Phenotypic ESBL and non-phenotypic ESBL isolates of Klebsiella pneumoniae exhibit differing responses to induced antimicrobials resistance and subsequent antibiotic cross-resistance. - Related Documents




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233901.0000Phenotypic ESBL and non-phenotypic ESBL isolates of Klebsiella pneumoniae exhibit differing responses to induced antimicrobials resistance and subsequent antibiotic cross-resistance. AIM: To investigate the effect of adapting Klebsiella pneumoniae clinical isolates harboring ESBL genes to cetrimide (CT) in terms of subsequent cross-resistance to other biocides and antibiotics, and to investigate changes to virulence markers, such as biofilm formation and efflux activity. The changes between phenotypic extended spectrum β-lactamases (pESBL) expressing isolates and non-phenotypic ESBL (npESBL) isolates was compared. METHODS AND RESULTS: Kl. pneumoniae isolates (14 pESBL and 17 npESBL) were adapted to increasing concentrations of CT, until 4 × MIC was reached. The MIC of the adapted isolates was tested against chloroxylenol (CX) and chlorhexidine. Disk diffusion techniques were used to determine the susceptibility of the isolates to different antibiotics. Biofilm formation was assessed for the isolates using the crystal violet method and efflux pump activity was studied using the ethidium bromide assay. After CT adaptation, 100% of npESBL isolates and 85.7% of pESBL isolates showed increase in CT MIC after CT adaptation. While 41.2% of npESBL and 57.1% of the pESBL isolates showed a cross-resistance with chlorhexidine. CT adaptation resulted in a significant decrease in the susceptibility of npESBL isolates to aztreonam and cefotaxime compared to pESBL isolates, which could be linked to the increase in efflux activity of npESBL compared to pESBL. Biofilm formation was significantly increased after CT adaptation regardless of the type of isolate. CONCLUSIONS: The extensive use of biocides in the environment can induce cross-resistance to other biocides and antibiotics, and can increase the ability of bacteria to form biofilms. The response of bacteria to biocide adaptation differs between pESBL and npESBL isolates, although the effect is strain specific.202336724268
233710.9995Klebsiella pneumoniae susceptibility to biocides and its association with cepA, qacΔE and qacE efflux pump genes and antibiotic resistance. BACKGROUND: Although antiseptics are some of the most widely used antibacterials in hospitals, there is very little information on reduced susceptibility to these biocides and its relationship with resistance to antibiotics. AIM: To determine the relationship between reduced susceptibility to biocides and the carriage of antiseptic resistance genes, cepA, qacΔE and qacE, as well as identifying the role of efflux pumps in conferring reduced susceptibility. METHODS: Susceptibility was assessed for five biocides: chlorhexidine, benzalkonium chloride, Trigene, MediHex-4, Mediscrub; and for 11 antibiotics against 64 isolates of Klebsiella pneumoniae. Susceptibility to all compounds was tested by the agar double dilution method (DDM) and the effect of efflux pumps on biocides determined by repeating the susceptibility studies in the presence of the efflux pump inhibitor carbonyl cyanide m-chlorophenyl hydrazone (CCCP). The presence of the cepA, qacΔE and qacE genes was identified by polymerase chain reaction. FINDINGS: The bacteria were not widely antibiotic resistant though a few showed reduced susceptibility to cefoxitin, chloramphenicol and rifampicin and later-generation cephalosporins but not to carbapenems. Biocide susceptibility, tested by DDM, showed that 50, 49 and 53 strains had reduced susceptibility to chlorhexidine, Trigene and benzalkonium chloride, respectively. The antiseptic resistance genes cepA, qacΔE and qacE were found in 56, 34 and one isolates respectively and their effects as efflux pumps were determined by CCCP (10 mg/L), which decreased the minimum inhibitory concentrations (MICs) of chlorhexidine and Medihex-4 by 2-128-fold but had no impact on the MICs of benzalkonium chloride, Trigene and Mediscrub. CONCLUSION: There was a close link between carriage of efflux pump genes, cepA, qacΔE and qacE genes and reduced biocide susceptibility, but not antibiotic resistance, in K. pneumoniae clinical isolates.201222498639
229920.9995Determining the resistance of carbapenem-resistant Klebsiella pneumoniae to common disinfectants and elucidating the underlying resistance mechanisms. INTRODUCTION: Carbapenem-resistant Klebsiella pneumoniae (CRKP) infection is a serious problem in hospitals worldwide, posing a particular risk to immunocompromised patients. Elimination strategies may prevent these drug-resistant bacteria from spreading within hospital environments. Here, the susceptibility of patient-derived CRKP strains to common chemical disinfectants and possible correlations between the presence of drug-resistance genes and increased resistance to disinfectants were investigated. METHODS: The minimum inhibitory (MIC) and the minimum bactericidal concentrations (MBC) of common chemical disinfectants against each CRKP strain were determined using agar dilution; K. pneumoniae ATCC700603 served as a standard. The presence of the drug-resistance genes qacΔE, qacA, acrA and qacE was determined using PCR. RESULTS: A total of 27 clinically isolated CRKP strains collected in our hospital from 2011 to 2013 exhibited sensitivity to the following common chemical disinfectants in decreasing order of sensitivity: 75% ethyl alcohol > 2% glutaraldehyde > "84" disinfectant > 0.2% benzalkonium bromide > 2% iodine tincture > 1% iodophor > 0.1% chlorhexidine acetate. Of the 27 strains, 59, 41, 19 and 15% contained qacΔE, qacA, acrA and qacE resistance genes; 15% carried acrA, qacΔE and qacA, and 26% carried both qacA and qacΔE. Comparative analysis indicated that drug-resistance genes were correlated with higher MIC values. CONCLUSION: These pan-resistant pathogenic CRKP strains contained various drug-resistance genes and exhibited relatively high resistance to ethyl alcohol, chlorhexidine acetate and iodophor. Monitoring the drug-resistance rates of CRKP strains displaying disinfectant resistance may facilitate appropriate and effective sterilisation and thus preventing the spread of these pan-resistant strains.201526184804
229730.9994Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South Africa. Acinetobacter baumannii (A. baumannii) has developed several resistance mechanisms. The bacteria have been reported as origin of multiple outbreaks. This study aims to investigate the use of efflux pumps and quinolone resistance-associated genotypic mutations as mechanisms of resistance in A. baumannii isolates at a tertiary hospital. A total number of 103 A. baumannii isolates were investigated after identification and antimicrobial susceptibility testing by VITEK2 followed by PCR amplification of bla (OXA-51) . Conventional PCR amplification of the AdeABC efflux pump (adeB, adeS, and adeR) and quinolone (parC and gyrA) resistance genes were performed, followed by quantitative real-time PCR of AdeABC efflux pump genes. Phenotypic evaluation of efflux pump expression was performed by determining the difference between the MIC of tigecycline before and after exposure to an efflux pump inhibitor. The Sanger sequencing method was used to sequence the parC and gyrA amplicons. A phylogenetic tree was drawn using MEGA 4.0 to evaluate evolutionary relatedness of the strains. All the collected isolates were bla (OXA-51) -positive. High resistance to almost all the tested antibiotics was observed. Efflux pump was found in 75% of isolates as a mechanism of resistance. The study detected parC gene mutation in 60% and gyrA gene mutation in 85%, while 37% of isolates had mutations on both genes. A minimal evolutionary distance between the isolates was reported. The use of the AdeABC efflux pump system as an active mechanism of resistance combined with point mutation mainly in gyrA was shown to contribute to broaden the resistance spectrum of A. baumannii isolates.202134659419
577240.9994Molecular evaluation of colistin-resistant gene expression changes in Acinetobacter baumannii with real-time polymerase chain reaction. BACKGROUND: Acinetobacter baumannii is an important human pathogen which has recently gained increased attention due to the occurrence of drug-resistant nosocomial infections in patients suffering from immune system disorders, and those in hospital intensive care units. The aim of this research was to identify and isolate A. baumannii strains resistant to colistin, determine antibiotic resistance pattern of this bacteria, investigate the presence of colistin-resistant genes, and finally assess the effect of expression changes in pmrA and pmrB genes resistant to A. baumannii against colistin via real-time polymerase chain reaction. METHODS: The samples were initially purified and isolated using biochemical tests and Micro-gen kit. Later, the resistance pattern evaluation of validated samples to different antibiotics and colistin was carried out using two methods viz., disc diffusion and E-test. This was followed by the assessment of genes resistant to colistin via polymerase chain reaction besides gene expression changes via real-time polymerase chain reaction. RESULTS: The results of this study indicated that eleven strains of A. baumannii isolated from Shahid Rajaee Trauma Hospital were resistant to colistin. However, in the resistance pattern evaluation of A. baumannii isolated from Ali Asghar Hospital, all the strains were sensitive to colistin. In the evaluation of genes resistant to pmrA and pmrB, most of the strains resistant to colistin were carriers of these genes. Besides, in the expression assessment of these genes, it was demonstrated that expression of pmrA in the strains resistant to colistin significantly increased in relation to sensitive strains, but the expression of pmrB increased at a lower rate in the strains resistant to colistin as compared to the sensitive strains. CONCLUSION: Thus, it can be safely mentioned that increased expression of pmrA was due to the resistance of A. baumannii to colistin.201729225477
565550.9994Study of Disinfectant Resistance Genes in Ocular Isolates of Pseudomonas aeruginosa. BACKGROUND: The prevalence of disinfectant resistance in Pseudomonas aeruginosa is on the rise. P. aeruginosa is the most common bacteria isolated from cases of microbial keratitis. Many multi-purpose contact lens disinfectant solutions are available to decontaminate contact lenses before use and to help reduce the incidence of infections. However, with increasing disinfectant resistance, the effect of multi-purpose disinfectant solutions may diminish. The goal of this study was to examine genes associated with disinfectant resistance in ocular isolates of P. aeruginosa and understand the strain's susceptibility to different multipurpose disinfectant solutions. METHODS: Seven potential disinfectant resistance genes were used in BLASTn searches against the whole genomes of 13 eye isolates of P. aeruginosa. A microdilution broth method was used to examine susceptibility to four different multipurpose disinfectant solutions. RESULTS: All strains possessed the sugE2, sugE3 and emrE (qacE) genes. The sugE1 and qacEdelta1 genes were present in 6/13 isolates. No strains contained the qacF or qacG genes. All tested disinfectant solutions had the ability to kill all test strains at 100% concentration, with some strains being susceptible at 1:8 dilutions of the disinfecting solutions. However, the presence of disinfectant resistance genes was not associated with susceptibility to multi-purpose disinfectants. CONCLUSION: All four tested contact lens disinfectant preparations are effective against P. aeruginosa isolates regardless of the presence of disinfectant resistance genes.201830326554
575460.9994Efflux pump inhibitor CCCP to rescue colistin susceptibility in mcr-1 plasmid-mediated colistin-resistant strains and Gram-negative bacteria. OBJECTIVES: Efflux in bacteria is a ubiquitous mechanism associated with resistance to antimicrobials agents. Efflux pump inhibitors (EPIs) have been developed to inhibit efflux mechanisms and could be a good alternative to reverse colistin resistance, but only CCCP has shown good activity. The aim of our study was to identify CCCP activity in a collection of 93 Gram-negative bacteria with known and unknown colistin resistance mechanisms including isolates with mcr-1 plasmid-mediated colistin resistance. METHODS: Colistin MIC was evaluated with and without CCCP and the fold decrease of colistin MIC was calculated for each strain. In order to evaluate the effect of this combination, a time-kill study was performed on five strains carrying different colistin resistance mechanisms. RESULTS: Overall, CCCP was able to reverse colistin resistance for all strains tested. The effect of CCCP was significantly greater on intrinsically colistin-resistant bacteria (i.e. Proteus spp., Serratia marcescens, Morganella morganii and Providencia spp.) than on other Enterobacteriaceae (P < 0.0001). The same was true for bacteria with a heteroresistance mechanism compared to bacteria with other colistin resistance mechanisms (P < 0.0001). A time-kill study showed the combination was bacteriostatic on strains tested. CONCLUSIONS: These results suggest an efflux mechanism, especially on intrinsically resistant bacteria and Enterobacter spp., but further analysis is needed to identify the molecular support of this mechanism. EPIs could be an alternative for restoring colistin activity in Gram-negative bacteria. Further work is necessary to identify new EPIs that could be used in humans.201829718423
231670.9994Clinical Klebsiella pneumoniae isolates and their efflux pump mechanism for antibiotic resistance challenge. BACKGROUND: Klebsiella pneumoniae is a serious pathogen that causes many disorders in humans and animals. Klebsiella pneumoniae, which is one of the most important pathogens in hospitals, often causes many clinical manifestations, including pneumonia, urinary tract infections, and meningitis. Interest in this bacterium has increased due to the increasing incidence of infection caused by it, as well as its high resistance to antibiotics, especially broad-spectrum antibiotics. AIM: This study showed the efflux pump mechanism of clinical K. pneumoniae isolates and antibiotic resistance in samples collected from sheep and human respiratory tract infection in southern Iraq. METHODS: Three hundred samples were collected, and the samples included: 150 nasal swabs from sheep and 150 sputum samples from humans. Through bacteriological and biochemical examinations. The isolates were identified K. pneumoniae isolates were also confirmed by 16S rRNA. Susceptibility testing of the antibiotics used in the study. To determine the phenotypic efflux pump activity, the agar ethidium bromide cartwheel method was used. RESULTS: Of 150 sputum human specimens and 150 nasal swabs from sheep were tested, 25 and 17 K. pneumoniae species isolates from patients and sheep, respectively, for the resistance of the bacteria isolated from humans to antibiotics. The highest rate of resistance was to piperacillin (88%), and the lowest rate was to antibiotics (36%), imipenem. The highest of bacterial susceptibility to the antibiotic imipenem was (44%) and (36%) for levofloxacin, respectively. For the bacterial isolates from sheep, the highest percentage of resistance to rifampin was (82.3%), and the highest percentage of sensitivity was to imipenem and Levofloxacin antibiotics. The results showed that most of the 39 bacterial isolates (92.8%) possessed an efflux pump mechanism. The result of genotyping to identify the efflux pump genes tolC and acrAB revealed that all isolates carried the genes. CONCLUSION: All the isolates were resistant to antibiotics, and the bacterial isolates under study most possess the efflux pump mechanism. All bacteria also have efflux pump genes, and this gives the bacteria more resistance against many antibiotics.202541036356
227580.9994Contribution of β-lactamase and efflux pump overproduction to tazobactam-piperacillin resistance in clinical isolates of Escherichia coli. INTRODUCTION: Tazobactam-piperacillin (TZP) is a mixture of a broad-spectrum penicillin and an irreversible β-lactamase inhibitor. TZP is effective against Gram-negative bacteria that produce extended-spectrum β-lactamases, and it is used as a first-line or second-line drug to treat serious infections. METHODS: This study identified three TZP-resistant and two TZP-intermediate strains among 514 clinical isolates of Escherichia coli. RESULTS: These five isolates possessed one or more β-lactamase genes, bla(TEM-1), bla(CTX-M-2), bla(CTX-M-14), and/or bla(CMY-8). The expression levels of β-lactamase genes and acrAB genes in the strains were examined by using real-time reverse transcription PCR. The total enzymatic piperacillin-degrading activity in cells was determined. Two TZP-resistance mechanisms were identified: hyperproduction of TEM-1 in the two resistant strains; and simultaneous high production of β-lactamase and efflux pump AcrAB in the two TZP-intermediate isolates. The latter are an international high-risk clone O25b:H4-ST131-H30R. CONCLUSION: TZP resistance is still rare in clinical isolates of E. coli. However, resistance can develop on high production and/or combinations of known antimicrobial resistance mechanisms in different ways.202032062000
575990.9993The Relationship between Antibiotic Susceptibility and pH in the Case of Uropathogenic Bacteria. Urinary tract infections (UTIs) are common bacterial infections caused mainly by enteric bacteria. Numerous virulence factors assist bacteria in the colonization of the bladder. Bacterial efflux pumps also contribute to bacterial communication and to biofilm formation. In this study, the phenotypic and genetic antibiotic resistance of clinical UTI pathogens such as Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis were determined by disk diffusion method and polymerase chain reaction (PCR). Following this, different classes of antibiotics were evaluated for their antibacterial activity at pH 5, 6, 7 and 8 by a microdilution method. Gentamicin (GEN) was the most potent antibacterial agent against E. coli strains. The effect of GEN on the relative expression of marR and sdiA genes was evaluated by quantitative PCR. The slightly acidic pH (pH 6) and GEN treatment induced the upregulation of marR antibiotic resistance and sdiA QS activator genes in both E. coli strains. Consequently, bacteria had become more susceptible to GEN. It can be concluded that antibiotic activity is pH dependent and so the artificial manipulation of urinary pH can contribute to a more effective therapy of multidrug resistant bacterial infections.202134943643
2340100.9993Binary CuO\CoO nanoparticles inhibit biofilm formation and reduce the expression of papC and fimH genes in multidrug-resistant Klebsiella oxytoca. BACKGROUND AND AIM: Binary copper-cobalt oxide nanoparticles (CuO\CoO NPs) are modern kinds of antimicrobials, which may get a lot of interest in clinical application. This study aimed to detect the effect of the binary CuO\CoO NPs on the expression of papC and fimH genes in multidrug-resistant (MDR) isolates of Klebsiella oxytoca to reduce medication time and improve outcomes. METHODS: Ten isolates of K. oxytoca were collected and identified by different conventional tests besides PCR. Antibiotic sensitivity and biofilm-forming ability were carried out. The harboring of papC and fimH genes was also detected. The effect of binary CuO\CoO nanoparticles on the expression of papC and fimH genes was investigated. RESULTS: Bacterial resistance against cefotaxime and gentamicin was the highest (100%), while the lowest percentage of resistance was to amikacin (30%). Nine of the ten bacterial isolates had the ability to form a biofilm with different capacities. MIC for binary CuO\CoO NPs was 2.5 µg/mL. Gene expression of papC and fimH was 8.5- and 9-fold lower using the NPs. CONCLUSION: Binary CuO\CoO NPs have a potential therapeutic effect against infections triggered by MDR K. oxytoca strains due to the NPs-related downregulation ability on the virulence genes of K. oxytoca.202337269387
2294110.9993Antimicrobial Resistance of Clinical Klebsiella pneumoniae Isolates: Involvement of AcrAB and OqxAB Efflux Pumps. BACKGROUND: Over the last several decades, the AcrAB and OqxAB efflux pumps have been found to cause multidrug resistance (MDR) in various bacteria, most notably Klebsiella pneumoniae. Antibiotic resistance surges with increased expression of the acrAB and oqxAB efflux pumps. METHODS: In accordance with CLSI guidelines, a disk diffusion test was carried out using 50 K. pneumoniae isolates obtained from various clinical samples. CT was computed in treated samples and compared to a susceptible ciprofloxacin strain (A111). The final finding is presented as the fold change in the target gene's expression in treated samples relative to a control sample (A111), normalized to a reference gene. As ΔΔCT = 0 and 2 to the power of 0 = 1, relative gene expression for reference samples is often set to 1 Results: The highest rates of resistance were recognized with cefotaxime (100%), cefuroxime (100%), cefepime (100%), levofloxacin (98%), trimethoprimsulfamethoxazole (80%), and gentamicin (72%), whereas imipenem (34%) had the lowest rates. Overexpression of acrA and acrB, oqxA and oqxB, regulators marA, soxS, and rarA were greater in ciprofloxacin-resistant isolates compared to the reference strain (strain A111). There was also a moderate connection between ciprofloxacin MIC and acrAB gene expression and a moderate connection between ciprofloxacin MIC and oqxAB gene expression. CONCLUSION: This work provides a deeper knowledge of the role of efflux pump genes, particularly acrAB and oqxAB, as well as transcriptional regulators marA, soxS, and rarA, in bacterial resistance to ciprofloxacin.202436999690
5758120.9993RND pump inhibition: in-silico and in-vitro study by Eugenol on clinical strain of E. coli and P. aeruginosa. Multidrug-resistant (MDR) gram-negative bacteria pose significant challenges to the public health. Various factors are involved in the development and spread of MDR strains, including the overuse and misuse of antibiotics, the lack of new antibiotics being developed, and etc. Efflux pump is one of the most important factors in the emergence of antibiotic resistance in bacteria. Aiming at the introduction of novel plant antibiotic, we investigated the effect of eugenol on the MexA and AcrA efflux pumps in Pseudomonas aeruginosa (P. aeruginosa) and Escherichia coli (E. coli). Molecular docking was performed using PachDock Server 1.3. The effect of eugenol on bacteria was determined by disk diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). A cartwheel test was also performed to evaluate efflux pump inhibition. Finally, the expression of the MexA and AcrA genes was examined by real-time PCR. The results of molecular docking showed that eugenol interacted with MexA and AcrA pumps at - 29.28 and - 28.59 Kcal.mol(-1), respectively. The results of the antibiogram test indicated that the antibiotic resistance of the treated bacteria decreased significantly (p < 0.05). The results of the cartwheel test suggested the inhibition of efflux pump activity in P. aeruginosa and E. coli. Analysis of the genes by real-time PCR demonstrated that the expression of MexA and AcrA genes was significantly reduced, compared to untreated bacteria (p < 0.001). The findings suggest, among other things, that eugenol may make P. aeruginosa and E. coli more sensitive to antibiotics and that it could be used as an inhibitor to prevent bacteria from becoming resistant to antibiotics.202337587975
1705130.9993Formation ability and drug resistance mechanism of Klebsiella pneumoniae biofilm and capsule for multidrug-resistant. This study was to explore the formation ability of biofilm and capsule and the drug resistance mechanism for multidrug-resistant Klebsiella pneumoniae. firstly, 55 strains of K. pneumoniae were screened out from the body fluid specimens of the laboratory. The strains were drug-resistant, and the characteristics of clinical infections of these strains were analyzed. Secondly, all strains were tested for the presence of biofilms and capsules, and then the deoxyribonucleic acid (DNA) genomes of the strains extracted were detected using polymerase chain reaction (PCR) technology. Finally, the serotype genes and virulence genes of the strains were screened, and the relationship between these two genes and the formation of capsules and biofilms was analyzed and compared. A new generation of sequencing technology was applied to analyze the genome structure of K. pneumoniae, comparative genomics technology was adopted to analyze the drug resistance plasmids, and molecular cloning and other methods were utilized to clone the drug resistance-related genes. of the 55 strains of K. pneumoniae isolated clinically, 61.8% came from blood with a total number of 34 strains; 8 strains were from secretion specimens (accounting for 14.5% of the total); and 7 strains were from drainage fluid (accounting for 12.7% of the total), including 2 strains from pus, bile, and pleural fluid, respectively. The strains were tested by PCR, of which iroN virulence genes were the most (34 strains), accounting for 61.8%, followed by wabG and fimH (33 strains, accounting for 60% of the total), followed by magA, K2, K20, K1, and K57. The positive rates of the two virulence genes (fimH and wabG) were higher in positive strains of biofilm. The drug susceptibility results showed that ampicillin and amoxicillin were more resistant to capsule-positive strains than the capsule-negative strains. K. pneumoniae had been able to form a complete capsule and biofilm, the formation rate of biofilm was higher than that of the capsule, and there was an increasing trend. The two serotype genes (K20 and K2) accounted for relatively high proportions, and K. pneumoniae carried relatively more virulence genes (wabG and fimH), which may be closely related to the capsule production of K. pneumoniae. In addition, resistance-related genes were also transferred horizontally in different strains of bacteria, forming a wide range of drug resistance, which brought great difficulties to clinical work.202337953580
2331140.9993Bacteriological and molecular study of fosfomycin resistance in uropathogenic Escherichia coli. The identification of genes associated with resistance has the potential to facilitate the development of novel diagnostic tests and treatment methods. The objective of this study was to examine the antibiotic resistance and Fosfomycin resistance genes in uropathogenic Escherichia coli (UPEC) in patients in Baghdad, Iraq. After analyzing 250 urine samples using various identification methods, including the examination of morphological characteristics, biochemical tests, and genetic detection, it was determined that E. coli was the most common bacteria present, accounting for 63.6% of the samples. Antibiotic susceptibility testing showed a significant prevalence of resistance to various antibiotics, with 99.3% of E. coli isolates exhibiting multiple drug resistance (MDR). Fosfomycin showed antibacterial properties against UPEC. The minimum inhibitory concentration (MIC) ranged from 512 to 1024 μg/mL, while the minimum bactericidal concentration (MBC) was 2048 μg/mL. In the time-kill assay, fosfomycin was effective against fosfomycin-resistant isolates within 8-12 h. The genetic determinants associated with fosfomycin resistance were examined through the utilization of polymerase chain reaction (PCR). The findings indicated that the genes murA, glpT, and cyaA were detected in all the isolates when genomic DNA was used as a template. However, all the tests yielded negative results when plasmid was used as a template. The genes fosA3 and fosA4 were detected in 8.6% and 5% of the isolates when genomic DNA was used as a template. When plasmid was used as a template, the genes fosA3 and fosA4 were found in 5.7% and 2.9% of the isolates, respectively. In conclusion, there is an increasing problem with antibiotic resistance in UPEC, with elevated rates of resistance to several antibiotics. The study also offers novel insights into the genetic foundation of fosfomycin resistance in UPEC.202438367167
2317150.9993Molecular Detection of Adefg Efflux Pump Genes and their Contribution to Antibiotic Resistance in Acinetobacter baumannii Clinical Isolates. BACKGROUND: Acinetobacter baumannii (A. baumannii) is one of the most important bacteria causing nosocomial infections worldwide. Over the past few years, several strains of A. baumannii have shown antibiotic resistance, which may be due to the activity of efflux pumps. This study was aimed to detect AdeFG efflux pump genes and their contribution to antibiotic resistance in A. baumannii clinical isolates. METHODS: A total of 200 A. baumannii clinical isolates were collected from clinical specimens of ulcers, pus, sputum, and blood. All isolates were identified using standard biochemical tests. After identifying and cleaving the genome by boiling, PCR was performed on samples using specific primers. The antimicrobial susceptibility patterns were determined by disk diffusion, with and without CCCP efflux pump inhibitor were determined according to CLSI guidelines. RESULTS: We identified 60 clinical isolates of A. baumannii using biochemical differential tests. Identification of all A. baumannii isolates was confirmed by blaOXA-51-like PCR. According to the results of our study, 98.37% of A. baumannii isolates were resistant to ciprofloxacin, norfloxacin, and levofloxacin. PCR results indicated that all 60 A. baumannii isolates contained the AdeF and 76.66% contained AdeG. CONCLUSION: the results of this study demonstrated that most of the A. baumannii isolates contained AdeF and AdeG efflux pump genes, and more than 98% of the isolates were resistant to ciprofloxacin, norfloxacin, and levofloxacin. This reflected the significant contribution of efflux pumps to the development of resistance to these antibiotics.202032582800
1760160.9993Proteomic analysis of clinical isolate of Stenotrophomonas maltophilia with blaNDM-1, blaL1 and blaL2 β-lactamase genes under imipenem treatment. The co-occurrence of L1 and AmpR-L2 with bla(NDM-1) gene with an upstream 250-bp promoter was detected in a clinical isolate of Stenotrophomonas maltophilia DCPS-01, which was resistant to all β-lactams and sensitive only to colistin and fluoroquinolones. To investigate expression of resistance genes and the molecular mechanisms of bacteria resistance to carbapenems, proteomic profiles of the isolate was passaged with and without the drug by using 2D-PAGE. The results showed that 33 genes exhibiting a ≥3-fold change were identified as candidates that may help S. maltophilia survive drug selection. Strikingly, L1 was expressed more highly in cells grown with imipenem, and the abundant NDM-1 further increased, while very little L2 was detected even following induction. Specific activities for β-lactamase revealed that L2 remained at constitutive low levels (10.6 U/mg), while L1 and NDM-1 showed clear activity (69.8 U/mg). Our data support that imipenem could specifically and reversibly induce L1 and NDM-1, which together played key roles in drug resistance in DCPS-01. Although NDM-1 mediated resistance to carbapenems has been found in very few cases, to our knowledge, this is the first proteomics research of S. maltophilia with NDM-1, giving very broad-spectrum antibiotic resistance profiles.201222702735
2314170.9993Imipenem resistance in aerobic gram-negative bacteria. A prospective study was undertaken to observe the emergence of resistance to imipenem, if any, among aerobic gram-negative bacteria. A total of 736 isolates were tested during 1994-95 and less than 1% of them were resistant to imipenem, whereas the next year ('95-'96) the rate increased to 11 of the 903 isolates tested. The resistant isolates during '94-'95 were all Stenotrophomonas maltophilia whereas the spectrum of resistant bacterial species increased in '95-'96 to include Pseudomonas aeruginosa, Burkholderia cepacia, Acinetobacter calcoaceticus, Enterobacter cloacae, Proteus mirabilis and Morganella morganii with a tendency to an increase in the minimum inhibitory concentration (MIC) in the later part of the year. A majority (72%) of the resistant isolates were from patients with burns, and burn wounds were most frequently infected with such organisms. These data suggest that over a period of time aerobic gram-negative bacteria may develop resistance to imipenem and the pool of such bacteria increases with extensive use of the drug. Non-fermentative aerobic bacteria tend to develop resistance faster with widespread dissemination than Enterobacteriaceae. Hospital Burn Units are a potential source of development of such resistance.19989603633
2296180.9993Multi-drug resistance profiles and the genetic features of Acinetobacter baumannii isolates from Bolivia. INTRODUCTION: Acinetobacter baumannii is opportunistic in debilitated hospitalised patients. Because information from some South American countries was previously lacking, this study examined the emergence of multi-resistant A. baumannii in three hospitals in Cochabamba, Bolivia, from 2008 to 2009. METHODOLOGY: Multiplex PCR was used to identify the main resistance genes in 15 multi-resistant A. baumannii isolates. RT-PCR was used to measure gene expression. The genetic environment of these genes was also analysed by PCR amplification and sequencing. Minimum inhibitory concentrations were determined for key antibiotics and some were determined in the presence of an efflux pump inhibitor, 1-(1-napthylmethyl) piperazine. RESULTS: Fourteen strains were found to be multi-resistant. Each strain was found to have the blaOXA-58 gene with the ISAba3-like element upstream, responsible for over-expression of the latter and subsequent carbapenem resistance. Similarly, ISAba1, upstream of the blaADC gene caused over-expression of the latter and cephalosporin resistance; mutations in the gyrA(Ser83 to Leu) and parC (Ser-80 to Phe) genes were commensurate with fluoroquinolone resistance. In addition, the adeA, adeB efflux genes were over-expressed. All 15 isolates were positive for at least two aminoglycoside resistance genes. CONCLUSIONS: This is one of the first reports analyzing the multi-drug resistance profile of A. baumannii strains isolated in Bolivia and shows that the over-expression of theblaOXA-58, blaADC and efflux genes together with aminoglycoside modifying enzymes and mutations in DNA topoisomerases are responsible for the multi-resistance of the bacteria and the subsequent difficulty in treating infections caused by them.201323592642
5752190.9993Cefoxitin inhibits the formation of biofilm involved in antimicrobial resistance MDR Escherichia coli. The study investigates the relationship between biofilm formation and antibiotic resistance in Escherichia coli (E. coli) isolated from calves. Using biochemical and molecular methods, we identified the isolates and assessed their biofilm-forming ability through an improved crystal violet staining method. The minimum inhibitory concentrations (MICs) of 18 antibiotics against the isolates were determined using the broth microdilution method. The impact of cefoxitin on biofilm formation was analyzed using laser scanning confocal microscopy (LSCM). Additionally, qRT-PCR was employed to evaluate the expression levels of biofilm-related genes (luxS, motA, fliA, pfs, and csgD) in response to varying cefoxitin concentrations. Results indicated a significant correlation between antimicrobial resistance (AMR) and biofilm formation ability. Cefoxitin effectively reduced biofilm formation of multidrug-resistant E. coli isolates at 1/2 and 1 MIC, with enhanced inhibition at higher concentrations. The QS-related genes luxS, pfs, motA, and fliA were downregulated, leading to decreased csgD expression. At 1/2 MIC, csgD expression was significantly reduced. In conclusion, cefoxitin inhibits biofilm formation in multidrug-resistant E. coli by down-regulating key genes, offering a potential strategy to mitigate resistance and control infections in calves caused by biofilm-positive E. coli isolates.202540122078