The collection of comprehensive data is crucial for monitoring antimicrobial resistance (AMR) patterns and implementing targeted interventions. However, there is a lack of data regarding the AMR status of pathogens in Libya, hinders the efforts to address the problem effectively. This retrospective study evaluates the prevalence of Escherichia coli AMR isolates to gentamycin, chloramphenicol, cefoxitin (or) ceftriaxone, meropenem, moxifloxacin (Mox), amoxicillin/clavulanate (AUG) or ampicillin-sulbactam (UNZ) among the visitors to the main pathology laboratories in Gharyan city. Reports from 107 AMR tests demonstrate that E. coli was mostly isolated from Urinary Tract Infections (UTIs, 85%) followed by vaginal infections (8.4%) and wound infections (6.5%). More than half of the isolates were multi-drug resistant which is a clear indicator of the need for continuous monitoring. The overall pattern demonstrates a high resistance rate to cephalosporins, and combinations of penicillins with beta-lactamase inhibitors (UNZ or AUG), followed by high resistance rates to Mox. In contrast, E. coli isolates showed high sensitivity to meropenem followed by gentamicin and chloramphenicol. However, the isolation site seems to affect the resistance patterns, most of the wound isolates were resistant to gentamycin. In contrast, all vaginal isolates were resistant to UNZ or AUG, yet both types showed high susceptibility to meropenem. 

 Human milk contains an abundant supply and diverse array of oligosaccharides that are known to impart significant health benefits to the nursing infant including establishment and maintenance of a healthy gut microflora, immune development and protection against gastro intestinal infections. When breastfeeding is not possible or insufficient, infant formulas are commonly used as an alternative. However, limited information is available about the presence of naturally occurring oligosaccharides in these infant formulas and their likely health benefits. The present study examined the presence of naturally occurring oligosaccharides in commercial goats’ milk-based stage 1 and stage 2 infant formulas and their prebiotic and anti-infection properties. LC/MS was used to detect and quantify oligosaccharides and their prebiotic potential was assessed by their ability, at concentrations present in reconstituted ready-to-use infant formula, to promote the growth of Bifidobacterium animalis subsp. lactis BB12, B. longum BB536, Lactobacillus acidophilus 4461 and L. casei 2607 in vitro. For anti-infection properties, the ability of goat milk oligosaccharides to prevent the adhesion of Escherichia coli NCTC 10418 and a Salmonella typhimurium isolate to Caco-2 cells was investigated. The results showed the presence off ourteen quantifiable oligosaccharides instage1andstage2goats’milk-basedinfantformula.Thiswassimilartothenumberofoligosaccharidesdetectedinthefreshgoats’milk.Of these, five were structurally similar to those foundinhumanmilk.Theseoligosaccharideswereshowntosignificantlyenhancethegrowthof bifidobacteria and lactobacilli and reduce the adhesion of E. coli NCTC 10418 and S. typhimurium to Caco-2 cells. Together, these results suggest that oligosaccharides naturally present in goats’ milk-based infant formula exhibit strong prebiotic and anti-pathogen adhesion properties and may confer gut health benefits to infants.

 The tetracycline promoter (tet) has been manipulated in several systems as an inducible regulator of gene expression. In this study the effect of doxycycline on C. glabrata viability has been examined in water and according to CLSI standard method. In addition, flow cytometry analysis was performed to detect SLT2 protein activation by sub-inhibitory concentrations of doxycycline. Our findings show that 100 µg/ml of doxycycline is the maximum tolerated dosage in media; however, C. glabrata was sensitive to 40-50µg/ml in water. 50 µg/ml of doxycycline induced SLT2p phosphorylation and our results support the idea of involvement of doxycycline in alteration of C. glabrata oxidative status. These results should be considered when the tet promoter system is used in yeast molecular studies

The antifungal effect of thymoquinone, a component of black seed essential oil, has been studied on different types of fungi. Its mechanism of action as an antifungal has not been described yet. This study demonstrates the fungicidal effect of thymoquinone on different Candida species with particular emphasis on C. glabrata planktonic cells and biofilms. Since cell death was induced via the generation of oxidative stress as evidenced by the abrogation of thymoquinone toxicity in cells incubated with antioxidants, a part of thymoquinone’s mechanism of action includes a direct involvement as a pro-oxidant. This was further confirmed by measuring the generation of reactive oxygen species, glutathione level reduction and decrease in mitochondrial membrane potential. The oxidative stress caused by thymoquinone was confirmed to be the cause of death and not a result of cell death.

The Calgary Biofilm Device (CBD) has been described as a technology for the rapid and reproducible assay of biofilm susceptibilities to antibiotics. In this study a simple and inexpensive alternative to the CBD was developed from polypropylene (PP) microcentrifuge tubes and pipette tip boxes. The utility of the device was demonstrated using Candida glabrata, a yeast that can develop antimicrobial-resistant biofilm communities. Biofilms of C. glabrata were formed on the outside surface of microcentrifuge tubes and examined by quantitative analysis and scanning electron microscopy. Growth of three C. glabrata strains, including a clinical isolate, demonstrated that biofilms could be formed on the microcentrifuge tubes. After 24 h incubation the three C. glabrata strains produced biofilms that were recovered into cell suspension and quantified. The method was found to produce uniform and reproducible results with no significant differences between biofilms formed on PP tubes incubated in various compartments of the device. In addition, the difference between maximum and minimum counts for each strain was comparable to those which have been reported for the CBD device.

 Candida glabrata cells suspended in water are under hypo-osmotic stress and undergo cell death in 1–2 days, unless they are at a density of more than 105 CFU mL 1. The dying cells exhibit FITC-annexin V staining, indicative of programmed cell death (apoptosis). In a higher cell density, cells are protected and survive at least for 4 days. Filtrates from cells at high density can protect those at lower density, indicating that cells release substances, amounting to c. 5mgL 1 of cell suspension, that protect each other against hypo-osmotic stress. In a concentrated form, the released materials can support growth, indi cating that the protective material includes carbon and nitrogen sources, as well as vitamins that are required by C. glabrata for growth. We conclude that cell death from hypo-osmotic stress can be alleviated by small amounts of nutrients.