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 byS. 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. Keywords: Streptococcus pneumoniae; neonatal meningitis; respiratory distress

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

Abstract 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

Rapid detection of Mycobacterium tuberculosis complex by real-time polymerase chain reaction (PCR) in pulmonary and extra-pulmonary samples in Casablanca, Morocco

Introduction: the laboratory diagnosis of tuberculosis (TB) relies mainly on conventional techniques. However, it either lacks sensitivity or it is time-consuming. This study aims to evaluate the use of real-time polymerase chain reaction (PCR) targeting IS6110 for Mycobacterium tuberculosis (TB) Complex (MTBC) in the routine diagnosis of TB in our laboratory. Methods: clinical samples were collected from the laboratory of bacteriology at Ibn Rochd University Hospital in Casablanca Morocco. Real-time polymerase chain reaction (PCR) results were compared to AFB smear and culture on Löwenstein-Jensen (LJ) solid media. Sensitivity, specificity, positive and negative predictive value (PPV and NPV) with 95% confidence intervals were calculated using GraphPad Prism. Results: on 171 clinical samples, the study showed positivity of microscopy, culture and real-time PCR for M. TB complex as 19%, 31%, and 32% respectively. Sensitivity, specificity, PPV and NPV for real-time PCR in pulmonary samples were 95.2%, 95.4%, 90.91% and 97.65% respectively. For extra-pulmonary samples, they were: 72.7%, 90.32%, 72.7%, and 90.3%. Conclusion: our study shows the effectiveness of using real-time PCR IS6110 in pulmonary and extra pulmonary samples. Future multicentric studies could seek to evaluate the place of this technique on routine diagnosis for better management of TB in Morocco

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

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 infection

Molecular characterization of penicillin non-susceptible Streptococcus pneumoniae isolated before and after pneumococcal conjugate vaccine implementation in Casablanca, Morocco

Abstract Background Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide, especially among children and the elderly. The ability to effectively treat pneumococcal infection has been compromised due to the acquisition of antibiotic resistance, particularly to β-lactam drugs. This study aimed to describe the prevalence and molecular evolution of penicillin non-susceptible S. pneumoniae (PNSP) isolated from invasive diseases before and after pneumococcal conjugate vaccine implementation in Casablanca, Morocco. Methods Isolates were obtained from the Microbiology Laboratory of Ibn Rochd University Hospital Centre of Casablanca. Serogrouping was done by Pneumotest Kit and serotyping by the Quellung capsular swelling. Antibiotic susceptibility pattern was determined by disk diffusion and E-test methods. The PNSP were analyzed by pulsed-field gel electrophoresis (PFGE) and by genotyping of pbp1a, pbp2b, and pbp2x genes. Results A total of 361 S. pneumoniae isolates were collected from 2007 to 2014. Of these isolates, 58.7% were obtained before vaccination (2007–2010) and 41.3% after vaccination (2011–2014). Of the 361 isolates, 80 were PNSP (22.2%). Generally, the proportion of PNSP between pre- and post-vaccination periods were 31 and 13% (p = 0.009), respectively. The proportion of PNSP isolated from pediatric and adult (age > 14 years) patients decreased from 34.5 to 22.9% (p = 0.1) and from 17.7 to 10.2% (p = 0.1) before and after vaccine implementation, respectively. The leading serotypes of PNSP were 14 (33 vs. 57%) and 19A (18 vs. 14%) before and after vaccination among children. For adults, serotypes 19A (53%) and 23F (24%) were the dominant serotypes in the pre-vaccination period, while serotype 14 (22%) was the most prevalent after vaccination. There were 21 pbp genotypes in the pre-vaccination period vs. 12 for post-vaccination period. PFGE clustering showed six clusters of PNSP grouped into three clusters specific to pre-vaccination period (clusters I, II and III), two clusters specific to post-period (clusters V and VI) and a cluster (IV) that contained clones belonging to the two periods of vaccination. Conclusion Our observations demonstrate a high degree of genetic diversity among PNSP. Genetic clustering among PNSP strains showed that they spread mainly by a restricted number of PNSP clones with vaccine serotypes. PFGE clustering combined with pbp genotyping revealed that vaccination can change the population structure of PNSP

Epidemiology of pertussis in Casablanca (Morocco): contribution of conventional and molecular diagnosis tools

Abstract Background Pertussis, a vaccine preventable disease, is still responsible of significant morbidity and mortality around the world, mostly in newborns. The aim of the present study was (1) to introduce pertussis surveillance in the major pediatric hospital of Casablanca (2) to analyze the prevalence of pertussis among children under 14 years of age and their entourage in Casablanca, Morocco. Methods This is a prospective and non-case controlled study, including children suspected of Pertussis admitted at the Abderrahim Harouchi Pediatric Hospital in Casablanca, from January 2013 to June 2015. Nasopharyngeal samples were obtained for Bordetella spp. culture and Real time PCR detection (RT-PCR) with specific primers of Bordetella spp., B. pertussis, B. parapertussis and B. holmesii. The detection of Bordetella spp. was also performed in some household contacts of the children suspected of pertussis. Results During the 2.5-years period, a total of 282 samples were collected from hospitalized children (156) and in some of their contacts (126). Among 156 samples from the children (from whom 57% were under 2 month of age), Bordetella DNA was detected in 61% (96/156) by RT-PCR. Among these positive samples, 91.7% (88/96) corresponded to B. pertussis DNA. Furthermore, in 39.5% (38/96) of the Bordetella positive samples, B. holmesii DNA was also detected. B. parapertussis DNA was detected in only one sample (1/156). Out of the 156 samples collected from the hospitalized children, only 48 were tested by culture, and 4 B. pertussis were isolated (8.3%). Among the 126 samples from the contacts of the children, mostly mothers (115 cases), Bordetella DNA was detected in 47% (59/126), 90% (53/59) being B. pertussis DNA. Moreover, B. holmesii DNA was also detected in 18.6% (11/59) of the Bordetella positive samples, and coexistence of B. pertussis and B. holmesii DNA in 36.5% (35/96). Two B. pertussis were isolated by culture performed on 43 samples of the contacts of the children (4.6%). Conclusions This study highlights the circulation of B. pertussis but also of B. holmesii in Casablanca-Morocco with a high proportion of co-infections B. holmesii/B. pertussis in infants and their mothers, indicate that infection of non-vaccinated infants could be more associated with young parents. Moreover, the RT- PCR provides a sensitive and specific diagnosis of B. pertussis infections and distinguishes it from other Bordetella species, and is therefore suitable for implementation in the diagnostic laboratory

Analysis of amino acid motif of penicillin-binding proteins 1a, 2b, and 2x in invasive Streptococcus pneumoniae nonsusceptible to penicillin isolated from pediatric patients in Casablanca, Morocco

Abstract Objectives This study aimed to investigate the nature of the amino acid motifs found in PBPs of Streptococcus pneumoniae isolates in invasive diseases from pediatric patients at Casablanca, Morocco. Five penicillin-susceptible (PSSP), ten penicillin-intermediate (PISP), and fifteen penicillin-resistant S. pneumoniae (PRSP) were studied by PCR–RFLP and DNA sequencing of the pbp1a, − 2b, and − 2x genes. Results There were no changes in the conserved motifs of PBP1a, PBP2b and PBP2x for PSSP strains. Substitution close to PBP1a conserved motifs were found in all PRSP isolates and six/five PISP. Analysis of PBP2b showed that all but one of the 10 PISP strains and all PRSP had substitutions. Substitution close to PBP2x motifs showed that all but three of the 10 PISP strains and all PRSP had substitutions in tow conserved motifs. A total of 6, 11 and 10 genotypes were found after analysis of pbp1a, pbp2b, and pbp2x, respectively. The penicillin-nonsusceptible S. pneumoniae isolated in Casablanca share most amino acid substitutions of those reported worldwide, but they occurred among pneumococci with low level resistance to b-lactams

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

International audienceBackground : 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