Genetic alterations in the pmrAB two-component system and lipid A biosynthesis genes of polymyxin-resistant Acinetobacter baumannii isolates.

The rate of pandrug-resistant Acinetobacter baumannii strains is on the rise in all continents. This bacterium can acquire resistance to all antibiotics, even to colistin. Alterations in the lipid A or/and the two-component pmrAB were earlier detected in colistin resistance. We investigated and analyzed two strains of A. baumannii (ABRC1 and ABRC2) isolated from two patients admitted to intensive care unit with a septic shock. Both strains were resistant to all tested antibiotics including colistin with a MIC >256 mg L-1. Colistin resistance genes (pmrA, pmrB, lpxA, lpxC, lpxD, and lpsB) of two strains (ABRC1 and ABRC2) were investigated by PCR and sequencing. Obtained nucleic acid sequences were aligned with reference sequences of ATCC 19606 and 17987. In this study two amino acid mutations, N287D in the lpxC gene and E117K in the lpxD gene, were detected in both ABRC1 and ABRC2 strains. ABRC1 had an additional H200L mutation in the pmrA gene. Both colistin resistant strains harbored the same A138T mutation in the pmrB gene. The ABRC2 strain also had an alteration in the kinase domain, specifically an R263S substitution of the histidine kinase domain. Three identical mutations were found in the lpsB gene of both A. baumannii strains: Q216K + H218G + S219E. As a result, a newly deduced protein sequence in both ABRC1 and ABRC2 strains differed from those described in ATCC 17978 and 19606 strains was determined. Colistin resistance is multifactorial in A. baumannii. In our study we detected novel mutations in colistin resistant A. baumannii clinical isolates.

Evaluation of GeneXpert MTB/RIF system performances in the diagnosis of extrapulmonary tuberculosis.

BACKGROUND: Tuberculosis represents a serious public health problem and a significant diagnostic and therapeutic challenge worldwide. Molecular diagnostic techniques are crucial in the World Health Organization's new tuberculosis control strategy. This study aims to evaluate the performance of GeneXpert MTB/RIF (Cepheid Sunnyvale, CA, United States) in diagnosis of extra-pulmonary tuberculosis then compare it's performance in detecting Rifampicin resistance to GenoType MTBDRplus (HAIN Life Sciences, Nehren, Germany). METHODS: Samples from pulmonary and/or extra-pulmonary origins were analysed in a 21 months retrospective study. Samples were sent to the bacteriology laboratory for Mycobacterium tuberculosis detection using conventional bacteriological and molecular methods (GeneXpert MTB/RIF and MTBDRplus). Sensitivity and specificity were calculated for the stained smear and GeneXpert according to culture (Gold Standard) as well as for GeneXpert MTB/RIF in both negative and positive microscopy tuberculosis cases. Data's statistical analysis was performed with SPSS13.0 software. RESULTS: Seven hundred fourteen patients' samples were analysed; the average age was 47.21 ± 19.98 years with a male predominance (66.4%). Out of 714 samples: 285 were from pulmonary and 429 were from extra-pulmonary origins. The positivity rates for microscopy, GeneXpert MTB/RIF and culture were 12.88, 20.59 and 15.82%, respectively. These rates were 18.9, 23.85 and 20.35% for pulmonary samples and 9.71, 18.41 and 12.82% for extra-pulmonary samples, respectively. The sensitivity and specificity of GeneXpert MTB/RIF were almost the same in both pulmonary and extra-pulmonary samples (78.2 and 90.4%) and (79,3 and 90.3%) respectively. Rifampicin resistance rate found by GeneXpert MTB/RIF was 0.84%. Comparison of Rifampicin resistance obtained by GeneXpert MTB/RIF and Genotype MTBDRplus, showed 100% agreement between the two techniques for studied samples. CONCLUSIONS: This confirms GeneXpert MTB/RIF advantage for tuberculosis diagnosis, particularly extra-pulmonary tuberculosis with negatively stained smear. The performance of GeneXpert and Genotype MTBDRplus are similar in detection of Rifampicin resistance. However, variability of detection performance according to tuberculosis endemicity deserves more attention in the choice of screening techniques of Rifampicin resistance, hence the interest of conducting comparative studies of detection performance under low and medium endemicity on large samples of tuberculosis populations.

Peritoneal tuberculosis, an underestimated diagnosis: a case report.

Tuberculosis is an infectious disease that most often affects the lungs, caused by human-to-human transmission of Mycobacterium tuberculosis. Peritoneal tuberculosis is an extra-pulmonary form of the disease that usually manifests as an ascitic syndrome, with or without fever, in a context of altered general condition, often in endemic areas. The diagnosis of peritoneal tuberculosis is not always easy, as the clinical signs are often insidious and unspecific. We report a case of peritoneal tuberculosis in an 18-year-old female, who had presented for 10 days with a progressive increase in abdominal volume associated with vomiting and diarrhoea.

Exceptional association of two species of bacteria causing acute appendicitis: Haemophilus influenzae and Enterobacter cloacae.

Appendicitis, typically caused by appendiceal lumen obstruction, is a prevalent abdominal surgical emergency worldwide. While most cases involve Enterobacterales, Haemophilus influenzae, primarily known for upper respiratory infections, is infrequently associated with gastrointestinal infections. This article presents an exceptional case of acute appendicitis caused by both Haemophilus influenza and Enterobacter cloacae in a 15-year-old child, highlighting the significance of recognizing uncommon pathogens in appendicitis and emphasizing the necessity for thorough microbiological investigations to refine diagnostic approaches.

Predominance of OXA-48 Carbapenemase-Producing Enterobacterales in a Moroccan Hospital.

Objective: The emergence of carbapenemase-producing Enterobacterales (CPE) is a major concern that is increasingly reported worldwide. Our study aimed at investigating the resistance of CPE isolates in a Moroccan teaching hospital using phenotypic and genotypic methods. Methods: Enterobacterales strains from March to June 2018 were collected from different clinical samples. The Enterobacterales isolates resistant to third-generation cephalosporins (3GC) and/or carbapenems were subjected to the Carba NP test and an immunochromatographic test for phenotypic detection. Detection of extended-spectrum ß-lactamases (ESBL) was also performed following standards. Molecular screening of carbapenemases genes (OXA-48, NDM, blaKPC, blaIMP, blaVIM, and blaOXA-24, blaOXA-23, OXA-51, OXA-58) using conventional multiplex PCR assays was also performed on 143 isolates. Results: Enterobacterales represented 52.7% with a proportion of 21.8% of bacteria resistant to 3GC and/or carbapenems. Within 143 isolates MDR to 3GC, K. pneumoniae, E. coli, and E. cloacae represent 53.1%, 40.6%, and 6.3%, respectively. These strains were isolated mainly from urinary samples (74.8%) in patients admitted to emergency and surgical units. 81.1% of strains are producing ESBL and 29% are carbapenemase producers as confirmed by the Carba NP test, immunochromatographic test, and molecular testing. OXA-48 carriers represent 83.3% of these strains, followed by NDM with 16.7%. blaKPC, blaIMP, blaVIM, and blaOXA-24, blaOXA-23, OXA-51, OXA-58 were not detected in any of these bacteria. Conclusions: A high rate of CPE carrying OXA-48 among Enterobacterales resistant to 3GC and/or carbapenems isolates was found. Strict observance of hospital hygiene measures and more rational use of antibiotics are mandatory. Implantation of carbapenemases detection should be encouraged in our hospital settings to estimate the true burden of the CPE.

Neutralising antibodies against SARS-CoV-2 give important information on Covid-19 epidemic evolution in Rabat, Morocco, March 2020-February 2021.

Background: The SARS-CoV-2 is an extremely contagious and acute viral disease mainly affecting humans. Objective: To estimate seroprevalence of SARS-CoV-2 neutralizing antibodies (NAbs) for illegible armed force individuals living in Rabat, Morocco. Method: A convenience sample (N = 2662) was conducted from May 2020 to February 2021. We used the standard neutralization assay to quantify the NAbs titers. A serum was positive when the titer was 1:4. High positive NAbs titers were defined when ≥ 1:32. Results: Demographic and socioeconomic status did not affect seroprevalence data. An overall seroprevalence of 24,9% was found. Sera from blood donors, young recruits and auto-immune population had lower NAbs titers. However, titers were above 1:16 in 9% of the population with high risk of SARS-CoV-2 exposure. Seropositivity increased over time with values reaching peaks after the epidemic waves (2.4% in May 2020; 16.2% in August 2020; 22.7% in December 2020 and 37% in February 2021). Conclusion: And increase of NAbs was observed over time and correlated with the post-epidemic waves of COVID-19 in Morocco.

Primary tuberculosis of the parotid gland: A forgotten diagnosis about a case!

The diagnosis of tuberculosis must be made in the face of any cervical swelling, and the treatment is essentially medical.

Intensive care unit-acquired Acinetobacter baumannii infections in a Moroccan teaching hospital: epidemiology, risk factors and outcome.

INTRODUCTION: The objective of this study was to examine the epidemiology, risk factors and outcome associated with Acinetobacter baumannii infections in the intensive care units (ICUs) in a Moroccan teaching hospital. METHODS: This is a matched case-control study conducted as a joint collaboration between the clinical Bacteriology department and the two ICUs of Mohammed V Military Teaching Hospital from January 2015 to July 2016. RESULTS: Among 964 patients hospitalized in the ICUs, 81 (8.4%) developed A. baumannii infections. Multivariate logistic regression analysis identified the following independent risk factors for ICU-acquired A. baumannii infections: ICU stay ≥14 days (odds ratio (OR)=6.4), prior use of central venous catheters (OR=18), prior use of mechanical ventilation (OR=9.5), duration of invasive procedures ≥7 days (OR=7.8), previous exposure to imipenem (OR=9.1), previous exposure to amikacin (OR=5.2), previous exposure to antibiotic polytherapy (OR=11.8) and previous exposure to corticotherapy (OR=5). On the other hand, the admission for post-operative care was identified as a protective factor. The crude mortality in patients with A. baumannii infection was 74.1%. Multivariate analysis showed that septic shock (OR=19.2) and older age (≥65 years) (OR=4.9) were significantly associated to mortality risk in patients with A. baumannii infection. CONCLUSION: Our results show that shortening the ICU stay, rational use of medical devices and optimizing antimicrobial therapy could reduce the incidence of these infections. Elderly patients and those with septic shock have a poor prognosis. These findings highlight the need for focusing on the high-risk patients to prevent these infections and improve clinical outcome.

Clonal diversity and detection of carbapenem resistance encoding genes among multidrug-resistant Acinetobacter baumannii isolates recovered from patients and environment in two intensive care units in a Moroccan hospital.

BACKGROUND: Carbapenem-resistant Acinetobacter baumannii has recently been defined by the World Health Organization as a critical pathogen. The aim of this study was to compare clonal diversity and carbapenemase-encoding genes of A. baumannii isolates collected from colonized or infected patients and hospital environment in two intensive care units (ICUs) in Morocco. METHODS: The patient and environmental sampling was carried out in the medical and surgical ICUs of Mohammed V Military teaching hospital from March to August 2015. All A. baumannii isolates recovered from clinical and environmental samples, were identified using routine microbiological techniques and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry. Antimicrobial susceptibility testing was performed using disc diffusion method. The carbapenemase-encoding genes were screened for by PCR. Clonal relatedness was analyzed by digestion of the DNA with low frequency restriction enzymes and pulsed field gel electrophoresis (PFGE) and the multi locus sequence typing (MLST) was performed on two selected isolates from two major pulsotypes. RESULTS: A total of 83 multidrug-resistant A. baumannii isolates were collected: 47 clinical isolates and 36 environmental isolates. All isolates were positive for the blaOXA51-like and blaOXA23-like genes. The coexistence of blaNDM-1/blaOXA-23-like and blaOXA 24-like/blaOXA-23-like were detected in 27 (32.5%) and 2 (2.4%) of A. baumannii isolates, respectively. The environmental samples and the fecally-colonized patients were significantly identified (p < 0.05) as the most common sites of isolation of NDM-1-harboring isolates. PFGE grouped all isolates into 9 distinct clusters with two major groups (0007 and 0008) containing up to 59% of the isolates. The pulsotype 0008 corresponds to sequence type (ST) 195 while pulsotype 0007 corresponds to ST 1089.The genetic similarity between the clinical and environmental isolates was observed in 80/83 = 96.4% of all isolates, belonging to 7 pulsotypes. CONCLUSION: This study shows that the clonal spread of environmental A. baumannii isolates is related to that of clinical isolates recovered from colonized or infected patients, being both associated with a high prevalence of the blaOXA23-like and blaNDM-1 genes. These findings emphasize the need for prioritizing the bio-cleaning of the hospital environment to control and prevent the dissemination of A. baumannii clonal lineages.

Whole-Genome Shotgun Sequences of Three Multidrug-Resistant Mycobacterium tuberculosis Strains Isolated from Morocco.

Tuberculosis is a contagious disease that usually attacks the lungs but sometimes attacks other parts of the body, such as the kidneys, glands, and bones. It is an endemic and major public health problem in Morocco. Tuberculosis is transmitted through the airways via the inhalation of microdroplets containing Mycobacterium tuberculosis We present here the whole-genome shotgun sequences of three multidrug-resistant M. tuberculosis strains isolated from Morocco.

First Whole-Genome Sequences of Two Multidrug-Resistant Acinetobacter baumannii Strains Isolated from a Moroccan Hospital Floor.

This report describes the whole-genome shotgun sequences of two multidrug-resistant Acinetobacter baumannii strains, ABE8_07 and ABE12_M, isolated from a Moroccan hospital floor. These two genome sequences will initiate the study and characterization of the Acinetobacter baumannii genome in Morocco.

Diabetic Foot Infection in Morocco: Microbiological Profile.

OBJECTIVE: The objective of this work is to describe the microbiology of diabetic foot infections (DFIs). MATERIALS AND METHODS: The authors included all samples of infected diabetic foot ulcers between January 2009 and June 2014 at the Mohammed Vth Military Teaching Hospital of Rabat, Morocco. RESULTS: The researchers collected 199 samples corresponding to 157 patients. The mean age of the patients was 59 years ± 12 years. Of the collected samples, deep samples represented 41% and swab samples 59%. Direct examination indicated anaerobic infection in 32% of the cases. There were 307 bacteria isolates from both deep and swab samples. There was no statistically significant association between the sampling method and isolate species (P = 0.237). Enterobacteriaceae, Staphylococcus aureus, Streptococcus sp, nonfermenting gram-negative bacilli (NFGNB), and Enterococcus sp represented 31.8%, 12.6 %, 12.3%, 11.7%, and 8.7% of the isolates, respectively. Methicillin-resistant S. aureus represented 4.7% of S. aureus isolates. Enterobacteriaceae and NFGNB-producing extended spectrum ß-lactamases represented 14.1% and 5.1%, respectively, with isolates producing carabapenemase representing 3.8% and 38.5%. Piperacillin-tazobactam, imipenem, and ciprofloxacin resistance concerned 7.5%, 4.7%, and 25.5%, respectively, of isolated Enterobacteriaceae, and 35.9%, 30.7%, and 35.9% of NFGNB. Low susceptibility to ß-lactams was found in 4.9% of Streptococcus sp isolates and 4.9% of Streptococcus sp isolates were resistant to moxifloxacin. CONCLUSION: Gram-negative bacilli are responsible for 43% of DFIs, and multidrug-resistant GNB is a challenging issue in DFI management. The sampling method doesn't seem to impact the bacteriological profile; however, this finding must be confirmed with further study.

Acinetobacter infections prevalence and frequency of the antibiotics resistance: comparative study of intensive care units versus other hospital units.

INTRODUCTION: This study aims to determine the Acinetobacter sp clinical isolates frequency and its antibiotic susceptibility pattern by comparing results obtained from the Intensive Care Units (ICUs) to that of other units at the Mohammed V Military Teaching Hospital in Rabat. METHODS: This is a retrospective study over a 2-years period where we collected all clinical isolates of Acinetobacter sp obtained from samples for infection diagnosis performed on hospitalized patients between 2012 to 2014. RESULTS: During the study period, 441 clinical and non-repetitive isolates of Acinetobacter sp were collected representing 6.94% of all bacterial clinical isolates (n = 6352) and 9.6% of Gram negative rods (n = 4569). More than a half of the isolates were from the ICUs and were obtained from 293 infected patients of which 65, 2% (191 cases) were males (sex ratio = 1.9) and the median age was 56 years (interquartile range: 42-68 years). Acinetobacter clinical isolates were obtained from respiratory samples (44.67%) followed by blood cultures (14.51%). The resistance to ciprofloxacin, ceftazidime, piperacillin / tazobactam, imipenem, amikacin, tobramycin, netilmicin, rifampicin and colistin was respectively 87%, 86%, 79%, 76%; 52%, 43%, 33% 32% and 1.7%. The difference in resistance between the ICUs and the other units was statistically significant (p <0.05) except for colistin, tetracycline and rifampicin. CONCLUSION: This paper shows that solving the problem of prevalence and high rate of multidrug resistant Acinetobacter infection which represents a therapeutic impasse, requires the control of the hospital environment and optimizing hands hygiene and antibiotics use in the hospital.