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Rajiha A Ibrahim,1– 3 Nega Berhe,1 Zelalem Mekuria,3,4 Eyasu T Seyoum,3 Joan-Miquel Balada-Llasat,3,5 Tamrat Abebe,6 Solomon H Mariam,1 Estifanos Tsige,2 Surafel Fentaw Dinku,2 Shu-Hua Wang3,7 1Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia; 2Bacterial, Parasitic and Zoonotic Research Directorate, Ethiopian Public Health Institute, Addis Ababa, Ethiopia; 3Global One Health Initiative (GOHi), The Ohio State University, Columbus, OH, USA; 4Veterinary Preventive Medicine, Colleges of Veterinary Medicine, The Ohio State University, Columbus, OH, USA; 5Department of Pathology, College of Medicine, The Ohio State University, Columbus, OH, USA; 6Department of Microbiology, Immunology, and Parasitology, Schools of Medicine, College of Health Sciences, Addis Ababa University, Addis Ababa, Ethiopia; 7Division of Infectious Diseases, Department of Internal Medicine, College of Medicine, The Ohio State University, Columbus, OH, USACorrespondence: Rajiha A Ibrahim, Bacterial, Parasitic and Zoonotic Research Directorate, Ethiopian Public Health Institute, PO Box: 1242, Addis Ababa, Ethiopia, Tel +251920602640, Email rajihabubeker@gmail.com; Ibrahim.258@osu.eduBackground: Staphylococcus aureus causes a wide range of infections from mild skin and soft tissue to severe life-threatening bacteremia. The pathogenicity of S. aureus infections is related to various bacterial surface components and extracellular proteins such as toxic-shock syndrome (TSS) toxin and Panton-Valentine leukocidin (PVL). In this study we determine the antimicrobial resistance of isolated strains and their virulence genes in Ethiopia.Methods: A total of 190 archived S. aureus isolates from four Ethiopia Antimicrobial Resistance (AMR) Surveillance sites were analyzed. The identification of S. aureus was done by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF Biotyper) and antimicrobial susceptibility test (AST) was done using VITEK® 2. Multiplex PCR was used to detect mecA, mecC, pvl and spa genes and super-antigens (sea, seb, sec, seh and sej staphylococcal enterotoxins).Results: A total of 172 isolates were confirmed as S. aureus, 9 (5.23%) were methicillin-resistant S. aureus (MRSA) and 163 (94.76%) were methicillin-susceptible S. aureus (MSSA). AST showed that 152 (88.4%) isolates were resistant to penicillin; 90 (52.32%) resistant to trimethoprim-sulfamethoxazole; and 45 (26.16%) resistant to tetracycline. A total of 66 (38.37%) isolates harbored at least one staphylococcal enterotoxin gene and 31 (46.96%) isolates had more than one. The most frequent enterotoxin gene encountered was seb 28 (16.28%). The TSST-1 gene was detected in 23 (13.37%). Presence of staphylococcal enterotoxin gene showed significant association with antibiotic resistance to cefoxitin, benzylpenicillin, oxacillin, erythromycin, clindamycin, tetracycline and SXT. The pvl gene was detected in 102 (59.3%) of isolates. Isolates from patients below 15 years of age showed significantly high numbers of pvl gene (P = 0.02). Presence of sej (P = 0.011) and TSST-1 (P < 0.001) genes were associated with the presence of pvl gene.Conclusion: In this study, isolates were highly resistant to oral antibiotics and the pvl, seb, sea and TSST-1 genes were prevalent.Keywords: MRSA, enterotoxin genes, Panton-Valentine leucocidin, PVL, Ethiopia

Mohamad S Yasin, Ghayda’a M Al-Labadi, Mohammad Ali Alshrouf, Bayan A AlRaie, Raneem A Ibrahim, Lana A AlRaie Department of Special Surgery, Division of Orthopedics, The University of Jordan, Amman, 11942, JordanCorrespondence: Lana A AlRaie, Department of Special Surgery, Division of Orthopedics, The University of Jordan, Amman, 11942, Jordan, Tel +962776129370, Email Lanaalraie@yahoo.comBackground: Flatfoot is commonly seen in the community and is a common cause of concern for parents because it could become symptomatic and lead to decreased quality of life. One of the most used management approaches is foot orthoses, although no clear evidence supports their use. We aimed to study flatfoot symptoms’ prevalence, effect on activities of daily living, and the use and effectiveness of orthoses.Methodology: This was a cross‐sectional study that included five countries from the Middle East and North Africa region (Jordan, Palestine, Syria, Egypt, and Iraq). Data were collected using an online questionnaire directed toward parents of children aged 0 to 16 from September to December 2020. The demographic factors were expressed as frequencies (percentages) using standard descriptive statistical parameters, and Pearson’s chi-square test was used to examine the relationship between study factors.Results: 1256 participants were recruited using this online survey. The majority (29.6%) of children were diagnosed in the age group of 0 to 2. The abnormal appearance of the foot was the most common (78.7%) complaint. Overall, 54.2% of patients were prescribed orthoses, of which 36.8% noticed improvement in flatness and 37.6% reported relief of symptoms.Conclusion: This study demonstrated that most participants have no or minimal symptoms and that there is a mismatch between participants’ expectations and the actual effectiveness of orthoses. Taking into consideration that there is no clear evidence to support the corrective effect of orthoses, we recommend that physicians prescribing them adhere more to their proper indications and spend more time and effort counseling and addressing patients’ and parents’ concerns about this developmental stage.Keywords: flatfoot, foot deformity, pediatric, foot orthoses, cross-sectional

Abdelazeem M Algammal,1 Khyreyah J Alfifi,2 Mahmoud Mabrok,3 Marfat Alatawy,2 Dalia A Abdel-moneam,4 Saad Alghamdi,5 Marwa M Azab,6 Reham A Ibrahim,7 Helal F Hetta,8 Reham M El-Tarabili1 1Department of Bacteriology, Immunology, and Mycology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, 41522, Egypt; 2Biology Department, Faculty of Science, Tabuk University, Tabuk, 71421, Saudi Arabia; 3Department of Fish Diseases and Management, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, 41522, Egypt; 4Department of Aquatic Animal Medicine and Management, Faculty of Veterinary Medicine, Cairo University, Cairo, 12613, Egypt; 5Laboratory Medicine Department, Faculty of Applied Medical Sciences, Umm Al-Qura University, Makkah, 21955, Saudi Arabia; 6Department of Microbiology and Immunology, Faculty of Pharmacy, Suez Canal University, Ismailia, 41522, Egypt; 7Marine Environmental Division- National Institute of Oceanography and Fisheries (NIOF), Suez, 43511, Egypt; 8Department of Medical Microbiology and Immunology, Faculty of Medicine, Assiut University, Assiut, 71515, EgyptCorrespondence: Abdelazeem M Algammal, Department of Bacteriology, Immunology and Mycology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, 41522, Egypt, Email abdelazeem.algammal@vet.suez.edu.eg Mahmoud Mabrok, Department of Fish Diseases and Management, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, 41522, Egypt, Email dr.mahmoudmabrok@yahoo.comBackground: Bacillus cereus is a common food poisoning pathogen in humans. This study aimed to investigate the prevalence, molecular typing, antibiogram profile, pathogenicity, dissemination of virulence and antibiotic resistance genes associated with natural B. cereus infection among Mugil seheli.Methods: Consequently, 120 M. seheli (40 healthy and 80 diseased) were obtained from private fish farms in Port-said Governorate, Egypt. Afterward, samples were processed for clinical, post-mortem, and bacteriological examinations. The recovered isolates were tested for antimicrobial susceptibility, phenotypic assessment of virulence factors, pathogeneicity, and PCR-based detection of virulence and antibiotic resistance genes.Results: B. cereus was isolated from 30 (25%) examined fish; the highest prevalence was noticed in the liver (50%). The phylogenetic and sequence analyses of the gyrB gene revealed that the tested B. cereus isolate displayed a high genetic similarity with other B. cereus strains from different origins. All the recovered B. cereus isolates (n =60, 100%) exhibited β-hemolytic and lecithinase activities, while 90% (54/60) of the tested isolates were biofilm producers. Using PCR, the tested B. cereus isolates harbor nhe, hbl, cytK, pc-plc, and ces virulence genes with prevalence rates of 91.6%, 86.6%, 83.4%, 50%, and 33.4%, respectively. Moreover, 40% (24/60) of the tested B. cereus isolates were multidrug-resistant (MDR) to six antimicrobial classes and carried the bla 1, bla 2, tetA, and ermA genes. The experimentally infected fish with B. cereus showed variable mortality in direct proportion to the inoculated doses.Conclusion: As far as we know, this is the first report that emphasized the existence of MDR B. cereus in M. seheli that reflects a threat to the public health and the aquaculture sector. Newly emerging MDR B. cereus in M. seheli commonly carried virulence genes nhe, hbl, cytK, and pc-plc, as well as resistance genes bla 1, bla 2, tetA, and ermA.Keywords: MDR B. cereus, virulence traits, antibiogram, virulence genes, antimicrobial resistance genes

Rehab Mahmoud Abd El-Baky,1,2 Reham Ali Ibrahim,1 Doaa Safwat Mohamed,2 Eman Farouk Ahmed,2 Zeinab Shawky Hashem1 1Department of Microbiology and Immunology, Faculty of Pharmacy, Minia University, Minia 61519, Egypt; 2Department of Microbiology and Immunology, Faculty of Pharmacy, Deraya University, Minia 11566, EgyptCorrespondence: Rehab Mahmoud Abd El-BakyDepartment of Microbiology and Immunology, Faculty of Pharmacy, Minia University Tel +20 1092487412Email rehab.mahmoud@mu.edu.egIntroduction: Escherichia (E.) coli can cause intestinal and extra-intestinal infections which ranged from mild to life-threatening infections. The severity of infection is a product of many factors including virulence properties and antimicrobial resistance. Objectives: To determine the antibiotic resistance pattern, the distribution of virulence factors and their association with one another and with some selected resistance genes.Methods: Virulence properties were analyzed phenotypically while antimicrobial susceptibility was tested by Kirby-Bauer agar disc diffusion method. In addition, 64 E. coli isolates were tested for 6 colicin genes, fimH, hlyA, traT, csgA, crl virulence genes and bla−CTX-M-15, bla−oxa-2, and bla−oxa-10 resistance genes by polymerase chain reaction (PCR).Results: Extra-intestinal pathogenic E. coli isolated from urine and blood samples represented a battery of virulence factors and resistance genes with a great ability to produce biofilm. Also, a significant association (P< 0.05) among most of the tested colicin, virulence and resistance genes was observed. The observed associations indicate the importance and contribution of the tested factors in the establishment and the progress of infection especially with Extra-intestinal E. coli (ExPEC) which is considered a great challenging health problem.Conclusion: There is a need for studying how to control these factors to decrease the rate and the severity of infections. The relationship between virulence factors and resistance genes is complex and needs more studies that should be specific for each area.Keywords: E. coli, virulence, resistance, colicin genes, ESBL, bla−CTX-M-15, bla−oxa-2, bla−oxa-10

Sara M Farhan,1 Reham A Ibrahim,2 Khaled M Mahran,3 Helal F Hetta4,5, Rehab M Abd El-Baky1,21Department of Microbiology and Immunology, Faculty of Pharmacy, Deraya University, Minia 11566, Egypt; 2Department of Microbiology and Immunology, Faculty of Pharmacy, Minia University, Minia 61519 Egypt; 3General Surgery and Laparoscopic Surgery, Faculty of Medicine, Minia University, Minia 61519 Egypt; 4Department of Medical Microbiology and Immunology, Faculty of Medicine, Assiut University, Assiut 71515, Egypt; 5Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, OH, USABackground: Pseudomonas aeruginosa (P. aeruginosa) represents a great threat to public health worldwide, due to its high ability to acquire resistance to different antibiotic classes. Carbapenems are effective against multidrug resistant (MDR) P. aeruginosa, but their widespread use has resulted in the emergence of carbapenem-resistant strains, which is considered a major global concern. This study aimed to determine the prevalence of carbapenem resistance among P. aeruginosa strains isolated from different sites of infection.Methods: Between October 2016 and February 2018, a total of 530 clinical specimens were collected from patients suffering from different infections, then processed and cultured. Isolates were tested for extended spectrum β-lactamase (ESBL) and metallo-β-lactamase (MBL) production using double-disk synergy test, modified Hodge tests, and disc potentiation test. PCR was used for the detection of selected OXA carbapenemases encoding genes.Results: Of 530 samples, 150 (28.3%) P. aeruginosa isolates were obtained. MDR strains were found in 66.6% (100 of 150) of isolates. Of 100 MDR P. aeruginosa isolates, 54 (54%) were ESBL producers and 21 (21%) carbapenem resistant P. aeruginosa. MBL production was found in 52.3% (eleven) carbapenem-resistant isolates. CTX-M15 was found among 55.5% of ESBL- producing P. aeruginosa. Carbapenemase genes detected were blaIMP (42.8%, nine of 21), blaVIM (52.3%, eleven of 21), blaGIM (52.3%, eleven of 21), blaSPM (38%, 8/21). In addition, isolates that were positive for the tested genes showed high resistance to other antimicrobials, such as colistin sulfate and tigecycline.Conclusion: Our study indicates that P. aeruginosa harboring ESBL and MBL with limited sensitivity to antibiotics are common among the isolated strains, which indicates the great problem facing the treatment of serious infectious diseases. As such, there is a need to study the resistance patterns of isolates and carry out screening for the presence of ESBL and MBL enzymes, in order to choose the proper antibiotic.Keywords: MDR; P. aeruginosa, ESBL, MBL, antimicrobial resistance

Barbara P Yawn1, Ibrahim Raphiou2, Judith S Hurley3, Anand A Dalal21Olmsted Medical Center, University of Minnesota, Rochester, Minnesota, USA; 2GlaxoSmithKline, Research Triangle Park, North Carolina, USA; 3Hurley Consulting, Placitas, New Mexico, USAAbstract: Exacerbations contribute significantly to the morbidity of COPD, leading to an accelerated decline in lung function, reduced functional status, reduced health status and quality of life, poorer prognosis and increased mortality. Prevention of exacerbations is thus an important goal of COPD management. In patients with COPD, treatment with a combination of the inhaled corticosteroid fluticasone propionate (250 μg) and the long-acting β2-agonist salmeterol (50 μg) in a single inhaler (250/50 μg) is an effective therapy option that has been shown to reduce the frequency of exacerbations, to improve lung function, dyspnea and health status, and to be relatively cost-effective as a COPD maintenance therapy. Importantly, results of various studies suggest that fluticasone propionate and salmeterol have synergistic effects when administered together that improve their efficacy in controlling symptoms and reducing exacerbations. The present non-systematic review summarizes the role of fluticasone propionate/salmeterol combination therapy in the prevention of exacerbations of COPD and its related effects on lung function, survival, health status, and healthcare costs.Keywords: Advair, COPD, disease exacerbation, fluticasone propionate, salmeterol, combination drug therapy

Background: Bacillus cereus is a common food poisoning pathogen in humans. This study aimed to investigate the prevalence, molecular typing, antibiogram profile, pathogenicity, dissemination of virulence and antibiotic resistance genes associated with natural B. cereus infection among Mugil seheli ., Methods: Consequently, 120 M. seheli (40 healthy and 80 diseased) were obtained from private fish farms in Port-said Governorate, Egypt. Afterward, samples were processed for clinical, post-mortem, and bacteriological examinations. The recovered isolates were tested for antimicrobial susceptibility, phenotypic assessment of virulence factors, pathogeneicity, and PCR-based detection of virulence and antibiotic resistance genes., Results: B. cereus was isolated from 30 (25%) examined fish; the highest prevalence was noticed in the liver (50%). The phylogenetic and sequence analyses of the gyr B gene revealed that the tested B. cereus isolate displayed a high genetic similarity with other B. cereus strains from different origins. All the recovered B. cereus isolates (n =60, 100%) exhibited β-hemolytic and lecithinase activities, while 90% (54/60) of the tested isolates were biofilm producers. Using PCR, the tested B. cereus isolates harbor nhe, hbl, cyt K, pc-plc , and ces virulence genes with prevalence rates of 91.6%, 86.6%, 83.4%, 50%, and 33.4%, respectively. Moreover, 40% (24/60) of the tested B. cereus isolates were multidrug-resistant (MDR) to six antimicrobial classes and carried the bla 1, bla 2, tet A, and erm A genes. The experimentally infected fish with B. cereus showed variable mortality in direct proportion to the inoculated doses., Conclusion: As far as we know, this is the first report that emphasized the existence of MDR B. cereus in M. seheli that reflects a threat to the public health and the aquaculture sector. Newly emerging MDR B. cereus in M. seheli commonly carried virulence genes nhe, hbl, cyt K, and pc-plc , as well as resistance genes bla 1, bla 2, tet A, and erm A., Competing Interests: The authors declare no conflict of interest., (© 2022 Algammal et al.)