BACKGROUNDUrinary tract infection is one of the commonest infectious diseases worldwide. This study was carried out to determine the antimicrobial susceptibility pattern of bacteria causing urinary ...tract infection visiting Kathmandu University Hospital. METHODSA total of 3,500 urine samples were processed and antibiotic resistance pattern was determined following Clinical Laboratory Standard Institute guidelines. Patients' information was obtained after informed consent. RESULTSTotal number of samples with positive growth was 434 (12.40%). 331 (76.27%) of the isolates were Escherichia coli followed by Klebsiella pneumoniae, Enterococcus spp., Pseudomonas aeruginosa, Staphylococcus saprophyticus, Proteus mirabilis, Enterobacter species, Klebsiella oxytoca, Citrobacter freundii, Proteus vulgaris, Staphylococcus aureus and Acinetobacter species. Over all 224 (51.61%) were multidrug resistant strains. All strains were sensitive to colistin, vancomycin and linezolid. Over all ampicillin and cefazolin had least sensitivity. Multidrug resistant strains were detected more among elderly patients with complicated urinary tract infection and diabetes which was 25 (83.33%) compared to elderly patients with uncomplicated urinary tract infection and having no diabetes or any other comorbid illnesses which was only 11(22.22%) (p-value<0.05). 21 (70.00%) of the pregnant females had multidrug resistant isolates and only 18 (36.73%) of pediatric age group patients had multidrug resistant isolates (p-value<0.05) Conclusion: Drug-resistant bacteria were observed in urine samples. Effective treatment and prevention of urinary tract infection need detailed microbiological diagnosis and drug susceptibility testing.
Resistance to antimicrobial agents has become a major source of morbidity and mortality worldwide. When antibiotics were first introduced in the 1900's, it was thought that we had won the war against ...microorganisms. It was soon discovered however, that the microorganisms were capable of developing resistance to any of the drugs that were used. Apparently most pathogenic microorganisms have the capability of developing resistance to at least some antimicrobial agents. The main mechanisms of resistance are: limiting uptake of a drug, modification of a drug target, inactivation of a drug, and active efflux of a drug. These mechanisms may be native to the microorganisms, or acquired from other microorganisms. Understanding more about these mechanisms should hopefully lead to better treatment options for infective diseases, and development of antimicrobial drugs that can withstand the microorganisms attempts to become resistant.
To evaluate the accuracy of beta-lactamase gene detection directly from urine samples by Nanopore sequencing.
DNA was extracted from bacterial pellets in spun urine. The purified DNA was then ...sequenced in native form by a Nanopore sequencer (MinION) to identify the organisms and beta-lactamase genes. Results were compared to routine urine cultures and standard antimicrobial susceptibility tests (AST).
We processed 60 urine samples of which routine cultures grew Enterobacteriaceae, including 28 carbapenem-resistant (CRE), 17 extended-spectrum beta-lactamase (ESBL) or AmpC producing, and 15 non-ESBL/AmpC phenotypes. We excluded 7 samples with extremely low DNA amounts (< 1 ng/μl) for a final case of 53 in total. The sensitivity of antimicrobial resistance gene detection within 6 hours, the optimal duration from real-time simulation, of Nanopore sequencing for the diagnosis of carbapenem-resistant and ceftriaxone-resistant phenotypes was 73.9% (95%CI 56.0–91.9%) and 81.1% (95%CI 68.5–93.7%), while the specificity was 96.7% (95%CI 90.2–100.0%) and 56.3% (95%CI 31.9–80.6%), respectively. The median times for MinION to generate DNA reads containing carbapenemase and ESBL/AmpC genes were 93 minutes (IQR 17–245.5) and 99 minutes (IQR 31.25–269.75) after sequencing commencement, respectively.
Nanopore sequencing can identify bacterial genotypic resistance in urine and may enable clinicians to adjust antimicrobial therapy earlier than routine AST.
•Nanopore sequencing rapidly reveals antimicrobial resistance genes in urine samples.•The accuracy of carbapenemase gene detection is acceptable (86.8%).•The accuracy is dependent on the quality of samples.•Optimal antimicrobial agents can be probably adjusted as soon as the second dose.
Background and Aim: One of the most important bacterial resistance mechanisms is production of Extended-Spectrum Beta-Lactamase (ESBL). In addition, ESBL strains usually have resistance to several ...antibiotic classes, because multiple resistance genes are often carried by a plasmid. One of these multiple resistances is the simultaneous resistance to quinolones and ESBL production. The aim of this study was to determine the prevalence of resistance to fluoroquinolones in ESBL-producing Klebsiella pneumoniae isolates in Sanandaj health centers in 2018. Materials and Methods: Evaluation of antibiotic susceptibility in all isolates (phenotypically and at Minimum inhibitory concentration (MIC) and identification of ESBL strains were performed by VITEK2. We evaluated the relationship between ESBL production and resistance to fluoroquinolones (ciprofloxacin and levofloxacin) by statistical tests. Results: According to the antibiotic susceptibility results of 67 isolates, 35 isolates (52.23%) were resistant to ciprofloxacin and levofloxacin. 25 isolates (37.31%) were identified as ESBL, of which 19 isolates (76%) were resistant to levofloxacin and ciprofloxacin and only 6 isolates (24%) were sensitive to these antibiotics. Statistical analysis showed that resistance to ciprofloxacin and levofloxacin was significantly increased in ESBL isolates compared to that in non-ESBL isolates (P-value = 0.003). Conclusions: In this study, simultaneous resistance to fluoroquinolones (ciprofloxacin and levofloxacin) and ESBL production was observed in Klebsiella pneumoniae isolates, which can limit treatment options. Also, the results of this study showed that the treatment of the ESBL Klebsiella pneumoniae infection with levofloxacin and ciprofloxacin antibiotics will be a challenge in these medical centers.
The use of antibiotics in veterinary and human treatment can cause the development of antibiotic resistance in bacteria for β-lactam antibiotics. Worldwide, this resistance has become a growing ...concern in public health. Limited data are currently available regarding Extended Spectrum β-Lactamase (ESBL) Escherichia coli in Indonesia. The current study determined the prevalence and characteristics of ESBL genes of E. coli in retail chicken meat and humans in Indonesia. Two hundred eighty retail chicken meat were randomly collected from various modern and traditional markets in Jakarta (70 retail sourced chicken meat from modern markets and 210 from traditional markets). The prevalence of E. coli from the chicken meat sold at traditional markets was 97.14%, which was significantly higher than those of the modern markets with 78.57 % (P < 0.05). The prevalence of ESBL-producing E. coli isolated from chicken meat sold at traditional market was 40.47% and the modern market was 35.71 % and the prevalence of ESBL-producing E. coli isolated from chicken meat was 38.09 %, which is significantly higher than those of the clinical sample (with average 5.57 %). The most predominant gene is blaTEM in 54.54 % as a single gene or mixed with other genes followed by blaCTX-M in 44.31 % and blaSHV gene was only found in three isolates in 1.13 %. This study found that isolates from both the broiler chicken meat and clinical samples were having the same molecular characteristics. It is speculated that there is a relationship between them. However, this needs to be substantiated further.
•This study demonstrates the important role of EVs in the transmission of ESBL-E. coli β-lactamase resistance.•This study demonstrated that the frequency of drug-resistant gene transfer mediated by ...EVs carrying different plasmids of blaCTX-M-55 varied somewhat but was generally transferable.•This study provides a theoretical basis for elucidating a new mechanism of EVs-mediated drug resistance transmission.
Extracellular vesicles (EVs) have become the focus of research as an emerging method of horizontal gene transfer. In recent years, studies on the association between EVs and the spread of bacterial resistance have emerged, but there is a lack of research on the role of EVs secreted by extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in the spread of β-lactam resistance. Therefore, the aim of this study was to investigate the role of EVs in the transmission of β-lactam resistance.
In this study, the role of EVs in the transmission of β-lactam resistance in E. coli was evaluated by the EVs-mediated bacterial resistance to β-lactam antibiotics test and the EVs-mediated blaCTX-M-55 transfer experiments using EVs secreted by ESBL-E. coli.
The results showed that ESBL-EVs were protective against β-lactam antibiotic-susceptible bacteria, and this protective effect was dependent on the integrity of the EVs and showed dose- and time-dependent effects. At the same time, ESBL-EVs can also mediate the horizontal transmission of blaCTX-M-55, and EVs-mediated gene transfer is selective, preferring to transfer in more closely related species.
In this study, we demonstrated the important role of EVs in the transmission of β-lactam resistance in chicken ESBL-E. coli, and evaluated the risk of EVs-mediated horizontal gene transfer, which provided a theoretical basis for elucidating the mechanism of EVs-mediated resistance transmission.
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Pathogenic ESBL-producing E. coli lineages occur frequently worldwide, not only in a human health context but in animals and the environment, also in settings with low antimicrobial pressures. This ...study investigated the fitness costs of ESBL-plasmids and their influence on chromosomally encoded features associated with virulence, such as those involved in the planktonic and sessile behaviors of ST131 and ST648 E. coli. ESBL-plasmid-carrying wild-type E. coli strains, their corresponding ESBL-plasmid-"cured" variants (PCV), and complementary ESBL-carrying transformants were comparatively analyzed using growth curves, Omnilog® phenotype microarray (PM) assays, macrocolony and biofilm formation, swimming motility, and RNA sequence analysis. Growth curves and PM results pointed toward similar growth and metabolic behaviors among the strains. Phenotypic differences in some strains were detected, including enhanced curli fimbriae and/or cellulose production as well as a reduced swimming capacity of some ESBL-carrying strains, as compared to their respective PCVs. RNA sequencing mostly confirmed the phenotypic results, suggesting that the chromosomally encoded csgD pathway is a key factor involved. These results contradict the hypothesis that ESBL-plasmid-carriage leads to a fitness loss in ESBL-carrying strains. Instead, the results indicate an influence of some ESBL-plasmids on chromosomally encoded features associated with virulence in some E. coli strains. In conclusion, apart from antibiotic resistance selective advantages, ESBL-plasmid-carriage may also lead to enhanced virulence or adaption to specific habitats in some strains of pandemic ESBL-producing E. coli lineages.
In cities across the globe, the majority of wastewater - that includes drug resistant and pathogenic bacteria among other contaminants - is released into streams untreated. This water is often ...subsequently used for irrigation of pastures and produce. This use of wastewater-contaminated streams allows antibiotic-resistant bacteria to potentially cycle back to humans through agricultural products. In this study, we investigated the prevalence of extended-spectrum β-lactamase (ESBL)-producing
isolated from produce and irrigation water across 17 provinces of Ecuador. A total of 117 vegetable samples, 119 fruit samples, and 38 irrigation water samples were analyzed. Results showed that 11% of the samples were positive for
including 11 irrigation water samples (29%), and samples of 13 vegetables (11%), and 11 fruits (9%). Among the 165
isolates cultured, 96 (58%) had the ESBL phenotype, and 58% of ESBL producing
came from irrigation water samples, 11% from vegetables, and 30% from fruits. The
,
, and
genes were the most frequently found gene associated with the ESBL phenotype and coincided with the
alleles associated with human infections in Ecuador. Three isolates had the mcr-1 gene which is responsible for colistin resistance. This report provides evidence of the potential role of irrigation water in the growing antimicrobial resistance crisis in Ecuador.
Respiratory tract infection due to Gram-negative bacteria is a common cause of morbidity and mortality worldwide. This study was carried out to determine the antimicrobial susceptibility pattern of ...Gram-negative bacteria from patients with lower respiratory tract infection visiting Kathmandu University Hospital.
A total of 3,403 respiratory samples including sputum and endotracheal aspirates were processed and antibiotic resistance pattern was determined following Clinical Laboratory Standard Institute guidelines. Patients' information was obtained after informed consent.
Growth of Gram-negative bacteria was 210 (6.17%). 83(39.52%) were Klebsiella pneumoniae followed by Escherichia coli, Pseudomonas aeruginosa, Acinetobacter species, Klebsiella oxytoca, Enterobacter species, Proteus mirabilis and Haemophilus influenzae. 151(71.90%) isolates were multidrug resistant. None of the strains were resistant to colistin. 51(24.29%) were resistant to carbapenem and out of these 3(1.43%) were metallo-? lactamase producers. 89(65.92%) of the patients having both pneumonia along with other respiratory illnesses were above 60 years of age indicating that old age might be a predisposing factor. (p value is less than 0.0001). 92(81.42%) of patients of age above 60 years had multidrug resistant isolates indicating that old age might be a predisposing factor for getting infection my multidrug resistant isolates. (p value equals 0.0012) Among 36 diabetic patients 33(91.67%) had multidrug resistant isolates. Whereas out of 174 non-diabetic patients only 118(67.82%) had multidrug resistant isolates (p value equals 0.0037).
Multidrug-resistant Gram negative bacteria were observed in respiratory samples. Effective treatment of lower respiratory tract infection need detailed microbiological diagnosis and drug susceptibility testing.
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