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This study was divided into two parts. The first part involved the isolation, and detection of the prevalence and antimicrobial resistance profile of Aeromonas hydrophila, Pseudomonas aeruginosa, and Vibrio species from Nile tilapia fish and marine aquatic water. One hundred freshly dead Nile tilapia fish were collected from freshwater aquaculture fish farms located in Al-Abbassah district, Sharkia Governorate, and 100 samples of marine aquatic water were collected from fish farms in Port Said. The second part of the study focused on determining the in vitro inhibitory effect of dual-combination of AgNPs-H2O2 on bacterial growth and its down regulatory effect on crucial virulence factors using RT-PCR. The highest levels of A. hydrophila and P. aeruginosa were detected in 43%, and 34% of Nile tilapia fish samples, respectively. Meanwhile, the highest level of Vibrio species was found in 37% of marine water samples. Additionally, most of the isolated A. hydrophila, P. aeruginosa and Vibrio species exhibited a multi-drug resistance profile. The MIC and MBC results indicated a bactericidal effect of AgNPs-H2O2. Furthermore, a transcriptional modulation effect of AgNPs-H2O2 on the virulence-associated genes resulted in a significant down-regulation of aerA, exoU, and trh genes in A. hydrophila, P. aeruginosa, and Vibrio spp., respectively. The findings of this study suggest the effectiveness of AgNPs-H2O2 against drug resistant pathogens related to aquaculture. [ABSTRACT FROM AUTHOR]

Endophytic fungi are very prolific organisms associated with a wide range of host plants. Although they contribute to tolerance of their hosts against environmental stress, their role in response of plants to metal toxicity has not been thoroughly illuminated. Therefore, in this study, endophytic fungi were isolated from Convolvulus arvensis and their potential to bioremediate heavy metals from liquid media was assessed. Furthermore, the effect of the endophytic fungi on heavy metal tolerance, accumulation, and translocation by host plants were evaluated. The results revealed that Fusarium falciforme (MW898136) and Fusarium sp. (MW898137) possess a significant ability for bio-removal of cadmium from aqueous solutions as their tolerance indexes were >1. The bioaccumulation capacity reached 276.5 and 101.6 μg/g with bioremediation percentages of 82.4% and 76.5%, respectively. Fusarium falciforme (MW898136) filtrates, in the presence of cadmium, stimulated the growth of Triticum aestivum, and dramatically decreased cadmium bioaccumulation from 30.18 to 7.06 μg/g. The presence of F. falciforme (MW898136) in cadmium-stressed soil assisted Phaseolus vulgaris to produce more biomass, uptake higher amounts of cadmium by roots, and decreased the translocation of cadmium from roots to shoots. These results suggest that F. falciforme (MW898136) has the ability to immobilize cadmium and decrease its bioavailability in both liquid media and soil through absorption and/or accumulation. [ABSTRACT FROM AUTHOR]

Purpose: Leukocyte adhesion deficiency (LAD) represents a rare group of inherited inborn errors of immunity (IEI) characterized by bacterial infections, delayed umbilical stump separation, and autoimmunity. This single-center study aimed at describing the clinical, immunological, and molecular characterizations of 34 LAD-I Egyptian pediatric patients. Methods: Details of 34 patients' personal medical history, clinical and laboratory findings were recorded; Genetic material from 28 patients was studied. Mutational analysis was done by Sanger sequencing. Results: Omphalitis, skin and soft tissue infections with poorly healing ulcers, delayed falling of the umbilical stump, and recurrent or un-resolving pneumonia were the most common presentations, followed by chronic otitis media, enteropathy, periodontitis; and recurrent oral thrush. Persistent leukocytosis and neutrophilia were reported in all patients, as well as CD18 and CD11b deficiency. CD18 expression was < 2% in around 90% of patients. Sixteen different pathological gene variants were detected in 28 patients who underwent ITGß2 gene sequencing, of those, ten were novel and six were previously reported. Three families received a prenatal diagnosis. Patients were on antimicrobials according to culture's results whenever available, and on prophylactic Trimethoprim-Sulfamethoxazole 5 mg/kg once daily, with regular clinical follow up. Hematopoietic stem cell transplantation (HSCT) was offered for 4 patients. However due to severity of the disease and delay in diagnosis, 58% of the patients passed away in the first 2 years of life. Conclusion: This study highlights the importance of early diagnosis and distribution of ITGß2 gene mutation in Egyptian children. Further molecular studies, however, remain a challenging necessity for better disease characterization in the region. [ABSTRACT FROM AUTHOR]

Aims: This work aimed to determine the occurrence, virulence, antibiogram, carbapenem resistance genes and susceptibility to disinfectants of Pseudomonas aeruginosa isolated from animals, environment and workers in intensive farms. Methods and Results: A total of 610 samples from intensive beef cattle and sheep farms in Kafr El Sheikh Governorate, Egypt were screened for the presence of P. aeruginosa using bacteriological assays. The isolates were characterized by PCR and tested for susceptibility to antibiotics using disk diffusion method and disinfectants by quantitative suspension test. In all, 60 P. aeruginosa isolates were recovered in this study and all isolates harboured at least one of the virulence genes tested. Human P. aeruginosa isolates were highly resistant to cephalosporins, fluroquinolones, aminoglycosides, carbapenems and penicillins+β‐lactamase inhibitors than non‐human isolates. Colistin resistance was higher in non‐human than human P. aeruginosa isolates, whereas low resistance to aztreonam was observed in non‐human and human isolates. Carbapenem‐resistant P. aeruginosa (CRPA) strains were recovered from workers (56.5%), sheep (8.3%) and cattle (8.3%). All CRPA harboured at least one of the carbapenem resistance genes tested and most of them showed multidrug resistance (MDR) or extensive drug resistance (XDR) phenotypes. Glutaraldehyde 1% and hydrogen peroxide 3% eliminated P. aeruginosa completely in the absence and presence of organic matter within short contact time compared with other disinfectants. Conclusions: This study reported the occurrence of CRPA in animals and workers in intensive farms. Glutaraldehyde and hydrogen peroxide were the most effective disinfectants against P. aeruginosa. Significance and Impact of the Study: The occurrence of CRPA in intensive livestock farms is a serious challenge that threatens animal and human health and increases the risk of P. aeruginosa infection in the community. Therefore, it is vital to control the spread of CRPA by banning or restricting the use of antibiotics and applying proper cleaning and disinfection protocols in livestock farms. [ABSTRACT FROM AUTHOR]

This study targeting the characterization of new lytic phages and studying their impact on Staphylococcus aureus biofilm and growth in milk. Two lytic phages (vB_SauM_ME18 & vB_SauM_ME126) were isolated using 40 Multidrug-Resistant (MDR) S. aureus isolates. Four isolates (10%) were susceptible to phages. Phages were related to Family Myoviridae, had icosahedral heads and long contractile tails. The genome size is 20 kbp for both phages and DNA was digested with hinf I, hind III and hae III. Phages had a latent period (~15 min), rise period (~30 min) and burst size (114 PFU/infected cell for ME18 and 140 PFU/infected cell for ME126). There was a significant reduction in biofilm formation and reduction in the established biofilm biomass in phage treated S. aureus compared with controls (P < 0.05). Phages showed high activity and eliminated MDR S. aureus from ultra-heat-temperature treated milk at 25 °C. However, at 37 °C, ME126 (MOI10) eradicated S. aureus and ME18 (MOI10) reduced the CFU/ml by 87.2% compared with controls (P < 0.05). In essence, lytic phages are promising natural antimicrobials for inhibiting the growth and biofilm of MDR S. aureus in milk, thereby, biocontrol of foodborne pathogens. • Lytic S. aureus phages with short latent period and high burst size were isolated. • Phages significantly reduced biofilm formation and the established biofilm biomass. • Both phages reduced S. aureus growth in UHT whole milk after 2–6 h incubation. • Phages are promising for controlling MDR S. aureus biofilm on food-contact surfaces. • Phages are valuable antibiotics alternatives for biocontrol of foodborne pathogens. [ABSTRACT FROM AUTHOR]

Background: Caspase recruitment domain family, member 11 (CARD11) is an important protein which plays a fundamental role in the activation of NF-κβ pathway in lymphocytes. CARD11 deficiency can be inherited in either autosomal dominant or autosomal recessive forms and present with different phenotypes including combined immunodeficiency, atopic dermatitis, and other variable manifestations. The present report describes clinical phenotypes and immunological defects of two unrelated patients with missense homozygous variants in CARD11 presenting with combined immunodeficiency (CID) and atopic skin disease resembling that reported in dominant negative CARD11 deficiency. The patients underwent next generation sequencing, immunophenotyping of T and B subsets by flow cytometry, T cell stimulation, and evaluation of CARD11 expression. Results: Both patients had features suggesting CID including repeated pneumoniae with ICU admissions, chronic diarrhea, and itchy atopic skin disease. Patient-1 has homozygous missense variant in the C terminal domain (c.2839G > A, p.Glu947Lys), and patient-2 has homozygous variant in the inhibitory domain (c.1073C > G, p.Pro568Arg). Both have profound defects in Tregs with normal recent thymic emigrants, memory, and naïve CD4+ T cells. However, in response to stimulation, T cells failed to upregulate the expression of CD25. CARD11 expression by flow cytometry was decreased rather than abolished as previously described in patients with autosomal recessive CARD11 deficiency. B cells showed marked deficiency of switched memory and increase in transitional B cells. Conclusion: Missense variants causing CARD11 deficiency may affect the protein function rather than the expression and can result in a phenotype combining the atopic skin disease and the features of CID. [ABSTRACT FROM AUTHOR]

This study targeted developing a novel Zinc oxide with alpha hematite nanoflowers (NFs)-loaded bee venom (Bv) (Bv-ZnO@αFe2O3 NFs) as a bio-natural product from bees to combine both the advantages of combination magnetic properties and the antimicrobial and anti-biofilm properties on isolated coliform bacteria from the effluent of wastewater treatment plants. About 24 isolates of treated wastewater isolates were multidrug resistant (MDR). The phylogenetic grouping of Escherichia coli (E. coli) and Klebsiella pneumonia (K. pneumonia) showed that the largest group was Group A, followed by Group B2 and Group B1. Fourier transform infrared (FTIR), The X-ray diffraction (XRD), and scanning electron microscopy-energy dispersive X-ray analysis (SEM− EDX) validated the coating operation's contact with Bv onto ZnO@αFe2O3 NFs. According to high-resolution transmission electron microscopy (TEM) and selected area electron diffraction (SAED), pattern analyses for prepared nanoformulations exhibited a spherical shape of αFe2O3 (~9–15 nm), and floral needle shapes with uniform distribution of size with aggregation of ZnOαFe2O3 and Bv-ZnO@αFe2O3 NFs around (~100–200 nm). The toxicity of Bv-ZnO@αFe2O3 NFs was comparable up to 125 µg mL−1, when it reached 64.79% (IC50, 107.18 µg mL−1). The antibacterial activity showed different zones of inhibition against different isolates. The biofilm inhibitory activity of NPs and NFs showed a highly significant reduction (p < 0.001) in treated biofilms with ZnO@αFe2O3 and Bv-ZnO@αFe2O3. In essence, ZnO@αFe2O3 and Bv-ZnO@αFe2O3 NFs are promising antimicrobials for inhibiting the growth and biofilm of MDR E. coli and K. pneumonia isolates, thereby, biocontrol of wastewater. [ABSTRACT FROM AUTHOR]

Purpose: Chronic granulomatous disease (CGD) is an inherited primary immunodeficiency disorder of phagocytes, characterized by recurrent fungal and bacterial infections. Our aim is to describe the different clinical presentations, non-infectious auto-inflammatory features, types and sites of infections, and to estimate the mortality among our large cohort. Methods: This is a retrospective study conducted at the Pediatric Department of Cairo University Children's Hospital in Egypt, including cases with a confirmed CGD diagnosis. Results: One hundred seventy-three confirmed CGD patients were included. AR-CGD was diagnosed in 132 patients (76.3%) including 83 patients (48%) with p47phox defect, 44 patients (25.4%) with p22phox defect, and 5 patients (2.9%) with p67phox defect. XL-CGD was diagnosed in 25 patients (14.4%). The most common recorded clinical manifestations were deep-seated abscesses and pneumonia. Gram-negative bacteria and Aspergillus were the most frequently isolated species. Regarding the outcome, 36 patients (20.8%) were lost from follow-up. Among patients with known outcome, 94/137 patients (68.6%) are living, while 43/137 patients (31.4%) died. Conclusion: AR-CGD is predominant in Egypt; CGD must always be ruled out in any patient presenting with typical or atypical mycobacterial or BCG-disease. [ABSTRACT FROM AUTHOR]

Background: Inborn errors of immunity (IEI) are a group of heterogeneous disorders with geographic and ethnic diversities. Although IEI are common in Egypt, genetic diagnosis is limited due to financial restrictions. This study aims to characterize the genetic spectrum of IEI patients in Egypt and highlights the adaptation of the molecular diagnostic methods to a resource-limited setting. Methods: Genetic material from 504 patients was studied, and proper diagnosis was achieved in 282 patients from 246 families. Mutational analysis was done by Sanger sequencing, next-generation sequencing (NGS) targeting customized genes panels, and whole-exome sequencing (WES) according to the patients' phenotypes and availability of genetic testing. Results: A total of 194 variants involving 72 different genes were detected with RAG1/2 genes being the most encountered followed by DOCK8, CYBA, LRBA, NCF1, and JAK3. Autosomal recessive (AR) inheritance was detected in 233/282 patients (82.6%), X-linked (XL) recessive inheritance in 32/282 patients (11.3%), and autosomal dominant (AD) inheritance in 18/282 patients (6.4%), reflecting the impact of consanguineous marriages on the prevalence of different modes of inheritance and the distribution of the various IEI disorders. Conclusion: The study showed that a combination of Sanger sequencing in selected patients associated with targeted NGS or WES in other patients is an effective diagnostic strategy for IEI diagnosis in countries with limited diagnostic resources. Molecular testing can be used to validate other nonexpensive laboratory techniques that help to reach definitive diagnosis and help in genetic counseling and taking proper therapeutic decisions including stem cell transplantation or gene therapy. [ABSTRACT FROM AUTHOR]

Cernunnos deficiency is a rare genetic disorder characterized by immunodeficiency, microcephaly, growth retardation, bird-like facies, sensitivity to ionizing radiation, few autoimmune manifestations, premature aging of hematopoietic stem cells at an early age, and occasional myeloproliferative disease. Herein we present five Egyptian Cernunnos patients from 3 different families. We describe the patients' clinical phenotypes, their immunological profile as well as genetic results. Sequence analysis revealed three different mutations in the NHEJ1 gene: a nonsense variant c.532C > T; p.(Arg178Ter), an intronic variant c.178-1G > A and a frameshift insertion variant c.233dup; p.(Asn78LysfsTer14). In conclusion, Cernunnos deficiency can have a wide range of clinical features. The characteristic immune profile including a decrease in recent thymic emigrants and naive T cells, markedly elevated memory T cells together with normal to high IgM, and a decrease in IgG and IgA. This immune profile is highly suggestive of Cernunnos deficiency in T-B-NK + SCID patients especially surviving for older ages. • This is the first Egyptian report of patients with mutations in NHEJ1 gene. • c.532C > T; p.(Arg178Ter), c.178-1G > A and c.233dup had been identified in NHEJ1 gene. • Cernunnos deficiency can have a wide range of clinical features. • Suspicion is raised in SCID with elevated memory and decrease naïve T cells. • Presence of BM aplasia and absence of malignancies are disease hallmarks. [ABSTRACT FROM AUTHOR]

Bacillus cereus is one of the foodborne pathogens that is incriminated in several outbreaks worldwide. The objectives of the current study were first to determine the prevalence of B. cereus -contaminated milk and dairy products retailed in Egypt. Secondly, the antimicrobial resistance profiling and virulence-associated genes of the recovered B. cereus isolates were assessed. Finally, natural and eco-friendly natural additives such as d -tryptophan and l -ascorbic acid were used for controlling the viability of Bacillus cereus spores. A total of 200 samples including market raw milk, Ultra-Heat Treated (UHT) milk, Kariesh cheese, soft white cheese (Damietta), and rice pudding, 50 of each were examined. The results of our study indicated that 31% of the examined dairy samples were contaminated with B. cereus. Surprisingly, 90% of the isolates showed multidrug-resistance against ampicillin, amoxicillin, oxacillin, and streptomycin. The molecular characterization revealed that 15 isolates harbored both hemolytic and non-hemolytic enterotoxin genes. Co-treatment of B. cereus spores-contaminated rice pudding with d -tryptophan at a concentration of 40 mM and l -ascorbic acid at a concentration of 3% revealed a significant reduction (P < 0.05) in the spore growth. Therefore, d -tryptophan and l -ascorbic acid are considered promising eco-friendly food preservatives with anti- B. cereus effects in the dairy industry. • Bacillus cereus could be isolated from Egyptian dairy products. • B. cereus isolates had multidrug resistance profiling. • B. cereus isolates had the ability to produce enterotoxins. • Ascorbic acid, and d -tryptophan had anti- Bacillus cereus effect. • The tested additives might act as promising tools against B. cereus growth. [ABSTRACT FROM AUTHOR]