca (Morocco). METHODS: The antibiotic resistance levels of 955 consecutive nonduplicate isolates were studied using E test and disc diffusion methods. Results were interpreted following Clinical and Laboratory Standards Institute guidelines (2005). Analysis was done according to three periods (1998-2001; 2002-2005; 2006-2008), age, and site of infection. RESULTS: Penicillin nonsusceptibility (PNS) increased significantly over time (15.6%, 17.8%, and 24.8%; p=0.003). Levels of PNS have changed as well: in 2006-2008, 9.1% of the isolates had an MIC ≥2 µg/ml versus 7.7% in 2002-2005 and 3.4% in 1998-2001. The PNS increase was particularly marked in pediatric isolates (21.4%, 25.5%, and 43.3%; p=0.001). There was no significant difference between the rates of PNS in invasive and noninvasive isolates from children, whereas in adults noninvasive isolates were more penicillin nonsusceptible. Amoxicillin and ceftriaxone nonsusceptible isolates were very rare. An increase of resistance rates was also noticed for erythromycin (9.4%, 12.2%, and 14.4%), tetracycline (20%, 18.6%, and 30.5%), and chloramphenicol (5.6%, 5.6%, and 8.1%). Trimethoprim-sulfamethoxazole resistance rates remained stable (22.8%, 20%, and 23.8%). Proportions of dual nonsusceptibility to penicillin and erythromycin, increased from 5.6% to 8.9%. Multiple drug resistance (resistance to 3 or more antibiotic classes) was found in 0%, 2.4%, and 7.7% of all isolates, respectively. CONCLUSION: The results reported here maybe useful for guiding update of treatment recommendations and suggest the need for continuous surveillance. Increase of antibiotic resistance correlated with antibiotic consumption, stressing the need for elaboration of antibiotic policy in Morocco.