Antibiotic-Resistance Genes in E. coli strains in GCC Countries: A Meta-Analysis.

Background: Antimicrobial resistance (AMR) in Escherichia coli is an alarming issue worldwide, including in the Gulf Cooperation Council (GCC) countries, yet the prevailing gene patterns have not recently been reviewed. This study was conducted to determine and report on the dominant E. coli antimicrobial resistant gene patterns in GCC countries. Method: A scoping review identified the predominant AMR genes in GCC countries: CTX M, TEM, SHV, NDM, OXA, and VIM genes. For the systematic review, two authors independently searched Scopus, PubMed, Google Scholar, Science Direct, and Web of Science for interventional, clinical, or observational studies on the chosen AMR-conferring genes in E. coli published from GCC countries between January 2013 and June 2019, when the last search was carried out. The search strategy followed the PRISMA guidelines. The risk of bias was assessed using a 6-item standardized checklist. Random-effects modeling was used for all analyses. Results: A total 32 studies were included in the final synthesis of evidence. Overall, CTX-M (53.8%) was the most prevalent gene in the region followed TEM (40.6%), NDM-1 (28.4%), OXA (24.3%), VIM (8.5%), and SHV (7.8%). Most included studies were from Saudi Arabia: CTX-M was again most common with a prevalence of 46.8% from 5442 isolates. Conclusion: The risk of bias analysis showed a mean quality score of 4.25 ± 0.75, indicating high-quality in studies included in this meta-analysis. This review found that CTX-M gene is the most common AMR-conferring gene in E. coli strains from most GCC countries.

Evaluation of Two Phenotypic Methods for the Detection of Plasmid-Mediated AmpC ß-Lactamases among Enterobacteriaceae Isolates.

Objectives AmpC ß-lactamases are cephalosporinases that confer resistance to cephalothin, cefazolin, cefoxitin, penicillin, and ß-lactamase inhibitor-ß-lactam combinations. Even though the AmpC resistance is reported, but the accurate occurrence of AmpC ß-lactamases in Enterobacteriaceae members is still unknown. Techniques to identify AmpC producers are still evolving but not yet optimized for the clinical laboratory. Here we aimed to compare the test performance of two different phenotypic methods, that is inhibitor-based assay using boronic acid and disk approximation test for AmpC detection in Enterobacteriaceae isolates from a tertiary hospital microbiology laboratory. Materials and Methods The study includes 137 nonrepeat Enterobacteriaceae strains. Bacterial isolates, that yielded a zone diameter of less than 18 mm for cefoxitin by disk diffusion method were considered potential AmpC producers and further confirmed by phenotype methods-inhibitor-based assay using boronic acid and disk approximation test. A multiplex polymerase chain reaction was used to detect the most common plasmid-mediated AmpC genes: ACC, FOX, MOX, DHA, CIT, and EBC. Results Of the 137 clinical isolates, 58 (42.33%) were cefoxitin resistant, while 53.4 and 18.9% of the cefoxitin-resistant isolates were positive by inhibitor-based assay and disk approximation test. Multiplex PCR detected 42 (30.6%) isolates with AmpC genes. Of the 42 isolates, the inhibitor-based assay detected 25 (59.5%) isolates, while the disk approximation test detected nine (21.4%) isolates. Conclusion Our findings suggest that inhibitor-based assay using boronic acid can be used for the detection of the isolates that harbor AmpC ß-lactamases. This method is cost-effective, simple to perform, and easy to interpret. Thus AmpC detection as a routine in clinical laboratories can help in appropriate therapeutic intervention and improved infection control.

Molecular detection of plasmid-derived AmpC ß-lactamase among clinical strains of Enterobacteriaceae in Bahrain.

BACKGROUND: Enterobacteriaceae with AmpC ß-lactamase are multidrug-resistant organisms and represent a significant challenge to patient care. This study aims to determine the prevalence of plasmid-derived AmpC ß-lactamase among extended spectrum ß-lactamases (ESBL)-producing Enterobacteriaceae strains in Bahrain. METHODS: It was a cross-sectional study. A total of 185 ESBL-producing Enterobacteriaceae isolates were recovered from clinically significant specimens from January 2018 to December 2019. The samples underwent initial screen for cefoxitin resistance by disc diffusion test and subsequent phenotypic confirmation of AmpC production with phenyl boronic acid assays as well as genotypic analysis by multiplex polymerase chain reactions for AmpC subtypes. Drug-resistant features of these clinical isolates were also examined. RESULTS: Twenty-nine ESBL-producing Enterobacteriaceae isolates were cefoxitin resistant. Phenotypic and genotypic analyses confirmed that 8 and 12 cefoxitin-resistant isolates are AmpC positive, respectively. These AmpC producers are multidrug resistant, and Escherichia coli is the dominant strain among them. CONCLUSIONS: Plasmid-mediated spread of AmpC is present in clinically relevant Enterobacteriaceae species in Bahrain. Rational antimicrobial therapy against these multidrug-resistant organisms and continued surveillance of antimicrobial resistance mechanisms among the clinical isolates are recommended for optimal patient care.

Identification of E484K and other novel SARS-COV-2 variants from the Kingdom of Bahrain.

The challenges imposed by the ongoing outbreak of severe acute respiratory syndrome coronavirus-2 affects every aspect of our modern world, ranging from our health to our socio-economic needs. Our existence highly depends on the vaccine's availability, which demands in-depth research of the available strains and their mutations. In this work, we have analyzed all the available SARS-COV2 genomes isolated from the Kingdom of Bahrain in terms of their variance and origin analysis. We have predicted various known and unique mutations in the SARS-COV2 isolated from Bahrain. The complexity of the phylogenetic tree and dot plot representation of the strains mentioned above with other isolates of Asia indicates the versatility and multiple origins of Bahrain's SARS-COV2 isolates. We have also identified two high impact spike mutations from these strains which increase the virulence of SARS-COV2. Our research could have a high impact on vaccine development and distinguishes the source of SARS-COV2 in the Kingdom of Bahrain.

Variant analysis of SARS-CoV-2 genomes in the Middle East.

BACKGROUND: Coronavirus (COVID-19) was introduced into society in late 2019 and has now reached over 88 million cases and 1.9 million deaths. The Middle East has a death toll of ~80,000 and over 35000 of these are in Iran, which has over 1.2 million confirmed cases. We expect that Iranian cases caused outbreaks in the neighbouring countries and that variant mapping and phylogenetic analysis can be used to prove this. We also aim to analyse the variants of severe acute respiratory syndrome coronavirus-2 (SARS -CoV-2) to characterise the common genome variants and provide useful data in the global effort to prevent further spread of COVID-19. METHODS: The approach uses bioinformatics approaches including multiple sequence alignment, variant calling and annotation and phylogenetic analysis to identify the genomic variants found in the region. The approach uses 122 samples from the 13 countries of the Middle East sourced from the Global Initiative on Sharing All Influenza Data (GISAID). FINDINGS: We identified 2200 distinct genome variants including 129 downstream gene variants, 298 frame shift variants, 789 missense variants, 1 start lost, 13 start gained, 1 stop lost, 249 synonymous variants and 720 upstream gene variants. The most common, high impact variants were 10818delTinsG, 2772delCinsC, 14159delCinsC and 2789delAinsA. These high impact variant ultimately results in 36 number of mutations on spike glycoprotein. Variant alignment and phylogenetic tree generation indicates that samples from Iran likely introduced COVID-19 to the rest of the Middle East. INTERPRETATION: The phylogenetic and variant analysis provides unique insight into mutation types in genomes. Initial introduction of COVID-19 was most likely due to Iranian transmission. Some countries show evidence of novel mutations and unique strains. Increased time in small populations is likely to contribute to more unique genomes. This study provides more in depth analysis of the variants affecting in the region than any other study.

Student Perception of Microbiology Laboratory Skills Learning Through a Problem-Based Learning Curriculum: Arabian Gulf University Experience.

INTRODUCTION: The curriculum at medical school at Arabian Gulf University is centered on small group learning and real-life problems provided to students and guiding students to learn actively. In microbiology, laboratory skills are taught in an innovative manner using mini cases and different lab sessions and are integrated with other basic sciences. This article describes the format and pattern of laboratory skills sessions conducted using PBL methods at Arabian Gulf University and discusses the perception of students towards PBL in laboratory skill learning and way forward for the same. METHODS: The study sample size was 110. The students' perception of the laboratory skills teaching methods was assessed through an exit survey at the end of each session. A semi-structured self-administered survey instrument was prepared, and the questions were arranged in two sessions and focused on identifying the relevance, timing, strengths, and weaknesses of the teaching method and recommendations to improve the same. RESULTS: We observed that more than 50% of the participants agreed or strongly agreed that the time given for PBL was adequate, topics discussed were relevant, presentations were clear, pre-session briefing and Case-Based Studies (team-based learning (TBL)) helped in their learning. The participants identified the demonstration of experiments and hands-on experience provided in the laboratory were most helpful. When enquired about the difficulty, among 48% of the participants, 80% observed that the slides used in the learning/teaching were lengthy. CONCLUSION: The use of PBL in a lab setting promotes active learning. In the heart of PBL, TBL is a powerful tool in the educational process offering the students deep comprehension and allowing them to gain practical and intellectual skills.

The Curriculum at the College of Medicine and Medical Sciences at Arabian Gulf University: A Way Forward to Meet the Future Medical Education Needs.

Arabian Gulf University (AGU) follows a curriculum based on Problem Based Learning (PBL). PBL is a learner-centered approach that empowers students for life-long learning. Students are taught through problems that are designed based on global health problems customized to the local needs. The classroom teaching is complemented through adjunct programs like community health activities and professional skills program. Medical education aims to meet the changing needs of society. Demographics, disease epidemiology and healthcare needs of the gulf countries have changed over 38 years since the inception of AGU. To keep pace with the changing demands, it is imperative that the curriculum is reviewed in the light of advances in technology and newer techniques of medical education.In the present article the curriculum at AGU is reviewed based on the predictors for future medical education and alternative teaching methods that can be integrated to optimize the student outputs are explored.

Detection of Overexpression of Efflux Pump Expression in Fluoroquinolone-Resistant Pseudomonas aeruginosa Isolates.

CONTEXT: Fluoroquinolones are the most effective antibiotics against Pseudomonas aeruginosa; many strains, however, have shown resistance due to mutations in DNA gyrase, topoisomerase IV, or in the efflux pumps. Little is known about P. aeruginosa efflux pump resistance mechanisms in the Kingdom of Bahrain. AIM: The aim was to study efflux pump-mediated fluoroquinolone resistance among P. aeruginosa isolates using phenotypic (E-test and agar dilution) and genotypic (real-time-polymerase chain reaction [RT-PCR]) methods. MATERIALS AND METHODS: Fifty ciprofloxacin-resistant P. aeruginosa isolates were included in this study. Genus and species of P. aeruginosa were confirmed by conventional PCR. The minimum inhibitory concentration (MIC) of ciprofloxacin with and without carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was determined by E-test and agar dilution test. The overexpression of genes MexB, MexD, MexF, and MexY was measured by RT-PCR. RESULTS: All isolates were confirmed as P. aeruginosa. Among the fifty isolates, four showed reduction in ciprofloxacin MIC after addition of CCCP. These four isolates showed upregulation of expression of at least one of the four genes by RT-PCR. The mean gene expression of MexB, MexD, MexF, and MexY increased by 1.6, 4.65, 3.4, and 3.68-fold, respectively. CONCLUSION: The results demonstrate the presence and type of efflux pump overexpression, mandating for large multicentric studies.

Detection of VIM and NDM-1 metallo-beta-lactamase genes in carbapenem-resistant Pseudomonas aeruginosa clinical strains in Bahrain.

INTRODUCTION: Carbapenem-resistant Pseudomonas aeruginosa has emerged as a life-threatening infectious agent worldwide. Carbapenemase genes are reported to be some of the most common mechanisms for carbapenem resistance in P. aeruginosa. No reports are available from the Kingdom of Bahrain about carbapenem resistance and the underlying cause. In this study, we determined to study the presence of the metallo-beta-lactamase (MßL) genes of VIM family and NDM-1 in carbapenem-resistant P. aeruginosa strains. METHODOLOGY: Fifty carbapenem-resistant P. aeruginosa isolates were obtained from three main hospitals of Bahrain. They were subjected to antimicrobial susceptibility testing by disc diffusion test. Subsequently, MßL was detected by imipenem-ethylene diamine tetraacetic acid (EDTA) combined disc test and conventional polymerase chain reaction. RESULTS: Among 50 P. aeruginosa strains, 40 (80%) were imipenem resistant. Among the 40 imipenem-resistant strains, 35 (87.5%) strains were positive for the imipenem-EDTA combined disc test, and 21 (52%) were carrying MßL genes. Nineteen (47.5%) strains were positive for the VIM gene; one (2.5%) strain was carrying the NDM-1 gene, while one strain was carrying both the VIM and NDM-1 genes. None of the imipenem sensitive strains carried the VIM or NDM-1 gene. CONCLUSION: This is the first study to report the presence of the VIM family gene and NDM-1 genes in imipenem-resistant P. aeruginosa isolates in the Kingdom of Bahrain. The study also confirms the multiple drug resistance by the MßL strains, attention should therefore from now on, be focused on prevention of further spread of such isolates by firm infection control measures, and to reduce its threat to public health.

Characterization of cephalosporin-resistant clinical Enterobacteriaceae for CTX-M ESBLs in Bahrain.

OBJECTIVE: To detect the presence of specific CTX-M class of extended spectyum ß-lactamases (ESBLs) in a collection of cephalosporin-resistant Enterobacteriaceae isolates from Bahrain. METHODS: A subset of 80 cephalosporin-resistant Enterobacteriaceae collected from Salmaniya Medical Complex, Bahrain, were characterized further for the presence of specific genogroups of CTX-M ß-lactamases by multiplex- and monoplex- PCRs. The primers used for the multiplex and monoplex PCRs were of genogroups- 1, 2, 8, 9 and 25. Sequencing of the representative isolates was performed to find the circulating CTX-M-types. RESULTS: A total of 93.8% (75/80) isolates showed the amplicons corresponding to any of the genogroups (1, 2, 8, 9, 25) and the remaining 6.2% isolates turned out negative in multiplex PCR. Some of the isolates demonstrated multiple bands corresponding to the sizes of different genogroups. Further confirmation with respective monoplex PCR on these 75 isolates demonstrated that 93.3% (70/75) harbored CTX-M genogroup-1 and 6.7% (5/75) harbored genogroup-9. We did not find the presence of genogroups 2, 8, and 25 in these isolates by monoplex PCR. Sequencing results of genogroup-1 isolates demonstrated the presence of CTX-M-15-like ESBL, however, discrepant results were noticed in genogroup-9 isolates, sequencing showed them as CTX-M-55-like ESBL. CONCLUSIONS: This is the first report from Bahrain characterizing the CTX-M genogroups of ESBLs and reporting the emergence of blaCTX-M-55-like gene in this region.

vacA genotypes in Helicobacter pylori strains isolated from patients with and without duodenal ulcer in Bahrain.

BACKGROUND: The vacuolating cytotoxin and the cytotoxinassociated protein, encoded by vacA and cagA, respectively, are important virulence determinants of Helicobacter pylori. OBJECTIVE: The aim of this study was to perform vacA genotyping and evaluate its association with cagA genotype and clinical outcome. METHODS: One hundred and twenty H. pylori strains were isolated from dyspeptic patients (29 with peptic ulcer, 91 with non-ulcer dyspepsia). Genotype was determined by PCR. RESULTS: Seventy-nine (66%) of 120 strains had the vacA signal sequence genotype s1 and 41 (34%) had the type s2. The vacA middle-region types m1 and m2 were detected in 56 (47%) and 64 (53%) strains, respectively. The combinations s1-m1 (n=56 [47%] and s2-m2 (41 [34%]) occurred more frequently than s1-m2 (23 [19.2%]; p=0.001). No strain with the combination s2-m1 was found. All patients with peptic ulcers harbored type s1 strains compared to 75 (82.4%) of 91 patients with non-ulcer dyspepsia (p=0.01). The vacA genotype s1 was associated with the presence of cagA (p <0.0001). The cagA gene was detectable in 38 (31.6%) of 120 isolates and present in all 29 patients with ulcer compared to nine of 91 with non-ulcer dyspepsia (p <0.001). CONCLUSION: Helicobacter pylori strains of vacA type s1 and the combination of s1-m1 were associated with peptic ulceration and the presence of cagA gene.

Prevalence of extended-spectrum beta-lactamase-producing Enterobacteriaceae in Bahrain.

OBJECTIVES: To determine the occurrence of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in Bahrain. METHODS: Retrospective analysis of records (January 2005-December 2006) at the Microbiology Laboratory of the Salmaniya Medical Complex, Bahrain which is the major national diagnostic laboratory. RESULTS: Out of a total of 11,886 member of family of Enterobacteriaceae isolated, 2695 (22.6%) were ESBL producers. Majority of ESBL isolates were from inpatients (n=2363; 87.7%). Escherichia coli (52.2%) and Klebsiella pneumoniae (24.3%) were predominant and distributed comparatively in the hospital wards while Proteus spp. (17.6%) was predominant in medical wards. Urine was the major source (52.2%) with low occurrence in blood cultures. No carbapenem resistant isolates was identified but resistance to three classes of antibiotics was exhibited by >25% of the isolated ESBL strains. Nitrofurantoin resistance was identified in 38.2% of urinary isolates. CONCLUSION: This is the first report from Bahrain and it indicates that the prevalence of ESBL-producing isolates is high. Carbapenems were the most active drug against the ESBL-producing isolates. We recommend strict infection control to prevent trafficking into the community.