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Nguyen Van An,1 Hai Thi Nguyen,2 Van Nguyen Le,1 Ha Thi Thu Van,1 Nguyen Minh Hai,1 Vu Huy Luong,3,4 Vinh Thi Ha Nguyen,4,5 Hoa Quynh Pham,6 Hung Van Le,4,6 Dinh Viet Hung,7 Hoang Trung Kien,8 Minh Nhat Le,9,10 Nguyen Hoang Viet,11 Luong Cong Thuc,12 Ta Ba Thang,13 Tran Viet Tien,14 Le Huy Hoang,15 Nguyen Thuy Tram,15 Tuan Dinh Le,16 Nguyen Tien Son,16 Hai Ha Long Le17,18 1Department of Microbiology, Military Hospital 103, Vietnam Military Medical University, Hanoi, Vietnam; 2Department of Central Laboratory, Bac Ninh Provincial General Hospital, Bac Ninh, Vietnam; 3Department of Laser and Skincare, National Hospital of Dermatology and Venereology, Hanoi, Vietnam; 4Department of Dermatology and Venereology, Hanoi Medical University, Hanoi, Vietnam; 5Department of General Planning, National Hospital of Dermatology and Venereology, Hanoi, Vietnam; 6Department of Microbiology, Mycology and Parasitology, National Hospital of Dermatology and Venereology, Hanoi, Vietnam; 7Department of Psychiatry, Military Medical 103, Vietnam Military Medical University, Hanoi, Vietnam; 8Department of Immunology, Vietnam Military Medical University, Hanoi, Vietnam; 9Tay Nguyen Institute of Science Research, Vietnam Academy of Science and Technology, VAST, Hanoi, Vietnam; 10Antimicrobial Resistance Research Center, National Institute of Infectious Disease, Tokyo, Japan; 11Molecular Pathology Department, Faculty of Medical Technology, Hanoi Medical University, Hanoi, Vietnam; 12Cardiovascular Center, Military Hospital 103, Vietnam Military Medical University, Hanoi, Vietnam; 13Respiratory Center, Military Hospital 103, Vietnam Military Medical University, Hanoi, Vietnam; 14Department of Infectious Diseases, Military Hospital 103, Vietnam Medical Military University, Hanoi, Vietnam; 15Department of Bacteriology, National of Hygiene and Epidemiology, Hanoi, Vietnam; 16Department of Rheumatology and Endocrinology, Military Hospital 103, Vietnam Medical Military University, Hanoi, Vietnam; 17Department of Clinical Microbiology and Parasitology, Faculty of Medical Technology, Hanoi Medical University, Hanoi, Vietnam; 18Department of Biochemistry, Hematology and Immunology, National Hospital of Dermatology and Venereology, Hanoi, VietnamCorrespondence: Hai Ha Long Le, Department of Clinical Microbiology and Parasitology, Faculty of Medical Technology, Hanoi Medical University, Hanoi, 100000, Vietnam, Tel +84978520055, Email lehalonghai@gmail.comPurpose: Staphylococcus aureus, including methicillin-resistant Staphylococcus aureus (MRSA) strain, can become resistant to all classes of clinically available antibiotics and causes skin infections and severe infections in the lungs, heart, and bloodstream. The study aimed to evaluate antimicrobial susceptibility patterns and MRSA exhibiting multidrug resistance obtained through a microbiological culture of clinical specimens at Bac Ninh Provincial General Hospital in Bac Ninh Province, Vietnam.Methods: We employed a cross-sectional analysis at Bac Ninh Provincial General Hospital in Vietnam. 15,232 clinical samples from inpatients were examined. S. aureus isolates were identified using established protocols and tested for MRSA and antibiotic susceptibility. Data was analyzed using R software, with statistical calculations to assess associations between variables.Results: Staphylococcus aureus was isolated from 417 samples (2.7%), with 77.2% being MRSA and 22.8% methicillin-susceptible Staphylococcus aureus (MSSA). Significant sources of MRSA were wounds (64.6%) and the surgical unit (50%) according to sample types and hospital wards, respectively. S. aureus showed high resistance rates, the highest being azithromycin (83.2%), and was fully susceptible to vancomycin. Among 294 multidrug-resistant (MDR) strains, the prevalence was 82.0% in MRSA and 18.0% in MSSA.Conclusion: The study highlights widespread antimicrobial resistance among MRSA isolates from a provincial hospital in Vietnam, emphasizing the urgent need for antibiotic surveillance, formulation of antibiotic policies, and preventive measures to tackle the increasing prevalence of multidrug-resistant MRSA.Keywords: Staphylococcus aureus, antibiotic resistance, MRSA, MDR, Vietnam

Yan Hu,1 Min Yu,1 Guoqing You,1 Jun Fan,1 Huiwen Zheng2 1Tuberculosis Reference Laboratory, Chongqing Tuberculosis Control Institute, Chongqing, People’s Republic of China; 2Laboratory of Respiratory Diseases, Beijing Key Laboratory of Pediatric Respiratory Infection Diseases, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, National Center for Children’s Health, Beijing, People’s Republic of ChinaCorrespondence: Jun Fan; Huiwen Zheng, Email zjwegg@163.com; hanhui820@126.comObjective: We compared the MeltPro assay to whole-genome sequencing (WGS) to investigate the molecular characterization of second-line injectable drug (SLID) resistance in multidrug-resistant tuberculosis (MDR-TB) isolates in Chongqing, China.Methods: A total of 122 MDR-TB patient isolates were collected between March 2019 and June 2020 from Chongqing Municipality, China. Conventional drug-susceptibility testing was performed using the proportion method, followed to generate minimum inhibitory concentrations (MICs) of SLIDs determined by microplate alamarblue assay. All strains were subjected to both MeltPro and WGS assays.Results: Among 122 MDR-TB isolates, 30 (24.6%), 22 (18.0%), and 14 (11.5%) were resistant to kanamycin (KM), amikacin (AM), and capreomycin (CM), respectively. Of the 31 SLID-resistant isolates, 24 (77.4%, 24/31) isolates harbored mutations in the rrs gene, with the most prevalent mutations in rrs A1401G (22/24, 91.7%). Mutation in rrs A1401G was associated with high levels of resistance to KM (MIC, ≥ 40 μg/mL) and AM (MIC, ≥ 64 μg/mL), but disparities in CM-resistance levels. Using phenotypic drug-susceptibility testing as gold standard, we found that the overall sensitivity of MeltPro and WGS was 87.1% and 90.32% and specificity 100% and 97.8%, respectively. Seven isolates had discordant results between phenotypic and genotypic resistance of SLIDs.Conclusion: MeltPro is a promising diagnostic tool for accurate identification of SLID-resistant MTB isolates with mutations in the rrs and eis genes. There was a disparity between MeltPro with WGS results in the proportion of heterogeneous drug-resistant bacteria with rrs mutation and limited probes. Resistance mechanisms other than genetic mutations will affect the consistency of MeltPro and WGS with phenotypic drug-susceptibility results.Keywords: MDR, MeltPro, resistance, genotype

Yue Jiang,1,2,* Huilin Yang,3,* Zhen-Yu Wang,1,2 Da-Chuan Lin,4 Xinan Jiao,1,2 Yunlong Hu,3,4 Jing Wang1,2 1Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, 225009, People’s Republic of China; 2Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, 225009, People’s Republic of China; 3Department of Clinical Laboratory, Peking University Shenzhen Hospital, Shenzhen, 518036, People’s Republic of China; 4Guangdong Key Laboratory of Regional Immunity and Diseases, Shenzhen University School of Medicine, Shenzhen University, Shenzhen, 518060, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yunlong Hu; Jing Wang, Email huyunlong@pkuszh.com; wj1231@yzu.edu.cnObjective: Salmonella enterica serovar Kentucky ST198 has emerged as a global threat to humans. In this study, we aimed to characterize the prolonged carriage of ciprofloxacin-resistant and extended-spectrum β-lactamase (ESBL)-producing S. Kentucky ST198 in a single patient with inflammatory bowel disease (IBD).Methods: Three S. Kentucky strains were collected from a single patient with IBD on 11th January, 23rd January, and 8th February, 2022, respectively. Antimicrobial susceptibility testing, whole-genome sequencing, and phylogenetic analysis with 38 previously described Chinese S. Kentucky ST198 strains from patients and food were performed.Results: All three S. Kentucky isolates belonged to ST198. They carried identical 16 resistance genes, such as blaCTX-M-55, tet(A), and qnrS1, and had identical mutations within gyrA (S83F and D87N) and parC (S80I). Therefore, they exhibited identical multidrug-resistant profiles, including the clinically important antibiotics cephalosporins (ceftazidime and cefepime), fluoroquinolones (ciprofloxacin and levofloxacin), and third-generation tetracycline (tigecycline). Our three S. Kentucky strains were classified into the subclade ST198.2– 2, and were genetically identical (2– 6 SNPs) to each other. They exhibited a close genetic similarity (15– 20 SNPs) to the isolate NT-h3189 from a patient and AH19MCS1 from chicken meat in China, indicating a possible epidemiological link between these S. Kentucky ST198 isolates from the patients and chicken meat.Conclusion: Long-term colonization of ciprofloxacin-resistant and ESBL-producing S. Kentucky ST198 in a single patient is a matter of concern. Due to the potential transfer of S. Kentucky ST198 from food sources to humans, ongoing surveillance of this particular clone in animals, animal-derived food products, and humans should be strengthened.Keywords: Salmonella, MDR, Kentucky, persistent colonization

Sandip Patil,1,2 Hongyu Chen,3 Yunsheng Chen,3 Shaowei Dong,2 Huirong Mai,1 Bruno Silvester Lopes,4,5 Sixi Liu,1 Feiqiu Wen1,2 1Department of Haematology and Oncology, Shenzhen Children’s Hospital, Shenzhen, Guangdong Province, People’s Republic of China; 2Paediatric Research Institute, Shenzhen Children’s Hospital, Shenzhen, Guangdong Province, People’s Republic of China; 3Department of Laboratory Medicine, Shenzhen Children’s Hospital, Shenzhen, Guangdong Province, People’s Republic of China; 4School of Health and Life Sciences, Teesside University, Middlesbrough, UK; 5National Horizons Centre, Teesside University, Darlington, UKCorrespondence: Feiqiu Wen; Sixi Liu, Department of Haematology and Oncology, Shenzhen Children’s Hospital, 7019 Yitian Road, Futian District, Shenzhen, Guangdong Province, People’s Republic of China, Tel +86-755-83009888, Email fwen62@163.com; tiger647@126.comPurpose: The emergence of multi-drug resistant ESBL-producing E. coli poses a global health problem. In this study, we aimed to investigate the prevalence of E. coli infections and their antibiotic susceptibility profiles in paediatric clinical cases in Shenzhen, China from Jan 1, 2014, to Jan 30, 2019, while also determining temporal trends, identifying ESBL-producing strains, and recommending potential empirical antibiotic therapy options.Methods: We isolated a total of 4148 E. coli from different specimens from a single paediatric healthcare centre. Additionally, we obtained relevant demographic data from the hospital’s electronic health records. Subsequently, we performed antimicrobial susceptibility testing for 8 classes of antibiotics and assessed ESBL production.Results: Out of the 4148 isolates, 2645 were from males. The highest burden of E. coli was observed in the age group of 0– 1 years, which gradually declined over the five-year study period. Antimicrobial susceptibility results indicated that 82% of E. coli isolates were highly resistant to ampicillin, followed by 52.36% resistant to cefazolin and 47.46% resistant to trimethoprim/sulfamethoxazole. Notably, a high prevalence of ESBL production (49.54%) was observed among the E. coli isolates, with 60% of them displaying a multi-drug resistance phenotype. However, it is worth mentioning that a majority of the isolates remained susceptible to ertapenem and imipenem. Our findings also highlighted a decrease in E. coli infections in Shenzhen, primarily among hospitalized patients in the 0– 1 year age group. However, this decline was accompanied by a considerably high rate of ESBL production and increasing resistance to multiple antibiotics.Conclusion: Our study underscores the urgent need for effective strategies to combat multi-drug resistant ESBL-producing E. coli Infections.Keywords: E. coli infections, antibiotic susceptibilities, young children, MDR

Selamyhun Tadesse,1 Alene Geteneh,1 Tilahun Hailu2 1Department of Medical Laboratory Science, College of Health Sciences, Woldia University, Woldia, Ethiopia; 2School of Public Health, College of Health Sciences, Woldia University, Woldia, EthiopiaCorrespondence: Selamyhun Tadesse, Email tselamyhun@gmail.comBackground: The emergence and spread of extended-spectrum β-lactamases (ESβLs) and carbapenemase (CP) producing gram negative non-fermenters are becoming a serious public health threat globally. Infections caused by these pathogens limit treatment options and contribute to the significant morbidity and mortality. Thus, to reduce their spread, early detection of these superbugs is very crucial. This study therefore aimed to assess the prevalence of ESβLs and CP producing gram negative non-fermenters at selected hospitals of North East Ethiopia.Methods: A cross-sectional study was conducted from January to June 2021. Acinetobacter baumannii (A. baumannii) and Pseudomonas aeruginosa (P. aeruginosa) were identified using standard bacteriological techniques. ESβL and CP production were detected by combined disk diffusion and modified carbapenem inhibitory methods, respectively. Data were collected via face-to-face interview and patient card review. Chi-squared and Fisher’s exact tests were calculated and p-value < 0.05 was considered statistically significant.Results: A total of 384 patients participated in this study. Overall, 30 (7.8%) patients had positive culture for A. baumannii and P. aeruginosa. The prevalence of A. baumannii was 20 (5.2%) and that of P. aeruginosa was 10 (2.6%). From the overall isolates, 16 (53.3%) were ESβL and the proportion of carbapenemase production was 4 (13.3%). ESβL production was 8 (40%) in A. baumannii and 8 (80%) in P. aeruginosa isolates. ESβL production infections were significantly associated with hospitalization (p=0.004). Intravenous catheterization, hospitalization, and surgery had significant association with ESβL production (p< 0.005). All isolates of A. baumannii and P. aeruginosa were MDR.Conclusion: ESβL and carbapenemase production among A. baumannii and P. aeruginosa were high in the selected hospitals. The treatment of such resistant infectious agents should be guided by antimicrobial susceptibility test in a study setting. Thus, restricted and wise use of antibiotics is highly recommended to contain the spread of these superbugs. Hospitals should develop infection prevention guidelines to prevent the spread of resistant pathogens in hospitalized patients.Keywords: ESβL, CP, MDR, A. baumannii, P. aeruginosa, Ethiopia

Panpan Fang,1,&ast; Kaijie Gao,1 Junmei Yang,1 Tiewei Li,1 Weihua Gong,1 Qianqian Sun,1 Yingyuan Wang2,&ast; 1Zhengzhou Key Laboratory of Children’s Infection and Immunity, Children’s Hospital Affiliated to Zhengzhou University, Henan Children’s Hospital, Zhengzhou Children’s Hospital, Zhengzhou, People’s Republic of China; 2Department of Neonatal Intensive Care Unit, Children’s Hospital Affiliated to Zhengzhou University, Henan Children’s Hospital, Zhengzhou Children’s Hospital, Zhengzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Panpan Fang; Yingyuan Wang, Email fang_panpan@163.com; yingyuanxse@163.comIntroduction: Sepsis is the most severe infectious disease with the highest mortality rate, particularly among neonates admitted to the neonatal intensive care unit (NICU). This study retrospectively analyzed the epidemiology, antibiotic resistance profiles, and prevalence of multidrug-resistant (MDR) bacteria isolated from blood or cerebrospinal fluid (CSF) cultures in order to evaluate the appropriateness of initial empirical therapy for neonatal sepsis.Methods: A retrospective study was conducted in the NICU from January 1, 2015, to December 31, 2022. Microbiological data from patients admitted to the NICU were anonymously extracted from the Laboratory of Microbiology database. Neonatal sepsis was classified into two types: early-onset sepsis (EOS), which occurs within the first 72 hours of life, and late-onset sepsis (LOS) for those begins later.Results: A total of 679 bacterial strains, 543 from blood and 136 from CSF, were detected in 631 neonates. Among these, 378 isolates (55.67%) were Gram-positive bacteria, and 301 isolates (44.33%) were Gram-negative bacteria. The most frequently isolated pathogens were Coagulase-negative staphylococci (CoNS) (36.52%), followed by Klebsiella pneumoniae (20.47%) and Escherichia coli (13.84%). In EOS, 121 strains were found, CoNS represented the majority (33.88%), followed by Klebsiella pneumoniae (23.97%) and Escherichia coli (8.26%). Early-onset septicemia exhibited 67 (55.37%) MDR bacteria. In LOS, 558 strains were isolated, CoNS represented the majority of pathogens (37.10%), followed by Klebsiella pneumoniae (19.71%) and Escherichia coli (15.05%). Late-onset septicemia showed 332 (59.50%) MDR bacteria. High rates of MDR were found in CoNS (76.21%), carbapenem-resistant Klebsiella pneumoniae (66.91%), and MRSA (33.33%).Conclusion: The study revealed an alarming prevalence of MDR strains isolated from neonatal sepsis, emphasizing the necessity of finding effective prevention and treatment measures. Colistin can be used for MDR Gram-negative bacteria, while vancomycin and teicoplanin can be considered treatment therapies for staphylococcal infections.Keywords: MDR, EOS, LOS, sepsis, bacterial strains

Shambel Araya,1 Zenebe Gebreyohannes,2 Getachew Tadlo,3 Gebreab Teklebirhan Gessew,1 Abebe Edao Negesso1 1Department of Medical Laboratory Science, College of Health Science Addis Ababa University Addis Ababa, Addis Ababa, 9086, Ethiopia; 2Department of Medical Microbiology, Parasitology and Immunology St. Paul Hospital Millennium Medical College, Addis Ababa, Ethiopia; 3Department of Medical Laboratory Science, St. Paul Hospital Millennium Medical College, Addis Ababa, EthiopiaCorrespondence: Shambel Araya, Tel +251 939459529, Email shambelaraya8@gmail.comBackground: A. baumannii and P. aeruginosa are important nosocomial pathogens in health-care settings. Both are intrinsically resistant to many drugs and are able to become resistant to the virtually most antimicrobial agents. An increasing prevalence of infections caused by multidrug-resistant isolates has been reported in many countries.Methods: An institutional-based cross-sectional five-year retrospective study was conducted to assess the antimicrobial resistance trend of P. aeruginosa and A. baumani. 893 A. baumani and 729 P. aeruginosa isolates were included in the study. Conventional method was used for identification and antimicrobial susceptibility was determined by Kirby-Bauer disc-diffusion method. The isolates were from suspected bloodstream infections, wound infections, urinary tract, or surgical site nosocomial infections. Socio-demographic and other variables of interest were collected using a structured check list from a patient record data. Data were analyzed using SPSS version 26 software. P value < 0.05 was considered statistically significant.Results: A total of 1622 A. baumanii and P. aeruginosa were isolated from various clinical specimens recorded from the year 2017– 2021. Out of which A. baumanni was 893 (60.6%) and P. aeruginosa was 729 (39.4%). Blood was the major source of the isolates (18.3%), followed by urine (16%), and tracheal aspirate (10.6%). Antimicrobial resistance among A. baumanni over the five years were; ampicillin (86% to 92%), ceftriaxone (66.7% to 82.2%), and ciprofloxacin (58.5% to 66.7%). In P. aeruginosa a significant increase in resistance was seen from 2017 to 2021 to Amoxicillin-clavulanate (74.1% to 84.2%), chloramphenicol (62% to 81.9%), and gentamicin (40% to 44.8%).Conclusion: A five-year antimicrobial resistance trend analysis of A. baumanni and P. aeruginosa showed increasing multi drug resistance and resistance to highly potent antimicrobial agents in Ethiopia. It should be addressed with infection control measures, surveillance, and alternative new therapeutic strategies to circumvent the spread of multi-drug resistance.Keywords: A. baumanii, antibiotic resistance, Ethiopia, MDR, P. aeruginosa

Abdelazeem M Algammal,1 Nada H Eidaroos,1 Khyreyah J Alfifi,2 Marfat Alatawy,2 Alhanouf I Al-Harbi,3 Yasmene F Alanazi,4 Madeha OI Ghobashy,2,5 Ahmed R khafagy,6 Aboelkheir M Esawy,7 Soha S El-Sadda,7 Helal F Hetta,8 Reham M El-Tarabili1 1Department of Bacteriology, Immunology, and Mycology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt; 2Department of Biology, Faculty of Science, University of Tabuk, Tabuk, Saudi Arabia; 3Department of Medical Laboratory, College of Applied Medical Sciences, Taibah University, Yanbu, Saudi Arabia; 4Department of Biochemistry, Faculty of Science, University of Tabuk, Tabuk, Saudi Arabia; 5Microbiology Department, Faculty of Science, Ain Shams University, Cairo, Egypt; 6Department of Microbiology, Faculty of Veterinary Medicine, Ain Shams University, Cairo, Egypt; 7Animal Health Research Institute, Mansoura, Egypt; 8Department of Medical Microbiology and Immunology, Faculty of Medicine, Assiut University, Assiut, 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.egBackground: Pseudomonas aeruginosa is incriminated in septicemia, significant economic losses in the poultry production sector, and severe respiratory infections in humans. This study aimed to investigate the occurrence, oprL sequencing, antimicrobial resistance patterns, virulence-determinant, Quorum sensing, and antibiotic resistance genes of P. aeruginosa retrieved from broiler chickens.Methods: Two hundred samples were collected from 120 broiler chickens from broiler farms at Ismailia Governorate, Egypt. Consequently, the bacteriological examination was conducted and the obtained P. aeruginosa strains were tested for oprL gene sequencing, antibiogram, and PCR screening of virulence, Quorum sensing, and antibiotic resistance genes.Results: The overall prevalence of P. aeruginosa in the examined birds was 28.3%. The oprL gene sequence analysis underlined that the tested strain expressed a notable genetic identity with various P. aeruginosa strains isolated from different geographical areas in the USA, India, China, Chile, and Ghana. PCR evidenced that the obtained P. aeruginosa strains, carrying virulence-related genes: oprL, toxA, aprA, phzM, and exoS in a prevalence of 100%, 100%, 42.5%, 33.3%, and 25.9%, respectively. Moreover, the recovered P. aeruginosa strains possessed the Quorum sensing genes: lasI, lasR, rhlI, and rhlR in a prevalence of 85.2%, 85.2%, 81.5%, and 81.5%, respectively. Furthermore, 40.7% of the isolated P. aeruginosa were XDR to seven antimicrobial classes, possessing sul 1, blaTEM, tetA, blaCTX-M, blaOXA-1, and aadA1 genes.Conclusion: As we can tell, this is the first report emphasizing the evolution of XDR P. aeruginosa strains from broiler chicken in Egypt, which is supposed to be a serious threat to public health. The emerging XDR P. aeruginosa in poultry frequently harbored the oprL, toxA, and aprA virulence genes, the lasI, lasR, rhlI, and rhlR Quorum sensing genes, and the sul 1, blaTEM, tetA, blaCTXM, blaOXA-1, and aadA1 resistance genes.Keywords: P. aeruginosa, MDR, XDR, oprL sequence analysis, Quorum sensing, virulence genes, resistance genes

Muhammad Qamar Saeed,1 Mubashar Aziz,1 Raja Kamran Afzal,2 Aamir Hussain,3 Hamid Manzoor,4 Sumaira Rasul,4 Tahira Ruby,5 Ahmed Akrem,5 Samrah Masud,5 Adnan Arshad,1 Ayesha Latif,1 Muhammad Yousif,1 Hamdan Aali,1 Menahil Fatima,1 Muhammad Mujahid Bhutta,1 Aleem Ahmed Khan5 1Dr. Ghulam Nabi Chaudhary Laboratory of Microbial Technologies, Department of Microbiology and Molecular Genetics, Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, Pakistan; 2Department of Microbiology, CMH Institute of Medical Sciences, Multan, Pakistan; 3Institute of Basic Medical Sciences, Khyber Medical University, Peshawar, Pakistan; 4Institute of Molecular Biology and Biotechnology, Bahauddin Zakariya University Multan, Multan, Pakistan; 5Institute of Pure and Applied Biology, Bahauddin Zakariya University Multan, Multan, PakistanCorrespondence: Muhammad Qamar Saeed, Dr. Ghulam Nabi Chaudhary Laboratory of Microbial Technologies, Department of Microbiology, Institute of Pure and Applied Biology, Bahauddin Zakariya University Multan, Pakistan, Tel +92-333-0231222, Email mqamarsaeed@bzu.edu.pkIntroduction: Antibiotics are being used in humans and animals for treatment and control of bacterial infections. Excessive use of antibiotics in the production of poultry is a popular practice, but it poses serious health issues by transferring resistance from farm to humans via food or direct exposure.Study Objective: The objective of this study was to carry out a comparison of the resistance and sensitivity profile of isolated isolates from sewage of toilets that were in use of workers inside the farm and from sewage of household toilets.Methodology: In this study, a total of 320 sewage samples were collected. The antibiotic susceptibility profile was checked by Kirby-Bauer disc diffusion method, and the statistical analysis was carried out by MS excel. Chi-square test was performed to determine whether the antibiograms from two sample types were statistically different from each other or not.Results: From 320 sewage samples, a total of 296 bacterial isolates were isolated among which the leading bacterium was E. coli. The proportion of resistance, ESBL production and MDR was significantly higher in bacteria isolated from sewage of toilets under use of poultry farm workers as compared to the sewage from domestic use toilets.Conclusion: Resistance significantly increased in the bacteria isolated from toilets under use of poultry farm workers as compared to the ones isolated from control sewage samples.Keywords: E. coli, antibiotic resistance, MDR, sewage, human microbiota

Abdirahman Mohamed Hassan Dirie,1 Sedat Çolako&gbreve;lu,1 Osman Muhyadin Abdulle,2 Bashir Mohamud Abdi,3 Mohamed Abdi Osman,4 Abdiaziz Mohamud Shire,5 Abdinafic Mohamud Hussein6 1Pulmonology Department, Somalia Mogadishu Turkey Recep Tayyip Erdogan Training and Research Hospital, Mogadishu, Banadir, Somalia; 2Manager of the Multi-Drug Resistant Center in Southern Somalia, Mogadishu, Banadir, Somalia; 3HIV Program manager, Ministry of Health, Somali Federal Government, Mogadishu, Banadir, Somalia; 4Infectious and Microbiology Department, Somalia Mogadishu Turkey Recep Tayyip Erdogan Training and Research Hospital, Mogadishu, Banadir, Somalia; 5National TB program Manager, Somali Federal Government, Ministry of Health, Mogadishu, Banadir, Somalia; 6Cardiovascular Surgery Department, Somalia Mogadishu Turkey Recep Tayyip Erdogan Training and Research Hospital, Mogadishu, Banadir, SomaliaCorrespondence: Abdirahman Mohamed Hassan Dirie, Tel +252615479778, Email dirie515@gmail.comBackground: Tuberculosis (TB) is an infectious disease that is the second most common cause of death from a single infectious agent. TB infection affects anyone, regardless of age, gender, and ethnicity. Drug-resistant TB is a serious public health problem, which needs treatment with a second-line anti-TB drug and it includes poly-drug resistance (PDR), multi-drug resistance (MDR), and extensive drug resistance (XDR). The goal of this research is to find out the prevalence of MDR TB among pulmonary TB patients in Banadir, Somalia.Methods: This was a multicenter retrospective review of data involving 1732 smear-positive pulmonary TB patients visiting Banadir TB centers between July 1, 2019 and June 30, 2020. Demographic, clinical, and drug susceptibility data were retrieved from TB treatment cards. The data were analyzed using Statistical Package for Social Sciences (SPSS) software (IBM SPSS Statistics version 26).Results: All 1732 pulmonary TB cases were previously diagnosed by the Gene Xpert MTB/RIF test. Among them, 70.4% (1219/1732) were males. The mean age was 31.77 years. Overall, the prevalence of drug resistance TB was 10.56% (183/1732). The MDR TB was 1.96%, poly-drug resistance (PDR) was 0.12%, and extensive drug resistance was 0.06%.Conclusion: This study showed a prevalence of MDR-TB among pulmonary TB patients, which is similar to some of the eastern African countries.Keywords: TB, drug-resistance, poly-drug resistance, MDR, XDR

Lamya Zohair Yamani, Nasreldin Elhadi Department of Clinical Laboratory Science, College of Applied Medical Sciences, Imam Abdulrahman Bin Faisal University, Dammam, Kingdom of Saudi ArabiaCorrespondence: Nasreldin Elhadi, Department of Clinical Laboratory Science, Imam Abdulrahman Bin Faisal University, Dammam, Kingdom of Saudi Arabia, Email nmohammed@iau.edu.saIntroduction: The Kingdom of Saudi Arabia has limited data on enteropathogenic Escherichia coli (EPEC). Therefore, this study was undertaken to contribute to EPEC surveillance and investigate the molecular epidemiology of EPEC strains that have been implicated in human infection in King Fahd Hospital of the University (KFHU) between 2013 and 2014 in the Eastern Province of Saudi Arabia.Methods: A total of 60 non-duplicate E. coli isolates associated with human gastroenteritis were included in this study. They were characterized using PCR to determine virulence genes, antimicrobial resistance patterns, and enterobacterial repetitive intergenic consensus (ERIC-PCR).Results: Among the 60 strains, 20% of those examined were positive for the intimin eae and bfpA genes and identified as typical EPEC (tEPEC). Furthermore, 44 of E. coli strains tested positive for the eae gene only and revealed a high occurrence rate of 73.3% of atypical EPEC (aEPEC) within the overall examined strains. All strains were positive for the EAST1 gene, and none tested positive for the stx gene. More than 70% of EPEC strains were multi-drug resistant (MDR) and aEPEC strains with the highest proportion of this feature of MDR. ERIC-PCR fingerprint revealed a total of 19 ERIC types with eight related distinct clusters and a similarity rate cut-off with ≥ 90% homology from the identified isolates.Conclusion: A high antibiotic resistance rate was reported for first-line antibiotics, such as ampicillin, tetracycline, nalidixic acid, trimethoprim-sulfamethoxazole, noroxin, and ciprofloxacin.Keywords: EPEC, virulence, antimicrobial resistance, MDR, ERIC-PCR

Bilal Aslam,1 Muhammad Hussnain Siddique,2 Abu Baker Siddique,1 Muhammad Shafique,1 Saima Muzammil,1 Mohsin Khurshid,1 Muhammad Hidayat Rasool,1 Moeed Ahmad,1 Tamoor Hamid Chaudhry,3 Afreenish Amir,3 Muhammad Salman,3 Zulqarnain Baloch,4 Norah A Alturki,5 Ahmad Alzamami6 1Department of Microbiology, Government College University Faisalabad, Faisalabad, Pakistan; 2Department of Bioinformatics & Biotechnology, Government College University Faisalabad, Faisalabad, Pakistan; 3Public Health Laboratories Division, National Institute of Health, Islamabad, Pakistan; 4Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, People’s Republic of China; 5Clinical Laboratory Science Department, College of Applied Medical Science, King Saud University, Riyadh, Saudi Arabia; 6Clinical Laboratory Science Department, College of Applied Medical Science, Shaqra University, AlQuwayiyah, Saudi ArabiaCorrespondence: Bilal Aslam, Department of Microbiology, Government College University Faisalabad, Faisalabad, Pakistan, Email drbilalaslam@gcuf.edu.pk Ahmad Alzamami, Clinical Laboratory Science Department, College of Applied Medical Science, Shaqra University, AlQuwayiyah, Saudi Arabia, Email aalzamami@su.edu.saBackground: The World Health Organization (WHO) has declared the multi-drug resistant (MDR) Klebsiella pneumoniae as one of the critical bacterial pathogens. The dearth of new antibiotics and inadequate therapeutic options necessitate finding alternative options. Bacteriophages are known as enemies of bacteria and are well-recognized to fight MDR pathogens.Methods: A total of 150 samples were collected from different clinical specimens through a convenient sampling technique. Isolation, identification, and antibiotic susceptibility testing (AST) of K. pneumoniae were done by standard and validated microbiological procedures. Molecular identification of virulence factors and antibiotic resistance genes (ARGs) was carried out through polymerase chain reaction (PCR) by using specific primers. For bacteriophage isolation, hospital sewage samples were processed for phage enrichment, purification, and further characterization ie, transmission electron microscopy (TEM) and stability testing, etc. followed by evaluation of the lytic potential of the phage.Results: Overall, a total of 41% of isolates of K. pneumoniae were observed as hypervirulent K. pneumoniae (hvKp). Among hvKp, a total of 12 (42%) were detected as MDR hvKp. A total of 37% of all MDR isolates were found resistant to colistin, and 66% of the colistin resistance isolates were recorded as mcr-1 positive. Isolated phage KpnM had shown lytic activity against 53 (79%) K. pneumoniae isolates. Remarkably, all 8 mcr-1 harboring MDR hvKp and non-hvKp isolates were susceptible to KpnM phage.Conclusion: Significant distribution of mcr-1 harboring hypervirulent Klebsiella pneumoniae was observed in clinical specimens, which is worrisome for the health system of the country. Characterized phage KpnM exhibited encouraging results and showed the lytic activity against the mcr-1 harboring hvKp isolates, which may be used as a prospective alternative control strategy to fight this ominous bacterium.Keywords: Klebsiella pneumoniae, bacteriophages, colistin, MDR, virulence

Dawei Shi,1,&ast; Qiulong Zhou,2,&ast; Sihong Xu,1 Yumei Zhu,3 Hui Li,4 Ye Xu2 1National Institutes for Food and Drug Control, Institute of Pathogen Biology at the Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China; 2Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, People’s Republic of China; 3Shenzhen Center for Chronic Disease Control, Shenzhen, People’s Republic of China; 4Tuberculosis Reference Laboratory, Henan Provincial Centers for Disease Control and Prevention, Zhengzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Hui Li, Tuberculosis Reference Laboratory, Henan Provincial Centers for Disease Control and Prevention, Zhengzhou, People’s Republic of China, Tel/Fax +86 371-68089049, Email lihui2005yan@126.com Ye Xu, Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, People’s Republic of China, Tel +86 592 2187992, Fax +86 592 2187363, Email xuye@xmu.edu.cnPurpose: Pyrazinamide (PZA) is a critical component of standardized chemotherapy for tuberculosis (TB) and is recommended for the treatment of multidrug-resistant (MDR) TB. We aimed to characterize mutations in pncA of M. tuberculosis and evaluate their diagnostic accuracy for PZA susceptibility in China. We also combined genotypic methods with phenotypic susceptibility testing and pyrazinamidase (PZAse) activity to confirm PZA-resistant M. tuberculosis isolates.Results: An evaluation of 82 MDR M. tuberculosis strains revealed that 28.0% (23/82) were phenotypically resistant to 100 mg/L PZA and 15.9% (13/82) showed resistance to 300 mg/L PZA. Mutations in pncA were detected at 33 unique sites, and the majority were point mutations. No evident mutation hotspots or mutations affecting multiple amino acids were found, but the association between pncA mutations and PZA resistance was significant under 100 and 300 mg/L. The sensitivity of pncA mutation detection for predicting PZA susceptibility was 82.6% (19/23), and the specificity was 61.0% (36/59), based on 100 mg/L PZA, whereas the sensitivity was 84.6% (11/13) and the specificity was 55.1% (38/69), based on 300 mg/L PZA. All mutations identified in the highly PZA-resistant (300 mg/L) strains had an 80% loss relative to PZAse activity. No evident PZAse activity loss was observed in one synonymous mutation strain and the loss exceed 60% in all other strains.Conclusion: The association between pncA mutation and PZA resistance was significant. Relatively, the molecular method have shown better reliability than the phenotypic method for the detection of PZA resistance. This provides a theoretical basis for the clinical diagnosis of drug-resistant TB.Keywords: MDR, PZA, pncA, DST, Beijing genotype, enzymatic activity

Nan Shi,1,2,&ast; Jianbang Kang,2,&ast; Shuyun Wang,2 Yan Song,2 Donghong Yin,2 Xiaoxia Li,2 Qian Guo,2 Jinju Duan,2 Shuqiu Zhang1 1Department of Pharmacy, Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of China; 2Department of Pharmacy, Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jinju Duan; Shuqiu Zhang, Email duanjinju@163.com; shuqiu.zhang@126.com; shuqiu.zhang@sxmu.edu.cnObjective: The aim of this study was to analyze the epidemiological of gram-negative bloodstream infection (GNBSI) and establish a risk prediction model for mortality and acquiring multidrug resistant (MDR), the extended spectrum beta-lactamases (ESBLs) producing and carbapenem-resistant (CR) GNBSI.Methods: This retrospective study covered five years from January 2015 to December 2019. Data were obtained from Hospital Information System (HIS) and microbiology department records. The risk factors for mortality and acquiring MDR, ESBLs-producing and CR GNBSI were analyzed by univariable and multivariable analysis.Results: A total of 1018 GNBSI cases were collected. A majority of GNBSI patients were in hematology ward (23.77%). There were 38.61% patients who were assigned in the 41– 60 age group. Escherichia coli was the most common gram-negative organism (49.90%). Among isolates of GNBSI, 40.47% were found to be MDR strains, 34.09% were found to be ESBLs-producing strains and 7.06% were found to be CR strains. Escherichia coli was the most common MDR (71.36%) and ESBLs-producing strain (77.81%). Acinetobacter baumannii was the most common CR isolate (46.15%). Multivariate analysis indicated that diabetes mellitus, solid organ tumor, non-fermentative bacteria, MDR strain, central venous cannula, urinary catheter, therapy with carbapenems or tigecycline prior 30 days of infection were independent mortality risk factors for GNBSIs. Over all, therapy with tigecycline prior 30 days of infection was the mutual predictor for mortality of GNBSI, acquiring MDR, ESBLs-producing and CR GNBSI (OR, 8.221, OR, 3.963, OR, 3.588, OR, 9.222, respectively, all p < 0.001).Conclusion: Collectively, our study implies that patients who were diagnosed as GNBSI had a younger age. Therapy with tigecycline was the mutual and paramount predictor for mortality of GNBSI, acquiring MDR, ESBLs-producing and CR GNBSI. Our investigation had provided a theoretical basis for the use of antibiotics and prevention and control of hospital infection in our region.Keywords: gram-negative bloodstream infection, epidemiology, risk prediction model, MDR, ESBL, carbapenem resistance

Mahmoud Mohamed Tawfick,1,2 Aliaa Ali Elshamy,2 Kareem Talaat Mohamed,3 Nagwan Galal El Menofy4 1Department of Microbiology and Immunology, Faculty of Pharmacy (For Boys), Al-Azhar University, Cairo, 11751, Egypt; 2Department of Microbiology and Immunology, Faculty of Pharmacy, Heliopolis University, Cairo, 11785, Egypt; 3Department of Microbiology and Immunology, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), 6th October City, Giza, 11787, Egypt; 4Department of Microbiology and Immunology, Faculty of Pharmacy (For Girls), Al-Azhar University, Cairo, 11751, EgyptCorrespondence: Mahmoud Mohamed Tawfick, Department of Microbiology and Immunology, Faculty of Pharmacy (For Boys), Al-Azhar University, 1 El-Mokhayam El-Daem Street, Nasr City, Cairo, 11751, Egypt, Tel +20 1157336676, Fax +20 238371543, Email mahmoud_tawfick@azhar.edu.egBackground: Escherichia coli (E. coli), the main human gut microorganism, is one of the evolved superbugs because of acquiring antimicrobial resistance (AMR) determinants via horizontal gene transfer (HGT).Purpose: This study aimed to screen isolates of gut commensal E. coli from healthy adult individuals for antimicrobial susceptibility and plasmid-mediated AMR encoding genes.Methods: Gut commensal E. coli bacteria were isolated from fecal samples that were taken from healthy adult individuals and investigated phenotypically for their antimicrobial susceptibility against diverse classes of antimicrobials using the Kirby Bauer disc method. PCR-based molecular assays were carried out to detect diverse plasmid-carried AMR encoding genes and virulence genes of different E. coli pathotypes (eaeA, stx, ipaH, est, elt, aggR and pCVD432). The examined AMR genes were β-lactam resistance encoding genes (blaCTX-M1, blaTEM, blaCMY-2), tetracycline resistance encoding genes (tetA, tetB), sulfonamides resistance encoding genes (sul1, sulII), aminoglycoside resistance encoding genes (aac(3)-II, aac(6′)-Ib-cr) and quinolones resistance encoding genes (qnrA, qnrB, qnrS).Results: PCR results revealed the absence of pathotypes genes in 56 isolates that were considered gut commensal isolates. E. coli isolates showed high resistance rates against tested antimicrobial agents belonging to both β-lactams and sulfonamides (42/56, 75%) followed by quinolones (35/56, 62.5%), tetracyclines (31/56, 55.4%), while the lowest resistance rate was to aminoglycosides (24/56, 42.9%). Antimicrobial susceptibility profiles revealed that 64.3% of isolates were multidrug-resistant (MDR). High prevalence frequencies of plasmid-carried AMR genes were detected including blaTEM (64%) sulI (60.7%), qnrA (51.8%), aac(3)-II (37.5%), and tetA (46.4%). All isolates harbored more than one gene with the most frequent genetic profile among isolates was blaTEM-blaCTX-M1-like-qnrA-qnrB-tetA-sulI.Conclusion: Results are significant in the evaluation of plasmid-carried AMR genes in the human gut commensal E. coli, suggesting a potential human health risk and the necessity of strict regulation of the use of antibiotics in Egypt. Commensal E. coli bacteria may constitute a potential reservoir of AMR genes that can be transferred to other bacterial species.Keywords: gut microorganisms, antibiotic resistance, plasmid, MDR, resistance genes

Patrycja Zalas-Wi&eogon;cek,1 Ma&lstrok;gorzata Pra&zdot;y&nacute;ska,1 &Lstrok;ukasz Pojnar,2 Anna Pa&lstrok;ka,2 Dorota &Zdot;abicka,3 Monika Orczykowska-Kotyna,3 Aleksandra Polak,3 Barbara Mo&zdot;ejko-Pastewka,4 Ewa Anna G&lstrok;owacka,4 Izabela Pieni&aogon;&zdot;ek,5 Ma&lstrok;gorzata Pawlik,5 Maciej Grys,5 Monika Bogiel4 1Department of Microbiology, Ludwik Rydygier Collegium Medicum; Department of Clinical Microbiology, University Hospital No. 1, Bydgoszcz, Poland; 2Department of Microbiology, University Hospital, Cracow, Poland; 3Department of Epidemiology and Clinical Microbiology, National Medicines Institute, Warsaw, Poland; 4Pfizer Polska Sp. z o.o., Warsaw, Poland; 5Arcana Institute, a Certara Company, Cracow, PolandCorrespondence: Monika Bogiel, Pfizer Polska sp. z o.o., &Zdot;wirki i Wigury 16B, Warszawa, 02-092, Poland, Tel +48 885557081, Fax +48 223356111, Email monika.bogiel@pfizer.comPurpose: Infections caused by resistant Gram-negative bacteria are becoming increasingly common and now pose a serious public health threat worldwide, because they are difficult to treat due to few treatment options and they are associated with high morbidity and mortality. The combination of ceftazidime with the beta-lactamase inhibitor avibactam – seems to be the right choice in this situation. The aim of the study was to evaluate the activity of ceftazidime/avibactam and other commonly used antibiotics against Enterobacterales and Pseudomonas aeruginosa strains isolated within last years in Poland.Patients and Methods: This study analyzed the antibiotic susceptibility of 1607 Enterobacterales isolates and 543 nonfermenting P. aeruginosa strains collected between 2015 and 2019 in 4 medical laboratories participating in the ATLAS (Antimicrobial Testing Leadership And Surveillance) program in Poland. Unduplicated clinically significant Enterobacterales and P. aeruginosa strains were collected from patients with respiratory, skin and musculoskeletal, genitourinary, abdominal, bloodstream or other infections (ear, eye).Results: The ceftazidime/avibactam combination demonstrates the highest activity against Enterobacterales (98.9%), in both adults and children, including strains presenting MDR (multidrug-resistant) (97.5%) and ESBL (extended spectrum β-lactamase) (96.3%) phenotypes. The activity of ceftazidime/avibactam increased to 100% when only MBL (metallo-β-lactamase)-negative subset of Enterobacterales was considered. This combination also achieved the second highest activity result (89.3%) after colistin in P. aeruginosa, including isolates of MDR (65.9%) and carbapenem-resistant (CR) phenotypes (54.8%). When MBL-positive isolates were excluded, susceptibility rate of P. aeruginosa increased to 94.7%. It is worth to note that susceptibility of the examined P. aeruginosa strains to ceftazidime/avibactam was very high in children (93.3%), especially in a pediatric intensive care unit (94.2%).Conclusion: Enterobacterales and P. aeruginosa included in this analysis presented high susceptibility rates to ceftazidime/avibactam. Ceftazidime/avibactam showed the highest activity against Enterobacterales strains among all antibiotics studied, both for the total population as well as for MDR phenotype and ESBL phenotype. Ceftazidime/avibactam also achieved the second highest activity result against P. aeruginosa strains (including MDR and CR phenotypes). These results are much higher when excluding MBL-positive isolates that exhibit intrinsic resistance to ceftazidime/avibactam.Keywords: ceftazidime/avibactam, drug resistance, Gram-negative rods, MDR, ESBL, carbapenem resistance

Yasabe Leykun,1 Chalachew Genet,1 Wondemagegn Mulu1,2 1Department of Medical Laboratory Science, College of Medicine and Health Sciences, Bahir Dar University, Bahir Dar, Ethiopia; 2Department of Biochemistry and Microbiology, Faculty of Sciences, Ghent University, Ghent, BelgiumCorrespondence: Wondemagegn Mulu Tel +251918706921Email wondem_32@yahoo.comBackground: Group B streptococci (GBS) is an important pathogen involved in stillbirths, neonatal deaths and neurological defects, and the occurrence of multi-drug resistance (MDR) is an alarming issue. This study determined the prevalence of GBS colonization in pregnant women and newborns, the proportion of vertical transmission, antimicrobial susceptibility profiles of isolates, and the factors associated with colonization and vertical transmission.Methods: A cross-sectional study was conducted from March 1, 2021 to June 30, 2021, at selected health facilities of Bahir Dar city. Vaginal-recto swabs from 292 pregnant women and pooled ear, nasal and umbilical swabs from 292 newborns were collected. GBS were identified following standard microbiological protocols. Antimicrobial susceptibility testing was performed using modified Kirby–Bauer disk diffusion method and interpreted by the accepted 2020 CLSI M100 guidelines. Logistic regression analysis was computed.Results: Overall, 54 (18.5%) of pregnant women and 22 (7.5%) of newborns had GBS colonization. The proportion of GBS vertical transmission was 22 (40.7%). Group B Streptococcus isolates scored susceptibility to penicillin, ampicillin and vancomycin with 88.9%, 90.7%, and 96.3% for pregnant women and 86.4%, 90.9% and 95.9% for newborns, respectively. A high percentage of non-susceptibility was found for clindamycin and erythromycin with 33.3% and 25.9% for pregnant women and 31.8% and 22.7% from newborns, respectively. Besides, 19 (35.2%) GBS from pregnant women and 8 (36.4%) from newborns were MDR. Group B streptococci colonization was significantly associated with delivery before 37th week of gestation (AOR=2.77, 95% CI 1.14– 6.68) and history of stillbirth (AOR=3.13, 95% CI 1.13– 8.70).Conclusion: Pregnant women vaginal-recto GBS colonization and transmission to newborns connected with MDR are a matter of concerns. Although non-susceptible GBS isolates are obtained, penicillin and vancomycin are relatively effective. The use of clindamycin, erythromycin and ceftriaxone should be guided by antimicrobial susceptibility testing. Genetic analysis is recommended to exactly identify the epidemiology of GBS strains, vertical transmission and antimicrobial resistance at the country level.Keywords: group B streptococci, vaginal-recto colonization, vertical transmission, pregnant women, newborns, MDR, Ethiopia

Liying Gong,1 Luwei Zhang,2 Xiaoli Liu,3 Bekzod Odilov,4 Shengnan Li,1 Zhao Hu,1 Xiaoyan Xiao1 1Department of Nephrology, Qilu Hospital of Shandong University, Jinan, People’s Republic of China; 2Department of Organ Transplantation, Qilu Hospital of Shandong University, Jinan, People’s Republic of China; 3Department of Kidney Transplantation, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China; 4Department of Endocrinology, Qilu Hospital of Shandong University, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of ChinaCorrespondence: Xiaoyan XiaoDepartment of Nephrology, Qilu Hospital of Shandong University, 107 Wenhua Xi Road, Jinan, 250012, Shandong Province, People’s Republic of ChinaTel/Fax +86-531-82169316Email xiaoyanxiao2007@163.comBackground: Infection ranks as the most common complication after kidney transplantation (KT) and threatens outcomes of kidney transplantation recipients (KTR). This study aimed to investigate the microbiological profile of infection, assess bacterial resistance and identify risk factors for multidrug-resistant (MDR) bacterial infection among KTR.Methods: During the study period, 866 recipients underwent kidney transplant surgery. We studied the distribution of pathogens, resistance rate of MDR bacteria and the risk factors of MDR bacterial infection.Results: Totally, 214 species of pathogens (110 species were MDR bacteria) were isolated in 119 KTR. Escherichia coli (E. coli) was the most common bacteria of the infection. MDR extended spectrum β-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-E) were most resistant to ampicillin, cefazolin, ciprofloxacin and complex sulfamethoxazole, while quite sensitive to imipenem, amikacin and piperacillin/tazobactam (PIT). All MDR gram-positive bacteria were quite sensitive to linezolid and vancomycin, except that MDR Staphylococcus was also susceptible to rifampicin. Female gender (OR = 3.497, 95% CI = 1.445– 8.467, P = 0.006), pathogen types > 1 (OR = 3.832, 95% CI = 1.429– 10.273, P = 0.008) and postoperative time < 3 months (OR = 0.331, 95% CI = 0.137– 0.799, P = 0.014) were independent risk factors for MDR bacterial infection.Conclusion: PIT and amikacin may be an alternative choice of ESBL-E infection. Rifampicin can also be prescribed for MDR Staphylococcus infection. MDR bacterial infection was associated with female gender, pathogen types more than 1 and 3 months postoperative period.Keywords: kidney transplantation, risk factor, MDR, ESBL, piperacillin/tazobactam, multidrug-resistant, KT

Amira El-Far,1 Safia Samir,2 Eman El-Gebaly,3,4 Maysa Omar,1 Heba Dahroug,1 Ahmed El-Shenawy,1 Noha Salah Soliman,5 Doaa Gamal1 1Microbiology Department, Theodor Bilharz Research Institute (TBRI), Cairo, Egypt; 2Biochemistry and Molecular Biology Department, Theodor Bilharz Research Institute (TBRI), Cairo, Egypt; 3Microbiology and Immunology Department, Faculty of Pharmacy, Beni-Suif University, Beni-Suif, Egypt; 4Department of Microbiology and Immunology, Faculty of Pharmacy, October 6 University, Beni-Suef, 12585, Egypt; 5Clinical and Chemical Pathology Department, Faculty of Medicine, Cairo University, Cairo, EgyptCorrespondence: Safia SamirTheodor Bilharz Research Institute, 1 Cornish El-Nile Street, Warrak El-Hadar, Giza, 12411, Cairo, EgyptTel +20 235401019Fax +20 235408125Email safiasamir0812@gmail.com; s.elnaggar@tbri.gov.egBackground: Multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains of Pseudomonas aeruginosa are the leading cause of healthcare-associated infections worldwide.Objective: The aim was to identify the resistant phenotypes among P. aeruginosa and to characterize different aminoglycosides and carbapenem resistance genes as major mechanisms of resistance in these isolates, in Theodor Bilharz Research Institute (TBRI), a tertiary care hospital in Cairo, Egypt.Methods: During a period of 11 months, 42 P. aeruginosa clinical isolates were collected from the microbiology laboratory by routine culture. Antimicrobial sensitivity testing to the aminoglycosides gentamicin and amikacin, and other classes of antibiotics, was performed by a disk diffusion method. Isolates were tested for aminoglycoside resistance genes, aac(6ʹ)-lb, aac-(3)-lla, rmtB, rmtC, armA, rmtD, and rmtF, and carbapenemase resistance genes blaNDM, blaVIM, and blaIMP, using conventional PCR.Results: Thirty-three (78.5%) of the clinical P. aeruginosa isolates showed MDR and XDR phenotypes at 42.4% and 57.65%, respectively, and these were included in the study. Aminoglycoside resistance was found in 97%, whereas carbapenem resistance was found in 81% of the isolates phenotypically. Only 59.4% (19/26) of the aminoglycoside-resistant isolates harbored resistance genes; none of the amikacin-susceptible isolates harbored any of the tested aminoglycoside resistance genes. Aminoglycoside resistance genes rmtB, armA, aac(6ʹ)-lb, and rmtF were found at rates of 17/33 (51.5%), 3/33 (9%), 2/33 (6%), and 2/33 (6%), respectively, whereas rmtD, acc(3)-II, and rmtC were not detected. Only 40.7% (11/27) of the carbapenem-resistant isolates harbored resistance genes. Carbapenem resistance genes, blaNDM andblaVIM, were found at rates of 7/33 (21.2%) and 6/33 (18.1%), respectively, and blaIMP was not detected.Conclusion: Rates of MDR and XDR P. aeruginosa and resistance to aminoglycosides and carbapenems in our setting are high. Methyltransferases and metallo-beta-lactamases are the main mechanisms of resistance to aminoglycosides and carbapenems, respectively. The presence of blaNDM and rmtF in the strains confirms their rapid dissemination in the Egyptian environment.Keywords: aminoglycosides, carbapenems, antibiotic resistance, Pseudomonas aeruginosa, MDR, XDR, rmtB, rmtF, NDM

Shaheena Kousar,1 Nikhar Rehman,1 Asghar Javed,2 Aamir Hussain,3 Muhammad Naeem,1 Samra Masood,1 Hafiz Arslan Ali,1 Amna Manzoor,1 Aleem Ahmed Khan,1 Ahmed Akrem,1 Fatima Iqbal,4 Aqsa Zulfiqar,1 Muhammad Bilal Jamshaid,1 Muhammad Waqas,1 Asma Waseem,1 Muhammad Qamar Saeed1 1Dr. Ghulam Nabi Chaudhry Laboratory of Microbial Technologies, Department of Microbiology and Molecular Genetics, Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, Pakistan; 2Pathology Department, Nishtar Medical University, Multan, Pakistan; 3Institute of Basic Medical Sciences, Khyber Medical University, Peshawar, Pakistan; 4MINAR, Multan Institute of Nuclear Medicine and Radiotherapy, Nishter Medical University, Multan, PakistanCorrespondence: Muhammad Qamar SaeedDr. Ghulam Nabi Chaudhry Laboratory of Microbial Technologies, Department of Microbiology, Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, PakistanTel +92-333-0231222Email mqamarsaeed@bzu.edu.pkBackground: The drastic increase in use of antibiotics as a mandatory part of production in poultry and livestock has led to the development of bacterial resistance against antibiotics. The spread of resistant bacteria from poultry to humans increases the risk of treatment failure by antibiotics because of resistance genes transfer.Study Objective: The objective of the study was to estimate and compare the P. aeruginosa resistance profile collected from areas around the poultry farm premises and areas at least 500 meters away from the nearest poultry farm. We studied the effect of antibiotic usage in farms on the bacterial profile present in the upper layer of soil.Methodology: A total of 1,200 moist soil samples were collected from areas within a 25 meters range of poultry farms and areas that had no poultry farms in its 500 meters vicinity. P. aeruginosa was cultured and isolated. The antibiotic susceptibility profile was carried out by Kirby-Bauer disc diffusion method and results were analyzed according to CLSI guidelines. Statistical analysis was carried out to check the significance of results.Results: A total of 300 P. aeruginosa isolates were isolated, among which 140 isolates were isolated from areas around the poultry farm premises and had higher prevalence of antibiotic resistance. A total of 160 isolates were isolated from areas outside the poultry farm range. Resistance was not as high as in the isolates from around the farm. The ESBL production was higher in the isolates that were in close contact with the poultry farm as compared to the isolates away from the farm.Conclusion: Use of antibiotics in the poultry farm for production significantly increases the resistance in bacterial strains present in the upper layer of soil around the poultry farm within at least a 25 meter range.Keywords: antibiotic resistance, P. aeruginosa, poultry, MDR

Rashmi Karki,1,2,* Samir Lamichhane,3,* Buddha Bahadur Basnet,4 Anuja Dahal,5 Bal Krishna Awal,2,6 Shyam Kumar Mishra7,8,* 1Janamaitri Foundation Institute of Health Sciences, Tribhuvan University, Nepal; 2National Public Health Laboratory, Kathmandu, Nepal; 3Department of Pharmacology, Maharajgunj Medical Campus, Institute of Medicine, Tribhuvan University, Nepal; 4Faculty of Science, Nepal Academy of Science and Technology, Lalitpur, Nepal; 5Department of Pharmacy, Maharajgunj Medical Campus, Institute of Medicine, Tribhuvan University, Nepal; 6Shahid Dharmabhakta National Transplant Center, Bhaktapur, Nepal; 7School of Optometry and Vision Science, Faculty of Medicine and Health, UNSW, Sydney, NSW, Australia; 8Department of Microbiology, Maharajgunj Medical Campus, Institute of Medicine, Tribhuvan University, Nepal*These authors contributed equally to this workCorrespondence: Shyam Kumar MishraSchool of Optometry and Vision Science, UNSW, Sydney, NSW, AustraliaTel +61415538329Email s.mishrabaishnab@unsw.edu.auPurpose: Inappropriate use of broad-spectrum antibiotics contributes to the emergence of multidrug-resistant (MDR) bacteria. Finding novel antimicrobial agents and strategies based on synergistic combinations are essential to combat MDR infections. This study was designed to determine in vitro synergy of different antimicrobials against extensively drug-resistant (XDR) Gram-negative clinical isolates.Methods: A descriptive, cross-sectional study was conducted at Human Organ Transplant Center, Nepal, for five months. Clinical isolates were checked for their drug-resistance properties including extended-spectrum beta-lactamase- (ESBL-) and metallo-beta-lactamase- (MBL-) production. The XDR isolates were further tested for antimicrobial synergy, and the results were interpreted as synergistic, additive, indifferent or antagonistic determining fractional inhibitory concentration of the antibiotics.Results: Out of total 1155 clinical samples, 308 showed significant growth. Escherichia coli was the most common isolate (n=142) followed by Klebsiella pneumoniae, Acinetobacter calcoaceticus baumannii (Acb) complex, Pseudomonas aeruginosa and miscellaneous bacteria. Out of the culture positive isolates, 21.4% were MDR and 10.06% were XDR. The XDR population comprised K. pneumoniae (18.42%), E. coli (9.86%), Acb complex (7.41%) and P. aeruginosa (4.17%). Among the culture positive isolates, 4.5% and 5.8% were ESBL- and MBL-producers, respectively. Colistin, polymyxin B, and tigecycline were the antibiotics effective in majority of MDR isolates as compared to carbapenems. The combination of antibiotics – meropenem and colistin showed the highest proportion of “synergy” among all XDR E. coli whereas the combination of amikacin and colistin showed synergistic effect in XDR K. pneumoniae.Conclusion: A significant proportion of isolates were MDR among which a large fraction was XDR. The combination of meropenem, amikacin and colistin with one another in pair showed beneficial activity in vitro. Such combinations can be utilized as effective therapy for XDR infections. Further studies are required to confirm these findings, and accordingly treatment protocols should be developed in the management of such infections.Keywords: antimicrobial synergy, ESBL, MBL, MDR, XDR

Liang Du,1 Yu Zhang,1 Xintong Lv,2 Yuxin Duan,1 Xiaoyan Shi,2 Haoqiang Ji,1 Ruiheng Wu,1 Jia Xu,1 Xu Chen,1 Yang Gao,2 Xiwei Lu,2 Ling Zhou1 1School of Public Health, Dalian Medical University, Dalian, Liaoning, 116044, People’s Republic of China; 2Dalian Tuberculosis Hospital, Dalian, Liaoning, 116031, People’s Republic of ChinaCorrespondence: Xiwei Lu; Ling Zhou 9 West Section, Lvshun South Road, Dalian, Liaoning Province, People’s Republic of ChinaTel +86 411 8611 0368Email tb_luxiwei@126.com; zhouling0609@163.comPurpose: Multidrug-resistant tuberculosis (MDR-TB) is the cause of serious health and economic burdens worldwide. The present study aimed to explore the initial and acquired drug-resistance rates among TB patients from 2012 to 2019 in Dalian, China. The effectiveness of MDR-TB prevention and control strategies were then evaluated.Patients and Methods: Drug susceptibility testing (DST) was performed for 6429 diagnosed, culture-positive, Mycobacterium tuberculosis (MTB) strains, including 4661 new cases and 1768 previously treated cases. Descriptive statistics were employed to calculate the frequencies and percentages of TB strains, and the average annual growth rates (AAGRs) for each strain were calculated. The Chi-square test was applied to examine the significance of linear drug-resistance trends over time during the study period.Results: Over the eight-year study period, the percentages of both initial (from 9.01% to 4.82%) and acquired (from 40.85% to 9.09%) MDR-TB cases decreased significantly, AAGRs of 8.55% and 19.32%, respectively. Among new and previously treated TB patients, significant downtrends were observed for the rates of both initial and acquired MDR-TB among young and middle-aged individuals (P < 0.05). Additionally, among both new and previously treated TB patients, the percentages of individuals with drug resistance against isoniazid (INH), rifampicin (RFP), ofloxacin (OFX), and amikacin (AMK) decreased significantly (P < 0.05) from 2012 to 2019 in Dalian, China.Conclusion: The initial and acquired multidrug resistance rates exhibited significantly decreasing trends from 2012 to 2019, suggesting that MDR-TB prevalence has been controlled effectively in Dalian, China. The MDR-TB epidemic was reversed in the short term by establishing feasible strategies for detection, diagnosis, treatment, and infection control.Keywords: TB, initial, acquired, MDR, epidemic

Ahmed Morad Asaad,1 Shamshul Ansari,2 Soma Elsayed Ajlan,3 Samah Mohammed Awad4 1Department of Medical Microbiology and Immunology, Faculty of Medicine, Zagazig University, Zagazig, Egypt; 2Department of Microbiology and Immunology, Chitwan Medical College School of Medicine, Bharatpur, 44200, Nepal; 3Department of Medical Microbiology and Immunology, Faculty of Medicine, Menoufia University, Menoufia, Egypt; 4Department of Clinical Microbiology and Immunology, Molecular Microbiology in Liver and GIT, National Liver Institute, Menoufia, EgyptCorrespondence: Shamshul AnsariDepartment of Microbiology, Chitwan Medical College School of Medicine, Bharatpur, 44200, Chitwan, NepalTel +977-9840656933Email shamshulansari483@yahoo.comObjective: This cross-sectional study aims to determine the prevalence and associated risk factors of biofilm-producing A. baumannii nosocomial isolates from a tertiary care hospital, as well as to investigate any possible association of biofilm formation with the distribution of biofilm-related genotypes and antibiotic resistance phenotypes.Methods: A total of 94 non-duplicate A. baumannii nosocomial isolates were identified, their biofilm formation was quantitatively detected using the modified microtiter plate assay, and their susceptibilities to different antibiotics were determined using the breakpoint method. Isolates were then subjected to PCR assays targeting bap, ompA and blaPER-1 genes.Results: The majority (70.1%) of isolates were biofilm producers. The most prevalent biofilm gene was ompA (63.8%), followed by bap (13.8%) and blaPER-1 (10.6%). The presence of multi- and extensive-drug resistance (MDR and XDR) was significantly associated with biofilm producers (p = 0.017 and 0.002, respectively). The length of hospital stay (aOR= 0.023), the presence of ompA gene (aOR = 0.286) or bap gene (aOR = 0.346), ampicillin/sulbactam resistance (aOR = 1), and the presence of MDR (aOR = − 0.329) or XDR (aOR = − 0.252) were considered significant risk factors associated with biofilm-producing isolates.Conclusion: The high prevalence of biofilm-producing MDR and XDR nosocomial isolates in this study is worrisome and alarming. Characterization of risk factors could help control the continuous selection and transfer of this serious A. baumannii phenotype inside hospitals and improve the quality of patients’ care.Keywords: Acinetobacter, biofilm, MDR, XDR, nosocomial infection

Samar S Mabrouk,1 Ghada R Abdellatif,1 Mona R El-Ansary,2 Khaled M Aboshanab,3 Yasser M Ragab4 1Department of Microbiology, Faculty of Pharmacy, Ahram Canadian University (ACU), 6th of October, Giza, Egypt; 2Department of Biochemistry, Modern University for Technology and Information (MTI), Cairo, Egypt; 3Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University (ASU), Cairo, Egypt; 4Department of Microbiology & Immunology, Faculty of Pharmacy, Cairo University (CU), Cairo, EgyptCorrespondence: Khaled M AboshanabDepartment of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University, Organization of African Unity St., PO: 11566, Abbassia, Cairo, EgyptTel +20 225082595Fax +20 224051107Email aboshanab2012@pharma.asu.edu.egPurpose: This study aimed to detect the prevalence of carbapenemase producers (CPs) among extensive drug-resistant (XDR)-carbapenemase producing Gram-negative bacteria (GNB) recovered from various clinical specimens of hospitalized neutrophilic febrile patients in two major tertiary care hospitals in Egypt.Methods: Standard methods were used to evaluate the antimicrobial susceptibility of clinical isolates according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI). Phenotypic and genotypic analysis of CPs were carried out and statistically analyzed using standard methods.Results: Three hundred and forty-two GNB were obtained from 342 clinical specimens during the period of the study, where 162 (47%) were enterobacterial isolates, including, 63 (18.4%) Escherichia coli, 87 (25.4%) Klebsiella spp., 5 (1.46%) Enterobacter cloacae, 5 (1.46%) Salmonella spp. and 2 (0.6%) Proteus and 180 (53%) were non-fermentative bacilli including, 129 (37.7%), Acinetobacter baumannii, and 51 (14.9%), Pseudomonas spp. Out of the 342 GNB, 188 (54.9%) isolates were multi-drug resistant (MDR). Of these, 52 (27.6%) were XDR as well as CPs as confirmed phenotypically. The MIC of imipenem against the XDR GNB against showed either low (11 isolates; 21.1%; MIC range =4– 32 μg/mL) or high levels of resistance (41 isolates; 78.8%; MIC range = 64-≥ 1024). The most prevalent carbapenem resistance (CR) genes were blaKPC (63.5%) followed by blaOXA-48 (55.7%) and blaVIM (28.8%). No significant association could be observed between the MIC level and the presence of CR genes (P value > 0.05).Conclusion: High prevalence of MDR (54.9%) and XDR (27.6%) GNB pathogens associated with high levels of resistance to carbapenems were observed. All XDR GNB were CPs and tested positive for at least one of the CR genes. However, most of them (78.8%) showed a high level of CR (MIC range = 64-≥ 1024) with no significant association with the CR genes.Keywords: carbapenem resistance, carbapenemases, MDR, XDR, Gram-negative pathogens

Leila Ahmadian,1,2 Mohammad Reza Haghshenas,1,2 Bahman Mirzaei,3 Zahra Norouzi Bazgir,1,2 Hamid Reza Goli1,2 1Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran; 2Department of Medical Microbiology and Virology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran; 3Department of Medical Microbiology and Virology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, IranCorrespondence: Hamid Reza GoliDepartment of Medical Microbiology and Virology, Faculty of Medicine, Mazandaran University of Medical Sciences, Farah Abad Blv, Khazar Square, Sari, Mazandaran, IranTel +98 1133543081Fax +98 1133543249Email goli59@gmail.comPurpose: The integrons, as the mobile exogenous elements, play a prominent role in the spreading of antimicrobial resistance genes from Pseudomonas aeruginosa clinical isolates to other bacteria. This study aimed to investigate the frequency of class 1 integrons andresistance gene cassettes carrying by them in clinical isolates as well as multidrug resistant P. aeruginosa.Materials and Methods: A total of 100 clinical isolates of P. aeruginosa were collected from 5 hospitals in Mazandaran province, north Iran. The antibiotic susceptibility pattern of the isolates was evaluated using the disk agar diffusion method. Genomic DNAs were extracted and then the presence of class 1 integrons was detected by the PCR test. All PCR products of the positive isolates were sequenced for the detection of resistance gene cassettes by the Sanger method.Results: Forty-one percent of the clinical isolates were multi-drug resistant. Also, 42% of the isolates were contained class 1 integron, and 61.9% of the integron positive isolates were detected as MDR. We detected 10 different gene cassettes sizing from 0.6 to 3.5 kb in the present study. The sequencing analysis of the internal variable regions of the class 1 integrons showed that the 0.75 kb gene cassette (aadB) was the most frequent resistance gene (54.76%) among all clinical isolates, as well as the MDR isolates. Other resistance genes detected in this study were included: aadA6-orfD (35.71%), aacA4-blaOXA-10 (21.42%), aadB-aacA4-blaOXA-10 (19.04%), blaOXA-10-aacA4-VIM 1 (11.9%), aacA4-catB10 (7.14%), aacA5-aadA1-cmlA5 (7.14%), blaOXA31-aadA2 (4.76%), and aac(3)-Ic-aacA5-cmlA5 (4.76%). To the best of our knowledge, blaOXA-10-aacA4-VIM1 cassette array is detected for the first time in this study.Conclusion: The treatment of infections caused by P. aeruginosa in this region of Iran is a major problem due to the high prevalence of class 1 integrons. It seems that the high prescription of beta-lactams and aminoglycosides for the treatment of these infections may be replaced by other combination therapy stewardships.Keywords: Pseudomonas aeruginosa, class 1 integrons, MDR, resistance gene cassettes

Minmin Lin,1,* Yongqiang Yang,2– 4,* Yanxian Yang,2,3,* Guanping Chen,5 Ruowen He,2,3 Yiping Wu,2,3 Lan-Lan Zhong,2,3 Mohamed Abd El-Gawad El-Sayed Ahmed,2,3,6 Siyuan Feng,2,3 Cong Shen,2,3 Xin Wen,2,3 Jin Huang,1 Hongyu Li,7 Xiaobin Zheng,1 Guo-Bao Tian2,3 1Department of Respiratory Medicine, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai 519000, People’s Republic of China; 2Department of Microbiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People’s Republic of China; 3Key Laboratory of Tropical Diseases Control (Sun Yat-sen University), Ministry of Education, Guangzhou 510080, People’s Republic of China; 4School of Pharmaceutical Sciences (Shenzhen), Sun Yat-sen University, Guangzhou 510006, People’s Republic of China; 5Sun Yat-sen University School of Medicine, Guangzhou 510006, People’s Republic of China; 6Department of Microbiology and Immunology, Faculty of Pharmaceutical Sciences and Drug Manufacturing, Misr University for Science and Technology (MUST), Cairo, 6th of October City, Egypt; 7Department of Laboratory, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, People’s Republic of China*These authors contributed equally to this workCorrespondence: Guo-Bao Tian; Xiaobin Zheng Email tiangb@mail.sysu.edu.cn; zhxbin@mail.sysu.edu.cnBackground: Bloodstream infection (BSI) caused by carbapenem-resistant Enterobacteriaceae are potentially life-threatening related to poorer outcomes. Colistin is considered one of the last-resort treatments against human infections caused by multidrug-resistant (MDR) Gram-negative bacteria. Therefore, emergence of strains from the blood that co-harboring mcr and carbapenem resistance genes were considered as a serious problem.Purpose: In this study, two mcr-9-harboring MDR Enterobacter cloacae isolates BSI034 and BSI072 recovered from BSI patients were identified, one of which co-harbored mcr-9 and blaNDM-1. The genetic characteristics of the MDR plasmid needed to be clarified.Methods: S1-PFGE and Southern blotting were conducted to determine the location of mcr-9. Whole-genome sequencing was performed to obtain the complete genome and plasmid sequences. The resistome and virulence genes of the strains, accompanied by the genetic characteristics of mcr-9- and blaNDM-1-harboring plasmids, were analyzed.Results: Whole-genome sequencing showed that BSI034 harbored mcr-9-carrying IncHI2-type pBSI034-MCR9 and blaNDM-1-carrying IncX3-type pBSI034-NDM1. The 278,517 bp pBSI034-MCR9 carried mcr-9 along with the other 19 resistance genes. mcr-9 was flanked by IS 903B (1057 bp) and IS 26 (820 bp) in the same orientation. In addition to resistance genes, strain BSI034 also carried a chromosome-located Yersinia high-pathogenicity island, which harbored genes of yersiniabactin biosynthesis operon ybtSXQPAUTE, irp1/2, and fyuA.Conclusion: We described the complete genome and mcr-9/blaNDM-1-co-harboring plasmid of E. cloacae from a BSI patient. Notable differences were observed within mosaic modules between pBSI034-MCR9 and other mcr-9-harboring plasmids due to extensive recombination via horizontal gene transfer.Keywords: MDR, mcr-9, colistin, virulence genes

Elnaz Abbasi,1 Mahdieh Mondanizadeh,2 Alex van Belkum,3 Ehsanollah Ghaznavi-Rad4,5 1Department of Microbiology & Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran; 2Department of Biotechnology and Molecular Medicine, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran; 3Data Analytics Department, BioMérieux, La Balme les Grottes, France; 4Molecular and Medicine Research Center, Faculty of Medicine Arak University of Medical Sciences, Arak, Iran; 5Department of Medical Laboratory Sciences, Arak School of Paramedicine, Arak University of Medical Sciences, Arak, IranCorrespondence: Ehsanollah Ghaznavi-Rad Tel/Fax +98-8634173526Email ghaznaviehs@yahoo.comBackground: Diarrheagenic Escherichia coli (DEC) is a significant cause of gastroenteritis and a major public health problem. This study investigates the prevalence and the antibiotic resistance patterns of DEC that were isolated from infectious diarrhea samples of pediatric patients from central Iran.Patients and Methods: Pediatric diarrhea samples were collected from 230 pediatric patients visiting the hospital. E. coli pathotypes were diagnosed by using conventional culture methods and PCR. Antibiotic resistance profiles, the frequency of multi-drug resistance (MDR), and the phenotypic and genotypic characteristics of extended spectrum-β-lactamase (ESBL), AmpC and integron-associated genes were analyzed.Results: Of the 230 samples of infectious diarrhea, 91 (39.5%) produced E. coli isolates. Of these, 32 cases (35.1%) were identified as DEC by culture and PCR. The frequency of the E. coli pathotypes obtained was as follows: EAEC 11/32 (34.3%), EPEC 9/32 (28.1%), ETEC 6/32 (18.7%), EIEC 3/32 (9.3%), and EHEC 3/32 (9.3%). The antibiotic resistance rates were greater for nalidixic acid (30/32; 93.7%), ampicillin (29/32; 90.6%), and tetracycline (25/32; 78.1%) than for any of the other tested antibiotics. High levels of MDR (25/32; 78.1%) and the presence of ESBL (18/32; 56.2%) and AmpC (9/32; 28.1%) were observed in the DEC isolates. The isolates showed a higher frequency of the ESBL genes [blaTEM (18/18; 100%), blaCTX-M15 (17/18; 94.4%)], and AmpC [blaCIT (4/9; 44.4%) and blaDHA (4/9; 44.4%)] than of the other ESBL and AmpC genes.Conclusion: Compared to the previous study, DEC appeared to be the second-most abundant agent of diarrhea in pediatric patients after Campylobacter jejuni, with frequent MDR and ESBL presence.Keywords: diarrheagenic Escherichia coli pathotypes, pediatrics, diarrhea, antibiotic resistance, MDR, Iran

Noha A Hassuna, 1 Sahar A Mandour, 2 Ebtisam Samir Mohamed 1 1Medical Microbiology and Immunology, Faculty of Medicine, Minia University, Minia, Egypt; 2Microbiology and Immunology Department, Faculty of Pharmacy, Deraya University, Minia, EgyptCorrespondence: Noha A HassunaMedical Microbiology and Immunology, Faculty of Medicine, Minia University, Minia 61111, EgyptTel +20 862342813Email nohaanwar@mu.edu.egPurpose: Ventilator-associated pneumonia caused by Pseudomonas aeruginosa (P. aeruginosa) is a major health-care problem. In this study, we explored the epidemiology of virulence determinants among multi-drug-resistant (MDR) clinical P. aeruginosa isolates from hospitalized patients with ventilator-associated pneumonia in intensive care units in Upper Egypt.Patients and Methods: MDR P. aeruginosa isolates were screened for the presence of eight virulence factors and typed by ERIC-PCR.Results: A total of 39 clinical MDR isolates were selected out of 173 isolated P. aeruginosa showing a combination of adhesion and cytotoxicity virulence patterns, with the detection of aprA, exoU, exoS, lasB, algD, toxA in 74.3%, 58.9%, 46.1%, 41.2%, 30.7%, 20.5% of the isolates, respectively. The MDR isolates were grouped into 13 different virulence profiles according to the pattern of virulence gene distribution. exoU genotype was more predominant among the P. aeruginosa isolates with more than 48% of the isolates harboring this gene alone, 7% harboring both exoU and exoS and 43.5% harboring exoS gene. An intermediate degree of diversity was detected by ERIC-PCR typing where the isolates were clustered in 7 major groups, indicating possible cross-infection within the hospital.Conclusion: Our results highlight the increased frequency of virulent P. aeruginosa isolates with a shift to the more virulent cytotoxic exoU genotype. Further hospital infection-control measures are mandatory to control the hospital cross-transmission of these highly virulent isolates. This study could vastly be a help to develop efficient treatment policies against P. aeruginosa induced ventilator-associated pneumonia.Keywords: P. aeruginosa, virulence, exoU, MDR

Rehab M Abd El-Baky, 1, 2 Salwa M Masoud, 1 Doaa S Mohamed, 2 Nancy GFM Waly, 1 Engy A Shafik, 3 Dina A Mohareb, 4 Azza Elkady, 5 Mohamed M Elbadr, 6 Helal F Hetta 7, 8 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, Egypt; 3Department of Clinical Pathology, South Egypt Cancer Institute, Assiut University, Assiut, Egypt; 4Department of Clinical Pathology, Faculty of Medicine, Assiut University, Assiut, Egypt; 5Sohag General Hospital, Sohag, Egypt; 6Department of Pharmacology, Faculty of Medicine, Assiut University, Assiut, Egypt; 7Department of Medical Microbiology and Immunology, Faculty of Medicine, Assiut University, Assiut, Egypt; 8Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, OH, USACorrespondence: Helal F HettaDepartment of Internal Medicine, University of Cincinnati College of Medicine, PO Box 670595, Cincinnati, OH 45267-0595, USAEmail helal.hetta@uc.eduBackground and Aim: The emergence of colistin-resistant strains is considered a great threat for patients with severe infections. Here, we investigate the prevalence and some possible mechanisms of colistin resistance among multidrug-resistant (MDR) and extensively drug-resistant (XDR) Pseudomonas aeruginosa (P. aeruginosa).Methods: Antimicrobial susceptibility was performed using disc diffusion methods while colistin resistance was detected by agar dilution method. Possible mechanisms for colistin resistance were studied by detection of mcr-1 and mcr-2 genes by conventional PCR, detection of efflux mechanisms using Carbonyl Cyanide 3-Chlorophenylhydrazone (CCCP), studying outer membrane protein profile and Lipopolysaccharide (LPS) profile of resistant isolates.Results: It was found that MDR and XDR represented 96% and 87% of the isolated P. aeruginosa, respectively, and colistin resistance represented 21.3%. No isolates were positive for mcr-2 gene while 50% of colistin-resistant isolates were positive for mcr-1. Efflux mechanisms were detected in 3 isolates. Protein profile showed the presence of a band of 21.4 KDa in the resistant strains which may represent OprH while LPS profile showed differences among colistin-resistant mcr-1 negative strains, colistin-resistant mcr-1 positive strains and susceptible strains.Conclusion: The current study reports a high prevalence of colistin resistance and mcr-1 gene in P. aeruginosa strains isolated from Egypt that may result in untreatable infections. Our finding makes it urgent to avoid unnecessary clinical use of colistin.Keywords: Pseudomonas aeruginosa, colistin resistance, mcr-1, mcr-2, toxA gene, XDR, MDR

Majid Taati Moghadam,1 Nour Amirmozafari,1 Aref Shariati,1,2 Masoumeh Hallajzadeh,1 Shiva Mirkalantari,1 Amin Khoshbayan,1,2 Faramarz Masjedian Jazi1,3 1Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran; 2Student Research Committee, Iran University of Medical Sciences, Tehran, Iran; 3Microbial Biotechnology Research Center, Iran University of Medical Science, Tehran, IranCorrespondence: Faramarz Masjedian JaziDepartment of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, IranTel +989123905926Email Masjedian.f@iums.ac.irAbstract: Nowadays the most important problem in the treatment of bacterial infections is the appearance of MDR (multidrug-resistant), XDR (extensively drug-resistant) and PDR (pan drug-resistant) bacteria and the scarce prospects of producing new antibiotics. There is renewed interest in revisiting the use of bacteriophage to treat bacterial infections. The practice of phage therapy, the application of phages to treat bacterial infections, has been around for approximately a century. Phage therapy relies on using lytic bacteriophages and purified phage lytic proteins for treatment and lysis of bacteria at the site of infection. Current research indicates that phage therapy has the potential to be used as an alternative to antibiotic treatments. It is noteworthy that, whether phages are used on their own or combined with antibiotics, phages are still a promising agent to replace antibiotics. So, this review focuses on an understanding of challenges of MDR, XDR, and PDR bacteria and phages mechanism for treating bacterial infections and the most recent studies on potential phages, cocktails of phages, and enzymes of lytic phages in fighting these resistant bacteria.Keywords: bacteriophage, drug resistant, MDR, XDR, PDR

Arezoo Mirzaei, Mehri Habibi, Saeid Bouzari, Mohammad Reza Asadi Karam Department of Molecular Biology, Pasteur Institute of Iran, Tehran 13164, IranCorrespondence: Mehri Habibi; Mohammad Reza Asadi KaramDepartment of Molecular Biology, Pasteur Institute of Iran, No. 69, Pasteur Ave, Tehran 1316943551, IranTel +9821 66953311Fax +9821 66492619Email m_habibi1362@yahoo.com; m_asadi12@yahoo.comPurpose: Proteus mirabilis is one of the most important agents of urinary tract infection (UTI). As there are limited data abou the pathogenicity P. mirabilis isolated from Iran, we investigated the virulence characteristics and antibiotic resistance in the isolates. Finally, the genotypic patterns were evaluated by Pulse field gel electrophoresis (PFGE).Methods: A total of 110 isolates of P. mirabilis causing UTIs were isolated from patients in Tehran, Iran. The virulence characteristics and antimicrobial susceptibility were assayed using phenotypic methods. Extended-spectrum β-lactamases (ESBLs) production was assayed by the combination disk diffusion test (CDDT). Presence of virulence genes and antimicrobial-resistant genes was detected by Polymerase chain reaction (PCR). Finally, thirty-three isolates were selected for PFGE.Results: All isolates showed the ability of biofilm and hemolysin formation. Antibiotic resistance ranged from 59.1% about cotrimoxazole to 2.7% about amoxicillin-clavulanic acid. Sixteen (14.5%) of the isolates were classified as multi-drug resistant (MDR). All isolates amplified mrpH, mrpA, pmfA, ureG and hpmA genes. Furthermore, the prevalence of zapA, fliC, ptaA, and ucaA genes was 98.2%, 98.2%, 95.5%, and 95.5%, respectively. The prevalence of plasmid-mediated quinolone resistance (PMQR) genes was 4.5% and 0.9% for aac(6ʹ)-Ib-cr and qnrA, respectively. Twenty-eight pulsotypes were detected among the 33 isolates by PFGE that pulsotypes 1, 2 and 4 with two isolates and pulsotype 3 with three isolates were the most prevalent ones.Conclusion: It was found that the P. mirabilis isolates had high frequency of virulence factors. In addition, antibiotic resistance to some antibiotics and also production of ESBLs is alarming and shows the need for hygienic procedures to prevent the dissemination of antibiotic resistance. Although PFGE showed genetic diversity among the isolates, finding of several pulsotypes among the isolates should be considered an alarm to prevent these infections in hospital environments.Keywords: Proteus mirabilis, urinary tract infection, virulence characteristics, antibiotic resistance, MDR, PFGE

Xue Li,1,* Qi Luo,1,* Xinyu Yu,1 Yanling Zhang,1 Xiaoyue Cao,1 Dan Li2,3 1Non-Coding RNA and Drug Discovery Key Laboratory of Sichuan Province, Chengdu Medical College, Chengdu, Sichuan 610500, People’s Republic of China; 2School of Medical Laboratory Science, Chengdu Medical College, Chengdu, Sichuan 610500, People’s Republic of China; 3Sichuan Provincial Engineering Laboratory for Prevention and Control Technology of Veterinary Drug Residue in Animal-Origin Food, Chengdu Medical College, Chengdu, Sichuan 610500, People’s Republic of China*These authors contributed equally to this workCorrespondence: Dan LiSchool of Medical Laboratory Science, Chengdu Medical College, Chengdu, Sichuan 610500, People’s Republic of ChinaTel/Fax +86 28 62739520Email lidan0811@cmc.edu.cnBackground: Escherichia coli strains are the most commonly isolated bacteria in hospitals. The normally harmless commensal E. coli can become a highly adapted pathogen, capable of causing various diseases both in healthy and immunocompromised individuals, by acquiring a combination of mobile genetic elements. Our aim was to characterize E. coli strains from a hospital in western China to determine their virulence and antimicrobial resistance potential.Methods: A total of 97 E. coli clinical isolates were collected from the First Affiliated Hospital of Chengdu Medical College from 2015 to 2016. Microbiological methods, PCR, and antimicrobial susceptibility tests were used in this study.Results: The frequency of occurrence of the virulence genes fimC, irp2, fimH, fyuA, lpfA, hlyA, sat, and cnf1 in the E. coli isolates was 93.81, 92.78, 91.75, 84.54, 41.24, 32.99, 28.86, and 7.22%, respectively. Ninety-five (97.9%) isolates carried two or more different virulence genes. Of these, 44 (45.4%) isolates simultaneously harbored five virulence genes, 24 (24.7%) isolates harbored four virulence genes, and 17 (17.5%) isolates harbored six virulence genes. In addition, all E. coli isolates were multidrug resistant and had a high degree of antimicrobial resistance.Conclusion: These results indicate a high frequency of occurrence and heterogeneity of virulence gene profiles among clinical multidrug resistant E. coli isolates. Therefore, appropriate surveillance and control measures are essential to prevent the further spread of these isolates in hospitals.Keywords: Escherichia coli, clinical isolates, virulence genes, antimicrobial resistance, MDR

Pallam Gopichand,1 Girija Agarwal,2 Mailan Natarajan,1 Jharna Mandal,1 Surendran Deepanjali,3 Sreejith Parameswaran,4 LN Dorairajan51Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education & Research (JIPMER), Pondicherry, India; 2Jawaharlal Institute of Postgraduate Medical Education & Research (JIPMER), Pondicherry, India; 3Department of Medicine, Jawaharlal Institute of Postgraduate Medical Education & Research (JIPMER), Pondicherry, India; 4Department of Nephrology, Jawaharlal Institute of Postgraduate Medical Education & Research (JIPMER), Pondicherry, India; 5Department of Urology, Jawaharlal Institute of Postgraduate Medical Education & Research (JIPMER), Pondicherry, IndiaBackground: Rising rates of resistance to antimicrobial drugs among Enterobacteriaceae limit the choice of therapeutic agents to treat urinary tract infections. In this context we assessed the in-vitro effect of fosfomycin against extended-spectrum beta-lactamases, AmpC beta-lactamases and carbapenemase-producing strains of Escherichia coli, Klebsiella pneumoniae, Enterobacter spp, and Pseudomonas aeruginosa isolated from the patients with urinary tract infection (UTI) and also studied the effect of fosfomycin on their biofilm formation.Materials and methods: A total of 326 multidrug-resistant (MDR) isolates comprising of Escherichia coli, Klebsiella pneumoniae, Enterobacter spp, and Pseudomonas aeruginosa from the urine samples of the patients with a diagnosis of UTI were included in the study. MIC 50 and MIC 90 were detected by agar dilution method and the capacity to form biofilm in the presence of fosfomycin by these MDR isolates was assessed by the tissue culture plate method.Results: The MIC50 for meropenem (0.5 μgm/mL) and nitrofurantoin (32 μgm/mL) was within the susceptible range only for E. coli. Fosfomycin was the only antibiotic that inhibited 100% E.coli, 70% Klebsiella spp, and 50% Pseudomonas spp and 40% Enterobacter spp which included the extended-spectrum beta-lactamases producers. It showed a similar effect on carbapenemase producers and AmpC producers. Fosfomycin disrupted biofilm in 67% (n=141) E.coli, 74% (n=50) Klebsiella spp, 88% (n=27) Pseudomonas spp and 36% (n=23) Enterobacter spp at 24 hrs of incubation with a concentration of 2 fold dilution lower than that of the MIC.Conclusion : Fosfomycin showed a good inhibitory effect on the biofilms produced by the MDR organisms studied here.Keywords: fosfomycin, MDR, UTI, MIC, biofilm

Elnaz Abbasi,1,2 Hamid Abtahi,3 Alex van Belkum,4 Ehsanollah Ghaznavi-Rad1,31Department of Microbiology & Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran; 2Department of Microbiology & Immunology, Khomein University of Medical Sciences, Khomein, Iran; 3Molecular and Medicine Research Center, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran; 4Data Analytics Department, BioMérieux, La Balme les Grottes, FranceBackground: Shigella spp. are primary pathogens of diarrhea in children worldwide. Emergence of resistance to fluoroquinolones and third-generation cephalosporins is crucial in the management of pediatric shigellosis. We determined the prevalence and the antibiotic resistance patterns of Shigella species isolated from pediatric patients in central Iran.Materials and methods: Pediatric diarrhea samples (n=230) were cultured on MacConkey and XLD agar media and in GN broth. Genus-specific PCR for ipaH was also used for detection directly from fecal specimens. Antibiotic resistance and the frequency of ESBL and AmpC genes were determined.Results: Out of the 230 samples, 19 (8.2%) cases of Shigella spp. were identified using culture. Twenty-six samples were positive by PCR (11.3%), S. flexneri (4/19; 21%) and S. sonnei (15/19; 78.9%) being the most detected. The highest antibiotic resistance rates were found for cotrimoxazole (19/19; 100%), ampicillin (16/19; 84.2%), cefixime (13/19; 68.4%) and ceftriaxone (12/19; 63.1%). Ten cases showed phenotypic ESBL presence and all these strains were positive for blaTEM, blaCTX-M-1, and blaCTX-M-15. Three strains were AmpC positive, all of which harbored blaCMY-2 and two contained blaCIT. Of the 19 Shigella isolates 5 (26.3%), 2 (10.5%), and 1 (5.2%) were phenotypically resistant to nalidixic acid, ciprofloxacin, and norfloxacin, respectively. Class 1 integron was found in 18 (94.7%) isolates whereas class 2 integron was found in 19 (100%) strains.Conclusion: We found a considerable presence of Shigella species with elevated antibiotic resistance levels. In particular, the resistance to third-generation cephalosporins (ESBL) and ciprofloxacin must be taken seriously.Keywords: Shigella, dysentery, antibiotic resistance, MDR, integrons, Iran

Hong Yan,1,* Rufeng Xu,2,* Xiangrong Zhang,3 Qian Wang,1 Jing Pang,2 Xia Zhang,3 Xiaoai Chang,2 Yaqin Zhang2 1Department of Laboratory Medicine, Nanjing Chest Hospital, Nanjing 210029, China; 2Department of Biochemistry and Molecular Biology, Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, Jiangsu 210029, China; 3Nanjing Public Health Clinical Center, Nanjing, Jiangsu 211133,China *These authors contributed equally to this work Purpose: The aim of this paper was to identify differentially expressed long non-coding RNAs (lncRNAs) in peripheral blood mononuclear cells (PBMCs) influenced by the infection of multidrug-resistant tuberculosis (MDR-TB). Materials and methods: IncRNA and mRNA expression profiles in PBMCs derived from healthy controls (HCs) and individuals with MDR-TB and drug-sensitive tuberculosis (DS-TB) were analyzed and compared by microarray assay. Six lncRNAs were randomly selected for validation by using real-time quantitative polymerase chain reaction (RT-qPCR). The biological functions and signaling pathways affected by the differentially expressed mRNAs were investigated by using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway-based approaches. Results: Compared with the HC group, 1,429 lncRNAs (983 mRNAs) and 2,040 lncRNAs (1,407 mRNAs) were identified to be deregulated in the MDR-TB group and in the DS-TB group, respectively, and 1,511 lncRNAs and 1,047 mRNAs were identified to be differentially expressed in both MDR-TB and DS-TB groups. Between the three groups, 22 lncRNAs and 38 mRNAs were found deregulated. Most deregulated lncRNAs were from intergenic regions (~55% of the total), natural antisense to protein-coding loci (~32% of the total), or intronic antisense to protein-coding loci (~5% of the total). Significantly enriched signaling pathways regulated by the deregulated mRNAs were mainly associated with natural killer cell-mediated cytotoxicity, antigen processing and presentation, graft-vs-host disease, the transforming growth factor-β signaling pathway, and the Hippo signaling pathway. Conclusion: This study is the first to report differentially expressed lncRNAs in PBMCs in response to MDR-TB infection. It revealed that some lncRNAs might be associated with regulating host immune response to MDR-TB infection. Further elucidation of the potential of these deregulated lncRNAs in MDR-TB and its reactivation requires further study. Keywords: mycobacterium tuberculosis, MDR, molecular characterization, lncRNA, infection

Xiuhong Yuan,1 Taohua Liu,2 Di Wu,3 Qiquan Wan3 1Psychosomatic Health Institute, the Third Xiangya Hospital of Central South University, Changsha, China; 2Clinical Medicine, Xiangya School of Medicine, Central South University, Changsha, China; 3Department of Transplant Surgery, the Third Xiangya Hospital of Central South University, Changsha, China Background: Multiple drug resistant/extensively drug resistant (MDR/XDR) Gram-negative urinary tract infections (UTIs) represent a growing threat to kidney transplant recipients. This retrospective study aimed to assess the incidence and microbiological profile of MDR/XDR Gram-negative UTIs, to identify drug susceptibility of MDR/XDR bacteria, and to determine the potential risk factors for MDR/XDR UTIs in kidney recipients. Materials and methods: During the study period, 1569 patients underwent consecutive kidney transplantation in two transplantation centers. We studied the demographics, clinical characteristics, and urine culture data from kidney recipients with MDR/XDR Gram-negative UTIs, and verified the risk factors associated with MDR/XDR infections. Results: Eighty-one kidney recipients yielded 88 episodes of MDR/XDR Gram-negative UTIs with five patients (6.2%) succumbing to all-cause in-hospital mortality. The most frequently isolated bacterium was Escherichia coli (62.5%). Almost all MDR/XDR Gram-negative bacteria were resistant to first- and second-generation cephalosporin, and monocyclic beta-lactam. They were relatively sensitive to meropenem, amikacin, and tigecycline. As for the 12 XDR bacteria, all of them were resistant to meropenem and 25% of them were resistant to tigecycline. All XDR Acinetobacter baumannii and E. coli were susceptible to tigecycline. Nosocomial infection (odds ratio [OR] = 11.429, 95% CI = 1.311–99.625, P = 0.027) was the only independent predictor of MDR/XDR Gram-negative UTIs. Non-fermenting bacterial infection (OR = 20.161, 95% CI = 3.409–119.240, P = 0.001), polycystic kidney disease (OR = 39.871, 95% CI = 1.979–803.384, P = 0.016), and serum creatinine level > 1.5 mg/dL (OR = 8.688, 95% CI = 1.354–55.747, P = 0.023) were significantly different between XDR and MDR Gram-negative UTIs. Conclusion: Meropenem, amikacin, and/or tigecycline can be prescribed for MDR/XDR Gram-negative infections. Tigecycline can also be prescribed for XDR A. baumannii and E. coli. Nosocomial infection was a risk factor for MDR/XDR Gram-negative UTIs, while XDR UTIs were associated with non-fermenting bacterial infection, polycystic kidney disease, and impaired renal function. Keywords: kidney transplantation, bacteria, urinary tract infections, risk factors, MDR, XDR

Background: Pseudomonas aeruginosa is incriminated in septicemia, significant economic losses in the poultry production sector, and severe respiratory infections in humans. This study aimed to investigate the occurrence, opr L sequencing, antimicrobial resistance patterns, virulence-determinant, Quorum sensing, and antibiotic resistance genes of P. aeruginosa retrieved from broiler chickens., Methods: Two hundred samples were collected from 120 broiler chickens from broiler farms at Ismailia Governorate, Egypt. Consequently, the bacteriological examination was conducted and the obtained P. aeruginosa strains were tested for opr L gene sequencing, antibiogram, and PCR screening of virulence, Quorum sensing, and antibiotic resistance genes., Results: The overall prevalence of P. aeruginosa in the examined birds was 28.3%. The opr L gene sequence analysis underlined that the tested strain expressed a notable genetic identity with various P. aeruginosa strains isolated from different geographical areas in the USA, India, China, Chile, and Ghana. PCR evidenced that the obtained P. aeruginosa strains, carrying virulence-related genes: opr L, tox A, apr A, phz M, and exo S in a prevalence of 100%, 100%, 42.5%, 33.3%, and 25.9%, respectively. Moreover, the recovered P. aeruginosa strains possessed the Quorum sensing genes: las I, las R, rhl I, and rhl R in a prevalence of 85.2%, 85.2%, 81.5%, and 81.5%, respectively. Furthermore, 40.7% of the isolated P. aeruginosa were XDR to seven antimicrobial classes, possessing sul 1, bla TEM , tet A, bla CTX-M , bla OXA-1 , and aad A1 genes., Conclusion: As we can tell, this is the first report emphasizing the evolution of XDR  P. aeruginosa  strains from broiler chicken in Egypt, which is supposed to be a serious threat to public health. The emerging XDR P. aeruginosa in poultry frequently harbored the opr L, tox A, and apr A virulence genes, the las I, las R, rhl I, and rhl R Quorum sensing genes, and the sul 1, bla TEM , tet A, bla CTXM , bla OXA-1 , and aad A1 resistance genes., Competing Interests: The authors declare no conflicts of interest in this work., (© 2023 Algammal et al.)

Background: Klebsiella pneumoniae ( K. pneumoniae ) is one of the most important pathogens in nosocomial infections. It has resistance to most antibiotics, even carbapenem, resulting in restricted therapeutic options., Purpose: We tried to assess the antimicrobial resistance and virulence fitness of carbapenem-resistant K. pneumoniae (CRKP) in addition to their phenotypic and genotypic diversity., Materials and Methods: The conventional methods, automated Vitek-32 system, and antimicrobial susceptibility pattern were used to detect CRKP isolates. Virulence and resistance genes profiles were created by using PCR technique. The correlation analysis was done by using R-program., Results: The antimicrobial resistance profile for all our K. pneumoniae isolates was shocking as the MDR and CRKP were the most prominent phenotypes. Unfortunately, high degrees of heterogeneity among our CRKP isolates were recorded, as 97.5% of them were differentiated into different clusters. We found a negative correlation between the existence of virulence and antimicrobial resistance genes. In contrast to sputum and urine CRPK isolates, the blood isolates showed high antimicrobial resistance and low virulence fitness. Finally, K. pneumoniae creates several outbreaks and crises in Egypt owing to the highly heterogeneity and the wide spread of multidrug-resistant (MDR) and multi-virulent CRKP phenotypes., Conclusion: Our results are significant and alarming to health organizations throughout the world for the severity and heterogeneity of K. pneumoniae infections. Therefore, the traditional method for treatment of CRKP infections must be renewed. Additionally, the treatment protocols must be well correlated with the site of infections, phenotypes, and genotypes of CRKP strains., Competing Interests: There is no conflict of interest with any authors., (© 2022 Elmanakhly et al.)