RESUMO
INTRODUCTION: Pseudomonas aeruginosa isolates producing metallo-ß-lactamase have caused nosocomial outbreaks, severe infections, and ineffective carbapenem therapy worldwide since 1991. Due to their prevalence, hospital infection control techniques are difficult. This study aimed to find out the prevalence of metallo-ß-lactamase among P. aeruginosa isolates from two tertiary care hospitals in Kathmandu. METHODS: A descriptive cross-sectional study was conducted at the Department of Microbiology and Department of Pathology of two tertiary care centres in Kathmandu from 7 December 2021 to 6 April 2023, after receiving ethical approval from the Ethical Review Board. Isolated strains were identified and tested for antibiotic susceptibility by modified Kirby-Bauer Methods. Metallo-ß-lactamase presence was confirmed using an imipenem-imipenem/ ethylenediaminetetraacetic acid disc. A convenience sampling method was used. The point estimate was calculated at 95% Confidence Interval. RESULTS: Among 255, Pseudomanas aeruginosa isolates, the distribution of metallo-ß-lactamase-producing Pseudomanas aeruginosa was 103 (40.39%) (34.32-46.69 at 95% Confidence Interval). Multidrug resistance categories included multidrug resistance 74 (71.80%), extensively drug resistance 32 (31.10%), P. aeruginosa difficult-to-treat 16 (15.53%) and carbapenem-resistant P. aeruginosa was determined to be 82 (79.60%). CONCLUSIONS: The study found a high prevalence of metallo-ß-lactamase-producing Pseudomanas aeruginosa isolates, requiring early identification, infection control measures, and an all-inclusive antimicrobial therapy protocol to reduce their spread in medical settings.
Assuntos
Antibacterianos , Infecções por Pseudomonas , Pseudomonas aeruginosa , Centros de Atenção Terciária , beta-Lactamases , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/efeitos dos fármacos , Nepal/epidemiologia , beta-Lactamases/metabolismo , Estudos Transversais , Humanos , Infecções por Pseudomonas/epidemiologia , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/tratamento farmacológico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Testes de Sensibilidade Microbiana , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Infecção Hospitalar/tratamento farmacológico , Farmacorresistência Bacteriana Múltipla , PrevalênciaRESUMO
The chronic airway infections with Pseudomonas aeruginosa are the major co-morbidity in people with cystic fibrosis (CF). Within CF lungs, P. aeruginosa persists in the conducting airways together with human mucins as the most abundant structural component of its microenvironment. We investigated the adhesion of 41 serial CF airway P. aeruginosa isolates to airway mucin preparations from CF sputa. Mucins and bacteria were retrieved from five modulator-naïve patients with advanced CF lung disease. The P. aeruginosa isolates from CF airways and non-CF reference strains showed a strain-specific signature in their adhesion to ovine, porcine and bovine submaxillary mucins and CF airway mucins ranging from no or low to moderate and strong binding. Serial CF clonal isolates and colony morphotypes from the same sputum sample were as heterogeneous in their affinity to mucin as representatives of other clones thus making 'mucin binding' one of the most variable intraclonal phenotypic traits of P. aeruginosa known to date. Most P. aeruginosa CF airway isolates did not adhere more strongly to CF airway mucins than to plastic surfaces. The strong binders, however, exhibited a strain-specific affinity gradient to O-glycans, CF airway and mammalian submaxillary mucins.
Assuntos
Aderência Bacteriana , Fibrose Cística , Mucinas , Infecções por Pseudomonas , Pseudomonas aeruginosa , Escarro , Fibrose Cística/microbiologia , Pseudomonas aeruginosa/metabolismo , Pseudomonas aeruginosa/isolamento & purificação , Mucinas/metabolismo , Humanos , Animais , Escarro/microbiologia , Infecções por Pseudomonas/microbiologia , Suínos , Bovinos , OvinosRESUMO
BACKGROUND: Epidemiological investigations have revealed an important association between infection, inflammation and prostate cancer. Certain bacterial species, such as Klebsiella spp, Escherichia coli, Pseudomonas spp, Proteus mirabilis, Chlamydia trachomatis have been linked to prostate cancer. This study aimed to examine the microbiota; specifically bacterial species that have been linked to prostate infections in the urine of individuals diagnosed with prostate cancer. RESULTS: Sixty-six prostate cancer patients and forty controls provided midstream urine samples. The urine samples were grown on suitable medium, and bacterial isolates were detected by standard microbiological methods. Additionally, the antibiotic sensitivity pattern of the bacterial isolates was analysed. A total of number of 72 bacterial isolates were obtained from the urine of study participants. The results showed the presence of Escherichia coli (50.0%), Pseudomonas aeruginosa (18.1%), Klebsiella spp (15.3%), Staphylococcus aureus (8.3%), Enterobacter spp (4.2%), and Proteus mirabilis (2.8%) in the urine. The most common bacterial species isolated from prostate cancer patients was Escherichia coli, which was susceptible to levofloxacin (100%), tobramycin (91.7%), and amikacin (62.5%). CONCLUSIONS: This study's findings established the presence of bacteria previously linked to prostatitis. This report indicates a high prevalence of pro-inflammatory bacteria and uropathogens in the urinary tract of men diagnosed with prostate cancer.
Assuntos
Antibacterianos , Bactérias , Testes de Sensibilidade Microbiana , Hiperplasia Prostática , Neoplasias da Próstata , Humanos , Masculino , Neoplasias da Próstata/microbiologia , Antibacterianos/farmacologia , Hiperplasia Prostática/microbiologia , Pessoa de Meia-Idade , Prevalência , Idoso , Nigéria/epidemiologia , Bactérias/efeitos dos fármacos , Bactérias/isolamento & purificação , Bactérias/classificação , Infecções Urinárias/microbiologia , Infecções Urinárias/epidemiologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/isolamento & purificação , Klebsiella/efeitos dos fármacos , Klebsiella/isolamento & purificação , Proteus mirabilis/efeitos dos fármacos , Proteus mirabilis/isolamento & purificação , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/isolamento & purificaçãoRESUMO
Infectious wounds on the skin surface are easily colonized by bacteria from pyogenic group that manifest as inflammation, such as Pseudomonas aeruginosa. P. aeruginosa is a Gram-negative bacterium and an opportunistic pathogen known for causing invasive state in critically ill and immunocompromised patients. The aim of this study was to detect the 16S rRNA and gyrB genes in P. aeruginosa using polymerase chain reaction (PCR) method. The sample in this study was pus isolate from a 5-year-old boy with leg wounds. The bacteria were isolated on brain heart infusion broth (BHIB) media and identified with molecular identification. Sequencing and BLAST analysis were carried out to determine the similarity of gene identity by comparing sample sequence with other isolate sequences on the Gene Bank. The results of molecular identification showed amplification DNA band of around 934 base pairs (bp) for 16S rRNA and 225 bp for gyrB gene. The BLAST program demonstrated that the sample had 99.89% similarity with P. aeruginosa strain XC4 (accession code ON795960.1) for the 16S rRNA gene. Meanwhile, the gyrB gene exhibited 99.10% similarity with the P. aeruginosa strain PSA-1.2 (accession code KP172300.1).
Assuntos
DNA Girase , Reação em Cadeia da Polimerase , Infecções por Pseudomonas , Pseudomonas aeruginosa , RNA Ribossômico 16S , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Masculino , Humanos , RNA Ribossômico 16S/genética , Reação em Cadeia da Polimerase/métodos , Pré-Escolar , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/diagnóstico , DNA Girase/genética , Indonésia , Infecção dos Ferimentos/microbiologia , Infecção dos Ferimentos/diagnóstico , Supuração/microbiologiaRESUMO
BACKGROUND: Several healthcare-associated infection outbreaks have been caused by waterborne Pseudomonas aeruginosa exhibiting its ability to colonize water systems and resist conventional chlorine treatment. This study aims to investigate the occurrence of Pseudomonas aeruginosa in hospital drinking water systems and the antimicrobial resistance profiles (antibiotic and chlorine resistance) of isolated strains. METHODS: We investigated the presence of Pseudomonas aeruginosa in water and biofilms developed in nine hospital water systems (n = 192) using culture-based and molecular methods. We further assessed the survival of isolated strains after exposure to 0.5 and 1.5 ppm concentrations of chlorine. The profile of antibiotic resistance and presence of antibiotic resistance genes in isolated strains were also investigated. RESULTS: Using direct PCR method, Pseudomonas aeruginosa was detected in 22% (21/96) of water and 28% (27/96) of biofilm samples. However, culturable Pseudomonas aeruginosa was isolated from 14 samples. Most of P. aeruginosa isolates (86%) were resistant to at least one antibiotic (mainly ß-lactams), with 50% demonstrating multidrug resistance. Moreover, three isolates harbored intI1 gene and two isolates contained blaOXA-24,blaOXA-48, and blaOXA-58| genes. Experiments with chlorine disinfection revealed that all tested Pseudomonas aeruginosa strains were resistant to a 0.5 ppm concentration. However, when exposed to a 1.5 ppm concentration of chlorine for 30 min, 60% of the strains were eliminated. Interestingly, all chlorine-resistant bacteria that survived at 30-minute exposure to 1.5 ppm chlorine were found to harbor the intI1 gene. CONCLUSIONS: The detection of antimicrobial resistant Pseudomonas aeruginosa in hospital water systems raises concerns about the potential for infections among hospitalized patients. The implementation of advanced mitigation measures and targeted disinfection methods should be considered to tackle the evolving challenges within hospital water systems.
Assuntos
Biofilmes , Cloro , Hospitais , Pseudomonas aeruginosa , Microbiologia da Água , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Cloro/farmacologia , Humanos , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/epidemiologia , Infecção Hospitalar/microbiologia , Infecção Hospitalar/epidemiologia , Antibacterianos/farmacologia , Doenças Transmitidas pela Água/microbiologia , Doenças Transmitidas pela Água/epidemiologia , Farmacorresistência Bacteriana Múltipla , Testes de Sensibilidade Microbiana , Desinfetantes/farmacologia , Água Potável/microbiologiaRESUMO
Carbapenem-resistant Pseudomonas aeruginosa is a common multidrug-resistant bacterium encountered in clinical practice. This pathogen causes pneumonia, which is difficult to treat owing to the limited choice of antimicrobial drugs, resulting in a relatively high mortality rate. Carrimycin is a new macrolide antibiotic with broad-spectrum antibacterial and potential immunomodulatory effects. Herein, we report a case of severe pneumonia caused by carbapenem-resistant Pseudomonas aeruginosa that presented with septic shock and Acute Respiratory Distress Syndrome (ARDS). Initially, we used piperacillin-tazobactam and ceftazidime-avibactam but without satisfactory results. Finally, we administered carrimycin in combination with piperacillin-tazobactam; the patient's condition improved, and he was successfully weaned off the ventilator. Therefore, the combined use of carrimycin should be considered for patients infected with carbapenem-resistant Pseudomonas aeruginosa who do not respond to conventional anti-infection treatments.
Assuntos
Antibacterianos , Carbapenêmicos , Infecções por Pseudomonas , Pseudomonas aeruginosa , Humanos , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/isolamento & purificação , Masculino , Carbapenêmicos/farmacologia , Carbapenêmicos/uso terapêutico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/microbiologia , Pessoa de Meia-Idade , Testes de Sensibilidade Microbiana , IdosoRESUMO
The indiscriminate use of antibiotics has resulted in a growing resistance to drugs in Pseudomonas aeruginosa. The identification of antibiotic resistance genes holds considerable clinical significance for prompt diagnosis. In this study, we established and optimized a Recombinase-Aided Amplification (RAA) assay to detect two genes associated with drug resistance, oprD and arr, in 101 clinically collected P. aeruginosa isolates. Through screening for the detection or absence of oprD and arr, the results showed that there were 52 Imipenem-resistant P. aeruginosa (IRPA) strains and 23 Rifampin-resistant P. aeruginosa (RRPA) strains. This method demonstrated excellent detection performance even when the sample concentration is 10 copies/µL at isothermal conditions and the results could be obtained within 20 minutes. The detection results were in accordance with the results of conventional PCR and Real-time PCR. The detection outcomes of the arr gene were consistently with the resistance spectrum. However, the antimicrobial susceptibility results revealed that 65 strains were resistant to imipenem, while 49 strains sensitive to imipenem with oprD were identified. This discrepancy could be attributed to genetic mutations. In summary, the RAA has higher sensitivity, shorter time, and lower-cost instrument requirements than traditional detection methods. In addition, to analyze the epidemiological characteristics of the aforementioned drug-resistant strains, we conducted Multilocus Sequence Typing (MLST), virulence gene, and antimicrobial susceptibility testing. MLST analysis showed a strong correlation between the sequence types ST-1639, ST-639, ST-184 and IRPA, while ST-261 was the main subtype of RRPA. It was observed that these drug-resistant strains all possess five or more virulence genes, among which exoS and exoU do not coexist, and they are all multidrug-resistant strains. The non-coexistence of exoU and exoS in P.aeruginosa is related to various factors including bacterial regulatory mechanisms and pathogenic mechanisms. This indicates that the relationship between the presence of virulence genes and the severity of patient infection is worthy of attention. In conclusion, we have developed a rapid and efficient RAA (Recombinase-Aided Amplification) detection method that offers significant advantages in terms of speed, simplicity, and cost-effectiveness (especially in time and equipment aspect). This novel approach is designed to meet the demands of clinical diagnostics.
Assuntos
Antibacterianos , Imipenem , Testes de Sensibilidade Microbiana , Técnicas de Amplificação de Ácido Nucleico , Infecções por Pseudomonas , Pseudomonas aeruginosa , Recombinases , Rifampina , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Imipenem/farmacologia , Rifampina/farmacologia , Humanos , Antibacterianos/farmacologia , Recombinases/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Infecções por Pseudomonas/microbiologia , Farmacorresistência Bacteriana/genética , Porinas/genética , Sensibilidade e Especificidade , Proteínas de Bactérias/genética , Técnicas de Diagnóstico Molecular/métodosRESUMO
RATIONALE: Infections due to multidrug-resistant (MDR) Pseudomonas aeruginosa are strongly associated with poor outcomes, including prolonged hospitalization and an increased risk of mortality. Antimicrobial options for the treatment of severe infections due to MDR P aeruginosa are quite limited, and treatment remains challenging. PATIENT CONCERNS: A 65-year-old woman presented to our orthopedic clinic with a 3-month history of progressive pain and stiffness in her left knee. Her primary care provider administered a hyaluronic acid injection, which unfortunately resulted in worsening symptoms. Subsequent treatment included a 1-month course of intravenous gentamicin and ceftriaxone, which failed to alleviate her symptoms. DIAGNOSIS: MDR P aeruginosa septic arthritis of the knee. The culture isolate was tested for susceptibility to multiple antibiotics. Magnetic resonance imaging evaluations were conducted, showing notable erosive and osteolytic changes around the joint surfaces that had progressed significantly. INTERVENTIONS: The patient underwent arthroscopic irrigation and synovectomy. The treatment regimen included a combination of intravenous colistin and piperacillin/tazobactam administered over a 6-week period. Total knee arthroplasty was performed 6 months later without additional antibiotic treatment. OUTCOMES: Patient's knee condition remained continuously stable without abnormal findings of inflammation. The patient's knee range of motion increased 0 to 125 degrees, her pain almost disappeared, and she was able to maintain activities of daily life. LESSONS: This case underscores the challenges of managing infections with MDR organisms in complex clinical scenarios, emphasizing the need for timely intervention and appropriate antibiotic therapy.
Assuntos
Antibacterianos , Artrite Infecciosa , Farmacorresistência Bacteriana Múltipla , Infecções por Pseudomonas , Pseudomonas aeruginosa , Humanos , Feminino , Idoso , Artrite Infecciosa/microbiologia , Artrite Infecciosa/tratamento farmacológico , Artrite Infecciosa/diagnóstico , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/isolamento & purificação , Antibacterianos/uso terapêutico , Antibacterianos/administração & dosagem , Articulação do Joelho/microbiologiaRESUMO
Outdoor decorative fountains usually attract residents to visit. However, opportunistic pathogens (OPs) can proliferate and grow in the stagnant fountain water, posing potential health risks to visitors due to the inhalation of spaying aerosols. In this study, the abundance of selected OPs and associated microbial communities in three large outdoor decorative fountain waters were investigated using quantitative PCR and 16S rRNA sequencing. The results indicated that Mycobacteria avium and Pseudomonas aeruginosa were consistently detected in all decorative fountain waters throughout the year. Redundancy analysis showed that OPs abundance was negatively correlated with water temperature but positively correlated with nutrient concentrations. The gene copy numbers of M. avium varied between 2.4 and 3.9 log10 (gene copies/mL), which were significantly lower than P. aeruginosa by several orders of magnitude, reaching 6.5-7.1 log10 (gene copies/mL) during winter. The analysis of taxonomic composition and prediction of functional potential also revealed pathogenic microorganisms and infectious disease metabolic pathways associated with microbial communities in different decorative fountain waters. This study provided a deeper understanding of the pathogenic conditions of the outdoor decorative fountain water, and future works should focus on accurately assessing the health risks posed by OPs in aerosols.
Assuntos
Mycobacterium avium , Pseudomonas aeruginosa , RNA Ribossômico 16S , Microbiologia da Água , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/genética , RNA Ribossômico 16S/análise , Mycobacterium avium/isolamento & purificação , Mycobacterium avium/genética , Microbiota , Monitoramento AmbientalRESUMO
BACKGROUND: In addition to antibiotic resistance, persistence is another cause of treatment failure in bacterial infections, representing a significant public health concern. Due to a lack of adequate data on clinical isolates, this study was initiated to investigate persistence in clinical isolates in Burkina Faso. METHODS: Eighty (80) clinical isolates, including 32 Pseudomonas aeruginosa, 41 Staphylococcus aureus, and 7 Salmonella sp. obtained from clinical laboratories in Burkina Faso, were analyzed to assess their susceptibility to ciprofloxacin and gentamicin, as well as to determine the presence of persistence genes. The effects of ciprofloxacin and gentamicin on persister formation were evaluated by conducting colony counts at 1, 3, 5, 7, and 20 h after exposing the bacteria to high concentrations of these antibiotics. RESULTS: Results showed high sensitivity to both antibiotics (72.5% for ciprofloxacin and 82.5% for gentamicin). Persister formation occurred in Staphylococcus aureus with gentamicin and in Salmonella sp. with ciprofloxacin, while Pseudomonas aeruginosa did not form persisters. The mazF gene was found in 28.13% of P. aeruginosa and 2.44% of S. aureus isolates, and the hipA gene in 28.57% of Salmonella sp. None of the relE1 or relE2 genes were detected. CONCLUSIONS: The study revealed high sensitivity in clinical bacterial isolates to ciprofloxacin and gentamicin. Staphylococcus aureus and Salmonella sp. showed persister formation under antibiotic stress, with low frequencies of the studied persistence genes. These findings enhance understanding of clinical bacterial behavior and inform strategies against antibiotic-resistant infections.
Assuntos
Antibacterianos , Ciprofloxacina , Gentamicinas , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa , Staphylococcus aureus , Burkina Faso , Humanos , Antibacterianos/farmacologia , Ciprofloxacina/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Gentamicinas/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Salmonella/efeitos dos fármacos , Salmonella/genética , Salmonella/isolamento & purificação , Farmacorresistência Bacteriana/genética , Infecções Bacterianas/microbiologia , Infecções Bacterianas/tratamento farmacológicoRESUMO
Targeted high-throughput sequencing (HTS) has revolutionized the way we look at bacterial communities. It can be used for the species-specific detection of bacteria as well as for the determination of the microbiome and resistome and can be applied to samples from almost any environment. However, the results of targeted HTS can be influenced by many factors, which poses a major challenge for its use in clinical diagnostics. In this study, we investigated the impact of the DNA extraction method on the determination of the bacterial microbiome and resistome by targeted HTS using principles from metrology and diagnostics such as repeatability and analytical sensitivity. Sputum samples spiked with Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa at three different concentrations (103-106 cells/mL) were used. DNA was extracted from each sample on 2 separate days in three replicates each using three different extraction methods based on cetrimonium bromide, magnetic beads, and silica membranes. All three spiked bacteria were detected in sputum, and the DNA extraction method had no significant effect on detection. However, the DNA extraction method had significant effects on the composition of the microbiome and the resistome. The sequencing results were repeatable in the majority of cases. The silica membrane-based DNA extraction kit provided the most repeatable results and the highest diversity of the microbiome and resistome. Targeted HTS has been shown to be a reliable tool for determining the microbiome and resistome; however, the method of DNA extraction should be carefully selected to minimize its impact on the results. IMPORTANCE: High-throughput sequencing (HTS) is one of the crucial new technologies that gives us insights into previously hidden parts of microbial communities. The DNA extraction method is an important step that can have a major impact on the results, and understanding this impact is of paramount importance for their reliable interpretation. Our results are of great value for the interpretation of sputum microbiome and resistome results obtained by targeted HTS. Our findings allow for a more rational design of future microbiome studies, which would lead to higher repeatability of results and easier comparison between different laboratories. This could also facilitate the introduction of targeted HTS in clinical microbiology for reliable identification of pathogenic bacteria and testing for antimicrobial resistance (AMR). As AMR is a major threat to public health, the improved methods for determining AMR would bring great benefits to both the healthcare system and society as a whole.
Assuntos
DNA Bacteriano , Sequenciamento de Nucleotídeos em Larga Escala , Microbiota , Escarro , Escarro/microbiologia , Humanos , Microbiota/genética , DNA Bacteriano/genética , DNA Bacteriano/análise , DNA Bacteriano/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/efeitos dos fármacos , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/efeitos dos fármacosRESUMO
Effective isolation and sensitive detection of Pseudomonas aeruginosa (P. aeruginosa) is crucial for the early diagnosis and prognosis of various diseases, such as urinary tract infections. However, efficient isolation and simultaneous detection of P. aeruginosa remains a huge challenge. Herein, we depict a novel fluorescence assay for sensitive, enzyme-free detection of P. aeruginosa by integrating DNAzyme cascade-induced DNA tweezers and magnetic nanoparticles (MNPs)-based separation. The capture probe@MNPs is capable of accurately identifying target bacteria and transporting the bacteria signal to nucleic acid signals. Based on the DNAzyme cascade-induced DNA tweezers, the nucleic acid signals are extensively amplified, endowing the method with a high sensitivity and a low detection limit of 1 cfu/mL. In addition, the method also exhibits a wide detection of six orders of magnitudes. The proposed method could be extended to other bacteria detection by simply changing the aptamer sequence. Taking the merit of the high sensitivity, greatly minimized detection time (less than 1.5 h), enzyme-free characteristics, and stability, the proposed method could be potentially applied to diagnosing and preventing diseases caused by pathogenic bacteria.
Assuntos
Técnicas Biossensoriais , DNA Catalítico , Limite de Detecção , Pseudomonas aeruginosa , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/genética , DNA Catalítico/metabolismo , Técnicas Biossensoriais/métodos , Aptâmeros de Nucleotídeos/química , Nanopartículas de Magnetita/química , DNA Bacteriano/genética , Humanos , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/microbiologiaRESUMO
BACKGROUND: Antimicrobial-resistant (AMR) bacterial infection is a significant global threat to the healthcare systems. Pseudomonas aeruginosa, the leading infectious agent in the healthcare setting is now one of the major threats due to AMR. A comprehensive understanding of the magnitude of AMR, particularly highly public health important pathogens such as P. aeruginosa, is necessary for the management of infections based on local information. OBJECTIVE: This systematic review and meta-analysis aimed to determine the country-wide AMR of P. aeruginosa. METHODS: Systematic searches were performed to retrieve articles from PubMed, Scopus, Web of Science, ScienceDirect electronic databases, Google Scholar search engine, and repository registrars from 2015 to 31st December 2023. Twenty-three studies that provided important data on AMR in P. aeruginosa were systematically reviewed and analyzed to determine the country-wide magnitude of P. aeruginosa AMR profile from healthcare-associated infections. AMR of P. aeruginosa to 10 different antibiotics were extracted separately into Microsoft Excel and analyzed using STATA 17.0. Cohen's kappa was computed to determine the agreement between reviewers, the Inverse of variance (I2) was used to evaluate heterogeneity across studies, and Egger's test to identify publication bias. A random effect model was used to determine the pooled resistance to each antibiotic. Subgroup analysis was performed by infection type and year of publication. RESULTS: This systematic review and meta-analysis revealed that the pooled prevalence of P. aeruginosa in clinical specimens associated with HAI was 4.38%(95%CI: 3.00-5.76). The pooled prevalence of AMR in P. aeruginosa for different antibiotics varies, ranging from 20.9% (95%CI: 6.2-35.8) for amikacin to 98.72% (95%CI: 96.39-101.4) for ceftriaxone. The pooled resistance was higher for ceftriaxone (98.72%), Trimethoprim-sulfamethoxazole (75.41), and amoxicillin-clavulanic acid (91.2). In contrast relatively lower AMR were observed for amikacin (20.9%) and meropenem (28.64%). The pooled multi-drug resistance (MDR) in P. aeruginosa was 80.5% (95%CI: 66.25-93.84). Upon subgroup analysis by infection types and year of publication, P. aeruginosa isolated from healthcare-associated infections exhibited higher resistance to ceftazidime (94.72%) compared to isolates from mixed types of healthcare-associated infections (70.84%) and surgical site infections (57.84%). Antimicrobial resistance in gentamicin was higher during the periods of 2018-2020 (73.96%), while comparatively lower during 2021-2023 (42.69%) and 2015-2017 (29.82%). CONCLUSIONS: Significantly high AMR and MDR were observed from this systematic review and meta-analysis. AMR obtained from this systematic review and meta-analysis urges the need for improved infection control, antimicrobial stewardship practices, and strengthened surveillance systems to control the spread of AMR and ensure effective treatment of P. aeruginosa infections. PROTOCOL REGISTRATION: This systematic review and meta-analysis was registered on PROSPERO (Registration ID: CRD42024518145).
Assuntos
Antibacterianos , Infecção Hospitalar , Infecções por Pseudomonas , Pseudomonas aeruginosa , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/isolamento & purificação , Humanos , Infecção Hospitalar/microbiologia , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/tratamento farmacológico , Etiópia/epidemiologia , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/epidemiologia , Farmacorresistência Bacteriana , Testes de Sensibilidade MicrobianaRESUMO
INTRODUCTION: Although rare, Malignant otitis externa is responsible for a high morbidity and could sometimes be fatal. The management of this condition is still challenging. AIM: To analyse the clinical, microbiological and radiological profile of malignant otitis externa, and the management of this condition. METHODS: A descriptive, cross-sectional study was conducted at ENT Department of Kairouan's hospital including 38 patients hospitalised and treated for malignant otitis externa from January 2013 to August 2021. RESULTS: The mean age of patients was 67.7 ± 12.9 years (35-98). All patients presented with continuous otalgia that resists to usual analgesics. Otorrhea was noticed in 76.3% of cases, facial palsy in 2 cases (5.3%) and dysphonia in one case (2.6%). Pseudomonas Aeruginosa was the main responsible pathogen (42%). Concomitant bacterial and fungal infection was noticed in 6.4% of the cases. First-line intravenous antibiotherapy used was mainly based on an association of Cephalosporins and Fluoroquinolones. Complete remission was noticed in 30 patients (79%). However, 8 cases of recurrences (21%) and 2 cases of deaths (5.2%) were noticed in our series. The mean follow-up was 4.6±6.3 (1-26 months). CONCLUSIONS: Pseudomonas Aeruginosa remains the main responsible pathogen for malignant otitis externa. Nevertheless, fungal infections are rising because of the overuse of antibiotics. Antibiotherapy should be adapted to culture results and resistance profile of pathogens in hospital. Practionners should be aware of the possibility of concomitant fungal infection, especially in the case of unfavorable evolution.
Assuntos
Antibacterianos , Otite Externa , Infecções por Pseudomonas , Pseudomonas aeruginosa , Humanos , Otite Externa/microbiologia , Otite Externa/diagnóstico , Otite Externa/epidemiologia , Pessoa de Meia-Idade , Idoso , Masculino , Feminino , Estudos Transversais , Idoso de 80 Anos ou mais , Adulto , Antibacterianos/uso terapêutico , Pseudomonas aeruginosa/isolamento & purificação , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/epidemiologiaRESUMO
BACKGROUND: P. aeruginosa, a biofilm-forming bacteria, is the main cause of pulmonary infection in CF patients. We applied ZnO-np as a therapeutic agent for eradicating multi-drug resistance and biofilm-forming P. aeruginosa isolated from young CF patients. METHODS: A total of 73 throat and sputum samples taken from young CF patients were inquired. ZnO-np was synthesized and characterized in terms of size, shape, and structure for anti-bacterial activity. The antibiotic susceptibility of isolates before and after the addition of 16 µg/ml of ZnO was evaluated using disc diffusion and microtiter methods, respectively. The gene expression level of QS genes was assessed after treatment with 16 µg/ml ZnO-np. RESULTS: The optimum concentration of ZnO-np with a higher inhibitory zone was 16 µg/ml (MIC) and 32 µg/ml (MBC). All isolates were resistant to applied antibiotics, and about 45 % of isolates were strong biofilm-forming bacteria. After treatment with 16 µg/ml ZnO-np, all strains became susceptible to the applied antibiotic except for amikacin, which confers an intermediate pattern. About 63 % and 20 % of isolates were, respectively, non-biofilm and weak biofilm-forming bacteria following the addition of ZnO-np. Relative gene expression of gacA, lasR, and rhlR genes were downregulated significantly (P < 0.001). Although the retS did not have a significant reduction (P = 0.2) CONCLUSION: ZnO-np at a concentration of 16 µg/ml could significantly reduce the P. aeruginosa infection by altering the antibiotic susceptibility pattern and inhibiting biofilm formation. Due to their photocatalytic properties and their ability to penetrate the extracellular polysaccharide layer, ZnO nanoparticles can produce ROS, which increases their susceptibility to antibiotics. Nasal delivery of ZnO-np in the form of aerosol can be considered a potential strategy to decrease the mortality rate in CF patients at an early age.
Assuntos
Antibacterianos , Biofilmes , Fibrose Cística , Testes de Sensibilidade Microbiana , Nanopartículas , Infecções por Pseudomonas , Pseudomonas aeruginosa , Escarro , Óxido de Zinco , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Humanos , Antibacterianos/farmacologia , Óxido de Zinco/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Fibrose Cística/microbiologia , Fibrose Cística/complicações , Infecções por Pseudomonas/microbiologia , Escarro/microbiologia , Nanopartículas/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Percepção de Quorum/efeitos dos fármacos , Transativadores/genética , Transativadores/metabolismo , Farmacorresistência Bacteriana Múltipla/genética , Faringe/microbiologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Amicacina/farmacologiaRESUMO
Rapid detection and classification of pathogenic microbes for food hygiene, healthcare, environmental contamination, and chemical and biological exposures remain a major challenge due to nonavailability of fast and accurate detection methods. The delay in clinical diagnosis of the most frequent bacterial infections, particularly urinary tract infections (UTIs), which affect about half of the population at least once in their lifetime, can be fatal if not detected and treated appropriately. In this work, we have fabricated aluminum (Al) foil integrated pegylated gold nanoparticles (AuNPs) as a potential surface-enhanced Raman scattering (SERS) substrate, which is used for the detection and classification of uropathogens, namely, E. coli, S. aureus, and P. aeruginosa directly from the culture without any pretreatment. The substrate is first drop cast with bacterial pellets and then pegylated AuNPs, and the interaction of two on Al foil base gives identifiable characteristic Raman peaks with good reproducibility. With the use of chemometric methods such as principal component analysis (PCA), the Al foil-based SERS substrate offers a quick, effective detection and classification of three strains of UTI bacteria with the least bacterial concentration (105 cells mL-1) necessary for clinical diagnosis. In addition, this substrate was able to detect E. coli positive clinical samples by giving SERS fingerprint information directly from centrifuged urine samples within minutes. The stability of pegylated AuNPs provides for its application at the point of care with rapid and easy detection of uropathogens as well as the possibility of advancement in healthcare applications.
Assuntos
Alumínio , Escherichia coli , Ouro , Nanopartículas Metálicas , Polietilenoglicóis , Análise Espectral Raman , Staphylococcus aureus , Ouro/química , Nanopartículas Metálicas/química , Alumínio/química , Polietilenoglicóis/química , Escherichia coli/isolamento & purificação , Staphylococcus aureus/isolamento & purificação , Tamanho da Partícula , Propriedades de Superfície , Teste de Materiais , Materiais Biocompatíveis/química , Pseudomonas aeruginosa/isolamento & purificação , Infecções Urinárias/diagnóstico , Infecções Urinárias/microbiologia , HumanosRESUMO
The opportunistic human pathogen Pseudomonas aeruginosa (P. aeruginosa) poses a significant threat to human health, causing sepsis, inflammation, and pneumonia, so it is crucial to devise an expeditious detection platform for the P. aeruginosa. In this work, bis (2- (3, 5- dimethylphenyl) quinoline- C2, N') (acetylacetonato) iridium (III) Ir (dmpq)2 (acac) with excellent electrochemiluminescence (ECL) and fluorescence (FL) and magnetic nanoparticles were encapsulated in silica spheres. The luminescent units exhibited equal ECL and FL properties compared with single iridium complexes, and enabled rapid separation, which was of vital significance for the establishment of biosensors with effective detection. In addition, the luminescent units were further reacted with the DNA with quenching units to obtain the signal units, and the ECL/FL dual-mode biosensor was employed with the CRISPR/Cas12a system to further improve its specific recognition ability. The ECL detection linear range of as-proposed biosensor in this work was 100 fM-10 nM with the detection limit of 73 fM (S/N = 3), and FL detection linear range was 1 pM-10 nM with the detection limit of 0.126 pM (S/N = 3). Importantly, the proposed dual-mode biosensor exhibited excellent repeatability and stability in the detection of P. aeruginosa in real samples, underscoring its potential as an alternative strategy for infection prevention and safeguarding public health and safety in the future.
Assuntos
Técnicas Biossensoriais , Sistemas CRISPR-Cas , Irídio , Limite de Detecção , Medições Luminescentes , Pseudomonas aeruginosa , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/genética , Técnicas Biossensoriais/métodos , Irídio/química , Humanos , Técnicas Eletroquímicas/métodos , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/microbiologia , Nanopartículas de Magnetita/química , Fluorescência , Complexos de Coordenação/químicaRESUMO
PURPOSE: To describe the prevalence and antibiotic resistance profiles of Pseudomonas aeruginosa isolated from the Asia Cornea Society Infectious Keratitis Study (ACSIKS). METHODS: All bacterial isolates from ACSIKS underwent repeat microbiological identification in a central repository in Singapore. Minimum inhibitory concentration (MIC) determination was conducted for isolates of P. aeruginosa against thirteen antibiotics from 6 different classes, and categorized based on Clinical Laboratory Standard Institutes' reference ranges. The percentage rates of resistance (non-susceptibility) to each antibiotic included isolates of both intermediate and complete resistance. Multi-drug resistance (MDR) was defined as non-susceptibility to at least one agent in three or more antimicrobial classes. RESULTS: Of the 1493 unique bacterial specimens obtained from ACSIKS, 319 isolates were of P. aeruginosa. The majority of isolates were from centers in India (n = 118, 37%), Singapore (n = 90, 28.2%), Hong Kong (n = 31, 9.7%) and Thailand (n = 30, 9.4%). The cumulative antibiotic resistance rate was the greatest for polymyxin B (100%), ciprofloxacin (17.6%) and moxifloxacin (16.9%), and lowest for cefepime (11.6%) and amikacin (13.5%). Isolates from India demonstrated the highest antibiotic resistance rates of all the centers, and included moxifloxacin (47.5%) and ciprofloxacin (39.8%). Forty-eight of the 59 MDR isolates also originated from India. Antibiotic resistance rates were significantly lower in the other ACSIKS centers, and were typically less than 10%. CONCLUSIONS: The antibiotic resistance profiles of P. aeruginosa varied between different countries. While it was low for most countries, substantial antibiotic resistance and a significant number of multi-drug resistant isolates were noted in the centers from India.
Assuntos
Antibacterianos , Infecções Oculares Bacterianas , Testes de Sensibilidade Microbiana , Infecções por Pseudomonas , Pseudomonas aeruginosa , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/isolamento & purificação , Humanos , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/epidemiologia , Infecções Oculares Bacterianas/tratamento farmacológico , Antibacterianos/farmacologia , Infecções por Pseudomonas/epidemiologia , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/tratamento farmacológico , Sociedades Médicas , Masculino , Feminino , Prevalência , Farmacorresistência Bacteriana , Úlcera da Córnea/microbiologia , Úlcera da Córnea/epidemiologia , Úlcera da Córnea/tratamento farmacológico , Ceratite/microbiologia , Ceratite/epidemiologia , Ceratite/tratamento farmacológicoRESUMO
Developing a simple and highly sensitive approach for Pseudomonas aeruginosa (P. aeruginosa) detection is crucial, as it is closely associated with various disorders, such as newborn infections. Nevertheless, few of techniques have the capability to accurately identify P. aeruginosa with a high level of sensitivity and significantly improved stability. The employment of the both-end blocked peroxidase-mimicking DNAzyme significantly diminished the interferences from background signals, so conferring the approach with a high degree of selectivity and reproducibility. The proposed method is demonstrated with exceptional discernment capacity in differentiating interfering microorganisms. The simplicity, elevated sensitivity and high discerning capability make the method a highly promising alternative instrument for pathogenic bacteria detection.
This research presents a novel method for detecting P. aeruginosa using a combination of a simple molecular beacon (MB), duplex-specific nuclease (DSN), and both-end blocked peroxidase-mimicking DNAzyme. The MB probe utilized in this method can be shielded from DSN hydrolysis without requiring any additional modifications by regulating the number of stem bases to five. This assay is simple yet precise in its ability to quantitatively detect P. aeruginosa with a high level of sensitivity and specificity. In addition, the beacon enabled the identification of P. aeruginosa without the need for labeling, exhibiting a higher sensitivity over the conventional hairpin fluorescence beacon based methods.
Assuntos
DNA Catalítico , Infecções por Pseudomonas , Pseudomonas aeruginosa , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/genética , DNA Catalítico/metabolismo , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/diagnóstico , Recém-Nascido , Humanos , Peroxidase/metabolismo , Técnicas Biossensoriais/métodos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: In the hospital environment, carbapenemase-producing Pseudomonas aeruginosa (CPPA) may lead to fatal patient infections. However, the transmission routes of CPPA often remain unknown. Therefore, this case study aimed to trace the origin of CPPA ST357, which caused a hospital-acquired pneumonia in a repatriated critically ill patient suffering from Guillain-Barré Syndrome in 2023. METHODS: Antimicrobial susceptibility of the CPPA isolate for 30 single and combination therapies was determined by disk-diffusion, Etest or broth microdilution. Whole-genome sequencing was performed for three case CPPA isolates (one patient and two sinks) and four distinct CPPA ST357 patient isolates received in the Dutch CPPA surveillance program. Furthermore, 193 international P. aeruginosa ST357 assemblies were collected via three genome repositories and analyzed using whole-genome multi-locus sequence typing in combination with antimicrobial resistance gene (ARG) characterization. RESULTS: A Dutch patient who carried NDM-1-producing CPPA was transferred from Kenya to the Netherlands, with subsequent dissemination of CPPA isolates to the local sinks within a month after admission. The CPPA case isolates presented an extensively drug-resistant phenotype, with susceptibility only for colistin and cefiderocol-fosfomycin. Phylogenetic analysis showed considerable variation in allelic distances (mean = 150, max = 527 alleles) among the ST357 isolates from Asia (n = 92), Europe (n = 58), Africa (n = 21), America (n = 16), Oceania (n = 2) and unregistered regions (n = 4). However, the case isolates (n = 3) and additional Dutch patient surveillance program isolates (n = 2) were located in a sub-clade of isolates from Kenya (n = 17; varying 15-49 alleles), the United States (n = 7; 21-115 alleles) and other countries (n = 6; 14-121 alleles). This was consistent with previous hospitalization in Kenya of 2/3 Dutch patients. Additionally, over half of the isolates (20/35) in this sub-clade presented an identical resistome with 9/17 Kenyan, 5/5 Dutch, 4/7 United States and 2/6 other countries, which were characterized by the blaNDM-1, aph(3')-VI, ARR-3 and cmlA1 ARGs. CONCLUSION: This study presents an extensively-drug resistant subclone of NDM-producing P. aeruginosa ST357 with a unique resistome which was introduced to the Netherlands via repatriation of critically ill patients from Kenya. Therefore, the monitoring of repatriated patients for CPPA in conjunction with vigilance for the risk of environmental contamination is advisable to detect and prevent further dissemination.