Emergence of Candida auris in intensive care units in Algeria.

BACKGROUND: Currently, Candida auris is among the most serious emerging pathogens that can be associated with nosocomial infections and outbreaks in intensive care units. Clinicians must be able to identify and manage it quickly. OBJECTIVE: Here, we report for the first time in Algeria seven cases of C. auris infection or colonisation. METHODS AND RESULTS: The strains were isolated from clinical sites including bronchial aspirates (n = 4), wound swabs (n = 1), urine sample (n = 1) and peritoneal fluid (n = 1), in patients admitted to the intensive care unit. Candida auris was identified both by MALDI-TOF and by sequencing the ITS region and the D1/D2 domain. Antifungal susceptibility testing was performed using the E-test method. Non-wildtype susceptibility was observed for five strains against fluconazole, itraconazole, voriconazole and caspofungin. Genotyping showed the presence of four clades (I-IV) in one hospital. CONCLUSIONS: Appropriate antifungal treatments with rapid and accurate microbial identification are the cornerstone for the management and control of C. auris infections.

Real-Time PCR Assay for Rapid and Simultaneous Detection of vanA and vanB Genes in Clinical Strains.

Here, we develop a robust and sensitive real-time PCR assay which allows the simultaneous detection of vanA and vanB genes using common primers. The system was designed using the Primer3 online software. The specificity of primers and probes was first checked by in silico PCR and by BlastN analysis. The genomic DNA of 255 bacterial isolates, including Enterococcus spp., Gram-negative, and Gram-positive strains, as well as a collection of 50 stool and 50 rectal swab samples, were tested to evaluate the specificity of the new real-time PCR (RT-PCR) system. The results of the designed RT-PCR were 100% specific and 100% positive on tested vancomycin resistant isolates harboring either the vanA or vanB gene. RT-PCR assays were negative for all other bacterial species tested including vancomycin-sensitive Enterococci and Enterococcus strains harboring vanC genes. The limit of detection of vanA and vanB genes by RT-PCR assay was 47 CFU/mL and 32 CFU/mL, respectively. The rapid and accurate detection of vancomycin-resistant Enterococci is the cornerstone for minimizing the risk of nosocomial transmissions and outbreaks. We believe that this assay will strengthen routine diagnostics and surveillance programs.

High frequency and diversity of Vancomycin-resistant Enterococci (VRE) in Algerian healthcare settings.

The spread of vancomycin-resistant Enterococci (VRE) in Algerian hospital settings is poorly reported. Since the first report in 2006, few data have been available on the molecular mechanism of this resistance across the country. In this study, we investigate the frequency and antibiotic resistance mechanisms of Enterococci strains isolated from hospitalised patients in the Tlemcen university hospital. 191 Enterococcus spp. strains were collected from various clinical samples and were identified using MALDI-TOF-MS. The presence of van genes was investigated by standard PCR and sequencing. Results revealed that E. faecium and E. faecalis strains are the main pathogens identified in the study. Antibiotic susceptibility testing revealed that the resistance rate was high for the majority of antibiotic classes, including glycopeptides, and only linezolid was effective on all strains. Molecular analysis revealed that 52.2% of strains from intensive care unit (ICU) were positive for the vanA gene, including 44.44% E. faecium, 5.55% E. faecalis and 2.22% E. avium. 25.5% of these isolates co-harboured both the vanA and vanC genes, including E. gallinarum (n = 16) and E. faecium (n = 6). In surgical wards (SW) 29.70% of strains harboured the van genes, including 4.90% of E. faecalis harbouring the vanB gene, and of the rest of strains, (24.80%) harboured the vanC genes. Indeed, 9.90% E. gallinarum and 4.90% E. faecalis were positive for vanC1 and 9.90% of E. casseliflavus were positive for the vanC2/C3 gene. The glycopeptide resistance rate was higher among strains from the ICU and was mainly composed by E. faecium strains compared with surgical wards where resistant E. faecalis strains were predominant.

Prevalence and Antimicrobial Resistance of Paeniclostridium sordellii in Hospital Settings.

(1) Background: The purpose of this study was to determine the prevalence of clostridia strains in a hospital environment in Algeria and to evaluate their antimicrobial susceptibility to antibiotics and biocides. (2) Methods: Five hundred surface samples were collected from surfaces in the intensive care unit and surgical wards in the University Hospital of Tlemcen, Algeria. Bacterial identification was carried out using MALDI-TOF-MS, and then the minimum inhibitory concentrations (MICs) of various antimicrobial agents were determined by the E-test method. P. sordellii toxins were searched by enzymatic and PCR assays. Seven products intended for daily disinfection in the hospitals were tested against Clostridium spp. spore collections. (3) Results: Among 100 isolates, 90 P. sordellii were identified, and all strains were devoid of lethal and hemorrhagic toxin genes. Beta-lactam, linezolid, vancomycin, tigecycline, rifampicin, and chloramphenicol all proved effective against isolated strains. Among all strains tested, the spores of P. sordellii exhibited remarkable resistance to the tested biocides compared to other Clostridium species. The (chlorine-based 0.6%, 30 min), (glutaraldehyde solution 2.5%, 30 min), and (hydrogen peroxide/peracetic acid 3%, 15 min) products achieved the required reduction in spores. (4) Conclusions: Our hospital's current cleaning and disinfection methods need to be optimized to effectively remove spores from caregivers' hands, equipment, and surfaces.

miRNA-23b as a biomarker of culture-positive neonatal sepsis.

BACKGROUND: Neonatal sepsis remains an important cause of morbidity and mortality. The ability to quickly and accurately diagnose neonatal sepsis based on clinical assessments and laboratory blood tests remains difficult, where haemoculture is the gold standard for detecting bacterial sepsis in blood culture. It is also very difficult to study because neonatal samples are lacking. METHODS: Forty-eight newborns suspected of sepsis admitted to the Neonatology Department of the Mother-Child Specialized Hospital of Tlemcen. From each newborn, a minimum of 1-2 ml of blood was drawn by standard sterile procedures for blood culture. The miRNA-23b level in haemoculture was evaluated by RT-qPCR. RESULTS: miR-23b levels increased in premature and full-term newborns in early onset sepsis (p < 0.001 and p < 0.005 respectively), but lowered in late onset sepsis in full-term neonates (p < 0.05) compared to the respective negative controls. miR-23b levels also increased in late sepsis in the negative versus early sepsis negative controls (p < 0.05). miR-23b levels significantly lowered in the newborns who died from both sepsis types (p < 0.0001 and p < 0.05 respectively). In early sepsis, miR-23b and death strongly and negatively correlated (correlation coefficient = - 0.96, p = 0.0019). In late sepsis, miRNA-23b and number of survivors (correlation coefficient = 0.70, p = 0.506) positively correlated. CONCLUSIONS: Lowering miR-23b levels is an important factor that favours sepsis development, which would confirm their vital protective role, and strongly suggest that they act as a good marker in molecular diagnosis and patient monitoring.

Biocide tolerance and antibiotic resistance of Enterobacter spp. isolated from an Algerian hospital environment.

OBJECTIVES: In this study, 77 Enterobacter spp. isolates from a collection of 175 Gram-negative bacilli isolated from Tlemcen University Hospital Center (North-West of Algeria) were tested for antibiotic resistance, biocide tolerance and genetic determinants of antimicrobial resistance. METHODS: The isolates were identified by 16S rDNA gene sequencing. Biocide tolerance was determined by broth microdilution, and antibiotic resistance was determined by disk diffusion. Genetic determinants of resistance were studied by PCR amplification using suitable primers. RESULTS: The most common Enterobacter species was Enterobacter cloacae (58.4%), followed by Enterobacter hormaechei (24.7%). The most common antibiotic resistance was to ticarcillin either alone or in combination with clavulanic acid (70.1%), followed by cefepime (68.8%), cefotaxime (63.6%), ceftazidime (54.5%) and gentamicin (54.5%). Tobramycin was active against 87.0% of the isolates. Levels of biocide tolerance were high for hexachlorophene and to a lesser extent for benzalkonium chloride. The extended-spectrum ß-lactamase genes blaTEM and blaCTX-M were detected in 44.2% and 36.4% of isolates, respectively. Other antimicrobial resistance genes (ARGs) frequently detected were aac(6')-Ib (57.1%) and sul2 (50.6%). Multidrug-resistant isolates carrying several ARGs were common. Significant positive correlations were detected for efflux pump genes with ARGs and also between ARGs. CONCLUSION: The results of this study reveal thatEnterobacter spp. isolates from hospital settings are both resistant to clinically-used antibiotics and tolerant to biocides. Biocide tolerance could be an advantage for antibiotic-resistant strains in hospitals.