Fenticonazole nitrate (FTN) is a potent antifungal drug adopted in the treatment of vaginal candidiasis. It has inadequate aqueous solubility hence, novel ultra-deformable liposomes ‘Terpesomes’ (TPs) were developed that might prevail over FTN poor solubility besides TPs might abstain the obstacles of mucus invasion. TPs were assembled by thin-film hydration then optimized by Box Behnken design utilizing terpenes ratio (X 1 ), sodium deoxycholate amount (X 2 ), and ethanol concentration (X 3 ) as independent variable, whereas their impact was inspected for entrapment efficiency (Y 1 ), particle size (Y 2 ), and polydispersity index (Y 3 ). Design Expert ® was bestowed to select the optimal TP for more studies. The optimal TP had entrapment efficiency of 62.18 ± 1.39%, particle size of 310.00 ± 8.16 nm, polydispersity index of 0.20 ± 0.10, and zeta potential of −10.19 ± 0.2.00 mV. Elasticity results were greater in the optimal TP related to classical bilosomes. Further, ex vivo permeation illustrated tremendous permeability from the optimal TP correlated to classical bilosomes, and FTN suspension. Besides, in vivo assessment displayed significant inhibition effect in rats from FTN-TPs gel compared to FTN gel. The antifungal potency with undermost histopathological variation was detected in rats treated with FTN-TPs gel. Overall, the acquired findings verified the potency of utilizing FTN-TPs gel for treatment of vaginal candidiasis.

Introduction: Pseudomonas aeruginosa is one of the most virulent nosocomial pathogens worldwide. Quorum sensing (QS) regulates the production of pathogenic virulence factors and biofilm formation in P. aeruginosa. The four genes lasR, lasI, rhlR,and rhlI were found to regulate this QS system. In this study, we aimed to assess the correlation between these four genes and QS-dependent virulence factors and to detect the inhibitory effect of clove oil on QS. Methodology: Fifty P. aeruginosa clinical isolates were collected. Susceptibility to different antibiotics was tested. Virulence factors including biofilm formation, pyocyanin production, and twitching motility were phenotypically detected. QS genes were amplified by polymerase chain reaction (PCR), and one strain subsequently underwent sequencing. The inhibitory effect of clove oil on virulence factors was also tested. Results: A positive correlation was found between biofilm formation and the presence of lasR and rhlI genes. Twitching motility was positively correlated with the presence of lasR, lasI, and rhlI genes. On the other hand, no correlation was found between pyocyanin production and any of the studied genes. Only one isolate amplified all the tested QS gene primers, but it did not express any of the tested virulence factors phenotypically. Sequence analyses of this isolate showed that the four genes had point mutations. Conclusions: Results emphasize the importance of QS in P. aeruginosa virulence; however, QS-deficient clinical isolates occur and are still capable of causing clinical infections in humans. Also, clove oil has an obvious inhibitory effect on QS, which should be clinically exploited.

Background: Klebsiella pneumoniae is a nosocomial pathogen in outbreaks of hospital infections. It is one of the major factors for morbidity and mortality in hospitalized patients especially those infected with colistin-resistant pathogens. Many plant essential oils have antimicrobial activities and have been investigated as natural sources to combat multiple antibiotic resistances. Moreover, recent advances in phytonanotechnology have created exciting opportunities for the management of many infections. Objective: This study aims at investigating the antimicrobial and antibiofilm effect of rosemary and ginger essential oil-based nano-sized formulations on colistin resistant K. pneumonia clinical isolates. Methods: Isolation and identification of 30 K. pneumonia isolates from different human samples were done followed by antibiotic susceptibility testing and detection of biofilm gene (mrkD). Examination of the activity of the tested essential oils and their chitosan nanoparticle formulations against the selected isolates was made by determination of their MICs using broth microdilution method followed by biofilm inhibition test and quantitative real-time PCR for the expression of mrkD gene in the presence of the oils and nanoparticles formulations compared to untreated bacterial isolates. Results: Our results showed that the minimum inhibitory concentration of rosemary and ginger oils was 1250 μg/ml, that of nanostructured lipid carrier-rosemary oil and nanostructured lipid carrier-ginger oil was 625 μg/ml and rosemary oil loaded chitosan nanoparticles and ginger oil loaded chitosan nanoparticles possessed minimum inhibitory concentration of 156 μg/ml. Results also revealed complete (100%) inhibition for mrkD gene expression when compared to untreated K. pneumonia. Conclusion: Oil loaded chitosan nanoparticles showed the highest antimicrobial and antibiofilm activity.

Introduction. Human bocavirus (HBoV) is a recently discovered parvovirus; it has been shown to be a common cause of respiratory infections and gastroenteritis in children. Since its identification, HBoV has been detected worldwide in nasopharyngeal swabs, serum and stool samples particularly those obtained from young children suffering from respiratory or gastrointestinal tract infections. Aim. The aim of this work was to determine HBoV prevalence among children with acute respiratory tract infection in Egypt, to detect the most prevalent HBoV genotype and to compare PCR and ELISA as diagnostic techniques for HBoV infection. Methods. Nasopharyngeal swabs and blood samples were obtained within the first day of admission from 75 children diagnosed with acute respiratory tract infection in El-Shatby University Hospital for Children in Alexandria, Egypt from October 2018 to March 2019. Conventional PCR was used to detect HBoV DNA, ELISA was used to detect HBoV IgM antibodies and sequencing of the VP1/2 genes was used for genotyping. Results. Seven (9.3%) of the 75 nasopharyngeal swabs obtained from patients with acute respiratory tract infection were positive for HBoV by PCR, while 5 (6.7 %) of the 75 serum samples were positive for HBoV IgM antibodies using ELISA. The correlation between PCR and ELISA results showed a highly significant association between PCR and ELISA techniques (X 2=52.041, P<0.01) and a highly significant agreement between the two methods (Kappa=81.9 %, P<0.01). Phylogenetic analysis showed that all positive samples were related to the HBoV-1 genotype. Conclusion. Human bocavirus was detected at 9.3 % prevalence in nasopharyngeal swabs obtained from children with acute respiratory tract infection. The HBoV-1 genotype was the only genotype detected, suggesting that a single genetic lineage of HBoV is circulating in Egypt. PCR and ELISA are two reliable methods for detection and diagnosis of HBoV.

Introduction Entamoeba histolytica-caused amoebiasis is a major cause of mortality worldwide. E. histolytica is morphologically indistinguishable from nonpathogenic species like E. dispar, E. moshkovskii, and E. hartmanni. Polymerase chain reaction (PCR) is the approved method by World Health Organization for diagnosis and differentiation of amoebiasis. This study aims to molecularly differentiate the four Entamoeba spp. using conventional PCR and correlate their prevalence with the patients' sociodemographic data. Methods We collected fecal samples of 175 patients with gastrointestinal diseases at Damanhour General Hospital (El-Behira, Egypt). All microscopically positive samples were subjected to conventional PCR. ResultsThe overall prevalence of Entamoeba infection was 65.7% (115/175). The differentiation by PCR was successfully attained in 102 samples. The species distribution was as follows: E. histolytica (14.7%), E. dispar (61.8%), E. moshkovskii (11.8%); besides, 11.8% of samples revealed mixed infection. Of note, the infection rate was higher in men, patients from rural areas and patients who did not have sanitation facilities for sewage disposal. Conclusion This study demonstrates a high prevalence of infections caused by the nonpathogenic Entamoeba spp. E. dispar, E. moshkovskii, and E. hartmanni along with the pathogenic E. histolytica. Hence, we recommend PCR assay as an accurate, rapid, and effective diagnostic method for the detection and differentiation of the four morphologically indistinguishable Entamoeba spp. in both routine diagnosis of amoebiasis and epidemiological surveys.

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Klebsiella pneumoniae is a highly drug-resistant human pathogen responsible for a variety of serious infections. Integrons, mobile genetic elements capable of integrating antibiotic resistance genes, and the capsule are important virulence factors that increase bacteria resistance to phagocytosis and antimicrobial agents. Molecular typing is an effective tool for identifying the likely etiology of infection. This study aimed to investigate the presence of the rmpA, wcaG, intI1, intI2, and intI3 virulence genes in clinical Klebsiella pneumoniae isolates, and explore their molecular genotypes by using ERIC-PCR. Fifty Klebsiella pneumoniae strains were isolated from various specimens. Antimicrobial resistance was evaluated by using the disc diffusion method. Five genes were amplified by conventional PCR. Genotyping was performed molecularly by using ERIC-PCR. Forty-seven isolates were multi-drug resistant. In all, 18%, 36%, and 98% of the 50 K. pneumoniae isolates were positive for rmpA, wcaG, and intI1 genes, respectively; however, all isolates were negative for intI2 and intI3 genes. Dendogram analysis of the ERIC-PCR results showed 49 distinct patterns, arranged in five clusters. Our study demonstrates high levels of antibiotic resistance and virulence among clinical isolates of K. pneumoniae. Such resistance reflects a growing problem for public health. Further, the presence of integrons increases the horizontal spread of antibiotic resistance and virulence genes among bacterial isolates. The ERIC-PCR technique is an effective method for molecular typing and epidemiological studies of hospital-acquired infections.

The emergence of multidrug-resistant Acinetobacter baumannii has partly increased treatment failure and patient mortality. Class D β-lactamases is an important mechanism of resistance to beta-lactam antibiotics in this species. This study aimed to investigate the relationship between the presence oxacillinase gene and genetic fingerprints of A. baumannii isolates from the intensive care unit of an Egyptian tertiary care hospital. One hundred and twenty A. baumannii clinical isolates were collected. Multiplex PCR was performed to detect genes encoding oxacillinases (OXA-23, OXA-24, OXA-51, OXA-58 and OXA-143). Molecular typing of all collected isolates was performed using random amplified polymorphic DNA (RAPD)-PCR assay. Out of 120 examined isolates, 92, 88 and 84% were resistant to ertapenem, imipenem and meropenem, respectively. The species-specific, commonly present OXA-51 gene was found in all isolates while OXA-23 showed a high prevalence of 88% of isolates. OXA-24 and OXA-143 genes were detected in 3% and 1% of isolates, respectively. No OXA-58 gene was detected. Five clusters consisting of 19 genotypes were detected using RAPD-PCR. Genotype A was the most prevalent, it was observed in 62% of the isolates followed by genotype B (12%). These results revealed that genotypes A and B are common in the hospital. Results also demonstrate that RAPD-PCR is a rapid and reliable method for studying the clonal similarity among A. baumannii isolated from different clinical specimens.