Acinetobacter baumannii Carbapenemase Producers in Morocco: Genetic Diversity.

INTRODUCTION:  Carbapenem resistance in Acinetobacter baumannii (A. baumannii) is a public health problem worldwide. Although carbapenem resistance is emerging in Morocco, few studies have shown the epidemiological profile of carbapenemase genes in Moroccan healthcare facilities. The aim of this study was to characterize the molecular profile of the carbapenemase enzyme in Acinetobacter baumannii from clinical isolates. METHODS: Clinical strains isolated in the laboratory from various samples were subjected to several phenotypic tests. Antibiotic susceptibility and identification were tested using Phoenix 100 (Becton Dickinson Co., Sparks, MD, USA) and Api 20 (bioMérieux, Marcy-l'Etoile, France). Simple phenotypic assays were used to detect carbapenemase oxacillinase (OXA) and metallo-ß-lactamase (MBL) production, including the modified Hodge test (MHT) and ethylenediaminetetraacetic acid (EDTA) test. The detection of carbapenemase genes was performed by multiplex and simple polymerase chain reaction (PCR). RESULTS: A total of 140 strains or 100% of isolates contained OXA-51 and ISbA1 sequences, 89% contained OXA-23 and OXA-58 sequences, and 1% contained OXA-24 sequence. The MBL genes were predominated by Verona integron-encoded metallo-ß-lactamase (VIM) (56%), followed by Seoul imipenemase (SIM) (39%), German imipenemase (GIM) (37%), São Paulo metallo-ß-lactamase (SPM) (13%), imipenemase (IMP) (11%), and New Delhi metallo-ß-lactamase (NDM) (4%). Guyana extended-spectrum ß-lactamase (GES) was not found in any isolation. CONCLUSION: Our study shows a high frequency of carbapenem resistance in Acinetobacter baumannii, as it reports a high molecular diversity of carbapenemase-encoding genes, mainly dominated by the carbapenemase ISaba1/OXA-23, which represents an emerging threat in our hospital.

Moroccan Children With Helicobacter pylori Infection: Demographics, Clinical Features, and Histological Findings.

BACKGROUND: Infesting nearly half of the world's population, Helicobacter pylori is thought to cause peptic ulcers and gastric adenocarcinoma. Several studies have examined the association between H. pylori and socioeconomic, clinical, and histological factors in pediatric populations. Similarly, this study aimed to describe the characteristics of H. pylori infection in Moroccan children. METHODS: Patients aged 1-17 years who underwent upper gastrointestinal endoscopy over a period of two years from January 2019 to January 2021 were included in this study. Gastric biopsies from the antrum and corpus of the stomach were collected. Detection of H. pylori infection was confirmed by Giemsa stain. Demographic data and clinical and endoscopic characteristics were collected and histopathological findings with gastritis scoring were recorded according to the Sydney System. RESULTS: In 213 children, 95 (45%) were found to be infected with H. pylori, and the infection rates increased as the children aged. While no significant relationship between the infection of H. pylori and all symptoms was founded, a significant association was found in nodular gastritis (p<0.05), and 98% of the infected children had chronic inflammation, which was active in 22% and atrophic in 47%. The atrophy and activity were absent or mild, and the inflammation was mild to moderate. CONCLUSION: According to this study, nodular gastritis and nonspecific symptoms were related to H. pylori infection in Moroccan children. In addition, the association between this disease and gastric atrophy in our study needs the monitoring of the mucosa of Moroccan children with gastritis and identifying factors that may contribute to gastric cancer.

The high sensitivity and specificity of rapid urease test in diagnosis of Helicobacter pylori infection in Moroccan children.

Background and Objectives: Infesting nearly 50% of the world's population, Helicobacter pylori are thought to cause peptic ulcers, as well as gastric adenocarcinoma. Several diagnostic methods are available to detect this bacterium; however, at least two must be used together for an accurate diagnosis. This study evaluated the use of rapid urease test for diagnosis of H. pylori infection in a pediatric population. Materials and Methods: Five gastric biopsies were taken from children during a 2-year period for the purpose of histological, molecular, bacteriological culture, and rapid urease testing. Results: Among 83 children, 38 were male, and 45 were female with an age ranging of 2 to 15 years. The infected group represented 31%. The rapid urease test had a sensitivity of 88.5%, a negative predictive value of 94%, a specificity of 84.2%, and a positive predictive value of 72%. Conclusion: A rapid urease test may be appropriate for ruling out H. pylori infection after a negative result. The positive results however, may be confirmed by a second invasive test.

Paediatric osteoarticular infections caused by staphylococcus aureus producing panton-valentine leucocidin in morocco: Risk factors and clinical features.

OBJECTIVE: We aimed to estimate the prevalence of Staphylococcus aureus producing Panton-Valentine leucocidin (PVL) isolated from children diagnosed with osteoarticular infections (OAIs), and to examine risk factors and clinical features. METHODS: This prospective study was conducted from January 2017 to December 2018. All hospitalised children diagnosed with S. aureus OAI are included. Blood cultures, articular fluids, synovial tissues and/or bone fragments were collected for bacteriological culture. Antimicrobial susceptibility tests were determined by disk diffusion method. Genes encoding methicillin resistance (mecA) and PVL virulence factors (luk-S-PV and luk-F-PV) were detected by multiplex polymerase chain reaction. The demographic, clinical, laboratory, radiographic and clinical features were reviewed prospectively from medical records. RESULTS: A total of 37 children with S. aureus OAIs were included, 46% of them have PVL-positive infection and 70.6% were male. The mean age was 8.12 years (±4.57), and almost were from rural settings (76.5%). Children with Staphylococcus aureus producing Panton-Valentine leucocidin (SA-PVL) were significantly associated with type of infection (P = 0.005), location of infection (P = 0.037) and abnormal X-ray (P = 0.029). All strains SA-PVL+ are sensitive to methicillin, but one strain SA-PVL negative was methicillin-resistant S. aureus, confirmed by gene mecA positive. CONCLUSION: The prevalence of S. aureus infections producing PVL toxin was high in OAIs amongst Moroccan children, mainly due to methicillin-susceptible S. aureus. Type and location of infections and abnormal X-ray were significantly associated with SA-PVL. Routine diagnostic testing of PVL-SA, continuous epidemiological surveillance and multidisciplinary management of OAI is essential to prevent serious complications.

First report of SPM metallo-ß-lactamases producing Acinetobacter baumannii isolates in Morocco.

Background and Objectives: Carbapenem-resistant Acinetobacter baumannii has recently been identified by the World Health Organization as a critical pathogen. We propose to characterize the molecular characteristics of clinical isolates of A. baumannii resistant to carbapenems collected in a Moroccan hospital. Materials and Methods: Seventy carbapenem-resistant A. baumannii isolates from various samples were received at the microbiology laboratory of the Hospital Center. Antibiotic susceptibility was tested by the diffusion disc method and molecular characterization of antimicrobial resistance was performed by PCR and sequencing. Results: Carbapenemase genes were detected in our isolates: the OXA-51 gene and the ISbA1 sequence were detected in all isolates (100%), the OXA-23 and OXA-58 genes were detected in 82.85% and 10% of isolates respectively, MBL genes were dominated by VIM 39 isolates (55.7%), followed by GIM 26 isolates (37%), SIM 20 isolates (28.5%), IMP 8 isolates (11, 4%), NDM 3 isolates (4%) and for the first time in Morocco SPM with 4 isolates (5.7%). Conclusion: The emergence of resistance of A. baumannii to carbapenems is a serious problem in our hospital which requires the establishment of a prevention strategy and strict respect for hygiene to minimize their dissemination.

First report of Kingella kingae diagnosed in pediatric bone and joint infections in Morocco.

BACKGROUND: The progress of diagnostic strategies and molecular methods improved the detection of Kingella kingae in bone and joint infections, and now, Kingella kingae is being increasingly recognized as the most frequent cause of bone and joint infection BJI in early childhood. The main objective of this prospective study is to report the frequency of Kingella Kingae in negative culture bone and joint pediatric infections, and to describe the clinical and biologic features of these children. METHODS: From December 2016 to June 2019, we selected all hospitalized patients with suspected BJI. When culture was negative on the fifth day, children under 10 years were subsequently included in the study, and PCR assay was performed systematically for researching K. kingae specific gene cpn60. Microbial culture and identification were made using standard bacteriological methods. The demographics, clinical, laboratory, radiographic and clinical features were reviewed from medical records. RESULTS: We enrolled 65 children with culture negative BJI, 46 of them having under 10 years old have been screened for the cpn60 gene. Thus, the gene encoding Kingella kingae was positive for 27 BJI cases (58.7%). The mean age of children was 3.02 years, 55.6% were aged 6 months-4 years and 29.6% of them were aged 5-10 years. The male to female ratio was 1.7 and 16 cases (59.26%) occurred during the fall-winter period. The most frequent BJI type was septic arthritis (77.8%) and the most affected sites were knee (51.9%) and hip (37.0%). We recorded a mild clinical picture with normal to mildly raised inflammatory markers. All patients had good clinical and functional outcomes, with no serious orthopedic sequelae.. CONCLUSION: K kingae is an important pathogen of culture-negative BJI in Moroccan children. PCR testing should be performed in culture-negative cases of children not only in the typical age range of 6 months to 4 years. When implemented in the routine clinical microbiology laboratory, a specific K. kingae PCR assay can provide a better diagnostic performance of BJI.

Detection of metallo-beta lactamases and oxacillinase genes in carbapenem-resistant Acinetobacter baumannii strains isolated in Morocco.

INTRODUCTION: resistance to carbapenem is widespread among Acinetobacter baumannii (A. baumannii) strains. Metallo-beta lactamases enzymes (MBL) are responsible for carbapenem resistance, as are oxacillinases (OXA). In recent years, MBL producing carbapenem-resistant strains have been reported in the world and Morocco at increasing rates. Our study aimed to investigate the presence of carbapenemases in acinetobacter strains isolated from hospitalized patients in CHU Fez. METHODS: a total of 58 imipenem-resistant A. baumannii strains isolated from clinical samples were investigated. The presence of MBL was described phenotypically by the double-disk synergy test (DDST), MBL E-test, and modified Hodge test. The blaIMP, blaVIM, genes, and blaOXA-23, blaOXA-51 genes were investigated by multiplex polymerase chain reaction (PCR). The blaNDM-1 gene was determined by simplex PCR. RESULTS: fifty-eight strains were resistant to imipenem (98%), the modified Hodge test (MHT) was positive for 58 strains (100%), 47 strains (82%) were found to be positive for MBL by the test of double-disk synergy (DDST), 58 strains (100%) were positive by E-test MBL. The OXA 51 gene was detected in all strains, and the OXA 23 gene was detected in 53 strains (91%). In addition, the MBL genes were not detected by genotypic methods. CONCLUSION: the OXA-23 and OXA-51 carbapenemases type are responsible for the resistance to carbapenems in A. baumannii resistant to carbapenems in our establishment. Resistance to carbapenems by MBL enzymes has been found by phenotypic tests, which must be confirmed by genotypic methods; and solicit other MBL genes.

Helicobacter pylori CagA EPIYA-C motifs and gastric diseases in Moroccan patients.

BACKGROUND: The pathogenicity of cagA-positive H. pylori strains is associated with the number and type of repeated sequences named EPIYA located in the C-terminal region of the CagA protein. The aim of this study is to determine the polymorphism of the H. pylori cagA 3' region circulating in Morocco and its association with different gastric pathologies. METHODS: A total of 1353 consenting patients, were recruited in this study. The gastric biopsies performed during endoscopy were used for histological examination and for molecular characterization of H. pylori. The study of the type and number of "EPIYA" motif was identified by PCR directly on H. pylori positive biopsies. RESULTS: Of all the biopsies, the infection rate was 61.1%. The cagA gene was amplified in 68.9% of the cases and the analysis of the 3' region of cagA showed the exclusive presence of the "Western CagA" type with a predominance of the EPIYA-ABC motif (71.4%). The number of EPIYA-C motif varies from 0 to 2. The multinomial analysis shows that the infection with strains of H. pylori having two EPIYA-C motifs is a factor that increases the risk of developing gastric cancer compared to gastritis cases with strains lacking this motif (OR = 11.64; CI: 3.34-45.15), whereas this risk is 6 fold higher in comparison with duodenal ulcer cases (OR = 6, CI: 1.29-27.76). CONCLUSIONS: The results of this study suggest that the number of EPIYA-C motifs might be useful as a predictive marker of the infection evolution and will help in the identification of patients at high risk of developing gastric cancer.

[Purulent pericarditis, an unusual complication of pneumococcal pneumonia: a case report]. / Péricardite purulente Complication inhabituelle de pneumonie à pneumocoque : à propos d'un cas chez une femme enceinte.

Purulent pericarditis has become rare since the advent of antibiotics. Among the involved germs, S. pneumoniae remains the most implicated pathogen to evoke in principle, especially that prescription of systematic antibiotics for any febrile condition can considerably mask the clinical picture. A 36-year-old pregnant woman was visiting the emergency department for dyspnea and flu-like syndrom that had been going on for a week. The chest X-ray showed a white lung on the left and the transthoracic ultrasound revealed a pericardial effusion, resulting in pericardial drainage and pleural puncture that allows the evacuation of a purulent fluid. S. pneumoniae was identified on the pericardial fluid. Antibiotic therapy and resuscitation measures have allowed a good evolution. Even if it has become exceptional, pneumococcal pericarditis must not be overlooked since the evolution is often favorable in triple conditions: early recognition, prompt institution of appropriate antibiotic therapy, and early surgical drainage.

VacA and CagA Status as Biomarker of Two Opposite End Outcomes of Helicobacter pylori Infection (Gastric Cancer and Duodenal Ulcer) in a Moroccan Population.

Helicobacter pylori (H. pylori) infection induces inflammation of the gastric mucosa, which may progress to precancerous lesions leading to gastric cancer. Pathological determinism is associated to some virulence genes of the bacterium, notably the vacA and cagA genes. The present study aimed to determine the H. pylori genotypes distribution and their association with sex, age and gastric diseases in a Moroccan population. Gastric biopsy was taken from 1079 consenting patients. The specimens were processed by PCR to identify H. pylori and to determine the genotypic profile by PCR characterizing vacA s, vacA m and vacA i regions directly from biopsies H. pylori positives. VacA genotyping revealed the predominance of vacA m2 (53.2%), vacA s2 (52.9%) and vacA i2 (52%). The most virulent vacA alleles (s1, i1 and m1) are more predominant in men (47.3%, 41.9% and 46.1% respectively) than in women (38.3%, 33.3% and 37% respectively). However, the association between vacA genotypes and age did not reach a statistical significant value. Logistic regression analysis results show that vacA i1m1 and vacA i1m2 genotypes were strongly associated with the risk of GC, the Odds Ratio (95% confidence interval) was 29.73 [5.08-173.73] and 9.17 [2.06-40.82] respectively, while vacAs1/cagA+ seems to be a risk factor for DU since it is inversely associated with GC (OR was 0.13 [0.02-0.75]. The results of this study suggest that vacA i1 genotype independently to vacAm status may be of a clinical usefulness and will help to identify patients at a high risk of GC development.

Detection of Helicobacter pylori urease antigen in saliva in patients with different gastric H. pylori status.

BACKGROUND: Finding a simple, accurate, and noninvasive diagnosis method is a substantial challenge for the detection of Helicobacter pylori. The aim of the present study was to compare the presence of H. pylori urease antigen in saliva with the presence of this bacterium in gastric mucosa. METHODS: Saliva samples and gastric biopsies were taken from 153 consenting Moroccan patients. Saliva samples were analyzed using an immunochromatographic test for urease antigen H. pylori detection. Thereafter, the gastric biopsies were analyzed by histology and polymerase chain reaction (PCR) to detect this bacterium. RESULTS: From a total of 153 recruited Moroccan patients, H. pylori was detected in 28 (18.30%), 87 (57.24%), and 69 (45.10%) cases by saliva test, histology, and PCR, respectively. A significant association was observed between the presence of H. pylori antigen in saliva and age. However, no association was found with sex, H. pylori virulence factors, gastric disease outcome, and density of the bacterium on the gastric mucosa. Considering that only 90 patients presented concordant results on H. pylori diagnosis (positive or negative) by both histology and PCR, the immunochromatographic test showed very low sensitivity (29.79%) and high specificity (90.70%). Of these two tests, the positive and negative predictive values were 77.78% and 54.17%, respectively. The accuracy of the test for salivary detection of urease antigen H. pylori was 58.89%. CONCLUSION: This study demonstrated a low detection rate of H. pylori antigens in saliva compared with the presence of this bacterium in gastric mucosa, suggesting that saliva cannot be used as a suitable sample for the diagnosis of H. pylori in our study population.

Helicobacter pylori genotypes associated with gastric histo-pathological damages in a Moroccan population.

H. pylori persistent infection induces chronic gastritis and is associated with peptic ulcer disease and gastric carcinoma development. The severity of these diseases is related to human's genetic diversity, H. pylori genetic variability and environmental factors. To identify the prevalence of histo-pathological damages caused by H. pylori infection in Moroccan population, and to determine their association to H. pylori genotypes, a prospective study has been conducted during 3 years on patients attending the gastroenterology department of Hassan II University Hospital (CHU) of Fez, Morocco. A total of 801 Moroccan adults' patients were recruited; H. pylori was diagnosed and genotyped by PCR in biopsy specimens and histological exam was performed. We found a high rate of glandular atrophy. Chronic inflammation, neutrophil activity and glandular atrophy showed statistically significant association with H. pylori infection. However, intestinal metaplasia was inversely associated to this infection and no association was observed with gastric cancer cases. A statistically significant association was found between intestinal metaplasia and vacAs1 and vac Am1 genotypes in patients aged 50 years and more but not in younger. This last genotype is also associated to gastric cancer. In this study, gastric cancer showed no significant association with H. pylori. Further studies are warranted to determine the role of other etiological agents such as Epstein-Barr virus, human papillomavirus and possibly environmental and dietetic factors in the occurrence of this pathology.

Prevalence of Helicobacter pylori and its recurrence after successful eradication in a developing nation (Morocco).

INTRODUCTION: In developed countries, reinfection after successful eradication of Helicobacter pylori appears unusual. High prevalences of H. pylori in developing countries may result in high reinfection rates. In Morocco, published studies regarding the prevalence of H. pylori are limited, and to our knowledge, there are no data regarding the reinfection rate of H. pylori after successful treatment. AIM: The aim of this study was to determine the prevalence of H. pylori in our area, and the reinfection rate at 6 months and 1 year of follow-up after successful eradication. METHODS: Consecutive patients with investigated ulcer or non-ulcer dyspepsia were prospectively enrolled in the Hassan II University Hospital of Fez. Patients with H. pylori infection were treated with a 1-week triple therapy or 10 day sequential therapy. Those tested urea breath test negative after 3 months of treatment were followed prospectively with repeated urea breath test at 6 months and 1 year. H. pylori status at endoscopic examination was determined by rapid urease test, histology, and culture. RESULTS: Four hundred and twenty-nine patients were enrolled in the study, among them 324 patients (75.5%) presented with H. pylori infection. Two hundred and fifty-six (83.3%) patients had successfully eradicated H. pylori following treatment, among them, two patients (0.8%) were reinfected with H. pylori over 12 months. The rate of reinfection was 0.42% in the first 6 months and of 0.45% in the first year of the study. CONCLUSION: The results of the present study demonstrate that firstly, prevalence of H. pylori is high (75.5%) in our area, secondly as in developed countries, there is a low (0.8%) but continuous risk of H. pylori infection in adulthood. A different approach for follow-up after H. pylori eradication is probably needed in patients of developing countries, since reinfection prevalence is different between countries.

Staphylococcus aureus nasal carriage in a Moroccan dialysis center and isolates characterization.

Staphylococcus aureus, which has its ecological niche in the anterior nares, has been shown to cause a variety of infectious diseases mainly for patients in hemodialysis units. We performed this study to evaluate the prevalence of nasal S. aureus carriage among hemodialysis outpatients, to determine the antimicrobial susceptibility of isolates, to characterize the virulence genes, and to identify associated risk factors. Nares swab specimens were obtained from 70 outpatients on hemodialysis between March and June 2010. Samples were plated immediately onto S. aureus specific media and pattern of antibacterial sensitivity was determined using disk diffusion method. Polymerase chain reaction was used to detect nuc, mecA, and genes encoding staphylococcal toxins. Medical record of patients was explored to determine S.aureus carriage risk factors. Nasal screening identified 42.9% S. aureus carriers with only one (3.3%) methicillin-resistant S. aureus isolate. Among the methicillin-susceptible S. aureus isolates, high rate of penicillin resistance (81.8%) has been detected. The identified risk factors were male gender and age ≤ 30 years. Research of virulence factors showed a high genetic diversity among the 30 S. aureus isolates. Twenty-one (70%) of them had at least one virulence gene, of which 3.3% were Panton-Valentine leukocidin (lukS/F-PV) genes. S. aureus carriage must be screened for at regular intervals in hemodialysis patients. Setting up a bacterial surveillance system is one of the strategies to understand the epidemiology of methicillin-resistant S. aureus, to guide local antibiotic policy and prevent spread of antibiotic-resistant S. aureus.

Sequential therapy versus standard triple-drug therapy for Helicobacter pylori eradication: result of the HPFEZ randomised study.

OBJECTIVE: The aim of this study was to assess and compare the efficacy and safety of sequential treatment with standard triple therapies in a located population in Morocco. METHODS: Consecutive H. pylori-positive patients with endoscopy-proven ulcer or non ulcer dyspepsia were prospectively randomized in the trial into one of three groups: AM and AC group were administered a tri-therapy for 7 days including PPI + amoxicillin + metronidazole (AM group)/clarithromycin (AC group) and SQ group was administered a sequential regimen consisting of PPI + amoxicillin for 5 days followed by PPI + clarithromycin + metronidazole for the remaining 5 days. Eradication was confirmed by 13C-urea breath test 3 months after the end of the treatment. RESULTS: Groups AM, AC and SQ included respectively 104, 115 and 104 patients. They were comparable in terms of age, sex, clinical and endoscopic presentation. The rate of H. pylori eradication with sequential therapy was found at 94.2% (n=98) in ITT and 96% (n=98) in PP. It was higher than those found in the AM group: 70% (n=73) in ITT and 70.8% (n=73) in PP and the AC group: 78.2% (n=90) in ITT and 79.6% (n=90) in PP (0.001). The prevalence of side effects following the sequential treatment was 9.6% (n=10) versus 22% (n=22) and 27.8% (n=32) in the AM and AC groups, respectively, (P=0.006). CONCLUSIONS: Sequential treatment was better tolerated and achieved a significantly higher eradication rate of H. pylori compared with standard triple therapies in this population.

Prevent infection linked to the dialysis water in a hemodialysis center in Fez city (Morocco).

BACKGROUND: Water treatment systems are a critical variable in dialysis therapy. Rigorous control of hemodialysis water quality is particularly important in order to guarantee a better quality of life of the hemodialysis patients. The objective of the study was to evaluate the chemical, microbiological quality and antimicrobial resistance of bacteria isolated from water and dialysate in a public HD center. METHODS: Fifty five samples of water and dialysate were collected weekly over a period of 4 months. The samples were collected from 4 points in the distribution loop. The microbiological and chemical analyses were performed according to our national standards. Antimicrobial susceptibilities patterns of isolated bacteria were determined by disk diffusion method. RESULTS: The chemical and microbiological parameters in all dialysis water and dialysate samples are in accordance with national standards. However, 70 Gram-negative bacteria were identified: Pseudomonas sp, Ochrobactrum antropi and Burkholderia cepacia, isolated at 52.8%, 12.8% and 17% simultaneously. Fourteen per cent of the isolates were resistant to three or more antibiotics. All resistant bacteria belong to the genus of Pseudomonas, 80% were resistant to tetracycline and to co-trimoxazole, 30% to ceftazidime. No colistin and imipenem resistance was observed. CONCLUSION: To avoid a health risk due to bacterial contamination, an adequate system for water treatment, disinfection of the hemodialysis system and microbiological monitoring of the water and dialysate are necessary.