Neonatal sepsis due to NDM-1 and VIM-2 co-producing Pseudomonas aeruginosa in Morocco.

BACKGROUND: Carbapenem-resistant Pseudomonas aeruginosa are being increasingly described worldwide. Here, we investigated the molecular mechanisms underlying carbapenem resistance in an extremely drug-resistant P. aeruginosa isolate from a neonatal intensive care unit in Morocco. MATERIALS AND METHODS: P. aeruginosa strain O82J1 was identified using MALDI-TOF-MS. Carba NP, immunochromatographic assay NG Carba5 and antimicrobial susceptibility testing using disc diffusion and microbroth were performed. Whole-genome sequencing using the Illumina and MinION technologies and different software packages available at the Center of Genomic Epidemiology were used to predict the resistome, sequence type and plasmid types. RESULTS: P. aeruginosa O82J1 co-expressed two metallo-ß-lactamases, blaNDM-1 and blaVIM-2, and was susceptible to colistin and apramycin only. It belonged to ST773 that is frequently reported worldwide as a high-risk P. aeruginosa clone. The blaVIM-2 gene was integron-borne on a IncP-2 465-kb plasmid, whereas the blaNDM-1 gene was chromosomally encoded and embedded in an integrative conjugative element, probably at the origin of its acquisition. A total of 23 antimicrobial resistance genes were detected including a blaPER-1 ESBL gene, and an 16S-rRNA methyltransferase gene rmtB. CONCLUSIONS: The isolation of XDR P. aeruginosa isolates expressing several carbapenemases in a neonatal intensive care unit is of great concern due to the reduced treatment options, relying only on colistin, but not recommended in neonates, and apramycin, not yet approved for human therapy. Concerns were further elevated due to the resistance to cefiderocol and ATM/AVI, two novel and last-resort antibiotics recommended to treat infections caused by Gram-negative bacteria, particularly XDR P. aeruginosa in adults.

Invasive Pneumococcal Infections Among Moroccan Children: Pneumococcal Vaccination Challenges in the Mature Vaccine Era.

BACKGROUND: Streptococcus pneumoniae, a major contributor to global morbidity and mortality, disproportionately affects children, the elderly, and immunocompromised individuals. Despite vaccination efforts, the challenge of serotype replacement highlights the ongoing struggle against invasive pneumococcal diseases (IPD) in Morocco, emphasizing the need for updated public health strategies and vaccine efficacy assessments. METHODS: This study was conducted at the Ibn Rochd University Hospital Center and the Mohammed VI University Hospital Center from 2019 to 2022, focusing on hospitalized children. It involved the analysis of 74 strains of IPD, assessing the distribution of pneumococcal serotypes and their antibiotic sensitivity in the post-vaccination era. RESULTS: The prevalence of meningitis or meningo-encephalitis was found to be 66% among the study subjects, with the most frequent serotypes being 3, 19A, 6B, 14, and 11. These serotypes varied significantly by age and location. Coverage rates for the pneumococcal conjugate vaccines, PCV-10 and PCV-13, were 20.27% and 56.75%, respectively. Notably, 43% of the strains were non-vaccine serotypes, with serotypes 3 and 19 accounting for 36% of the infections in children, indicating a lack of vaccine efficacy against these types. Additionally, 31.3% of the strains were Penicillin non-susceptible Streptococcus pneumoniae (PNSP), with 81.25% associated with non-vaccine serotypes. CONCLUSIONS: This study highlights the persistence of IPD in Moroccan children, revealing significant challenges despite vaccination efforts. With the reintroduction of PCV-13, concerns about the efficacy against non-vaccine serotypes, particularly 3 and 19A, remain. Continuous surveillance and adaptable vaccination strategies are essential to combat these serotype replacements and ensure the effectiveness of future preventive measures.

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.

In Vitro Activity of Ceftazidime-Avibactam Against Enterobacteriaceae and P. aeruginosa Isolated at the Ibn Rochd University Hospital of Casablanca.

BACKGROUND: The aim of this study is to determine the in vitro activity of ceftazidime-avibactam (CZA) on Entero-bacteriaceae and P. aeruginosa strains isolated in the bacteriology-virology laboratory of the Ibn Rochd University of Casablanca. METHODS: This is a prospective descriptive longitudinal study conducted from May 28 through June 25, 2022, on Enterobacteriaceae and P. aeruginosa isolated from diagnostic samples received at the Bacteriology-Virology and Hospital Hygiene Laboratory of the Ibn Rochd University Hospital of Casablanca. The isolation and identification of the strains were carried out using standard bacteriological techniques. The study of sensitivity to ceftazidime-avibactam was done by diffusion susceptibility testing on agar medium according to EUCAST 2022 recommendations. RESULTS: During the study period, 271 strains of Enterobacteriaceae were isolated. The sensitivity rate to ceftazidime-avibactam was 91% vs. 74% for ceftazidime alone. R. terrigena was the most resistant strain to CZA with a rate of 69%, followed by E. cloacae (14%), then K. pneumoniae (6%), and finally E. coli (5%). Among the strains isolated, 24% (n = 66) produced ESBL, of which 29% (n = 19) were resistant to CZA, and 10.7% (n = 29) were re-sistant to imipenem, including 44, 8% (n = 13) that were resistant to imipenem and CZA. Regarding P. aeruginosa, 92 strains were isolated. The CZA resistance rate was 33.6% (n = 31). Among the strains isolated, 30.4% (n = 29) were resistant to imipenem, of which 82.7% (n = 24) were resistant to CZA. CONCLUSIONS: The in vitro evaluation of the ceftazidime-avibactam activity on the strains isolated, mainly: E. Coli, K. Pneumoniae, and E. Cloacae, showed a good inhibitory activity of this molecule which can constitute a therapeutic alternative for the treatment of infections due to these microorganisms.

Evaluation of Antibiograms Interpretation Training Needs of Medical Biology Residents at Ibn Rochd University Hospital.

BACKGROUND: In order to determine domains with the most difficulties and plan a relevant training, the aim of this study was to assess training needs of medical biology residents at the Ibn Rochd University Hospital of Casablanca, concerning antibiograms interpretation. METHODS: A descriptive and comparative study was carried out in February 2022, with two resident groups. The first one included all residents in training in the bacteriology-virology laboratory and having spent at least 2 months. The 2nd one included residents who had validated their course (9 months). Data were collected from collective interviews and a self-administered questionnaire with a qualitative and quantitative grid analysis. RESULTS: Residents expressed needs in the three learning domains (knowledge, know-how, interpersonal skills) but they were low to moderate (scores from 0.2/4 to 2.4/4). Priority topics for training were: interpretation of observed phenotypes (5.85/8), followed by interpretative reading according to EUCAST recommendations (5.05/8). The two groups' responses were similar, except for the know-how domain of natural resistance mechanisms where the needs were greater in G1 (p = 0.008). CONCLUSIONS: In the light of these results, additional training with relevant aims should be planned.

Nosocomial Bacterial Infections in Covid-19 ICU at Ibn Rochd University Hospital Casablanca Morocco.

BACKGROUND: The aim is to determine the prevalence of bacterial nosocomial infections (NI) and its associated factors in COVID-19 ICU. METHODS: A descriptive retrospective study, from April to August 2020, was carried on patients with respiratory distress following SARS-Cov-2 infection presenting a sign of bacterial infection. RESULTS: A total of 29 patients (33% of hospitalized patients) contracted a NI, their age was 65 years ± 13.90 and sex-ratio M/F was 2.62. All patients had an invasive device: central line (65%), bladder catheter (72%), and mechanical ventilation (76%). The time of NI occurring was 5 to 8 days. Charlson comorbidity index was 4 - 6 in 18 patients (62%). Of the 35 samples taken, 45 bacteria were isolated: multi drug resistant A. baumannii and P. aeruginosa accounted for 24.4%. Of all, 16 deaths were recorded (55, 17%). CONCLUSIONS: NI is common in COVID-19 ICU and can be predicted by risk factors that should be managed.

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.

Phytochemical constituents, antioxidant and antistaphylococcal activities of Evernia prunastri (L.) Ach., Pseudevernia furfuracea (L.) Zopf. and Ramalina farinacea (L.) Ach. from Morocco.

The purpose of this work was to assess chemical composition, antibacterial activity against Staphylococcus aureus isolates from catheter-associated infections and antioxidant activity of methanol extracts of three lichens collected from Morocco. The phytochemical analysis of the methanol extracts of these lichens was performed by HPLC-UV method, the predominant phenolic compounds were evernic acid, physodalic acid and usnic acid for Evernia prunastri, Pseudevernia furfuracea and Ramalina farinacea, respectively. Total phenolic compounds and total flavonoid content of all extracts were also determined. As a result, Pseudevernia furfuracea extract had the strongest effect and the highest phenolic compounds content. All extracts showed antibacterial activity against all tested strains (MIC values ranging from 0.078 to 0.625 mg/mL), the strongest inhibition was obtained with the extract of Evernia prunastri.

Pertussis in infants, in their mothers and other contacts in Casablanca, Morocco.

BACKGROUND: In recent decades, there has been a marked increase in the number of reported cases of pertussis around the world, and pertussis continues to be a frequently occurring disease despite an effective childhood vaccination. This study aims to determine the role of household contacts of children diagnosed with pertussis in Casablanca Morocco. METHODS: From November 2015 to October 2017, children suspected of whooping cough that consulted Ibn Rochd University hospital at Casablanca with their household contacts were enrolled in the study. Nasopharyngeal (NP) samples of the suspected children were analyzed by culture and RT-PCR. For the household contacts, NP and blood samples were collected and analyzed by RT-PCR and specific detection of pertussis toxin antibodies by ELISA, respectively. RESULTS: During the study period, the survey was carried out on 128 infants hospitalized for pertussis suspicion and their families (N = 140). B. pertussis DNA was specifically detected in 73 (57%) samples, coexistence of B. pertussis and B. parapertussis DNA in 3 (2.3%) samples, coexistence of B. pertussis and B. holmesii DNA in 10 (7.81%) and only one (0.78%) sample was IS 481 RT-PCR positive without the possibility of determining the Bordetella species with the diagnostic tools used. Confirmations of Pertussis infection in household contacts by culture, RT- PCR and serology were 10, 46 and 39%, respectively. B. pertussis DNA was confirmed in the infants as well in their mothers in 38% of the cases. Co detection of B. pertussis and B. parapertussis DNA in 2% and co-detection of B. pertussis and B. holmesii DNA in 4%. B. holmesii DNA alone was detected in 5 NP samples of index cases and their mothers. CONCLUSIONS: The results of this study confirm that B. pertussis is still circulating in children and adults, and were likely a source of pertussis contamination in infants still not vaccinated. The use of RT-PCR specific for B. pertussis in the diagnosis of adults is less sensitive and should be associated with serologic tests to improve diagnosis of pertussis and contributes to preventing transmission of the disease in infants.

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.

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

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.

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

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 ß-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.

A case report of parapneumonic pleural effusion caused by Streptococcus pneumoniae serotype 19A in a child immunized with 13-valent conjugate pneumococcal vaccine.

BACKGROUND: Simple parapneumonic effusion is a pleural effusion associated with lung infection (i.e., pneumonia). Streptococcus pneumoniae remains the most common pathogen causing parapneumonic effusions. In Morocco, the pneumococcal conjugate vaccine 13-valent (PCV13) was introduced in the national immunization program in October 2010 in 2 + 1 schedule for prevention of pneumococcal disease, and replaced by the PCV10 in July 2012 in the same schedule. We report a case of parapneumonic pleural effusions caused by S. pneumoniae serotype 19A in a child immunized with 3 doses of PCV13. CASE PRESENTATION: This is a 2.5 years old previously healthy Moroccan female, fully vaccinated by PCV13 and immunocompetent, admitted to a private medical clinic with a six months history of persistent asthma. On arrival (7 February 2015), she was febrile to 40.3 °C with a brutal flu syndrome, chills, dry cough and serous rhinitis, for which she received symptomatic treatment. A biological assessment was done that confirmed the clinical diagnosis of flu. Seven days after, she presented a progressive deterioration of its general condition and the onset of severe abdominal pain. She was hospitalized and a second biological assessment, computed tomography scans and chest radiography were done that confirmed a diagnosis of a pneumococcal parapneumonia with abscess of the left lower lobe with encysted empyema. Microbiological analysis of the pleural fluid showed a S. pneumoniae serotype 19A with susceptibility intermediate to penicillin. The patient was treated by antibiotics including amoxicillin, cefixime ceftriaxone and vancomycin. CONCLUSIONS: We reported a case of parapneumonic pleural effusions caused by a vaccine serotype pneumococcal 19A occurring in an immunocompetent child immunized with 3 doses of PCV13.

Genotypic characterization of quinolone resistant-Escherichia coli isolates from retail food in Morocco.

This study was conducted to assess the retail food as a possible vehicle for antimicrobial resistant, particularly quinolones resistant and pathogenic Escherichia coli. We determined the prevalence and characteristics of nalidixic acid (Nal) resistant E. coli isolates from diverse retail food samples. In all, 70 (28%) of 250 E. coli isolates studied were Nal-resistant E. coli and 91% of these were multi-drug resistant. Plasmid mediated quinolone resistance genes were identified in 32 isolates, including aac(6')-Ib-cr (n = 16), qnrS1 (n = 11) and qnrB19 (n = 7). Mutations in gyr A and par C genes were detected among 80% of the isolates, and the isolates showed substitution Ser83-Leu and Asp87-Asn in gyrA and Ser80-Ile in parC. In addition, three different gene cassettes were identified (aadA1, aadA7, aac(3)-Id) in 18%. Virulence-associated genes stx1, eae, sfa, hlyA and stx2 were found in six (8%), three (4%), two (3%), three (4%) and three (4%) isolates, respectively. E. coli isolates of phylogenetic group A were dominant (64%, 45/70). Pulsed field gel electrophoresis revealed none epidemiological relationship between these isolates. The results of this work report the higher frequency of Nal-resistant E. coli isolates from Moroccan retail food samples including MDR and pathogenic isolates.

[Healthcare-associated bacteraemia in intensive care units of Ibn Rochd University Hospital, Casablanca]. / Les bactériémies associées aux soins en réanimation au Centre hospitalier universitaire Ibn Rochd, Casablanca, Maroc.

Objective: This study was designed to determine the incidence of healthcare-associated bacteraemia in intensive care units (ICU) of Ibn Rochd University Hospital, Casablanca, Morocco, the frequency of the bacterial strains isolated and their antibiotic resistance patterns. Methods: A prospective longitudinal study was conducted between May and July 2015 on patients admitted to the Ibn Rochd University Hospital ICU. The Comité technique des infections nosocomiales et des infections liées aux soins (CITNILS) definition of bacteraemia was used. Micro-organisms were isolated and identified according to standard bacteriology techniques. Antibiotic susceptibility testing was performed according to CLSI 2013 guidelines. Results: A total of 720 patients were hospitalised during the study period; 48 (6.7%) acquired healthcare-associated bacteraemia with an incidence ranging from 7.3/1,000 HD to 13.7/1,000 HD. Fifty-four bacteria were isolated: A. baumannii and K. pneumoniae each accounted for 18.5% of isolates. 66.7% of A. baumannii strains were resistant to imipenem and 20% of K. pneumoniae strains were ESBL+ Conclusion: Healthcare-associated bacteraemia raises treatment problems due to multidrug-resistant bacteria, which is why regular surveillance is necessary to evaluate the time-course of these strains and the efficacy of prevention measures.

[Healthcare-associated infections in a paediatric haematology/oncology unit in Morocco]. / Infections associées aux soins dans une unité d'hématologie-oncologie pédiatrique au Maroc.

OBJECTIVE: HAI cause considerable morbidity and mortality and are associated with prolonged hospital stay and increased health care costs. To describe the incidence of HAI in paediatric cancer patients as the first step towards improving infection control policies. METHODS: A prospective surveillance study was performed in the Casablanca university hospital paediatric haematology/oncology unit over an 8-month period from January to August 2011. Data including extrinsic risk factors associated with HAI were recorded. RESULTS: The incidence of HAI was 28 per 1000 patient-days. The median age was 9.6 years and the most frequent diagnosis was acute myeloid leukaemia (32%). Neutropenia at diagnosis was significantly correlated with the risk of HAI. 55.7% of HAls were nosocomial fever of unknown origin. Gram-negative bacteria were the main pathogens (60%), gram-positive cocci were responsible for 26% of HAI and Candida for 14% of HAI. The length of hospital stay for patients with and without infection were 16.5 and 5 days, respectively (P < 0.001). Six of the 11 deaths were related to HAI. CONCLUSION: These findings suggest the need to evaluate infection control measures in order to reduce morbidity and mortality in paediatric haematology/oncology units.

Prevalence of Healthcare-Associated Infections in a Tertiary Hospital in Casablanca, Morocco, 2021.

BACKGROUND: During the COVID-19 pandemic, healthcare professionals experienced an increased workload, which may have affected infection prevention and control (IPC) programs and consequently healthcare-associated infection (HAI) rates. The objective of this study was to estimate the prevalence of HAI in Ibn Rochd University Hospital Center (IRUHC) and identify associated factors. METHODS: A survey was conducted on November 30, 2021 at IRUHC, including all patients hospitalized for at least 48 hours. Data was collected using a questionnaire, and analyzed using SPSS IBM software version 16. The significance level was set at 0.05. RESULTS: Among 887 patients, the prevalence of HAI was 9.7% (7.7%; 11.6%). The highest prevalence was observed in intensive care units (ICUs) (44.2%). Nosocomial pneumonia was the most common site (26.8%). The main isolated microorganisms were Acinetobacter baumannii (18.0%) and Escherichia coli (16.0%). All Acinetobacter baumannii isolated strains were imipenem-resistant. The presence of HAI was significantly associated with the presence of an invasive medical device (p<0.001), a higher physical status score of American Society of Anesthesiologists (ASA) (p<0.001), and a longer hospital stay (p<0.001). Conclusion : The emergence of imipenem-resistant Acinetobacter baumannii (IRAB) represents a serious therapeutic and epidemiological problem requiring the establishment of a system for monitoring the microbial environment and the application of strict hygiene measures.

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.

Carbapenem- and colistin-resistant Enterobacterales in intensive care unit patients in Mediterranean countries, 2019.

Introduction: The colonization of patients by carbapenemase-producing Enterobacterales (CPE) has been associated with heightened mortality, especially in vulnerable individuals within intensive care units (ICUs). Our study aimed to comprehensively assess CPE prevalence among ICU patients across the Mediterranean region pre-COVID-19, conducting a multicenter prevalence study in the first quarter of 2019. Methods: We collected clinical data and rectal or fecal samples from 256 ICU patients for CPE testing. Additionally, we performed whole-genome sequencing on 40 representative CPE strains to document their molecular characteristics. Results: Among the 256 patients, CPE was detected in 73 samples (28.5%), with prevalence varying from 3.3 to 69.0% across participating centers. We observed 13 colistin-resistant CPE strains, affecting three ICUs. Genetic analysis revealed highly diverse E. coli and K. pneumoniae strains, predominantly from international high-risk clones. Notably, blaOXA-48 and blaNDM-1 were the most prevalent carbapenemase genes. Molecular typing uncovered potential patient clusters in six centers. Significantly, longer hospital stays were associated with increased CPE carriage (p < 0.001). Nine centers across Morocco, Tunisia, Egypt, and Lebanon voluntarily participated. Discussion: Our study provides CPE prevalence in Mediterranean ICUs and reaffirms established CPE presence in this setting but also provides updates on the molecular diversity of CPE strains. These findings highlight the imperative of reinforcing infection control measures in the participating ICUs to curtail escalated mortality rates, and of strictly applying isolation measures around patients originating from the Mediterranean region when transferred to other healthcare institutions.

Prevalence and types of extended spectrum ß-lactamases among urinary Escherichia coli isolates in Moroccan community.

This study was designed to characterize extended-spectrum-ß-lactamases (ESBL) produced by Escherichia coli isolates causing community urinary tract infections over a 2-year period (2010 and 2011) in a Moroccan large geographical region. Molecular characterization was done by using PCR and sequencing of the ß-lactamases genes and plasmid-mediated quinolone resistance determinants. Among 1174 isolates, 49 (4.1%) were ESBL producers. The blaCTx-M-15 (n = 31) was the most frequent ESBL gene detected, followed by blaCTx-M-1 (n = 5), blaSHV-12 (n = 6), blaPER-2 (n = 3), then blaTEM-3, blaTEM-20, blaTEM-158, blaSHV-27, blaSHV-28, blaSHV-36, blaSHV-125, blaCTx-M-14 and blaCTx-M-27 with one isolate for each. The non-ESBL genes detected were blaTEM-70 (n = 1), blaTEM-176 (n = 1), blaTEM-104 (n = 6), blaTEM-1 (n = 15) and blaOxA-1 (n = 12). Plasmid mediated AmpC ß-lactamases genes; blaACT-5 (n = 1), blaDHA-1(n = 2) and blaCMY-2 (n = 4) were detected in seven isolates (14.2%). The blaOxA-48 (n = 1) and blaIMP-1 (n = 1) carbapenemases genes were detected among five carbapenem-resistant E. coli. Five isolates (10.2%) harboured qnr genes, qnrB1 (n = 3), qnrB2 (n = 1) and qnrS1 (n = 1) type were detected. Thirty isolates (61.2%) were positive for aac(6')-Ib-cr gene. The class 1 integron was detected in twenty two (44.8%) isolates. Phylogenetic grouping revealed that 22 (44.8%) isolates belonged to group A, while 15 (30.6%), 11 (22.4%) and 1 (2%) belonged to B2, D and B1. Results of conjugation experiments indicated that blaCTx-M-15, blaTEM-1, blaOxA-1, aac(6')-Ib-cr and qnrB1 genes were co-transferred and that these genes were carried by a conjugative plasmid of high molecular weight. The results of this work reports the genetic diversity of ESBL genes, with the CTX-M-15 enzyme being the most common among ESBL-producing E. coli in Moroccan community.