Background Emerging worldwide in the past decade, there has been a significant increase in multidrug-resistant bacteria from serious nosocomial infections, especially carbapenemase-producing Gram-negative bacilli that have emerged worldwide. The objective of this study is to investigate carbapenem resistance in Gram-negative bacilli bacteria using phenotypic detection, antimicrobial resistance profiles and genotypic characterisation methods. Methods 200 Gram-negative bacilli isolates were collected from different clinical specimens. All clinical samples were exposed to isolation and identification of significant pathogens applying bacteriological examination and an automated Vitek-2 system. The isolates were subjected to susceptibility tests by the Vitek-2 automated system and those isolates that were resistant to beta-lactam drugs, including carbapenems, third-generation cephalosporines or cefoxitin, were selected for phenotyping using Carba plus disc system assay for detection of carbapenemase-producing isolates. These isolates were further confirmed by molecular detection. PCR was used for the detection carbapenem-resistant genes (OXA-48, IMP, NDM, VIM, and KPC). Results 110 (55%) of 200 Gram-negative bacilli were identified as beta-lactam-resistant isolates. The frequency of carbapenem-resistant isolates was calculated to be 30.9% (n = 34/110). A collection totalling 65/110 (59%) isolates were identified as carbapenemase producers by phenotypic method. Moreover, among the 65 carbapenemase-producing Gram-negative isolates with a positive phenotype-based result, 30 (46%), 20 (30%) and 18 (27%) isolates were positive for OXA-48, KPC and MBL enzymes, respectively, as well as the production of 27% of AmpC with porin loss. Tigecycline was the most effective antibiotic that affected 70% of MDR isolates, but high rates of resistance were detected to other tested antimicrobials. Of interest, a high incidence of MDR, XDR and PDR profiles were observed among all carbapenemase-producing isolates. 36% (24/65) of the tested isolates were MDR to 3 to 5 antimicrobial classes. 29% (17/65) of the recovered isolates were XDR to 6 to 7 antimicrobial classes. Alarmingly, 24% (16/65) of isolates displayed PDR to all the tested 8 antimicrobial classes. Genotype assay, including 53 phenotypically confirmed carbapenemase-producing isolates of Gram-negative bacilli, found 51(96%) isolates were harbouring one or more genes. The most common carbapenemase gene was bla NDM 83% (44/53) followed by bla OXA-48 75% (40/53), bla VIM 49% (26/53) and bla IMP 43% (23/53), while the gene bla KPC was least frequent 7% (4/53). 92% (46/51) of isolates were involved in the production of more than one carbapenemase gene. Conclusion This study demonstrated the emergence of carbapenemase-producing Gram-negative pathogens implicated in healthcare-related infections. Accurate identification of carbapenem-resistant bacterial pathogens is essential for patient treatment, as well as the development of appropriate contamination control measures to limit the rapid spread of pathogens. Tigecycline exhibited potent antimicrobial activity against MDR, XDR and PDR-producing strains that establish a threatening alert which indicates the complex therapy of infections caused by these pathogens.

Bifidobacterium selectively colonizes the infants' intestinal tract, and the relevant coliform bacteria in adults are particularly beneficial because of their enhanced capability to prevent pathogens of gastro intestine by direct antimicrobial action and relieve infection, which led to their intensification, the antibacterial activities of titanium nanoparticles producing by some bacteria, makes them attractive as a new agent against pathogenic bacteria. In our present study, we used a probiotic bacteria Bifidobacterium bifidum which was isolated from the commercial market capsule to produce TiO2 nanoparticles and study the biologically characterized nanoparticle using various techniques like Scanning Electron Microscopic (SEM), atomic force microscopy (AFM), and study its antimicrobial activity against a bacteria isolated from the stool of patients suffering from acute diarrhea. The results showed that the morphological characteristics of nanoparticles were found to have a spherical shape and mean size of 81 nm by AFM while scanning electron microscope viewed as an oval shape with anatase form synthesized by B. bifidum. TiO2-NP synthesized by B. bifidum had an inhibitory effect against P. aeruginosa, A. baumanii, K. pneumonia at a concentration 16 mg/ml and 32 mg/ml towards E. coli and S. typhi, the minimum inhibitory concentration (MIC) against pathogenic bacteria isolated from acute diarrhea included Pseudomonas aeruginosa, Acinetobacter baumanii, Klebsiella pneumonia, E.coli and salmonella typhi was utilized to determine the antibacterial impact of the synthesized TIO2 nanoparticles. Our biologically synthesized titanium nanoparticles were effective against all the tested pathogenic bacteria at various degrees and had a probable role in significantly greater antimicrobial efficacy against all isolates under study. This trial may have considerable significance for the prevention of antibiotic resistance associated diarrhea in hospitals.

Increasing prevalence of infections caused by multidrug resistant Pseudomonas aeruginosa due to production of Extend spectrum and Metallo-β-lactamases is a major concern throughout the World,is a serious problem and calls for an effective infection control measure to curb their dissemination,the prevalence among clinical isolates varies greatly worldwide and they are rapidly changing over time.The aim of the present study is to investigate the presence of the bla CTX-M in clinically isolates of P.aeruginosa strains producing Extend spectrum and Metallo-β-lactamases.A total of 227 samples were collected from different clinical specimens (sputum, urine, wound, burn, Bronchial wash) attending public hospitals in Erbil provincebetween (September 2017 -March 2018.isolated microorganisms were processed and identified using standard bacteriological conventional culture methods, biochemical tests, Species identification and antibiograms for pathogens were performed using Vitek 2 system against 15clinically important antibiotics. ESBL producing Pseudomonas aeruginosa isolates were detected using Vitek 2 system and also the screened isolates were further phenotypically studied for MBL production was detected by performing combined disc test by using imipenem discs with and without ethylenediaminetetraacetic acid (EDTA), which chelates zinc required for Bacterial genomic DNA was extracted, and the presence of the bla CTX-M were detected by PCR.Out of 227 samples 40 (17.6%)isolates were positive for P. aeruginosa,the highest isolation rate (2.6%) was observed in burn specimens, high rate (> 90%) of antibiotic resistant P.aeruginosa isolates were observed for penicillin, amikacin, piperacillin, lincomycin, vancomycin, chloramphenicol, rifampicin ,ciprofloxacin, Erythromycin. The prevalence rate of ESBL in our study 32 (80.6%) isolates, 72.5% MBL production by P. aeruginosa was observed and co-resistance was recorded in 21(52.5%). Our results demonstrated that

Nanotechnology is being investigated for its potential to improve nanomedicine for human health. The purpose of this study was to isolate carbapenemase-producing Gram-negative bacilli (CPGB), investigate the presence of carbapenemase resistance genes, determine their antibiogram and ability to biosynthesise silver nanoparticles (Ag NPs), and estimate the antibacterial activity of Acinetobacter baumannii-biosynthesised Ag NPs on CPGB alone and in combination with antibiotics. A total of 51 CPGBs were isolated from various specimens in the study. The automated Vitek-2 system was used to identify and test these strains' antimicrobial susceptibilities. The carbapenemase resistance genes were identified using a polymerase chain reaction (PCR). Under the CPGB, A. baumannii could biosynthesise Ag NPs. X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), and field emission scanning electron were used to characterise Ag NPs. The antibacterial activity of Ag NP alone and in combination with antibiotics against CPGB was determined using the broth microdilution method, and their synergistic effect was determined using the checkerboard assay. blaNDM and blaOXA-48 were the most commonly reported, and 90% of the isolates produced multiple carbapenemase genes. Tigecycline proved to be the most effective anti-CPGB antibiotic. Isolates with more resistance genes were more resistant to antibiotics, and isolates with three genes (42%) had the most extensively drug-resistant patterns (38%). A significant relationship was discovered between genetic and antibiotic resistance patterns. Only A. baumannii produced Ag NPs out of all the isolates tested. Ag NPs with a size of 10 nm were confirmed by UV–visible spectroscopy, FT-IR, XRD, and TEM analysis. The Ag NPs were effective against CPGB, with minimum inhibitory concentrations ranging from 64 to 8 μg/ml on average. Surprisingly, the combination of Ag NPs and antibiotics demonstrated synergistic and partial synergistic activity (fractional inhibitory concentration between 0.13 and 0.56) against CPGB, as well as a significant reduction in antibiotic concentrations, particularly in the case of A. baumanii versus ceftriaxone (1024 to 4 μg/ml). The notable synergistic activity of Ag NPs with antibiotics represents a valuable nanomedicine that may find clinical application in the future as a combined remedy.

Pseudomonas aeruginosa is a ubiquitous microorganism that is difficult to treat due to the increasing prevalence of multidrug resistance patterns. A total of 227 samples were taken from different clinical samples during the study period from January 2018 to December 2018. The isolates were identified with antibiotic sensitivity testing with ESBL by the Vitek-2 automated method. MDR, XDR, and PDR were determined. 40 (17.6%) isolates were positive for P. aeruginosa, maximum of ESBL and MBL. Positive isolates were detected in the burn, coexisting ESBL + MBL enzymes in 21 (52.5%) of our isolates. Imipenem followed by Meropenem were found to be effective against ESBL and MBL producers. Resistance was reached between 72-100% to 5 antibiotics. The frequency of PDR, MDR, and XDR were 5%, 50%, and 45%, respectively. The frequency of co-production between MDR, XDR, and PDR with MBL, ESBL, and Biofilm was 35%, 12.5% and 5%, respectively. Among the ESBLs, the frequency of distribution of bla VEB-1gene and blaGES-5 gene was 50% and 40 %, respectively. Bacterial isolates simultaneously carrying blaVEB-1 gene with multiple ?-lactamases of different classes of biofilm, MDR, PDR, and XDR as same as a coexisting blaGES-5 gene. One isolate was detected as new isolates registered in global gene bank as locally P. aeruginosa isolates in Erbil city (LOCUS MN900953). The phylogenetic trees of the blaVEB gene isolates were demonstrated a genotype closely related to others, deposited in GenBank similar to the P. aeruginosa gene; gene sequencing revealed a 99% similarity with other isolates deposited in GenBank.