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1.
J Water Health ; 22(5): 825-834, 2024 May.
Article in English | MEDLINE | ID: mdl-38822462

ABSTRACT

Hospital wastewater has been identified as a hotspot for the emergence and transmission of multidrug-resistant (MDR) pathogens that present a serious threat to public health. Therefore, we investigated the current status of antibiotic resistance as well as the phenotypic and genotypic basis of biofilm formation in Pseudomonas aeruginosa from hospital wastewater in Dhaka, Bangladesh. The disc diffusion method and the crystal violet assay were performed to characterize antimicrobial resistance and biofilm formation, respectively. Biofilm and integron-associated genes were amplified by the polymerase chain reaction. Isolates exhibited varying degrees of resistance to different antibiotics, in which >80% of isolates showed sensitivity to meropenem, amikacin, and gentamicin. The results indicated that 93.82% of isolates were MDR and 71 out of 76 MDR isolates showed biofilm formation activities. We observed the high prevalence of biofilm-related genes, in which algD+pelF+pslD+ (82.7%) was found to be the prevalent biofilm genotypic pattern. Sixteen isolates (19.75%) possessed class 1 integron (int1) genes. However, statistical analysis revealed no significant association between biofilm formation and multidrug resistance (χ2 = 0.35, P = 0.55). Taken together, hospital wastewater in Dhaka city may act as a reservoir for MDR and biofilm-forming P. aeruginosa, and therefore, the adequate treatment of wastewater is recommended to reduce the occurrence of outbreaks.


Subject(s)
Anti-Bacterial Agents , Biofilms , Drug Resistance, Multiple, Bacterial , Hospitals , Pseudomonas aeruginosa , Wastewater , Biofilms/drug effects , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/physiology , Pseudomonas aeruginosa/isolation & purification , Pseudomonas aeruginosa/genetics , Wastewater/microbiology , Bangladesh/epidemiology , Anti-Bacterial Agents/pharmacology , Integrons , Microbial Sensitivity Tests
2.
Food Microbiol ; 122: 104568, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38839227

ABSTRACT

The plasmid of emerging S. Infantis (pESI) or pESI-like plasmid in Salmonella enterica Infantis are consistently reported in poultry and humans worldwide. However, there has been limited research on these plasmids of S. Infantis isolated from eggs. Therefore, this study aimed to analyze the prevalence and characteristics of S. Infantis carrying the pESI-like plasmid from eggs in egg grading and packing plants. In this study, the pESI-like plasmid was only detected in 18 (78.3%) of 23 S. Infantis isolates, and it was absent in the other 9 Salmonella serovars. In particular, S. Infantis isolates carrying the pESI-like plasmid showed the significantly higher resistance to ß-lactams, phenicols, cephams, aminoglycosides, quinolones, sulfonamides, and tetracyclines than Salmonella isolates without the pESI-like plasmid (p < 0.05). Moreover, all S. Infantis isolates carrying the pESI-like plasmid were identified as extended-spectrum ß-lactamase (ESBL) producer, harboring the blaCTX-M-65 and blaTEM-1 genes, and carried non-ß-lactamase resistance genes (ant(3'')-Ia, aph(4)-Ia, aac(3)-IVa, aph(3')-Ic, sul1, tetA, dfrA14, and floR) against five antimicrobial classes. However, all isolates without the pESI-like plasmid only carried the blaTEM-1 gene among the ß-lactamase genes, and either had no non-ß-lactamase resistance genes or harbored non-ß-lactamase resistance genes against one or two antimicrobial classes. Furthermore, all S. Infantis isolates carrying the pESI-like plasmid carried class 1 and 2 integrons and the aadA1 gene cassette, but none of the other isolates without the pESI-like plasmid harbored integrons. In particular, D87Y substitution in the gyrA gene and IncP replicon type were observed in all the S. Infantis isolates carrying the pESI-like plasmid but not in the S. Infantis isolates without the pESI-like plasmid. The distribution of pulsotypes between pESI-positive and pESI-negative S. Infantis isolates was clearly distinguished, but all S. Infantis isolates were classified as sequence type 32, regardless of whether they carried the pESI-like plasmid. This study is the first to report the characteristics of S. Infantis carrying the pESI-like plasmid isolated from eggs and can provide valuable information for formulating strategies to control the spread of Salmonella in the egg industry worldwide.


Subject(s)
Anti-Bacterial Agents , Eggs , Plasmids , beta-Lactamases , Plasmids/genetics , Republic of Korea , Anti-Bacterial Agents/pharmacology , Eggs/microbiology , Animals , beta-Lactamases/genetics , Salmonella/genetics , Salmonella/isolation & purification , Salmonella/classification , Salmonella/drug effects , Microbial Sensitivity Tests , Drug Resistance, Multiple, Bacterial/genetics , Chickens/microbiology , Humans , Salmonella enterica/genetics , Salmonella enterica/isolation & purification , Salmonella enterica/drug effects , Salmonella enterica/classification
3.
Microbes Environ ; 39(2)2024.
Article in English | MEDLINE | ID: mdl-38839365

ABSTRACT

Shigella species are a group of highly transmissible Gram-negative pathogens. Increasing reports of infection with extensively drug-resistant varieties of this stomach bug has convinced the World Health Organization to prioritize Shigella for novel therapeutic interventions. We herein coupled the whole-genome sequencing of a natural isolate of Shigella flexneri with a pangenome ana-lysis to characterize pathogen genomics within this species, which will provide us with an insight into its existing genomic diversity and highlight the root causes behind the emergence of quick vaccine escape variants. The isolated novel strain of S. flexneri contained ~4,500 protein-coding genes, 57 of which imparted resistance to antibiotics. A comparative pan-genomic ana-lysis revealed genomic variability of ~64%, the shared conservation of core genes in central metabolic processes, and the enrichment of unique/accessory genes in virulence and defense mechanisms that contributed to much of the observed antimicrobial resistance (AMR). A pathway ana-lysis of the core genome mapped 22 genes to 2 antimicrobial resistance pathways, with the bulk coding for multidrug efflux pumps and two component regulatory systems that are considered to work synergistically towards the development of resistance phenotypes. The prospective evolvability of Shigella species as witnessed by the marked difference in genomic content, the strain-specific essentiality of unique/accessory genes, and the inclusion of a potent resistance mechanism within the core genome, strengthens the possibility of novel serotypes emerging in the near future and emphasizes the importance of tracking down genomic diversity in drug/vaccine design and AMR governance.


Subject(s)
Anti-Bacterial Agents , Genome, Bacterial , Genomics , Shigella flexneri , Wastewater , Shigella flexneri/genetics , Shigella flexneri/isolation & purification , Shigella flexneri/classification , Shigella flexneri/drug effects , Genome, Bacterial/genetics , Wastewater/microbiology , Anti-Bacterial Agents/pharmacology , Phylogeny , Whole Genome Sequencing , Drug Resistance, Multiple, Bacterial/genetics , Virulence/genetics
4.
J Coll Physicians Surg Pak ; 34(6): 677-681, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38840350

ABSTRACT

OBJECTIVE: To compare the susceptibility of colistin by two methods in extensive drug-resistant (XDR) Gram-negative isolates from ICU patients. STUDY DESIGN: Cross-sectional comparative analysis. Place and Duration of the Study: Department of Microbiology, Combined Military Hospital Karachi, Pakistan, from August 2022 to February 2023. METHODOLOGY: A total of 100 clinical specimens received from the intensive care unit yielded growth of extensively drug-resistant gram-negative bacteria, which were evaluated for polymyxin E susceptibility. The agar dilution method was compared with the reference broth microdilution (BMD) method. Minimum inhibitory concentration (MIC) was noted for both methods. RESULTS: Comparison of the MIC method by agar dilution showed a 90% correlation with the reference method of broth microdilution. With MICs within the acceptable range of the clinical and laboratory standards institute (CLSI) recommendations, 89 isolates were susceptible to colistin, whereas only 11 remained resistant. Polymyxin E's MIC 50 and MIC 90 were determined to be 1 and 2 µg/ml, respectively, with 97% susceptibility. CONCLUSION: Agar dilution susceptibility method can be used for screening purposes for the susceptibility testing of polymyxin E. This method is reliable and can easily identify the heteroresistance. KEY WORDS: Extensively drug-resistant, Broth microdilution, Multidrug-resistant, Agar dilution, Minimum inhibitory concentration, Colony forming unit.


Subject(s)
Anti-Bacterial Agents , Colistin , Drug Resistance, Multiple, Bacterial , Gram-Negative Bacteria , Gram-Negative Bacterial Infections , Intensive Care Units , Microbial Sensitivity Tests , Colistin/pharmacology , Humans , Microbial Sensitivity Tests/methods , Anti-Bacterial Agents/pharmacology , Cross-Sectional Studies , Gram-Negative Bacteria/drug effects , Gram-Negative Bacteria/isolation & purification , Pakistan , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/drug therapy
5.
Pak J Biol Sci ; 27(5): 268-275, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38840467

ABSTRACT

<b>Background and Objective:</b> Urinary tract infections from the use of an indwelling urinary catheter are one of the most common infections caused by <i>Proteus mirabilis</i>. Due to their biofilm-producing capacity and the increasing antimicrobial resistance in this microorganism, this study aimed to determine the prevalence, biofilm-producing capacity, antimicrobial resistance patterns, multidrug resistance and plasmid mediated resistance of the recovered isolates. <b>Materials and Methods:</b> A total of 50 urinary samples were collected from May to August, 2018 from patients on indwelling urinary catheters. Using routine microbiological and biochemical methods, 37 <i>P. mirabilis</i> were isolated. Biofilm forming capability was determined among the isolates using the tube method while antimicrobial susceptibility and plasmid curing were also performed. <b>Results:</b> All isolates were biofilm producers with 17(46%) being moderate producers while 20(54%) were strong biofilm formers. The study isolates exhibited a high resistance rate to empiric antibiotics, including ceftazidime (75.8%), cefuroxime (54.5%), ampicillin (69.7%) and amoxicillin-clavulanic acid (51.5%). Low resistance was seen in the fluoroquinolones, gentamicin and nitrofurantoin. Plasmid curing experiment revealed that most isolates lost their resistance indicating that resistance was borne on plasmids. Plasmid carriage is likely the reason for the high MDR rate of 56.8% observed. <b>Conclusion:</b> These findings necessitate the provision of infection control programs which will guide and implement policies.


Subject(s)
Anti-Bacterial Agents , Biofilms , Catheters, Indwelling , Microbial Sensitivity Tests , Proteus mirabilis , Biofilms/drug effects , Biofilms/growth & development , Proteus mirabilis/drug effects , Proteus mirabilis/genetics , Proteus mirabilis/isolation & purification , Catheters, Indwelling/microbiology , Catheters, Indwelling/adverse effects , Humans , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Urinary Tract Infections/microbiology , Urinary Tract Infections/drug therapy , Urinary Tract Infections/diagnosis , Plasmids/genetics , Urinary Catheters/microbiology , Urinary Catheters/adverse effects , Drug Resistance, Bacterial , Proteus Infections/microbiology , Proteus Infections/drug therapy , Catheter-Related Infections/microbiology , Catheter-Related Infections/diagnosis , Catheter-Related Infections/drug therapy , Female , Male , Drug Resistance, Multiple, Bacterial/genetics
6.
J Med Microbiol ; 73(6)2024 Jun.
Article in English | MEDLINE | ID: mdl-38842435

ABSTRACT

Introduction. Colistin (polymyxin E) has emerged as a last-resort treatment option for multidrug-resistant infections.Hypothesis/Gap Statement. Studies on the use, safety and efficacy of colistin in South Africa are limited.Aim. This study aims to describe the use of colistin and its clinical outcomes at a tertiary public hospital in South Africa.Methodology. We conducted a retrospective review of adult and paediatric patients who received parenteral colistin between 2015 and 2019.Results. A total of 69 patients (26 adults, 13 children and 30 neonates) were reviewed. Acinetobacter baumannii was the most common causative pathogen isolated (70.1 %). Colistin was predominately used to treat septicaemia (75.4 %). It was primarily administered as definitive therapy (71.0 %) and as monotherapy (56.5 %). It was used in 11.5 % of adults with infections susceptible to other antibiotics. Loading doses of intravenous colistin were administered in only 15 (57.7 %) adult patients. Neurotoxicity and nephrotoxicity occurred in 5.8 % and 43.5 % of patients, respectively. Clinical cure was achieved in 37 (53.6 %) patients. On multivariate logistic regression analysis, adults [adjusted odds ratio (aOR), 25.54; 95 % CI, 2.73-238.65; P < 0.01] and children (aOR, 8.56; 95 % CI, 1.06-69.10; P < 0.05) had higher odds of death than neonates.Conclusion. The study identified significant stewardship opportunities to improve colistin prescription and administration. Achieving optimal patient outcomes necessitates a multidisciplinary approach and vigilant monitoring of colistin use.


Subject(s)
Anti-Bacterial Agents , Antimicrobial Stewardship , Colistin , Tertiary Care Centers , Humans , Colistin/administration & dosage , Colistin/therapeutic use , Tertiary Care Centers/statistics & numerical data , South Africa , Retrospective Studies , Female , Adult , Male , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/administration & dosage , Infant , Middle Aged , Infant, Newborn , Child , Child, Preschool , Acinetobacter baumannii/drug effects , Adolescent , Young Adult , Aged , Drug Resistance, Multiple, Bacterial , Acinetobacter Infections/drug therapy , Sepsis/drug therapy , Sepsis/microbiology
7.
Nat Commun ; 15(1): 4731, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38830889

ABSTRACT

Major antibiotic groups are losing effectiveness due to the uncontrollable spread of antimicrobial resistance (AMR) genes. Among these, ß-lactam resistance genes -encoding ß-lactamases- stand as the most common resistance mechanism in Enterobacterales due to their frequent association with mobile genetic elements. In this context, novel approaches that counter mobile AMR are urgently needed. Collateral sensitivity (CS) occurs when the acquisition of resistance to one antibiotic increases susceptibility to another antibiotic and can be exploited to eliminate AMR selectively. However, most CS networks described so far emerge as a consequence of chromosomal mutations and cannot be leveraged to tackle mobile AMR. Here, we dissect the CS response elicited by the acquisition of a prevalent antibiotic resistance plasmid to reveal that the expression of the ß-lactamase gene blaOXA-48 induces CS to colistin and azithromycin. We next show that other clinically relevant mobile ß-lactamases produce similar CS responses in multiple, phylogenetically unrelated E. coli strains. Finally, by combining experiments with surveillance data comprising thousands of antibiotic susceptibility tests, we show that ß-lactamase-induced CS is pervasive within Enterobacterales. These results highlight that the physiological side-effects of ß-lactamases can be leveraged therapeutically, paving the way for the rational design of specific therapies to block mobile AMR or at least counteract their effects.


Subject(s)
Anti-Bacterial Agents , Escherichia coli , Microbial Sensitivity Tests , beta-Lactamases , beta-Lactamases/genetics , beta-Lactamases/metabolism , Escherichia coli/genetics , Escherichia coli/drug effects , Anti-Bacterial Agents/pharmacology , Drug Collateral Sensitivity/genetics , Plasmids/genetics , Azithromycin/pharmacology , Colistin/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , beta-Lactam Resistance/genetics
8.
BMC Res Notes ; 17(1): 151, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38831376

ABSTRACT

Staphylococcus aureus is a pathogen with high epidemic potential frequently involved in nosocomials and communities infections. The pathogenicity of Staphylococcus aureus is due to both its ability to resist antibiotics and to Produce toxins. This work aims at studying the resistance and Molecular Epidemiology of Staphylococcus aureus. Antibiotic susceptibility of the 70 strains isolates of Staphylococcus aureus was determined by agar diffusion while Multiplex PCR and MLST were used to search toxin-coding genes and MRSA typing, respectively. 14.28% of isolates were multidrug resistant. Staphylococcus aureus showed high susceptibility to aminoglycoside and Macrolides familly. lukS-PV/lukF-PV and sea genes were detected in 45% and 3% of Staphylococcus aureus respectively. Ten (10) sequence types including ST5710, ST2430, ST5289, ST5786, ST6942, ST6943, ST6944, ST6945, ST6946, ST6947 have been reported. The study showed a diversity of antibiotic resistance phenotypes and a great diversity of MRSA clones causing infections.


Subject(s)
Anti-Bacterial Agents , Microbial Sensitivity Tests , Staphylococcal Infections , Staphylococcus aureus , Humans , Staphylococcus aureus/genetics , Staphylococcus aureus/drug effects , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/pathogenicity , Staphylococcal Infections/microbiology , Staphylococcal Infections/epidemiology , Burkina Faso/epidemiology , Anti-Bacterial Agents/pharmacology , Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/pathogenicity , Multilocus Sequence Typing , Drug Resistance, Multiple, Bacterial/genetics
9.
PLoS One ; 19(6): e0299416, 2024.
Article in English | MEDLINE | ID: mdl-38833431

ABSTRACT

OBJECTIVE: To evaluate the association of multidrug-resistant bacteria (MDRB) and adverse clinical outcomes in patients with diabetic foot infection (DFI) in a Peruvian hospital. MATERIALS AND METHODS: This retrospective cohort study evaluated patients treated in the Diabetic Foot Unit of a General Hospital in Lima, Peru. MDRB was defined by resistance to more than two pharmacological groups across six clinically significant genera. The primary outcome was death due to DFI complications and/or major amputation. Other outcomes included minor amputation, hospitalization, and a hospital stay longer than 14 days. Relative risks were estimated using Poisson regression for all outcomes. RESULTS: The study included 192 DFI patients with a mean age of 59.9 years; 74% were males. A total of 80.8% exhibited MDRB. The primary outcome had an incidence rate of 23.2% and 5.4% in patients with and without MDRB, respectively (p = 0.01). After adjusting for sex, age, bone involvement, severe infection, ischemia, diabetes duration, and glycosylated hemoglobin, MDRB showed no association with the primary outcome (RR 3.29; 95% CI, 0.77-13.9), but did with hospitalization longer than 14 days (RR 1.43; 95% CI, 1.04-1.98). CONCLUSIONS: Our study found no association between MDRB and increased mortality and/or major amputation due to DFI complications, but did find a correlation with prolonged hospitalization. The high proportion of MDRB could limit the demonstration of the relationship. It is urgent to apply continuous evaluation of bacterial resistance, implement a rational plan for antibiotic use, and maintain biosafety to confront this threat.


Subject(s)
Anti-Bacterial Agents , Diabetic Foot , Drug Resistance, Multiple, Bacterial , Humans , Male , Female , Middle Aged , Diabetic Foot/microbiology , Diabetic Foot/drug therapy , Retrospective Studies , Peru/epidemiology , Aged , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology , Amputation, Surgical/statistics & numerical data , Treatment Outcome , Bacterial Infections/drug therapy , Bacterial Infections/epidemiology , Bacterial Infections/microbiology , Bacterial Infections/mortality , Length of Stay , Hospitalization
10.
PLoS One ; 19(6): e0304621, 2024.
Article in English | MEDLINE | ID: mdl-38833480

ABSTRACT

Salmonella enteritidis is a main pathogen responsible for sporadic outbreaks of gastroenteritis, and therefore is an important public health problem. This study investigated the drug resistance and genomic characteristics of S. enteritidis isolated from clinical and food sources in Huzhou, Zhejiang Province, China, from February 1, 2021, to December 30, 2023. In total, 43 S. enteritidis strains isolated during the study period were subjected to virulence gene, drug resistance gene, genetic correlation, antibiotic resistance, and multilocus sequence typing analyses. All 43 isolates were identified as ST11, and contained 108 virulence-related genes. Drug sensitivity analysis of the 43 isolates showed resistance rates of 100% to nalidixic acid and 90.70% to ampicillin and ampicillin/sulbactam. Multidrug resistance is a serious issue, with 81.40% of strains resistant to three or more antibacterial drugs. Genome sequencing indicated that S. enteritidis possessed 23 drug resistance genes, of which 14 were common to all 43 isolates. Phylogenetic analysis based on core genome single-nucleotide polymorphisms divided the 43 S. enteritidis strains into three clusters, with the 10 samples from an outbreak forming an independent branch located in cluster 3.


Subject(s)
Anti-Bacterial Agents , Genome, Bacterial , Phylogeny , Salmonella enteritidis , Salmonella enteritidis/genetics , Salmonella enteritidis/drug effects , Salmonella enteritidis/isolation & purification , China/epidemiology , Anti-Bacterial Agents/pharmacology , Humans , Multilocus Sequence Typing , Microbial Sensitivity Tests , Salmonella Infections/microbiology , Salmonella Infections/epidemiology , Salmonella Infections/drug therapy , Drug Resistance, Multiple, Bacterial/genetics , Polymorphism, Single Nucleotide , Drug Resistance, Bacterial/genetics , Whole Genome Sequencing
11.
Arch Microbiol ; 206(7): 292, 2024 Jun 07.
Article in English | MEDLINE | ID: mdl-38849633

ABSTRACT

In recent years, the evolution of antibiotic resistance has led to the inefficacy of several antibiotics, and the reverse of resistance was a novel method to solve this problem. We previously demonstrated that matrine (Mat) and berberine hydrochloride (Ber) had a synergistic effect against multidrug-resistant Escherichia coli (MDREC). This study aimed to demonstrate the effect of Mat combined with Ber in reversing the resistance of MDREC. The MDREC was sequenced passaged in the presence of Mat, Ber, and a combination of Mat and Ber, which did not affect its growth. The reverse rate was up to 39.67% after MDREC exposed to Mat + Ber for 15 days. The strain that reversed resistance was named drug resistance reversed E. coli (DRREC) and its resistance to ampicillin, streptomycin, gentamicin, and tetracycline was reversed. The MIC of Gentamicin Sulfate (GS) against DRREC decreased 128-fold to 0.63 µg/mL, and it was stable within 20 generations. Furthermore, the susceptible phenotype of DRREC remained stable within 20 generations, as well. The LD50 of DRREC for chickens was 8.69 × 109 CFU/mL. qRT-PCR assays revealed that the transcript levels of antibiotic-resistant genes and virulence genes in the DRREC strain were significantly lower than that in the MDREC strain (P < 0.05). In addition, GS decreased the death, decreased the bacterial loading in organs, alleviated the injury of the spleen and liver, and decreased the cytokine levels in the chickens infected by the DRREC strain. In contrast, the therapeutic effect of GS in chickens infected with MDREC was not as evident. These findings suggest that the combination of Mat and Ber has potential for reversing resistance to MDREC.


Subject(s)
Alkaloids , Anti-Bacterial Agents , Berberine , Chickens , Drug Resistance, Multiple, Bacterial , Escherichia coli Infections , Escherichia coli , Gentamicins , Matrines , Microbial Sensitivity Tests , Poultry Diseases , Quinolizines , Animals , Gentamicins/pharmacology , Escherichia coli/drug effects , Escherichia coli/genetics , Berberine/pharmacology , Anti-Bacterial Agents/pharmacology , Quinolizines/pharmacology , Escherichia coli Infections/veterinary , Escherichia coli Infections/drug therapy , Escherichia coli Infections/microbiology , Alkaloids/pharmacology , Poultry Diseases/microbiology , Poultry Diseases/drug therapy , Virulence/drug effects , Drug Synergism
12.
Sci Rep ; 14(1): 12902, 2024 06 05.
Article in English | MEDLINE | ID: mdl-38839922

ABSTRACT

Bacterial biofilms are highly complex communities in which isogenic bacteria display different gene expression patterns and organize in a three-dimensional mesh gaining enhanced resistance to biocides. The molecular mechanisms behind such increased resistance remain mostly unknown, also because of the technical difficulties in biofilm investigation at the sub-cellular and molecular level. In this work we focus on the AcrAB-TolC protein complex, a multidrug efflux pump found in Enterobacteriaceae, whose overexpression is associated with most multiple drug resistance (MDR) phenotypes occurring in Gram-negative bacteria. We propose an optical method to quantify the expression level of the AcrAB-TolC pump within the biofilm volume at the sub-cellular level, with single-molecule sensitivity. Through a combination of super-resolution PALM with single objective light sheet and precision genome editing, we can directly quantify the spatial distribution of endogenous AcrAB-TolC pumps expressed in both planktonic bacteria and, importantly, within the bacterial biofilm volume. We observe a gradient of pump density within the biofilm volume and over the course of biofilm maturation. Notably, we propose an optical method that could be broadly employed to achieve volumetric super-resolution imaging of thick samples.


Subject(s)
Biofilms , Biofilms/growth & development , Escherichia coli/genetics , Escherichia coli/metabolism , Escherichia coli Proteins/metabolism , Escherichia coli Proteins/genetics , Membrane Transport Proteins/metabolism , Membrane Transport Proteins/genetics , Drug Resistance, Multiple, Bacterial/genetics , Bacterial Proteins/metabolism , Bacterial Proteins/genetics , Carrier Proteins
13.
Euro Surveill ; 29(23)2024 Jun.
Article in English | MEDLINE | ID: mdl-38847120

ABSTRACT

BackgroundThe war in Ukraine led to migration of Ukrainian people. Early 2022, several European national surveillance systems detected multidrug-resistant (MDR) bacteria related to Ukrainian patients.AimTo investigate the genomic epidemiology of New Delhi metallo-ß-lactamase (NDM)-producing Providencia stuartii from Ukrainian patients among European countries.MethodsWhole-genome sequencing of 66 isolates sampled in 2022-2023 in 10 European countries enabled whole-genome multilocus sequence typing (wgMLST), identification of resistance genes, replicons, and plasmid reconstructions. Five bla NDM-1-carrying-P. stuartii isolates underwent antimicrobial susceptibility testing (AST). Transferability to Escherichia coli of a bla NDM-1-carrying plasmid from a patient strain was assessed. Epidemiological characteristics of patients with NDM-producing P. stuartii were gathered by questionnaire.ResultswgMLST of the 66 isolates revealed two genetic clusters unrelated to Ukraine and three linked to Ukrainian patients. Of these three, two comprised bla NDM-1-carrying-P. stuartii and the third bla NDM-5-carrying-P. stuartii. The bla NDM-1 clusters (PstCluster-001, n = 22 isolates; PstCluster-002, n = 8 isolates) comprised strains from seven and four countries, respectively. The bla NDM-5 cluster (PstCluster-003) included 13 isolates from six countries. PstCluster-001 and PstCluster-002 isolates carried an MDR plasmid harbouring bla NDM-1, bla OXA-10, bla CMY-16, rmtC and armA, which was transferrable in vitro and, for some Ukrainian patients, shared by other Enterobacterales. AST revealed PstCluster-001 isolates to be extensively drug-resistant (XDR), but susceptible to cefiderocol and aztreonam-avibactam. Patients with data on age (n = 41) were 19-74 years old; of 49 with information on sex, 38 were male.ConclusionXDR P. stuartii were introduced into European countries, requiring increased awareness and precautions when treating patients from conflict-affected areas.


Subject(s)
Anti-Bacterial Agents , Drug Resistance, Multiple, Bacterial , Enterobacteriaceae Infections , Microbial Sensitivity Tests , Multilocus Sequence Typing , Plasmids , Providencia , Whole Genome Sequencing , beta-Lactamases , Humans , Ukraine/epidemiology , beta-Lactamases/genetics , Enterobacteriaceae Infections/microbiology , Enterobacteriaceae Infections/epidemiology , Enterobacteriaceae Infections/drug therapy , Drug Resistance, Multiple, Bacterial/genetics , Providencia/genetics , Providencia/isolation & purification , Providencia/drug effects , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Europe/epidemiology , Plasmids/genetics , Male , Adult , Female , Middle Aged , Aged , Young Adult
14.
Sci Rep ; 14(1): 13235, 2024 Jun 09.
Article in English | MEDLINE | ID: mdl-38853154

ABSTRACT

The study investigated the economic concerns associated with livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in livestock (cow), examining its connection to severe infections, antimicrobial resistance (AMR), and virulence factors. The research, conducted in Edo State, Nigeria, analyzed 400 samples (200 rectal and 200 nasal swabs) collected between March 2018 and February 2019. MRSA prevalence was identified using conventional culture-based methods and polymerase chain reaction (PCR) techniques, revealing 63.5% (n = 254) for Staphylococcus aureus and 55% (n = 220) for MRSA. Of the 76 mecA-positive MRSA isolates, 64.5% (n = 49) exhibited multidrug resistance (MDR) while the remaining were sensitive to specific antimicrobials. Key virulence genes, such as PVL (81.6%; n = 62) and tsst-1 (44.7%; n = 34), were prevalent, along with AMR genes like mecC, tetM, ermA, ermC, vanA, and vanC. Staphylococcal chromosomal cassette mec (SCCmec) typing identified different types, notably II, IVa, and IVb. Biofilm formation, a crucial virulence factor varied in strength, is associated with icaA and icaB genes (p < 0.01). The findings highlighted substantial AMR and biofilm-forming capacity within LA-MRSA isolates, emphasizing the importance of ongoing surveillance for informed treatment strategies, AMR policies, and control measures against MDR staphylococcal infections.


Subject(s)
Biofilms , Livestock , Methicillin-Resistant Staphylococcus aureus , Staphylococcal Infections , Virulence Factors , Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/pathogenicity , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Animals , Virulence Factors/genetics , Staphylococcal Infections/microbiology , Staphylococcal Infections/veterinary , Livestock/microbiology , Cattle , Biofilms/drug effects , Biofilms/growth & development , Microbial Sensitivity Tests , Anti-Bacterial Agents/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Nigeria/epidemiology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism
15.
AMA J Ethics ; 26(6): E494-501, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38833425

ABSTRACT

Overprescription of antibiotics in cases in which bacterial infection is clinically uncertain contributes to increased prevalence of multidrug-resistant bacteria. Ethically, merits and drawbacks of stricter prescription practice oversight should be weighed against risks of untreatable bacterial infections to patients and communities. This article considers how to balance this set of ideas and values.


Subject(s)
Anti-Bacterial Agents , Patient-Centered Care , Humans , Anti-Bacterial Agents/therapeutic use , Patient-Centered Care/ethics , Bacterial Infections/drug therapy , Drug Resistance, Multiple, Bacterial , Inappropriate Prescribing/prevention & control , Drug Resistance, Bacterial
16.
PLoS Pathog ; 20(6): e1012235, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38843111

ABSTRACT

Amikacin and piperacillin/tazobactam are frequent antibiotic choices to treat bloodstream infection, which is commonly fatal and most often caused by bacteria from the family Enterobacterales. Here we show that two gene cassettes located side-by-side in and ancestral integron similar to In37 have been "harvested" by insertion sequence IS26 as a transposon that is widely disseminated among the Enterobacterales. This transposon encodes the enzymes AAC(6')-Ib-cr and OXA-1, reported, respectively, as amikacin and piperacillin/tazobactam resistance mechanisms. However, by studying bloodstream infection isolates from 769 patients from three hospitals serving a population of 1.2 million people in South West England, we show that increased enzyme production due to mutation in an IS26/In37-derived hybrid promoter or, more commonly, increased transposon copy number is required to simultaneously remove these two key therapeutic options; in many cases leaving only the last-resort antibiotic, meropenem. These findings may help improve the accuracy of predicting piperacillin/tazobactam treatment failure, allowing stratification of patients to receive meropenem or piperacillin/tazobactam, which may improve outcome and slow the emergence of meropenem resistance.


Subject(s)
Anti-Bacterial Agents , DNA Transposable Elements , Humans , Anti-Bacterial Agents/pharmacology , DNA Transposable Elements/genetics , Drug Resistance, Multiple, Bacterial/genetics , Piperacillin/pharmacology , Amikacin/pharmacology , Microbial Sensitivity Tests , Enterobacteriaceae Infections/microbiology , Enterobacteriaceae Infections/drug therapy , Enterobacteriaceae Infections/genetics , Enterobacteriaceae/genetics , Enterobacteriaceae/drug effects , Integrons/genetics , Bacteremia/microbiology , Bacteremia/drug therapy , Bacteremia/genetics
17.
Front Cell Infect Microbiol ; 14: 1375872, 2024.
Article in English | MEDLINE | ID: mdl-38846355

ABSTRACT

Introduction: Pseudomonas aeruginosa is notorious for its multidrug resistance and its involvement in hospital-acquired infections. In this study, 20 bacterial strains isolated from soil samples near the Hindan River in Ghaziabad, India, were investigated for their biochemical and morphological characteristics, with a focus on identifying strains with exceptional drug resistance and pyocyanin production. Methods: The isolated bacterial strains were subjected to biochemical and morphological analyses to characterize their properties, with a particular emphasis on exopolysaccharide production. Strain GZB16/CEES1, exhibiting remarkable drug resistance and pyocyanin production. Biochemical and molecular analyses, including sequencing of its 16S rRNA gene (accession number LN735036.1), plasmid-curing assays, and estimation of plasmid size, were conducted to elucidate its drug resistance mechanisms and further pyocynin based target the Candida albicans Strain GZB16/CEES1 demonstrated 100% resistance to various antibiotics used in the investigation, with plasmid-curing assays, suggesting plasmid-based resistance gene transmission. The plasmid in GZB16/CEES1 was estimated to be approximately 24 kb in size. The study focused on P. aeruginosa's pyocyanin production, revealing its association with anticandidal activity. The minimum inhibitory concentration (MIC) of the bacterial extract against Candida albicans was 50 µg/ml, with a slightly lower pyocyanin-based MIC of 38.5 µg/ml. Scanning electron microscopy illustrated direct interactions between P. aeruginosa strains and Candida albicans cells, leading to the destruction of the latter. Discussion: These findings underscore the potential of P. aeruginosa in understanding microbial interactions and developing strategies to combat fungal infections. The study highlights the importance of investigating bacterial-fungal interactions and the role of pyocyanin in antimicrobial activity. Further research in this area could lead to the development of novel therapeutic approaches for combating multidrug-resistant infections.


Subject(s)
Antifungal Agents , Candida albicans , Drug Resistance, Multiple, Bacterial , Microbial Sensitivity Tests , Plasmids , Pseudomonas aeruginosa , Pyocyanine , RNA, Ribosomal, 16S , Soil Microbiology , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/genetics , Pseudomonas aeruginosa/metabolism , Pyocyanine/metabolism , Drug Resistance, Multiple, Bacterial/genetics , Antifungal Agents/pharmacology , Candida albicans/drug effects , Candida albicans/genetics , Candida albicans/growth & development , RNA, Ribosomal, 16S/genetics , India , Plasmids/genetics , Anti-Bacterial Agents/pharmacology , Antibiosis
18.
Arch Microbiol ; 206(7): 294, 2024 Jun 08.
Article in English | MEDLINE | ID: mdl-38850339

ABSTRACT

Antimicrobial resistance is a prevalent problem witnessed globally and creating an alarming situation for the treatment of infections caused by resistant pathogens. Available armaments such as antibiotics often fail to exhibit the intended action against resistant pathogens, leading to failure in the treatments that are causing mortality. New antibiotics or a new treatment approach is necessary to combat this situation. P. aeruginosa is an opportunistic drug resistant pathogen and is the sixth most common cause of nosocomial infections. P. aeruginosa due to its genome organization and other factors are exhibiting resistance against drugs. Bacterial biofilm formation, low permeability of outer membrane, the production of the beta-lactamase, and the production of several efflux systems limits the antibacterial potential of several classes of antibiotics. Combination of phytoconstituents with antibiotics is a promising strategy to combat multidrug resistant P. aeruginosa. Phytoconstituents such as flavonoids, terpenoids, alkaloids, polypeptides, phenolics, and essential oils are well known antibacterial agents. In this review, the activity of combination of the phytoconstituents and antibiotics, and their corresponding mechanism of action was discussed elaborately. The combination of antibiotics and plant-derived compounds exhibited better efficacy compared to antibiotics alone against the antibiotic resistance P. aeruginosa infections.


Subject(s)
Anti-Bacterial Agents , Biofilms , Phytochemicals , Pseudomonas aeruginosa , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/genetics , Anti-Bacterial Agents/pharmacology , Phytochemicals/pharmacology , Phytochemicals/chemistry , Biofilms/drug effects , Drug Resistance, Multiple, Bacterial/drug effects , Humans , Pseudomonas Infections/drug therapy , Pseudomonas Infections/microbiology , Microbial Sensitivity Tests
19.
World J Microbiol Biotechnol ; 40(8): 235, 2024 Jun 08.
Article in English | MEDLINE | ID: mdl-38850338

ABSTRACT

Lactobacillus delbrueckii, a widely used lactic acid bacterium in the food industry, has been studied for its probiotic properties and reservoir of antibiotic-resistant genes, raising safety concerns for probiotic formulations and fermented products. This review consolidates findings from 60 articles published between 2012 and 2023, focusing on the global antibiotic resistance profile and associated genetic factors in L. delbrueckii strains. Resistance to aminoglycosides, particularly streptomycin, kanamycin, and gentamicin, as well as resistance to glycopeptides (vancomycin), fluoroquinolones (ciprofloxacin), and tetracyclines was predominant. Notably, although resistance genes have been identified, they have not been linked to mobile genetic elements, reducing the risk of dissemination. However, a significant limitation is the insufficient exploration of responsible genes or mobile elements in 80% of studies, hindering safety assessments. Additionally, most articles originated from Asian and Middle Eastern countries, with strains often isolated from fermented dairy foods. Therefore, these findings underscore the necessity for comprehensive analyses of new strains of L. delbrueckii for potential industrial and biotherapeutic applications and in combating the rise of antibiotic-resistant pathogens.


Subject(s)
Anti-Bacterial Agents , Lactobacillus delbrueckii , Probiotics , Probiotics/pharmacology , Lactobacillus delbrueckii/genetics , Lactobacillus delbrueckii/drug effects , Anti-Bacterial Agents/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Food Industry , Food Microbiology , Fermented Foods/microbiology
20.
Arch Virol ; 169(7): 142, 2024 Jun 08.
Article in English | MEDLINE | ID: mdl-38851653

ABSTRACT

Uropathogenic Escherichia coli (UPEC) is the most common causative agent of urinary tract infections, and strains that are resistant to antibiotics are a major problem in treating these infections. Phage therapy is a promising alternative approach that can be used to treat infections caused by polyresistant bacterial strains. In the present study, 16 bacteriophages isolated from sewage and surface water were investigated. Phage host specificity was tested on a collection of 77 UPEC strains. The phages infected 2-44 strains, and 80% of the strains were infected by at least one phage. The susceptible E. coli strains belonged predominantly to the B2 phylogenetic group, including strains of two clones, CC131 and CC73, that have a worldwide distribution. All of the phages belonged to class Caudoviricetes and were identified as members of the families Straboviridae, Autographiviridae, and Drexlerviridae and the genera Kagunavirus, Justusliebigvirus, and Murrayvirus. A phage cocktail composed of six phages - four members of the family Straboviridae and two members of the family Autographiviridae - was prepared, and its antibacterial activity was tested in liquid medium. Complete suppression of bacterial growth was observed after 5-22 hours of cultivation, followed by partial regrowth. At 24 hours postinfection, the cocktail suppressed bacterial growth to 43-92% of control values. Similar results were obtained when testing the activity of the phage cocktail in LB and in artificial urine medium. The results indicate that our phage cocktail has potential to inhibit bacterial growth during infection, and they will therefore be preserved in the national phage bank, serving as valuable resources for therapeutic applications.


Subject(s)
Drug Resistance, Multiple, Bacterial , Host Specificity , Phylogeny , Uropathogenic Escherichia coli , Uropathogenic Escherichia coli/virology , Uropathogenic Escherichia coli/drug effects , Bacteriophages/classification , Bacteriophages/physiology , Bacteriophages/genetics , Bacteriophages/isolation & purification , Sewage/virology , Phage Therapy/methods , Humans , Anti-Bacterial Agents/pharmacology , Escherichia coli Infections/microbiology , Escherichia coli Infections/therapy
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