Platelets Can Associate with SARS-Cov-2 RNA and Are Hyperactivated in COVID-19.

Rationale: In addition to the overwhelming lung inflammation that prevails in COVID-19, hypercoagulation and thrombosis contribute to the lethality of subjects infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Platelets are chiefly implicated in thrombosis. Moreover, they can interact with viruses and are an important source of inflammatory mediators. While a lower platelet count is associated with severity and mortality, little is known about platelet function during COVID-19. Objective: To evaluate the contribution of platelets to inflammation and thrombosis in COVID-19 patients. Methods and Results: Blood was collected from 115 consecutive COVID-19 patients presenting non-severe (n=71) and severe (n=44) respiratory symptoms. We document the presence of SARS-CoV-2 RNA associated with platelets of COVID-19 patients. Exhaustive assessment of cytokines in plasma and in platelets revealed the modulation of platelet-associated cytokine levels in both non-severe and severe COVID-19 patients, pointing to a direct contribution of platelets to the plasmatic cytokine load. Moreover, we demonstrate that platelets release their alpha- and dense-granule contents in both non-severe and severe forms of COVID-19. In comparison to concentrations measured in healthy volunteers, phosphatidylserine-exposing platelet extracellular vesicles were increased in non-severe, but not in severe cases of COVID-19. Levels of D-dimers, a marker of thrombosis, failed to correlate with any measured indicators of platelet activation. Functionally, platelets were hyperactivated in COVID-19 subjects presenting non-severe and severe symptoms, with aggregation occurring at suboptimal thrombin concentrations. Furthermore, platelets adhered more efficiently onto collagen-coated surfaces under flow conditions. Conclusions: Taken together, the data suggest that platelets are at the frontline of COVID-19 pathogenesis, as they release various sets of molecules through the different stages of the disease. Platelets may thus have the potential to contribute to the overwhelming thrombo-inflammation in COVID-19, and the inhibition of pathways related to platelet activation may improve the outcomes during COVID-19.

Indeterminate genotypes of hepatitis C virus by the Abbott RealTime HCV Genotype II assay in Morocco. About eight cases resolved by a sequencing method.

The early detection and genotyping of the hepatitis C virus (HCV) are important in the management of this infection. The genotype is the major factor influencing treatment decisions. That's why it is necessary to use fast and accurate methods in its determination. This study reports, over a period of 3 years (from May 2012 to July 2015), the percentage of indeterminate genotypes by the Abbott RealTime HCV Genotype II test and their results using a sequencing technique. Of 309 samples of 309 patients tested, 11 were indeterminate (4.4%). There were three cases of cross-reactivity (1b/4 in one case, 2/5 in two cases) and a possible co-infection 1 + 4. Among those indeterminate genotypes, cross-reactivities and co-infections, ten samples were tested by sequencing. The results were for four of them a 1d subtype, five were a 2i subtype and one was a 2l subtype. These results support the thesis of complementarity between the two methods: genotyping for the detection of mixed reactions and sequencing for resolving indeterminate cases by genotyping.

Enterobacteriaceae producing extended-spectrum ß-lactamases (ESBLs) colonization as a risk factor for developing ESBL infections in pediatric cardiac surgery patients: "retrospective cohort study".

BACKGROUND: Children with cardiac defects need many hospitalizations and repetitive antibiotic therapies, with an increasing risk of colonization with multidrug-resistant bacteria (MDRB) such as extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBL-E) Post-operative infections with these bacteria in paediatric cardiac surgery are life threatening. This article aims to study the prevalence of ESBL colonization among paediatric cardiac surgery patients, and to compare occurrence of post-operative infections with and without ESBL colonization. We also aim to study the correlation between the onset of postoperative infection and other parameters such as age, length of stay and preoperative antibiotic therapy. METHODS: A retrospective cohort study included paediatric cardiac surgery patients in Cheikh Zaid hospital in Rabat, Morocco, between the 1st of January 2011 and 31 December 2014. A screening for ESBL colonization was requested for children who had a risk factor (previous hospitalization and/or taking antibiotics) at admission. Swabs were collected from three sites (throat, nose and anus). Two groups were compared - patients colonized and not colonized with ESBLs. Statistical analysis was performed using R software. RESULTS: ESBL colonization screening was performed in 111 patients. Positive colonization was detected in 17 cases (15%). Klebsiella pneumoniae (KP): 9 (53%) was the most frequently isolated species. Among the 17 patients, 23.5% (4/17) developed a postoperative infection due to ESBLs versus only one patient without colonization (1%). There was a statically significant difference in terms of occurrence of postoperative infection between the two groups (p = 0.001). Relative risk of developing a postoperative infection with positive colonization was 22 (95% CI, 8.37-58.5). CONCLUSIONS: The analysis of colonization with multidrug-resistant bacteria and the prevention of nosocomial infections appear to be important challenges for paediatric cardiac surgery. Systematic screening of ESBL colonization for cardiac surgery could have a significant contribution, on one hand to guide prophylactic antibiotic therapy of patients, and on the other, to prevent spread of those infections.

Bacteraemia in Intensive Care Unit: Clinical, Bacteriological, and Prognostic Prospective Study.

Objectives. We conducted a one-year observational study from December 2012 to November 2013 to describe the epidemiology of bacteraemia in intensive care units (ICU) of Mohammed V Military Teaching Hospital of Rabat (Morocco). Methods. The study consisted of monitoring all blood cultures coming from intensive care units and studying the bacteriological profile of positive blood cultures as well as their clinical significance. Results. During this period, a total of 46 episodes of bacteraemia occurred, which corresponds to a rate of 15,4/1000 patients. The rate of nosocomial infections was 97% versus 3% for community infections. The most common source of bacteraemia was the lungs in 33%, but no source was identified in 52% of the episodes. Gram negative organisms were isolated in 83,6% of the cases with Acinetobacter baumannii being the most frequent. Antibiotic resistance was very high with 42,5% of extended-spectrum beta-lactamases (ESBLs) in Enterobacteriaceae and 100% of carbapenemase in Acinetobacter baumannii. The antibiotherapy introduced in the first 24 hours was adequate in 72% of the cases. Conclusions. Bloodstream infections in ICU occur most often in patients over 55 years, with hypertension and diabetes. The bacteria involved are mainly Gram negative bacteria multiresistant to antibiotics. Early administration of antibiotics significantly reduces patients mortality.

Evaluation of the MAScIR SARS-CoV-2 M Kit 2.0 on the SARS-CoV-2 Infection.

SARS-CoV-2 is a major public health problem worldwide. Since its emergence, several diagnostic kits have been developed to ensure rapid patient management. The aim of our study is to check the performance of the new Moroccan SARS-CoV-2 detection kit: MAScIR SARS-CoV-2 M 2.0. The following parameters were studied: repeatability, reproducibility, analytical specificity, analytical sensitivity, and comparison with the GeneFinder™ COVID-19 Plus RealAmp Kit. In addition, an external quality evaluation comprising five specimens was carried out as part of an international program for the external quality evaluation of sublaboratories of the WHO and the Laboratory Office of the National Institute of Hygiene of Morocco. The results of all parameters studied showed an analytical performance that complied with the requirements of the method verification/validation protocol adopted by the Central Laboratory of Virology and met the recommendations of COFRAC (French Accreditation Committee). During the current study, the sequencing of some randomly selected positive samples was performed, among which the carriers of the Alpha variant, the Delta variant, and the Omicron variant were detected. These results allowed us to deduce that this kit was valid for detecting these three variants.

First Report of a SARS-CoV-2 Genome Sequence with a Spike His69-Val70 Deletion and an Asn439Lys Mutation in Morocco.

We report the nearly complete genome sequence and the genetic variations of a clinical sample of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) collected from a nasopharyngeal swab specimen from a male patient from Harhoura-Rabat, Morocco. The sequence, which was obtained using Ion Torrent technology, is valuable as it carries a recently described deletion (His69-Val70) and substitution (Asn439Lys).

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.

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.

Drug resistance mutations in HIV type 1 isolates from patients failing antiretroviral therapy in Morocco.

The emergence of viral-resistant strains is a major problem for the medical management of HIV-infected individuals. The aim of this study was to characterize viral subtypes and drug-resistance mutations (DRMs) in HIV-1 isolates from patients failing antiretroviral therapy (ART). A total of 45 HIV-1-infected patients failing ART were enrolled. The viral RT and Prot genes were amplified and sequenced to determine subtypes and potential DRMs. The subtype distribution was 74% subtype B, 11% subtype A, 9% CRF02-AG, 4% subtype G, and 2% subtype C. Virus samples from 34% of the patients had no DRM while 53%, 27%, and 2% of samples carried at least one DRM conferring resistance to drugs of one, two, or three classes, respectively. DRMs were observed in 50% of the patients infected with non-B strains. The prevalence of nucleoside transcriptase inhibitor (NRTI) mutations was 48%, M184V being largely predominant. The prevalence of nonnucleoside reverse transcriptase inhibitor (NNRTI) mutations was 13%, with K103N present in 57% of samples from NNRTIs-exposed patients. The prevalence of protease inhibitor (PI) mutations was 22%, with major mutations V82A and M46I seen in 16% and 11% of viruses from PI-exposed individuals, respectively. Our study shows the emergence of DRMs in HIV-1 isolates from Moroccan patients failing ART. Although not surprising, the data plead for longitudinal surveys of the dynamics of emergence of DRMs (with a focus on multidrug resistance) in treated patients and circulation of resistant HIV-1 strains in this country.

Factors influencing uptake of influenza vaccine amongst healthcare workers in a regional center after the A(H1N1) 2009 pandemic: lessons for improving vaccination rates.

BACKGROUND: Vaccination of health-care workers (HCWs) against seasonal influenza has been consistently recommended worldwide in order to prevent nosocomial transmission and ensure delivery of health-care services during outbreaks. Overall, immunization rates were low across all nation, including among HCWs. Little is known about the acceptability and compliance with seasonal influenza vaccine among HCWs after the A(H1N1) 2009 pandemic. PARTICIPANTS AND SETTING: Between 1st and 31 January 2011, we conducted a questionnaire-based survey at the Ibn Sina regional center (Rabat, Morocco). Seven hundred twenty one HCWs have answered about their influenza immunization during the 2010/2011 season, as well as the reasons for accepting or declining this vaccine. Finally, we compare our results with previous moroccan survey. RESULTS: A total of 122 HCWs (17%) reported having received the 2010/2011 seasonal vaccine; "self-protection" and "protection of the patient" were the most frequently adduced reasons for acceptance of the influenza vaccination, whereas media controversy during the pandemic was the main argument for refusal. DISCUSSION: The post pandemic seasonal influenza vaccination coverage among the HCWs in our institution was very low. The role of media, specific attitudinal barriers and misconceptions about immunization in a global pandemic scenario is clear. The nearly constant media coverage of the A (H1N1) 2009 pandemic, reported with varying degrees of accuracy, and sometimes portraying dramatic scenarios caused some to question whether unnecessary alarm and public panic resulted. We suggest that international or national health authorities have a clear speech over looked media and to own these institutions, which will air fair and real time information about the disease.