Aims: Carbapenemase-producing bacteria make infections of the urinary tract (UTIs) challenging to cure with last-resort treatment like carbapenem. Carbapenemase-producing E. coli and K. pneumoniae implicated in UTI must be detected molecularly since their ability to spread broadly among patients is rising ( Nomeh et al., 2022). Methodology: Ten non-repeated clinical isolates of Escherichia coli (Ecoli1, Ecoli2, Ecoli3, Ecoli4, and Ecoli5) and Klebsiella pneumoniae (Kp6, Kp7, Kp8, Kp9, Kp10) were selected from urinary tract infection patients at Institute of Kidney Disease Peshawar, Pakistan, based on their in vitro phenotypic carbapenem antibiotic resistance. These isolates were confirmed using standard routine microbiological techniques. PCR-specific primers screened E. coli and K. pneumoniae strains for Carbapenemase-producing genes. Result: Molecular Detection of Carbapenemase-producing Gene in UTI Patients with Uropathogenic Escherichia coli and Klebsiella pneumoniae. The higher proportion of Carbapenemase-producing genes in all the bacterial isolates in this study was blaKPC 15(100 %), followed by blaNDM 12.3(90.1 %), blaIMP 6(60.2 %) and blaVIM 3(30.6 %). The most common Carbapenemase gene in Escherichia coli 8 (80%) was blaKPC, followed by blaNDM 7 (70%) and blaOXA 45 (4.5%), which was the least common. Klebsiella pneumoniae had more blaNDM and blaKPC than blaOXA. Both had a percentage of 4.5 (40.9%). Conclusion:These results are consistent with the rapid spread of genes responsible for generating Carbapenemases in E. coli and Klebsiella pneumoniae that cause urinary tract infections. Despite the lack of blaVIM in K. pneumoniae, the pathogenic function of Carbapenemase-producing genes in UTI in this study should not be underestimated because of the potential they have to cause treatment failure and the subsequent persistence of UTI in patients.

The opportunistic bacteria Pseudomonas aeruginosa is commonly linked to skin infections. When dealing with P. aeruginosa, carbapenem is your best bet. It is of international concern because beta-lactamase synthesis might lead to carbapenem resistance. Among all the beta-lactamases, Metallo-Beta lactamases are the most versatile. The purpose of this study was to isolate metallolactamase-producing P. aeruginosa from wound infections. One hundred and twenty samples from patients with burn wound infections were collected and tested using conventional microbiological methods for the presence of P. aeruginosa. Using species-specific primers against the oprL and Oprl genes of P. aeruginosa, a polymerase chain reaction was used to conduct a molecular characterisation of P. aeruginosa. Antimicrobial susceptibility testing was conducted using the Kirby Bauer disc diffusion technique. Certain primers were used in a polymerase chain reaction to detect P. aeruginosa carrying the carbapenemase gene. Using the modified carbapenem inactivation technique and the EDTA carbapenem inactivation approach, we were able to phenotypically identify metallo-beta-lactamase-expressing genes. Carbapenemase-encoding genes' sensitivities and specificities were determined. Forty-six out of a possible one hundred P. aeruginosa (38%) were successfully recovered, with their identity validated by a polymerase chain reaction (PCR) experiment. The highest level of resistance was discovered against Cefepime (87%) and the lowest level of resistance was recorded against colistin (33%). A total of 25 (54%) were multidrug-resistant isolates. Out of 46, 35(76%) were confirmed for carbapenemase production by performing PCR. The prevalence of carbapenemase encoding genes was as follows: blaSPM (14%), blaVIM (25.7%), blaNDM (40%), blaKPC (2.85%) and blaIMP (17%). The modified carbapenem inactivation method showed 91.42% positive results and eCIM showed 90.62% positive results. Phenotypic detection showed more sensitivity and less specificity. Results concluded that mCIM and eCIM were considered a less expensive, more sensitive and suitable method to distinguish class A and class B of carbapenemaseproducing P. aeruginosa.

Infections caused by Pseudomonas aeruginosa may be either acquired in the community or contracted in a healthcare setting. Multidrug-resistant (MDR) P. aeruginosa is a growing problem; a new treatment approach is required to tackle this. Combination therapy of antibiotics and nanoparticle is thus applied to overcome this problem. Therefore, this study was planned to evaluate the synergistic effect of AgNPs along with different antibiotics against MDR P. aeruginosa. A total of 120 surgical or burn wound samples were collected from a tertiary care hospital. The plates containing the samples cultivated on cetrimide agar were then heated to 37°C. Isolates were identified based on colony shape, Gram staining, and several biochemical tests. A Kirby-Bauer disc diffusion technique antibiogram was conducted following CLSI 2022 recommendations. A minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were also determined. The Agar well diffusion technique and the microdilution method were used to test the antibacterial activity of the AgNPs, respectively. The synergistic effect of antibiotics and AgNPs was estimated by the Checkerboard method. Out of 120 samples, 46 (38.8%) were confirmed positive for P. aeruginosa, and out of that, 33 were confirmed as MDR P. aeruginosa. Seven representative isolates proceeded for further procedures. Antibacterial activity of AgNPs revealed a maximum zone of inhibition of 12 mm at 4 mg/ml and a minimum of 2.5 mm at 1 mg/ml by agar well diffusion method. MIC and MBC of AgNPs showed that all the isolates were inhibited at 250 mg/ml. The FIC index of checkerboard results showed that colistin and gentamicin exhibited complete synergism with AgNPs, while ciprofloxacin showed partial synergism with AgNPs.

The misuse of antibiotics is one of the primary causes of the rapidly expanding problem of multidrug resistance. Fungi are responsible for the production of a variety of potent metabolites (Akhtar, et al., 2019). Formation of nanoparticles of silver (AgNPs) is a simple non-toxic, and environmentally friendly method of the preparation and development of nanoparticles. Which considered a crucial step in nanotechnology. Producing AgNPs from Aspergillus fumigatus samples involved the use of X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM) (Al-Abdullah, et al., 2023). The effect of synthesized AgNPs and crude extract on several bacterial pathogens was observed. Both fungal crude extract and (AgNPs) showed the greatest antibacterial efficacy against bacterial isolates. The ethyl acetate crude extract showed the highest possible antibacterial activity, according to the reports against E. coli was seen at 16 mm at a 50µl concentration (12mg/1ml DMSO). Conversely ethyl crude extract has the least antibacterial action against S.typhi at 50µl concentration was (14mm) (Bala, M., et al., 2013). The maximum activity of the ethyl acetate crude extract was observed against E. coli at 100 µl, which showed a zone of inhibition measuring 21 mm, while an inhibition zone of 18 mm was observed against S.typhi. Surface Plasmon Resonance (SPR) at 432 nm was found during UV-visible spectroscopy, confirming the production of AgNPs (Guilger, et al., 2019). The spherical shape of AgNPs was seen in the SEM micrograph. The reduction of Ag+ ions into AgNPs was largely mediated by phenolic, carboxyl, and hydroxyl groups, according to the results of FTIR investigation (Farjana, et al., 2014). The stabilization of AgNPs was accomplished through amino acid linkage. The produced peak of AgNPs' XRD revealed information about their nature, including their phase purity, size, and internal crystalline structure (Pena et al., 2010). It is possible that the pharmaceutical and medical fields will find a great use for the AgNPs that are produced from the extract of Aspergillus fumigatus. Silver AgNPs and crude extract Aspergillus fumigatus enhance antibacterial activity, outlining their potential in future research.