Background: Biofilm formation is an essential virulence factor that creates a highly protected growth mode for Staphylococcus aureus (S. aureus) to survive in any hostile environment. Antibiotic sub-minimal inhibitory concentration (sub-MIC) may modulate the biofilm formation ability of bacterial pathogens, thereby affecting bacterial pathogenesis and infection outcomes. Intense antimicrobial therapy to treat biofilm-associated infections can control the pathogenic infection aggravation but cannot guarantee its complete eradication. Objective: This study aimed to assess the sub-MICs effect of 5 different antimicrobial classes on biofilm-forming capacity among Staphylococcus aureus clinical isolates using three different biofilm quantitation techniques. Methods: In this study, the effects of 5 different antimicrobial agents, namely, azithromycin, gentamicin, ciprofloxacin, doxycycline, and imipenem, at sub-MICs of 12.5%, 25%, and 50% were tested on 5 different clinical isolates of S. aureus. The biofilms formed in the absence and presence of different antimicrobial sub-MICs were then assessed using the following three different techniques: the crystal violet (CV) staining method, the quantitative PCR (qPCR) method, and the spread plate method (SPM). Results: Biofilm formation was significantly induced in 64% of the tested conditions using the CV technique. On the other hand, the qPCR quantifying the total bacterial count and the SPM quantifying the viable bacterial count showed significant induction only in 24% and 17.3%, respectively (Fig. 1). The difference between CV and the other techniques indicates an increase in biofilm biomass without an increase in bacterial growth. As expected, sub-MICs did not reduce the viable cell count, as shown by the SPM. The CV staining method revealed that sub-MICs of imipenem and ciprofloxacin had the highest significance rate (80%) showing an inductive effect on the biofilm development. On the other hand, doxycycline, azithromycin, and gentamicin displayed lower significance rates of 73%, 53%, and 47%, respectively. Conclusion: Exposure to sub-MIC doses of antimicrobial agents induces the biofilm-forming capacity of S. aureus via increasing the total biomass without significantly affecting the bacterial growth of viable count.

Background Many infectious and noninfectious triggers lead to inflammation of the vagina. Aim We investigated the prevalence of causative vaginitis microorganisms in 516 pregnant and nonpregnant female volunteers. Vaginal samples were examined microscopically, cultured and tested for different pathogens. Results Of the participants, 310 (60.1%) were pregnant, whereas 206 (39.9%) were nonpregnant. Using Amsel's criteria and Nugent's scores, bacterial vaginosis (BV) was diagnosed in 59.1%, and the prevalence of vulvovaginal candidiasis (VVC) was 50.2% in the population. Candida infections were significantly higher in nonpregnant females (p value ≤ 0.01), and 24% of females had mixed infections. The most common mixed infection was BV and Candida spp., detected in 21% of the cases. Conclusions Bacterial vaginosis is the most common cause of vaginitis. We observed that 24% of females experienced mixed infections, and Candida albicans was the most common fungal species causing VVC. Trichomonas vaginalis prevalence was underestimated using wet mounts.

Background: Human immunodeficiency virus type 1 (HIV-1) is characterized by high genetic diversity due to its high mutation and recombination rates. Although, there is an increasing prevalence of Circulating Recombinant Forms (CRFs) worldwide, subtype B is still recognized as the predominant subtype in the Middle East and North Africa (MENA) region. There is a limited sampling of HIV in this region due to its low prevalence. The main purpose of this study is to provide a summary of the current status of the resident HIV subtypes and their distribution among Egyptian patients. Methodology: Forty-five HIV-1 patients were included in this study. Partial pol gene covering the protease (PR) and Reverse Transcriptase (RT) was successfully amplified in 21 HIV patients using nested PCR of cDNA of the viral genomic RNA, then sequenced. The sequence data were used for viral HIV-1 subtyping by 5 online subtyping tools: NCBI viral genotyping tool, Stanford University HIV database (HIVDB) subtyping program, REGA tool, Context-Based Modeling for Expeditious Typing (COMET) tool, and Recombinant Identification Program (RIP) tool. The final subtype assignment was based on molecular phylogenetic analysis. Results: Unexpectedly, non-B subtypes are dominating, with the most common circulating one is CRF02_AG (57.1%) followed by subtype B (14.3%), subtype BG recombinant (9.5%), CRF35_ AD (9.5%), subtype A1 and CRF06_cpx (4.8% each). Conclusion: To the best of our knowledge, this is the first study to tackle HIV-1 subtyping among the group of HIV-1 patients in Egypt. CRF02_AG is the most prevalent subtype in Egypt.

Background: Antibiotics at sub-inhibitory concentrations can rewire bacterial regulatory networks, impacting virulence. Objective: The way that exposure to selected antibiotics (ciprofloxacin, amikacin, azithromycin, ceftazidime, and meropenem) below their minimum inhibitory concentration (sub-MIC) modulates the physiology of Pseudomonas aeruginosa is examined in this study using growth-phase-resolved analysis. Methods: Standard P. aeruginosa strain cultures were exposed to ¼ and ½ MIC to determine the growth kinetics under antibiotic stress. The study measured protease and pyocyanin production and the expression level of important quorum sensing and virulence genes (lasI/R, rhlI/R, pqsR/A, and phzA) at different growth phases. Results: Meropenem produced the most noticeable growth suppression at ½ MIC. Sub-MIC antibiotics did not completely stop growth, but caused distinct, dose-dependent changes. Azithromycin eliminated protease activity in all phases and had a biphasic effect on pyocyanin. Ciprofloxacin consistently inhibited both pyocyanin and protease in all phases. The effects of amikacin varied by phase and dose, while β-lactams markedly increased pyocyanin production during the log phase. In contrast to the plateau phase, when expression was often downregulated or unchanged, most quorum-sensing- and virulence-associated genes showed significant upregulation during the death phase under sub-MIC exposure. Conclusions: These findings indicate that sub-MIC antibiotics act as biochemical signal modulators, preserving stress-adapted sub-populations that, in late growth phases, activate quorum sensing and stress tolerance pathways.

Background:This study aimed at analyzing the microbiological profile of neonatal sepsis in Egypt and to determine the antibiotic susceptibility patterns of the isolated microorganisms. Methods: Two thousand and four hundred blood samples were collected from neonates showing symptoms suggestive of septicemia, inoculated into BACT/ALERT culture bottles. Positive blood culture samples were identified and tested for antimicrobial susceptibility. Results: Among the neonates included in our study, 457 (19%) neonates were positively diagnosed with sepsis. early onset neonatal sepsis (EOS) was detected in 181 (39.6%), while late-onset neonatal sepsis (LOS) in 276 (60.4%) cases. Klebsiella pneumoniae was the most commonly isolated microorganism. Staphylococcus aureus and Enterococcus spp. were the most common isolated Gram-positive bacteria. Candida spp. was more encountered in LOS.An alarming feature of the present study is the high incidence of multidrug resistant microorganisms (65%). Among Gram negative isolates (56%) of were extended spectrum beta-lactamase (ESBL) producers and (70.5%) were carbapenem resistant. In Gram positive bacteria, resistance to methicillin in S. aureus and coagulase negative staphylococci were detected in (50%) and (41%) of isolates respectively. Additionally, 17% of Enterococcus isolates were vancomycin resistant. Coclusion: Our bacteriological profile of neonatal sepsis showed that Gram negative bacteria represented the majority of isolates. Klebsiella pneumoniae was the predominant isolate. In our study, both EOS & LOS share a nosocomial infection profile, as high antimicrobial resistance was observed among our isolates. The susceptibility profiles of the isolates may urge for the change of the current used empirical therapies.

Background: Antimicrobial stewardship aims to stop or slow the emergence of antimicrobial-resistant strains. Following the launch of Egypt's National Action Plan on Antimicrobial Resistance, numerous hospitals took steps to adopt ASP, but no formal assessments were conducted. Objectives: The goal of this study is to determine the extent to which ASP is used, as well as stewardship understanding and implementation, in several Egyptian hospitals. Methodology: A cross-sectional study was carried out to measure stewardship knowledge and execution, from June to July 2020, in various Egyptian hospitals. The hospital description, ASP committee, ASP activities, reporting antibiotic use, antibiogram, information technology support, ASP restriction techniques, and future ASP application were all covered in the survey. Results: The study covered a variety of hospitals, the majority of which (61.8%) adopted an ASP program while others (38.2%) did not; at the time of the study, 71.4 % of the latter were planning to implement an ASP program in the future. Twenty-two institutions (48.5 %) said they routinely use facility-specific antibiograms. Antimicrobial reports were received on a regular basis by thirteen hospitals (37.1%). Fifteen hospitals (42.9%) said they were already using one or more computer applications. Resistance trends, infection rates, duration of stay, mortality rate, and reported adverse drug reactions were all monitored at the hospitals. Only 20% of the hospitals in this study had an ASP training program. Conclusion: Egyptian hospitals have a good ASP implementation; however, IT support programs, cooperation between healthcare members, monitoring outcomes, and infectious disease consultations are still needed.