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.

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.

Investigations of carbapenem resistant and hypervirulent Klebsiella pneumoniae.

Hypervirulent Klebsiella pneumoniae is an emerging pathogen that has gained attention due to its increased ability to cause infections even in healthy individuals. The aim of this study is to investigate virulence factors in K. pneumoniae strains isolated from clinical specimens and their association with carbapenem resistance. The study was conducted on 260 isolates identified between 2018 and 2023 at the Mohammed V Military Teaching Hospital in Rabat, Morocco. The isolates were categorized based on their susceptibility to antibiotics. The hypermucoviscosity was determined by a string test, while the presence of capsular serotypes and virulence genes were identified by PCR. Among our strains, 6.2% (n = 16) exhibited hypervirulent characteristics, 56% were resistant to carbapenem. Notably, 5.7% (n = 6) of carbapenem-resistant isolates expressed the hypermucoviscous phenotype, while 1.5% (n = 2) of carbapenem-susceptible K. pneumoniae isolates exhibited the same trait. In our study, we found that a total of 10 isolates (3.8%) had virulent capsular serotypes, with K2 being the most prevalent 40% (n = 4) and K20 in 30% (n = 3). Furthermore, we detected the presence of the Aerobactin gene in 1.5% (n = 4) of the isolates examined. Based on our findings, it appears that there was no correlation between the presence of virulence factors and carbapenem resistance. In conclusion, identifying hypervirulent K. pneumoniae in clinical specimens and assessing their antibiotic resistance profiles are crucial to ensure effective therapy and to prevent outbreaks.

The occurrence of a fatal tuberculous pancreatic abscess simulating a pancreatic tumor in an immunocompromised patient.

Introduction: Tuberculosis is currently undergoing a worrying recovery in Morocco. It is becoming a tropical disease again and can take deceptive clinical forms and involve unusual localizations. We report a rare case of pancreatic abscess due to Mycobacterium tuberculosis in an immunocompromised patient. Case report: The patient was 48 years old and was diagnosed with HIV infection 16 months previously during a systematic check-up. He had no notable pathological history, no notion of tuberculosis contagion and no signs of tuberculosis impregnation, and was admitted for the management of epigastric pain associated with an altered general condition. Abdominal CT scan showed a bulbar perforation and multiple deep necrotic adenopathies of infectious or tumoral origin. Direct examination of the pus with Ziehl Neelsen stain was positive (1-10 BAAR/field). Molecular study using the GeneXpert MTB/RIF technique revealed M. tuberculosis complex without rifampicin resistance. The patient was put on antibacillary treatment based on isoniazid, rifampicin, ethambutol and pyrazinamide. The patient died of septic shock with multiple organ failure. Conclusions: The diagnosis of a tuberculous pancreatic abscess may be overlooked because of its rarity and its clinical state simulating a pancreatic tumor, so it should be considered especially in endemic countries like ours.

A very rare case of bacteraemia in a 4-year-old girl with osteopetrosis with probable Leuconostoc lactis infection.

Leuconostoc lactis (LLac) is a Gram-positive coccus of the family Leuconostocaceae . It can be found in a variety of vegetables and dairy products. LLac is an opportunistic pathogen with intrinsic resistance to vancomycin and teicoplanin. In this case report, we discuss a rare case of LLac-associated bacteraemia in a patient with osteopetrosis. A 4-year-old girl was admitted to the paediatric emergency department with acute fever without other signs. Blood culture revealed an infection with LLac. Using the streptococcus antibiogram, the isolate was resistant to vancomycin, teicoplanin, rifampicin and sulfamethoxazole-trimethoprim but sensitive to ß-lactams, gentamicin, streptomycin, azithromycin, clarithromycin, lincomycin, clindamycin and erythromycin. The patient was treated with intravenous ceftriaxone and gentamicin, and subsequently with oral amoxicillin. After a favourable course, she was discharged from the hospital on the 10th day. The modes of transmission and physiopathology of LLac remain unknown. Factors associated with this infection include compromised immunity, previous antibiotic therapy especially with vancomycin, and application of a central venous catheter. In our patient, the risk factors for infection were pancytopenia and multiple transfusions used to treat bone marrow failure. The source of the bacteraemia could have been the cutaneous route, but it could also have been digestive due to the reservoir of the bacteria. LLac is known as an opportunistic bacterium. Further studies on its pathogenesis and other risk factors are needed to understand the true prevalence of this potentially fatal bacterium in compromised individuals, such as the case of our patient.

Osteoarticular tuberculosis of the ankle, a rare localization: a case report.

Tuberculosis poses a considerable public health problem in countries where the disease is endemic. Osteoarticular tuberculosis represents 3-5 % of all tuberculosis cases and 10-15 % of extra-pulmonary tuberculosis cases. Involvement of the foot and ankle is rarer. We report a case of osteoarticular tuberculosis of the ankle in a 71-year-old patient with type 2 diabetes and hypertension who presented to the trauma department of the Mohammed V Military Hospital with a painful swelling of the ankle. Standard X-rays and computed tomography scans of the ankle showed inflammatory involvement of the bone and joints. Antitubercular therapy was instituted. Given the context of endemicity, any atypical presentation of lingering bone lesions should raise the suspicion of an osteoarticular tuberculosis in order to ensure early therapeutic management.

Prediction of the bacterial shape in urinary tract infections with the Sysmex UF-1000i analyser: technical note.

Background: The aim of our study was to explore the utility of the Sysmex UF-1000i analyzer as a rapid screening tool for urinary tract infections (UTI) and its ability to predict bacterial shape in order to help physicians choose the appropriate empiric treatment. Materials and methods: This is a retrospective study, including 1023 urine cytobacteriological examinations. Urines were processed according to the recommendations of the medical microbiology reference system (REMIC). Using the Sysmex Uf-1000i analyzer, the authors evaluated bacteria forward scatter (B_FSC) and fluorescent light scatter (B_FLH) in a preliminary discrimination step for UTI caused by bacilli or cocci bacteria. Results: The authors got 1023 positive samples. Comparing baccili and cocci bacteria, the authors observed a statistically significant difference for B_FSC but not for B_FLH. The values of B_FLH are very close for the four categories of microorganisms compared (bacilli, cocci, bacilli-cocci association, and yeasts). For these same categories, tests show different values for the B_FSC. A separate analysis of the B_FSC values for bacilli shows that their distribution is relatively homogeneous and exhibits a peak between 20 and 30 ch. Conclusion: Dimensional parameters of bacteria generated by UF-1000i could be a rapid and useful tool for predicting the bacterial shape causing UTI.

Risk factors for acquiring Acinetobacter baumannii infection in the intensive care unit: experience from a Moroccan hospital.

Introduction: Acinetobacter species are non-fermenting and ubiquitous Gram-negative coccobacilli, which in recent years have become the leading cause of healthcare-associated infections worldwide. Our objective here was to study the epidemiology and risk factors associated with Acinetobacter baumannii infections in the intensive care unit (ICU). Methods: This retrospective case-control study was conducted collaboratively between the Medical Bacteriology Department and the two ICUs of the Military Hospital of Instruction Mohammed V-Rabat over a 3 month period. Results: We included 180 patients, of whom 60 had A. baumannii infection. We observed a male predominance in both matched groups, with a sex ratio of 1.6. The median age was 67 years [interquartile range (IQR) 59.5-77]. The median length of stay in the ICU before infection was 8.5 days (IQR 5-14). Multivariate logistic regression analysis identified the risk factors statistically associated with A. baumannii infection at the ICU level as follows: duration of invasive procedures >7 days [odds ratio (OR)=1.02], parenteral nutrition (OR=3.514), mechanical ventilation (OR=3.024), imipenem (OR=18.72), colistin (OR=5.645), probabilistic antibiotic therapy >4 days (OR=9.063) and neoplastic pathology (OR=5.727). Conclusion: Based on our results, it can be inferred that shortening the duration of stay in the resuscitation setting, implementing rational use of medical devices and optimizing antibiotic therapy could decrease the incidence of these infections.

Urinary tract infections older adults at Mohammed VI University Hospital of Oujda: case series.

Due to its incidence, clinical polymorphism and severity, urinary tract infection is an important problem in elderly. The objectives of the authors' work were to establish the bacteriological profile of urinary tract infection and/or colonization in the elderly and then to study drug resistance of bacterial strains isolated. Materials and methods: This is a 36 months retrospective study from 22 March 2016 to 11 May 2019. The study included urinary specimens of persons aged 65 years or over, hospitalized or consulting at the authors' hospital. Urines were processed according to the recommendations of the medical microbiology reference system and European Committee on Antimicrobial Susceptibility Testing. Results: The authors collected 6552 requests for cytobacteriological examination of urine. Most of the specimens was collected in the middle stream (n=5503; 84%). Cultures were sterile in 49.77% of cases. Positive in 50.22% of cases. Among positive samples we had 53.41% polymorphic cultures, 32.75% urinary tract infection, and 13.82% urinary tract colonization. Gender distribution showed a sex ratio at 0.62. Gram-negative bacilli, with Escherichia coli as the main species, dominated the isolated bacteria. Resistance rates of E. coli strains that we isolated were 70% for amoxicillin, 36.31% for amoxicillin-clavulanate and 25% for ciprofloxacin. A high resistance rate was seen for third generation cephalosporins. Least resistance recorded to nitrofurantoin. Conclusion: ITU in the elderly is diverse and significantly different from that of younger patients, through its high contamination rate, difficulty in acquiring clinical information, high rate of asymptomatic bacteriuria, and high proportion of multidrug resistant bacteria.

Coding-Complete Genome Sequences of an Omicron Subvariant (BA.5.2.20) of SARS-CoV-2.

Here, we present the complete coding sequences of two severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains that were recovered from a nasopharyngeal swab from a female patient and the second viral passage in cell culture. After testing, both strains were identified as BA.5.2.20, a subvariant of Omicron.

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.

Pre-analytical non-conformities of cytobacteriological examination of urine in the microbiology laboratory of Oujda's Mohammed VI University Hospital.

INTRODUCTION: The pre-analytical step of cytobacteriological examination of urine (CBEU) is one of the most critical in microbiology. AIM: To analyze quantitatively and qualitatively the pre-analytical non-conformities related to the CBEU in order to propose reliable corrective measures. METHOD: This was a 76-month retrospective study from March 2016 to June 2022. The study included all CBEU referred to our laboratory. The conformity of the requests was evaluated according to the requirements of the medical microbiology standard (REMIC). It concerned the CBEU request, the urine sample and its packaging. RESULT: We collected 66631 CBEU requests. The urine was not conform in 1646 (2.47%) cases. The majority of non-conformities came from the emergency department (n= 653; 39.67%). The predominant non-conformities were (i) deteriorated sample (53.53%; n=878), (ii) delayed transport (28.55%; n=469) and (iii) damaged equipment (4.62%; n= 76). CONCLUSION: In our study, pre-analytical non-conformities of CBEU were frequent and affected all steps of the pre-analytical process. They had a direct clinical and economic impact on the patient. Continuous improvement of the pre-analytical phase of the CBEU is necessary in our institution.

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.

Bacteriological aspects of bacteremia in the intensive care unit of the Mohammed V Military Hospital: 10 months prospective study.

Introduction: Bacteremia is responsible for high rates of morbidity and mortality. The increasing prevalence of multidrug-resistant (MDR) bacteria in intensive care units (ICU) is a growing concern. Hence, prior knowledge of bacterial epidemiology and resistance phenotypes is required to optimize these infections' management. The objective of this study was to determine the epidemiological profile of bacteremia in ICU settings, as well as the place occupied by MDR bacteria in these infections. Methods: It is a prospective study carried out over 10 months on episodes of bacteremia in the ICU of Mohammed V Military Teaching Hospital (Rabat, Morocco). Microorganism growth was detected using fluorescent technology, species identification was based on morphological and biochemical characteristics. Antimicrobial susceptibility testing was performed following the recommendations of the Antibiogram Committee of the French Society of Microbiology (CA-SFM) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Results: Among 504 hospitalized patients, sixty-one (12.1%) presented at least one episode of bacteremia. Forty patients (65.6% of bacteremic patients) presented at least one episode of bacteremia due to MDR bacteria. Male gender, cardiovascular diseases, diabetes and previous hospitalization were significant risk factors for the acquisition of MDR bacteremia. Isolated bacteria were mainly Gram-negative bacilli (GNB) (n = 62; 68.9%) dominated by Acinetobacter baumannii (n = 19; 21.1%) and Klebsiella pneumoniae (n = 16; 17.8%). MDR bacteria were represented by multi-resistant Acinetobacter baumannii (n = 19; 44.2%), extended-spectrum beta-lactamases-producing Enterobacterales (n = 9; 20.9%) and carbapenem-resistant Enterobacterales (n = 7; 16.3%). Carbapenems (n = 40; 65.6%), Aminoglycosides (n = 32; 52.5%) and Polypeptides (n = 24; 39.3%) were the most used antimicrobials. Mortality rates were 66.6% (n = 40) and 85% (n = 43) in patients with non MDR bacteremia and MDR bacteremia respectively. Conclusion: Limiting the spread of MDR bacteria and improving the management of bacteremic patients require continuous monitoring of bacteremia as well as adapting the therapeutic and preventive strategy.

Characterization of imipenem-resistant Acinetobacter baumannii and Pseudomonas aeruginosa clinical isolates in a Moroccan hospital.

Acinetobacter baumannii and Pseudomonas aeruginosa are two pathogens with an important power of adaptation to antibiotics thus, both pose a real public health problem. Our study investigated epidemiological characteristics, antibiotic sensitivity profile and resistance genes of imipenem resistant A. baumannii and P. aeruginosa. This was a retrospective study carried out in the bacteriology laboratory of Mohammed V military training hospital, spanning from January 2018 to April 2021. Antibiotic susceptibility was studied by Mueller Hilton agar diffusion method with OXOID® type antibiotic discs and interpreted according to the recommendations of EUCAST 2021. Carbapenemase detection was performed by CarbaNP-test®. The molecular study was performed using conventional PCR. During the study period, we collected 1,072 imipenem-resistant isolates namely, 820 A. baumannii and 252 P. aeruginosa. The molecular study showed that out of 108 A. baumannii isolates 102 carried the bla OXA-51 and 100 isolates carried the bla OXA-23 gene. The coexistence of bla OXA-23 and bla NDM genes was detected in only 4 isolates. Altogether 50% of P. aeruginosa strains carried bla VIM-2. All investigated A. baumannii and P. aeruginosa strains were colistin susceptible in this study. Multiresistant bacterial infections are associated with longer hospitalization, higher hospital costs and higher mortality rates. Therefore, a collective action including the different actors of the healthcare system is necessary.

Coding-Complete Sequence of a SARS-CoV-2 Strain from an Omicron (B.1.1.529+BA.1) Variant Detected in Morocco.

Here, we describe the coding-complete sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strain HM36, identified as a strain of concern of B.1.1.529+BA (Omicron).

Shewanella putrefaciens: a rare cause of purulent otorrhoea.

Shewanella putrefaciens is a Gram-negative, non-fermenting, motile and oxidase-positive bacillus. Its incrimination in human pathology is very rare, although there has been a resurgence in Shewanella infections in recent years. We report the first case in Morocco of a purulent otorrhoea caused by S. putrefaciens , resistant to conventional treatment, occurring in a 25-year-old female, afebrile, without deterioration of the general state and possibly acquired during sea bathing. We also describe the bacteriological characteristics of and antibiotic susceptibility results for the isolate.

Leclercia adecarboxylata invasive infection in a patient with Hirschsprung disease: A case report.

INTRODUCTION: Leclercia adecarboxylata is a ubiquitous aerobic, motile, gram-negative bacilli. The human gastro-intestinal tract is known to harbor this rarely opportunistic microorganism. We describe a rare case of invasive infection with a gastrointestinal starting point due to L. adecarboxylata in a patient with Hirschsprung disease. CASE REPORT: It is about a newborn female who was admitted on the 3rd day of life to the neonatal intensive care unit for intestinal obstruction. On the 9th day of life, while managing the neonatal obstruction, the patient developed febrile peaks. Cytobacteriological examination of cerebrospinal fluid, blood cultures and culture of umbilical vein catheter allowed the exclusive isolation of Leclercia adecarboxylata. It was producing extended spectrum beta-lactamase and was treated with intravenous imipenem. After favourable evolution, the patient was transferred to the pediatric surgery department. There, she was diagnosed with Hirschsprung disease. DISCUSSION: Knowledge of the route of transmission of L. adecarboxylata is limited and the possible source of the infection is unclear. However, the authors describe three hypotheses of contamination of our propositus. In our patient, one or more of these routes of contamination would be possible. Indeed, bacteremia could occur as a result of a bacterial translocation across the mucosal barrier of the colon altered by Hirschsprung disease, antibiotic use and feeding practices. CONCLUSION: Infection with L. adecarboxylata revealed a wide range of infection. It has only recently been acknowledged as an emerging pathogen. Further studies of the pathogenesis and risk factors are required.

The novel biphasic medium for transport, culture and conservation at an ambient temperature of Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae.

Bacterial meningitis remains a very important disease worldwide, and the major causative pathogens were Neisseria meningitidis (N. meningitidis), Streptococcus pneumoniae (S. pneumoniae) and Haemophilus influenzae (H. influenzae). In our context, the technical difficulties encountered in the routine practice were associated with the fragility of these bacteria, the high rates of negative culture and the demanding transport conditions. That's why the need to look for a solution to its technical problems and to propose a new proper solution with the local situation. The aim of this study was to develop, perform and evaluate a novel biphasic medium used for the transport, culture and conservation at an ambient temperature of N. meningitidis, S. pneumoniae and H. influenzae. The results showed that this biphasic medium provided more, novels and easy nutriments through the addition of liquid phase and solid phase medium and it was found to be conducive to the growth and conservation of N. meningitidis, S. pneumoniae and H. influenzae at an ambient temperature of a minimum of 40 days. And the ingredients used in the medium are readily available at a low cost as well as the components prepared in large quantities, they could be stored at + 4 ± 1 °C for 2 years without significantly altering their growth and conservation supporting their potential. The survival and recovery for the fastidious bacteria on the biphasic medium and the other media used for comparison in this study were significantly different (P < 0.05). In addition, the Sensitivity, Specificity, Positive and Negative Predictive Value of biphasic medium showed highest among the three bacteria at least 40 days of storage at room temperature in this study. In conclusion, we found the biphasic medium to be low cost and suitable for previously mentioned bacteria from suspected meningitis patients, offering an optimal condition and an increase in the viability of the isolates at ambient temperature. And it was concluded that this biphasic medium could be used as a technical solution in laboratories for the management of meningitis.