The Rising Tide of Antibiotic Resistance: A Study on Extended-Spectrum Beta-Lactamase and Carbapenem-Resistant Escherichia coli and Klebsiella pneumoniae.

BACKGROUND: The global spread of extended-spectrum beta-lactamase (ESBL)-producing and carbapenem-resistant Enterobacterales (CRE) poses a significant concern. Acquisition of antimicrobial resistance genes leads to resistance against several antibiotics, limiting treatment options. We aimed to study ESBL-producing and CRE transmission in clinical settings. METHODS: From clinical samples, 227 ESBL-producing and CRE isolates were obtained. The isolates were cultured on bacterial media and confirmed by VITEK 2. Antibiograms were tested against several antibiotics using VITEK 2. The acquired resistance genes were identified by PCR. RESULTS: Of the 227 clinical isolates, 145 (63.8%) were Klebsiella pneumoniae and 82 (36.1%) were Escherichia coli; 76 (33.4%) isolates were detected in urine, 57 (25.1%) in pus swabs, and 53 (23.3%) in blood samples. A total of 58 (70.7%) ESBL-producing E. coli were resistant to beta-lactams, except for carbapenems, and 17.2% were amikacin-resistant; 29.2% of E. coli isolates were resistant to carbapenems. A total of 106 (73.1%) ESBL-producing K. pneumoniae were resistant to all beta-lactams, except for carbapenems, and 66.9% to ciprofloxacin; 38 (26.2%) K. pneumoniae were resistant to carbapenems. Colistin emerged as the most effective antibiotic against both bacterial types. Twelve (20.6%) E. coli isolates were positive for blaCTX-M, 11 (18.9%) for blaTEM, and 8 (33.3%) for blaNDM. Forty-six (52.3%) K. pneumoniae isolates had blaCTX-M, 27 (18.6%) blaTEM, and 26 (68.4%) blaNDM. CONCLUSION: This study found a high prevalence of drug-resistant ESBL-producing and CRE, highlighting the need for targeted antibiotic use to combat resistance.

MicroRNA-155, a double-blade sword regulator of innate tuberculosis immunity.

Tuberculosis (TB) is a chronic, life-threatening disease caused by unusual facultative intracellular bacteria, Mycobacterium tuberculosis. This bacterium has unique resistance to many antimicrobial agents and has become a major global health concern due to emerging multidrug-resistant strains. Additionally, it has developed multiple schemes to exploit host immune signaling and establish long-term survival within host tissues. Thus, understanding the pathways that govern the crosstalk between the bacterium and the immune system could provide a new avenue for therapeutic interventions. MicroRNAs (miRs) are short, noncoding, and regulator RNA molecules that control the expression of cellular genes by targeting their mRNAs post-transcriptionally. MiR-155 is one of the most crucial miR in shaping the host immune defenses against M. tuberculosis. MiR-155 is remarkably downregulated in patients with clear clinical TB symptoms in comparison with latently infected patients and/or healthy individuals, thereby implicating its role in controlling M. tuberculosis infection. However, functional probing of miR-155 suggests dual effects in regulating the host's innate defenses in response to mycobacterial infection. This review provides comprehensive knowledge and future perspectives regarding complex signaling pathways that mediated miR-155 expression during M. tuberculosis infections. Moreover, miR-155-targeting signaling orchestrates inflammatory mediators' production, apoptosis, and autophagy.

Molecular characterization and antibiogram of the carbapenemase gene variants in clinical strains of Pseudomonas aeruginosa.

BACKGROUND: Carbapenemase-producing Pseudomonas aeruginosa (CPPA) is a substantial clinical concern because it jeopardizes therapeutic choices. This study characterizes the gene variants of CPPA and report its antibiogram. METHODS: CPPA was isolated prospectively from diverse clinical sources in a tertiary care setting using a routine microbiological approach. Carbapenem-resistant P. aeruginosa strains were phenotypically identified using the modified carbapenem inactivation (mCIM) method. Minimum inhibitory concentration (MIC) breakpoints of several antibacterial drug groups were determined using broth microdilution methods and the MicroScan WalkAway plus system. Carbapenemase gene variants blaNDM, blaVIM, blaOXA,blaGES, and blaIMP were amplified using polymerase chain reaction (PCR), and the purified gene products were sequenced. RESULTS: Seventy-one P. aeruginosa-infected cases were found, with 47 (66.2%) carrying CPPA; 46.8% of the latter were significantly associated with intensive care units (p = 0.03). CPPA was frequently detected in wound swabs (13; 27.7%), sputum (11; 23.4%), and blood (9; 19.1%). All strains were multidrug-resistant (MDR), and several were extensively drug-resistant. MIC50 and MIC90 breakpoints of all antibiotics, except colistin, were within the resistance range. MIC90 breakpoints of aztreonam, amikacin, cefepime, and piperacillin-tazobactam were > 512 µg/mL. The multiple antibiotic resistance index (MARI) was remarkably high, with a range of 0.38-0.92. The most commonly detected carbapenemase genes were blaVIM (74%), blaNDM-1 (19%), blaOXA-23 (14.9%), and blaGES (10.6%), while 12 of 47 strains co-harbored different combinations of carbapenemase gene variants. CONCLUSION: A large proportion of CPPA strains carried the blaVIM gene variant, indicating intimidating health problems and emphasizing the need for extensive surveillance and antibiotic stewardship.

Adenosine A2A receptor as a potential target for improving cancer immunotherapy.

The adenosine nucleoside performs a wide range of actions on various human tissues by activating four cell surface receptors. Adenosine A2A receptors (A2ARs) are widely expressed in the striatum, olfactory bulb, platelets, leukocytes, spleen, and thymus. They promote vasodilatation, platelet antiaggregatory effect, protection from ischemic damage, and regulation of sensorimotor neurons in basal ganglia. Adenosine signaling plays a vital part in modulating in vivo pathophysiological responses. A2ARs are potent negative regulators of the antitumor and proinflammatory actions of activated T cells. This axis offers several therapeutic targets, the most important of which are A2ARs, HIF-1α, and CD39/CD73. Downregulation of this axis increases the effectiveness of modern immunotherapeutic approaches against cancer, such as αCTLA-4/αPD-1. These discoveries have led to a promising novel role of antagonists of A2AR in blocking angiogenesis in immunotherapy of cancer. A small molecule, AZD4635, strongly inhibits A2AR, lowering cancer volume and increasing anticancer immunity. Deletion of A2AR with CRISPR/Cas9 in both human and murine CAR T cells produces a substantial increase in the efficiency of these cells. This review asserts that inhibition of the adenosinergic pathway can boost antitumor immunity, and this axis should be a target for future immunotherapeutic strategies.

pH Responsive Abelmoschus esculentus Mucilage and Administration of Methotrexate: In-Vitro Antitumor and In-Vivo Toxicity Evaluation.

The rapid progression in biomaterial nanotechnology apprehends the potential of non-toxic and potent polysaccharide delivery modules to overcome oral chemotherapeutic challenges. The present study is aimed to design, fabricate and characterize polysaccharide nanoparticles for methotrexate (MTX) delivery. The nanoparticles (NPs) were prepared by Abelmoschus esculentus mucilage (AEM) and chitosan (CS) by the modified coacervation method, followed by ultra-sonification. The NPs showed much better pharmaceutical properties with a spherical shape and smooth surface of 213.4-254.2 nm with PDI ranging between 0.279-0.485 size with entrapment efficiency varying from 42.08 ± 1.2 to 72.23 ± 2.0. The results revealed NPs to possess positive zeta potential and a low polydispersity index (PDI). The in-vitro drug release showed a sustained release of the drug up to 32 h with pH-dependence. Blank AEM -CS NPs showed no in-vivo toxicity for a time duration of 14 days, accompanied by high cytotoxic effects of optimized MTX loaded NPs against MCF-7 and MD-MBA231 cells by MTT assay. In conclusion, the findings advocated the therapeutic potential of AEM/CS NPs as an efficacious tool, offering a new perspective for pH-responsive routing of anticancer drugs with tumor cells as a target.

Synthesis and Characterization of Dihydrouracil Analogs Utilizing Biginelli Hybrids.

Dihydrouracil presents a crucial intermediate in the catabolism of uracil. The vital importance of uracil and its nucleoside, uridine, encourages scientists to synthesize novel dihydrouracils. In this paper, we present an innovative, fast, and effective method for the synthesis of dihydrouracils. Hence, under mild conditions, 3-chloroperbenzoic acid was used to cleave the carbon-sulfur bond of the Biginelli hybrids 5,6-dihydropyrimidin-4(3H)-ones. This approach led to thirteen novel dihydrouracils synthesized in moderate-to-high yields (32-99%).

Design and Synthesis of Some New Furan-Based Derivatives and Evaluation of In Vitro Cytotoxic Activity.

New furan-based derivatives have been, designed, synthesized, and evaluated for their cytotoxic and tubulin polymerization inhibitory activities. DNA flow cytometric study of pyridine carbohydrazide 4 and N-phenyl triazinone 7 demonstrated G2/M phase cell cycle disruptions. Accumulation of cells in the pre-G1 phase and positive annexin V/PI staining, which may be caused by degeneration or fragmentation of the genetic components, suggested that cell death occurs via an apoptotic cascade. Furthermore, compounds 4 and 7 had a strong pro-apoptotic impact through inducing the intrinsic mitochondrial mechanism of apoptosis. This mechanistic route was verified by an ELISA experiment that indicated a considerable rise in the levels of p53 and Bax and a drop in the level of Bcl-2 when compared with the control.

Estimation of serum iron, serum lipids and serum liver enzymes in celiac disease patients of Saudi Arabia.

Objectives: To evaluate the serum biochemical levels in celiac disease (CD) patients. Methods: It was a cross-sectional study carried out on 70 subjects, including 40 patients with CD and 30 healthy controls. This study was conducted at Jouf University from November, 2020 to October, 2021. The collected blood specimens were used to perform serum iron, serum lipids, liver enzymes, and human tissue transglutaminase IgA antibodies (anti-HTTG). The hematological parameters including hematocrit and MCV were determined to establish the diagnosis of iron deficiency. Results: Serum iron was significantly lower in patients as compared to the controls. Serum iron, serum HDL, blood hematocrit and MCV were significantly lower in patients than in controls (p = 0.000). Serum levels of liver enzymes (ALT and AST) and serum human tissue transglutaminase antibodies (anti-HTTG) were significantly higher in patients than in controls (p = 0.000). The correlation studies established the negative correlation of anti-HTTG IgA with serum iron (r = -0.991, p = 0.000), hematocrit (r = -0.967, p = 0.000) and MCV (r = -0.946, p = 0.000) in patients. Conclusion: The serum iron was remarkably reduced in CD patients. A negative correlation was found between anti-HTTG IgA and serum iron, while a positive serum iron was correlated with hematocrit and MCV in CD patients.

Determination of serum tumor necrosis factor-alpha (TNF-α) levels in metabolic syndrome patients from Saudi population.

OBJECTIVES: To detect the relationship between serum tumor necrosis factor-alpha (TNF-α) and metabolic syndrome (MetS) components in patients of the Saudi population. METHODS: This cross-sectional study was carried out at Jouf University Saudi Arabia from September 2019 to August 2020 and comprised of 183 individuals (91 cases and 92 controls). The blood samples were drawn from the patients visiting two tertiary care settings of Al Jouf province. Biochemical analysis was conducted on various instruments, and serum TNF-α was measured by the ELISA method. RESULTS: The levels of serum glucose fasting, lipid profile, HbA1c and body mass index (BMI) were raised significantly in cases of MetS than controls (p = 0.001). Serum TNF-α was significantly higher in patients (58.04 ± 15.44) than controls (48.81 ± 10.30). It was correlated with the BMI, blood HbA1c, serum fasting glucose (SFG) and serum high density lipoprotein (HDL). The weak positive correlation was found with BMI (r = 0.18; p = 0.01), serum glucose (r = 0.21; p = 0.007) and HbA1c (r = 0.14; p = 0.04), but found negative association with serum HDL (r = -0.18; p = 0.01). CONCLUSION: The serum TNF-α was raised in metabolic syndrome patients than the healthy controls. It was positively associated with high BMI, serum fasting glucose, and HbA1c and found linked and negatively linked to low HDL levels in MetS patients in the Saudi population.

Antibacterial efficacy of silver nanoparticles against metallo-ß-lactamase (blaNDM, blaVIM, blaOXA) producing clinically isolated Pseudomonas aeruginosa.

Carbapenem resistance in Pseudomonas aeruginosa is a major concern in the public health sector, primarily in developing countries such as Pakistan. Therefore, novel approaches such as Silver nanoparticles (AgNPs) can be used to address emerging concerns. Clinical isolates (n=200) were reconfirmed using selective media and API 20NE kit. The antibiogram was determined according to the CLSI 2016 guidelines. Molecular detection was carried out by PCR. Antibacterial activity in AgNPs was achieved by dilution method. Of 200 P. aeruginosa, mostly (n=82; 41%) were isolated from pus samples. Of 110 MDR P. aeruginosa, 70 (63%) were carbapenemase and 58 (52%) were MBL producers. Antimicrobial profile of MBL producing P. aeruginosa reported that all isolates were resistant to ß-lactams, and 89% to levofloxacin and ciprofloxacin except colistin. Of 25 (35.7%) blaNDM producing P. aeruginosa, 12 isolates (48%) had MIC 16µg/mL to imipenem. Of 23 (32%) blaVIM producing P. aeruginosa, 12 (52%) contained MIC 16µg/mL to imipenem. However, 12 (17.1%) blaOXA-48 producing P. aeruginosa, 4 (33%) contained MIC 16µg/mL to imipenem. In vitro AgNPs activity inhibited and killed MBL producing isolates at 1 mg/mL and 2 mg/mL, respectively. AgNPs may be used as an alternative therapy followed by multiple clinical trials.

Covid-19 Pandemic: Through the Lens of Science, a Painstaking Review.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has imperiled human lives and global infrastructure since the emergence of SARS-CoV-2 in China. The current review meticulously summarizes the COVID-19 pandemic situation through the lens of science from the inception of the outbreak to the current progression, which is valuable to mitigate the current pandemic situation. METHODS: We reviewed all the relevant literature available on PubMed, Web of Sciences, Google Scholar, and World Health Organization (WHO) website related to COVID-19 from the inception of the outbreak to 18 June 2020. We selected ninety different scientific studies and reports to compile the current review. RESULTS: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a betacoronavirus with four major structural proteins encoded by S, M, E, and N genes and distinct in morphology. The potential provenance of SARS-Cov-2 is zoonotic, and it binds to the host cell receptors by spike protein. The SARS-CoV-2 infectious cycle carries on through direct contact, air, inanimate objects, and contaminated surfaces. The reproductive number (R0) of SARS-CoV-2 is 2 to 3.5, representing that one infected patient can spread this virus to two to three people. An expeditious laboratory diagnosis has a pivotal role in patient management and prevention. Due to the lack of definitive treatment, symptomatic medication regimen and supportive organ therapies are adapted for debilitated patients. CONCLUSIONS: Nucleoside analogs and protease-inhibitors have approved to attenuate the viral infection until the discovery of a specific drug. The other treatment strategies comprise antimalarial drugs, monoclonal antibodies, and glucocorticoids. The use of alcoholic scrubs, sodium hypochlorite, masks, social distancing, and quarantine the affected individual is inevitable to eradicate the infection vector and to break the transmission path.

Molecular Analysis of the Antibiotic Resistant NDM-1 Gene in Clinical Isolates of Enterobacteriaceae.

BACKGROUND: The emergence of the New Dehli metallo-beta-lactamase (NDM) gene in Enterobacteriaceae is responsible for multidrug resistance responsible for severe infections and serious morbidity in patients. Our study aimed to define the molecular characteristics and antibiogram of the NDM-1 producing Enterobacteriaceae. METHODS: We isolated 370 individual enterobacteria from the clinical specimens collected from the two tertiary hospitals in Sakaka, Saudi Arabia. Bacterial isolation was performed using standard microbiological techniques and the Phoenix and Microscan WalkAway Plus automated analyzers. Bacterial strains were characterized by phenotypic methods and PCR, and DNA sequencing was used for the molecular characterization of NDM genes. RESULTS: The blaNDM gene was detected among the 68 members of the Enterobacteriaceae including a single case of rarely reported Cedecea lapagei. Of these 68, 43 isolates (63.2%) were blaNDM-1 and 25 (36.8%) were blaNDM variants. A statistically significant relationship between the NDM-1 and Klebsiella pneumoniae (p = 0.004) was seen, and the relationship between the NDM variants was significantly associated with Citrobacter freundii (p = 0.02) and Escherichia coli (p = 0.03). The in vitro minimum inhibitory concentrations (MICs) of NDM-producing Enterobacteriaceae revealed a very high rate of antibiotic resistance against several groups of antibiotics. These bacterial strains were less resistant to two aminoglycosides, gentamicin (39; 57.3%) and amikacin (27; 39.7%), and showed minimum resistance to tigecycline (25; 36.8%). CONCLUSIONS: The emergence of a large number of NDM-1 enterobacteria in our study identifies a substantial public concern, both within hospitals and the wider community, and leaves us a narrow choice of therapeutic options: the aminoglycosides, co-trimoxazole, and tigecycline.

Antibiogram and recent incidence of multi-drug resistant carbapenemase producing Escherichia coli isolated from paediatric patients.

OBJECTIVE: To gauge the recent breadth of MDR E. coli along with antibiogram of carbapenemase producing (CP) E. coli among children from an institute which receives patients from all over Punjab. METHODS: The bacterial strains of E. coli isolated from various specimens of patients were collected from April 2017 to August 2018 and processed using standard biochemical tests and API 20E system (bioMerieux). Phenotypic screening for CP E. coli was done by the modified Hodge test, whereas antibiotic susceptibility testing was done with Kirby-Bauer disc diffusion technique. RESULTS: Total of 6,468 bacterial strains were isolated, out of which 1,552 (24%) were E. coli. Carbapenem resistance was observed in 245 (16%) strains, amongst which 113 (46%) were confirmed to be CP. E. coli isolated from males were higher as compared to females (p<0.05). Majority of the organisms were isolated from blood (37.2%) samples. The hospital discharged about 65% of patients, while 23% left against medical advice. Overall MDR amongst E. coli was 93.26%. Colistin sulphate (15.9%) and nitrofurantoin (16.8%) showed the most efficacy followed by amikacin (15%) and fosfomycin (10.6%). CONCLUSION: The isolation of high number of MDR E. coli amongst the paediatric patients is worrisome, which could serve as a potential source of horizontal genes transfer to other genera.

Molecular Detection of blaIMP Genes in Metallo-Beta-Lactamase Producing Clinical Gram-Negative Isolates.

BACKGROUND: The use of carbapenem antibiotics in the treatment of serious Gram-negative bacterial infections is under threat due to the emergence of the blaIMP gene amongst the bacterial pathogens. METHODS: The present descriptive cross-sectional study aimed to determine the occurrence of the blaIMP gene and minimum inhibitory concentrations (MICs) of the bacterial pathogens. The carbapenem-resistant isolates were screened out for the detection of MBLs by using the modified Hodge test and disk potentiation method. MBL producing strains were tested for the presence of the blaIMP gene by using PCR technique. The MICs of the blaIMP gene positive bacterial isolates were detected on the Vitek 2 system (bioMerieux). RESULTS: The primary source of MBLs and blaIPM gene carrying bacterial pathogens was blood (38.5%). We isolated 104 bacterial isolates in the initial screening of carbapenem resistance. Metallo-beta-lactamases (MBLs) were detected in 76 (73%) of the isolates which predominantly included 27 (26%) Klebsiella pneumoniae, 20 (19.2%) Acinetobacter baumannii, 16 (15.4%) Pseudomonas aeruginosa, and 11 (10.6%) E. coli while the other Gram-nega-tive MBL producing bacteria were few in number. The blaIPM gene was detected in 1 (1.3%) case of Acinetobacter baumannii and 1 (1.3%) case of Stenotrophomonas maltophilia. These strains were found to be multi-drug resistant with high MICs (≥ 8 to ≥ 256 µg/mL) against the majority of the drugs. CONCLUSIONS: The emergence of the blaIPM gene is a matter of serious concern as it left us with limited treatment options of minocycline, tigecycline, and levofloxacin. The horizontal transfer of blaIPM gene in other Gram-negative isolates can lead the epidemics of multidrug resistance.

Determination of minimum inhibitory concentrations of various antifungal agents in clinical isolates of Candida species.

OBJECTIVE: To evaluate the occurrence and in vitro minimum inhibitory concentrations of different Candida species against nine antifungal agents in blood and urine. METHODS: The prospective descriptive study was conducted at The Children's Hospital, Lahore, Pakistan, from June 2015 to May 2016. Identification of the Candid aspecies was done by API (Analytical Profile Index) Candida (bioMerieux) and in vitro minimum inhibitory concentration break points were reported by using Sensititre Yeast One. Data was analysed using SPSS 20. RESULTS: Of the 87 samples, 68(78.2%) were isolated from blood and 19(21.8%) from urine specimens. Also, 66 (75.9%) samples were non-albicans Candida, 31(35.6%) Candida parapsilosis and 21(24.1%) were Candida albicans. Besides, 83(95.4%) strains exhibited excellent susceptibility pattern with the three echinocandins at minimum inhibitory concentration endpoint of ≤2µg/ml and all (100%) the strains inhibited by ≤1µg/ml of voriconazole. None (0%) of the C. albicans isolates was resistant to an antifungal agent. CONCLUSIONS: Excellent susceptibility pattern against antifungal agents was found with an increasing resistance trend towards non-albicans Candida species.

Emerging resistance of van genotype in enterococci: A potential menace for therapeutic failure.

OBJECTIVES: Emerging cases of vancomycin-resistant enterococci (VRE) are detrimental for the patients. The current study aimed to ascertain the occurrence of VRE, their antibiogram and the van genotype responsible for vancomycin resistance. METHODS: A total number of 2,958 clinical specimens were processed at Microbiology Department of the Alrazi Health Care, Lahore during the one year (2016-2017) using microbiological culture media, biochemical and serology. Antibiogram of enterococcal strains was performed using disc diffusion and E-test. ATCC Enterococcus faecalis 29212 was used as a quality control strain. The detection of van genotypes was accomplished by multiplex PCR assay. RESULTS: Out of the 147 enterococci, 139 (94.6%) were E. faecalis, and 8 (5.4%) were E. faecium. Statistically significant associations of urine (p < 0.001), pus (p < 0.001) and wound swabs (p = 0.001) were observed with E. faecalis. A significant correlation of enterococcal infections (p = 0.05) was seen with female patients. Four (2.9%) strains of E. faecalis found to be VRE with vanB (75%) and vanA (25%) genotypes. CONCLUSION: The emerging strains of VRE (vanB and vanA genotype) in the current study are a potential menace for therapeutic failure, which left the physicians with only linezolid as a therapeutic option.

AmpC beta-lactamases in Klebsiella pneumoniae: An emerging threat to the paediatric patients.

OBJECTIVE: To determine the burden of AmpC beta-lactamase producing Klebsiella pneumoniae and its antimicrobial profile among paediatric patients. METHODS: This cross-sectional study was conducted at the Microbiology Department of The Children's Hospital and the Institute of Child Health in Lahore, Pakistan, from May 2014 to April 2015, in which isolates of Klebsiella pneumoniae were screened by using the cefoxitin disc. Confirmation was done by inhibitor-based method using 400 micro grams of boronic acid dispensed on the cefoxitin discs. The zone sizes of cefoxitin with and without the boronic acid were compared. The antimicrobial susceptibility testing was performed using Kirby Bauer disc diffusion method. RESULTS: Positive cultures yielded 585 Klebsiella pneumoniae out of which 220(37.6%) strains were AmpC beta-lactamase-positive on the basis of cefoxitin screening and 126(21.53%) were positive on the basis of inhibitor-based confirmatory method. Most of the infected patients 73(57.9%) were neonates. All AmpC beta-lactamase-producing strains were resistant to cephalosporins. They also exhibited resistance to ciprofloxacin 109(86.5%), amikacin 98(77.8%), levofloxacin 8(77.8%), cefoperazone-sulbactam 81(64.3%), piperacillin-tazobactam 82(65.1%), meropenem, 56(44.4%) and imipenem 32(25.4%). CONCLUSIONS: Prompt identification of AmpC beta-lactamases using inhibitor-based confirmatory test can help reduce the burden of these pathogens.

Bacterial contamination of Pakistani currency notes from hospital and community sources.

OBJECTIVE: We determined the bacterial contamination and antibiotic resistance profile of circulating Pakistani currency notes collected from hospital and community sources. METHODS: This prospective study was organized from July to December 2015 in the Microbiology Department of The Children's Hospital and The Institute of Child Health Lahore. It was done on one hundred currency notes of four different denominations collected from various groups of people in sterile polythene bags. Gram staining, colony morphology and various biochemical tests were used to identify the bacterial isolates. Kirby Bauer disc diffusion method was used to observe the antibacterial drug resistance. RESULTS: There were 11 different types of bacterial species which contaminated 97 (97%) currency notes. The bacterial isolates discovered from paper currency notes included Klebsiella spp. (26.0%), Coagulase-negative Staphylococci (CoNS) (18.3%), E. coli (14.5%), Pseudomonas spp. (13.7%), Citrobacter spp. (11.5%), Enterobacter spp. (5.3%), Acinetobacter spp. (5.3%), Streptococcus spp. (2.3%), Shigella spp. (1.5%), Salmonella spp. (0.8%) and Pantoea spp. (0.8%). Most of the Gram-positive isolates were resistant to penicillin and ampicillin. None of the Gram-negative isolates found to be resistant to amikacin, cefoperazone-sulbactam and piperacillin-tazobactam. CONCLUSION: The currency notes circulating in hospital and community are contaminated with highly pathogenic and some multi-drug resistant bacteria. These currency notes could be a potential source of nosocomial and community-acquired infections.

Phylogenetic Analysis of Klebsiella pneumoniae from Hospitalized Children, Pakistan.

Klebsiella pneumoniae shows increasing emergence of multidrug-resistant lineages, including strains resistant to all available antimicrobial drugs. We conducted whole-genome sequencing of 178 highly drug-resistant isolates from a tertiary hospital in Lahore, Pakistan. Phylogenetic analyses to place these isolates into global context demonstrate the expansion of multiple independent lineages, including K. quasipneumoniae.

Phenotypic characterization of extended-spectrum-beta-lactamase producing E. coli from healthy individuals, patients, sewage sludge, cattle, chickens and raw meat.

OBJECTIVE: The present study aimed to determine the frequency and antimicrobial profile of ESBL-producing isolates of E. coli in different environments. METHODS: This cross-sectional study was conducted at The Children's Hospital and The Institute of Child Health, Lahore from July to December 2015. The faecal specimens from healthy individuals, patients, sewage sludge, cattle, chickens and raw meat (n = 122) were processed for microbiological analysis using MacConkey agar supplemented with cefotaxime. The identification of organisms was confirmed by API 10S and antimicrobial resistance profile was recorded by Kirby-Bauer disc diffusion method. RESULTS: On the basis of screening, 77 (63.0%) specimens were found to be positive for ESBL production. The confirmation of 74 (60.0%) ESBL producing E. coli was done using double disc synergy test (DDST). The frequency of ESBL producing E. coli was found to be 17 (57.0%) in healthy individuals, 15 (53.0%) in patients, 10 (66.0%) in cattle faeces, 5 (71.0%) in sewage sludge, 14 (70.0%) in raw meat and 13 (59.0%) in chicken faeces. All of these isolates were resistant to cephalosporins and some of these were resistant to fluoroquinolones and meropenem. None of the isolates showed resistance to cefoperazone-sulbactam, imipenem, piperacillin-tazobactam and amikacin. CONCLUSION: The prevalence of ESBL-producing E. coli was recorded in all the environments, suggesting a global expansion of these enzymes.