ABSTRACT
Antibiotic resistance has emerged as a global threat to public health, generating a growing interest in investigating the presence of antibiotic-resistant bacteria in environments influenced by anthropogenic activities. Wastewater treatment plants in hospital serve as significant reservoirs of antimicrobial-resistant bacteria, where a favorable environment is established, promoting the proliferation and transfer of resistance genes among different bacterial species. In our study, we isolated a total of 243 strains from 5 hospital wastewater sites in Mexico, belonging to 21 distinct Gram-negative bacterial species. The presence of ß-lactamase was detected in 46.9% (114/243) of the isolates, which belonging to the Enterobacteriaceae family. We identified a total of 169 ß-lactamase genes; blaTEM in 33.1%, blaCTX-M in 25.4%, blaKPC in 25.4%, blaNDM 8.8%, blaSHV in 5.3%, and blaOXA-48 in 1.1% distributed in 12 different bacteria species. Among the 114 of the isolates, 50.8% were found to harbor at least one carbapenemase and were discharged into the environment. The carbapenemase blaKPC was found in six Citrobacter spp. and E. coli, while blaNDM was detected in two distinct Enterobacter spp. and E. coli. Notably, blaNDM-1 was identified in a 110 Kb IncFII conjugative plasmid in E. cloacae, E. xiangfangensis, and E. coli within the same hospital wastewater. In conclusion, hospital wastewater showed the presence of Enterobacteriaceae carrying a high frequency of carbapenemase blaKPC and blaNDM. We propose that hospital wastewater serves as reservoirs for resistance mechanism within bacterial communities and creates an optimal environment for the exchange of this resistance mechanism among different bacterial strains. IMPORTANCE: The significance of this study lies in its findings regarding the prevalence and diversity of antibiotic-resistant bacteria and genes identified in hospital wastewater in Mexico. The research underscores the urgent need for enhanced surveillance and prevention strategies to tackle the escalating challenge of antibiotic resistance, particularly evident through the elevated frequencies of carbapenemase genes such as blaKPC and blaNDM within the Enterobacteriaceae family. Moreover, the identification of these resistance genes on conjugative plasmids highlights the potential for widespread transmission via horizontal gene transfer. Understanding the mechanisms of antibiotic resistance in hospital wastewater is crucial for developing targeted interventions aimed at reducing transmission, thereby safeguarding public health and preserving the efficacy of antimicrobial therapies.
Subject(s)
Bacterial Proteins , Citrobacter , Enterobacter , Hospitals , Wastewater , beta-Lactamases , Wastewater/microbiology , beta-Lactamases/genetics , Bacterial Proteins/genetics , Citrobacter/genetics , Citrobacter/enzymology , Citrobacter/drug effects , Citrobacter/isolation & purification , Enterobacter/genetics , Enterobacter/drug effects , Enterobacter/isolation & purification , Enterobacter/enzymology , Anti-Bacterial Agents/pharmacology , MexicoABSTRACT
The species included in the ESKAPE group (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and the genus Enterobacter) have a high capacity to develop antimicrobial resistance (AMR), a health problem that is already among the leading causes of death and could kill 10 million people a year by 2050. The generation of new potentially therapeutic molecules has been insufficient to combat the AMR "crisis", and the World Health Organization (WHO) has stated that it will seek to promote the development of rapid diagnostic strategies. The physicochemical properties of metallic nanoparticles (MNPs) have made it possible to design biosensors capable of identifying low concentrations of ESKAPE bacteria in the short term; other systems identify antimicrobial susceptibility, and some have been designed with dual activity in situ (bacterial detection and antimicrobial activity), which suggests that, in the near future, multifunctional biosensors could exist based on MNPs capable of quickly identifying bacterial pathogens in clinical niches might become commercially available. This review focuses on the use of MNP-based systems for the rapid and accurate identification of clinically important bacterial pathogens, exhibiting the necessity for exhaustive research to achieve these objectives. This review focuses on the use of metal nanoparticle-based systems for the rapid and accurate identification of clinically important bacterial pathogens.
Subject(s)
Biosensing Techniques , Klebsiella pneumoniae , Metal Nanoparticles , Staphylococcus aureus , Metal Nanoparticles/chemistry , Humans , Klebsiella pneumoniae/drug effects , Staphylococcus aureus/drug effects , Acinetobacter baumannii/drug effects , Pseudomonas aeruginosa/drug effects , Enterococcus faecium , Drug Resistance, Bacterial , Anti-Bacterial Agents/pharmacology , Bacteria/drug effects , Early Diagnosis , Enterobacter/drug effectsABSTRACT
BACKGROUND: Currently, the Enterobacteriaceae species are responsible for a variety of serious infections and are already considered a global public health problem, especially in underdeveloped countries, where surveillance and monitoring programs are still scarce and limited. Analyses were performed on the complete genome of an extensively antibiotic-resistant strain of Enterobater hormaechei, which was isolated from a patient with non-Hodgkin's lymphoma, who had been admitted to a hospital in the city of Manaus, Brazil. METHODS: Phenotypical identification and susceptibility tests were performed in automated equipment. Total DNA extraction was performed using the PureLink genomic DNA mini-Kit. The genomic DNA library was prepared with Illumina Microbial Amplicon Prep and sequenced in the MiSeq Illumina Platform. The assembly of the whole-genome and individual analyses of specific resistance genes extracted were carried out using online tools and the Geneious Prime software. RESULTS: The analyses identified an extensively resistant ST90 clone of E. hormaechei carrying different genes, including blaCTX-M-15, blaGES-2, blaTEM-1A, blaACT-15, blaOXA-1 and blaNDM-1, [aac(3)-IIa, aac(6')-Ian, ant(2â³)-Ia], [aac(6')-Ib-cr, (qnrB1)], dfrA25, sul1 and sul2, catB3, fosA, and qnrB, in addition to resistance to chlorhexidine, which is widely used in patient antisepsis. CONCLUSIONS: These findings highlight the need for actions to control and monitor these pathogens in the hospital environment.
Subject(s)
Drug Resistance, Multiple, Bacterial , Enterobacter , Genome, Bacterial , Lymphoma, Non-Hodgkin , Whole Genome Sequencing , Humans , Enterobacter/genetics , Enterobacter/drug effects , Enterobacter/isolation & purification , Lymphoma, Non-Hodgkin/genetics , Lymphoma, Non-Hodgkin/microbiology , Lymphoma, Non-Hodgkin/drug therapy , Drug Resistance, Multiple, Bacterial/genetics , Whole Genome Sequencing/methods , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Enterobacteriaceae Infections/microbiology , Enterobacteriaceae Infections/drug therapy , Enterobacteriaceae Infections/genetics , Microbial Sensitivity Tests , BrazilABSTRACT
WHAT IS KNOWN AND OBJECTIVE: The widespread use of antibiotics as therapeutic agents caused an increase of multidrug resistant bacteria (MDR) appearance. Regarding MDRs, we highlight the Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.., which are the ESKAPE group. COMMENT: New treatment alternatives for infections caused by ESKAPE are under current scientific research. The main suggestions are the use of actinomycetes that produce promising substances with antibiotic activity, the synergistic effect between antimicrobials and peptides, photoinactivation, peptide rich in cationic histidine, association of new antimicrobials; besides the repositioning of drugs already approved for the treatment of other diseases. WHAT IS NEW AND CONCLUSION: These selected studies showed that researchers from many countries are focused on the development of effective alternative strategies for the treatment of infections caused by these microorganisms.
Subject(s)
Bacterial Infections/drug therapy , Acinetobacter baumannii/drug effects , Enterobacter/drug effects , Enterococcus faecium/drug effects , Humans , Klebsiella pneumoniae/drug effects , Pseudomonas aeruginosa/drug effects , Staphylococcus aureus/drug effectsABSTRACT
CTX-M-type extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae have become identified in marine ecosystem constituting a serious ecological issue. In this respect, although contamination of coastal waters and seafood, and even colonization of seabirds and fishes have been increasingly reported, molecular data are lacking to elucidate the clinical impact of ESBL producers in infected marine animals. In this study, using a genomic approach, we have analysed the genetic background of CTX-M-15-producing Enterobacter hormaechei (belonging to the international human clone ST114) and Citrobacter freundii (ST265) co-infecting a free-living green turtle (Chelonia mydas) suffering from septic arthritis, which progressed to generalized coelomitis and death. Wide resistome of these pathogens contributed to treatment failure and death of the animal.
Subject(s)
Citrobacter freundii/genetics , Coinfection/veterinary , Enterobacter/genetics , Enterobacteriaceae Infections/veterinary , Turtles/microbiology , beta-Lactamases/genetics , Animals , Anti-Bacterial Agents/pharmacology , Citrobacter freundii/drug effects , Coinfection/microbiology , Drug Resistance, Multiple, Bacterial , Enterobacter/drug effects , Enterobacteriaceae Infections/microbiologyABSTRACT
We report contemporary (2014-2016) Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) global data on activity of tigecycline and comparators against WHO 'priority pathogens', and global trends (2004-2016) in antimicrobial resistance. MICs were determined using CLSI broth microdilution methodology. Antimicrobial resistance was determined using CLSI breakpoints (FDA breakpoints for tigecycline). Data are reported for Africa, Asia, Europe, North America and South America. From 2014-2016, Africa, Asia and South America reported highest resistance rates among Acinetobacter baumannii; North America lowest (all antimicrobials tested). The tigecycline MIC90 against A. baumannii was 2 mg/L in all regions except South America (1 mg/L). Among Enterobacteriaceae, meropenem resistance was low and tigecycline resistance was ≤1.3% in all regions (Escherichia coli, 0.0-0.3%; Klebsiella pneumoniae 0.0-1.3%; Enterobacter spp. 0.5-1.1%; Serratia marcescens 0.0-1.3%). Ceftriaxone resistance among E. coli ranged from 14.5% (North America) to 54.7% (Asia), and among K. pneumoniae from 9.1% (North America) to 54.0% (South America). North America reported highest rates of vancomycin-resistant Enterococcus faecium (64.6%); Europe lowest (17.7%). The tigecycline MIC90 against methicillin-resistant Staphylococcus aureus (MRSA) ranged from 0.12 mg/L (Africa and North America) to 0.5 mg/L (Asia). From 2004-2016, carbapenem resistance increased among A. baumannii (all regions), reaching 92.3% in Africa and 85.7% in South America (2016). Rates of ceftriaxone-resistant E. coli increased in all regions except Asia. Ceftriaxone resistance in K. pneumoniae increased in Europe. Rates of vancomycin-resistant E. faecium and MRSA were highest in North America and South America (and Asia for MRSA); lowest in Europe.
Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Multiple, Bacterial , Epidemiological Monitoring , Gram-Negative Bacterial Infections/epidemiology , Gram-Positive Bacterial Infections/epidemiology , Tigecycline/pharmacology , Acinetobacter baumannii/drug effects , Acinetobacter baumannii/growth & development , Africa/epidemiology , Asia/epidemiology , Carbapenems/pharmacology , Ceftriaxone/pharmacology , Enterobacter/drug effects , Enterobacter/growth & development , Enterococcus faecium/drug effects , Enterococcus faecium/growth & development , Escherichia coli/drug effects , Escherichia coli/growth & development , Europe/epidemiology , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/microbiology , Gram-Positive Bacterial Infections/drug therapy , Gram-Positive Bacterial Infections/microbiology , Humans , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/growth & development , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/growth & development , Microbial Sensitivity Tests , North America/epidemiology , Serratia marcescens/drug effects , Serratia marcescens/growth & development , South America/epidemiologyABSTRACT
Abstract The production of KPC (Klebsiella pneumoniae carbapenemase) is the major mechanism of resistance to carbapenem agents in enterobacterias. In this context, forty KPC-producing Enterobacter spp. clinical isolates were studied. It was evaluated the activity of antimicrobial agents: polymyxin B, tigecycline, ertapenem, imipenem and meropenem, and was performed a comparison of the methodologies used to determine the susceptibility: broth microdilution, Etest® (bioMérieux), Vitek 2® automated system (bioMérieux) and disc diffusion. It was calculated the minimum inhibitory concentration (MIC) for each antimicrobial and polymyxin B showed the lowest concentrations for broth microdilution. Errors also were calculated among the techniques, tigecycline and ertapenem were the antibiotics with the largest and the lower number of discrepancies, respectively. Moreover, Vitek 2® automated system was the method most similar compared to the broth microdilution. Therefore, is important to evaluate the performance of new methods in comparison to the reference method, broth microdilution.
Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , beta-Lactamases/metabolism , Enterobacteriaceae Infections/microbiology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Microbial Sensitivity Tests/methods , Bacterial Proteins/genetics , beta-Lactamases/genetics , beta-Lactams/pharmacology , Drug Resistance, Bacterial , Enterobacter/drug effects , Enterobacter/genetics , Enterobacter/isolation & purification , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Polymyxin B/pharmacologyABSTRACT
The production of KPC (Klebsiella pneumoniae carbapenemase) is the major mechanism of resistance to carbapenem agents in enterobacterias. In this context, forty KPC-producing Enterobacter spp. clinical isolates were studied. It was evaluated the activity of antimicrobial agents: polymyxin B, tigecycline, ertapenem, imipenem and meropenem, and was performed a comparison of the methodologies used to determine the susceptibility: broth microdilution, Etest® (bioMérieux), Vitek 2® automated system (bioMérieux) and disc diffusion. It was calculated the minimum inhibitory concentration (MIC) for each antimicrobial and polymyxin B showed the lowest concentrations for broth microdilution. Errors also were calculated among the techniques, tigecycline and ertapenem were the antibiotics with the largest and the lower number of discrepancies, respectively. Moreover, Vitek 2® automated system was the method most similar compared to the broth microdilution. Therefore, is important to evaluate the performance of new methods in comparison to the reference method, broth microdilution.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Enterobacteriaceae Infections/microbiology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Microbial Sensitivity Tests/methods , beta-Lactamases/metabolism , Bacterial Proteins/genetics , Drug Resistance, Bacterial , Enterobacter/drug effects , Enterobacter/genetics , Enterobacter/isolation & purification , Ertapenem , Humans , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Polymyxin B/pharmacology , beta-Lactamases/genetics , beta-Lactams/pharmacologyABSTRACT
BACKGROUND: Carbapenems resistance in Enterobacter spp. has increased in the last decade, few studies, however, described the mechanisms of resistance in this bacterium. This study evaluated clonality and mechanisms of carbapenems resistance in clinical isolates of Enterobacter spp. identified in three hospitals in Brazil (Hospital A, B and C) over 7-year. METHODS: Antibiotics sensitivity, pulsed-field gel electrophoresis (PFGE), PCR for carbapenemase and efflux pump genes were performed for all carbapenems-resistant isolates. Outer-membrane protein (OMP) was evaluated based on PFGE profile. RESULTS: A total of 130 isolates of Enterobacter spp were analyzed, 44/105 (41, 9%) E. aerogenes and 8/25 (32,0%) E. cloacae were resistant to carbapenems. All isolates were susceptible to fosfomycin, polymyxin B and tigecycline. KPC was present in 88.6% of E. aerogenes and in all E. cloacae resistant to carbapenems. The carbapenems-resistant E. aerogenes identified in hospital A belonged to six clones, however, a predominant clone was identified in this hospital over the study period. There is a predominant clone in Hospital B and Hospital C as well. The mechanisms of resistance to carbapenems differ among subtypes. Most of the isolates co-harbored blaKPC, blaTEM and /or blaCTX associated with decreased or lost of 35-36KDa and or 39 KDa OMP. The efflux pump AcrAB-TolC gene was only identified in carbapenems-resistant E. cloacae. CONCLUSIONS: There was a predominant clone in each hospital suggesting that cross-transmission of carbapenems-resistant Enterobacter spp. was frequent. The isolates presented multiple mechanisms of resistance to carbapenems including OMP alteration.
Subject(s)
Bacterial Proteins/genetics , Enterobacter/genetics , Membrane Proteins/genetics , beta-Lactam Resistance/genetics , beta-Lactamases/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/pharmacology , Brazil , Carbapenems/pharmacology , DNA, Bacterial/genetics , Drug Resistance, Bacterial , Electrophoresis, Gel, Pulsed-Field/methods , Enterobacter/drug effects , Enterobacter/isolation & purification , Enterobacter/pathogenicity , Enterobacteriaceae Infections/microbiology , Female , Fosfomycin/pharmacology , Genes, Bacterial , Hospitals , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Minocycline/analogs & derivatives , Minocycline/pharmacology , Polymerase Chain Reaction , Polymyxin B/pharmacology , Tigecycline , Young AdultABSTRACT
This study assessed the antimicrobial efficacy and surface tension of established irrigating solutions with a new experimental chelating solution in infected dentin tubes. Twenty-five specimens were randomly assigned to each of the irrigating solutions. Twenty specimens were used as negative and positive controls. After 21 days of contamination with E. faecalis, the irrigating solutions MTAD, QMiX and Tetraclean NA were delivered into each infected root canal. The solutions were removed and dentin samples were withdrawn from the root canals with sterile low-speed round burs with increasing ISO diameters. The dentin powder samples obtained with each bur were immediately collected in separate test tubes containing 3 mL of BHI broth. After that, 100 µL from each test tube was cultured on blood agar. The grown colonies were counted and recorded as colony-forming units (CFU). The surface tension of the irrigants was measured using a Cahn DCA-322 Dynamic Contact Angle Analyzer. A Kruskal Wallis nonparametric ANOVA and a Friedman test were used (p<0.05). Tetraclean NA showed lower surface tension and CFU values than MTAD and QMiX. Better antibacterial action and low surface tension were observed for Tetraclean NA, probably due to the improved penetration into the root canal and dentinal tubes.
Subject(s)
Anti-Infective Agents/pharmacology , Chelating Agents/chemistry , Surface Tension , Surface-Active Agents/chemistry , Animals , Cattle , Enterobacter/drug effects , Root Canal IrrigantsABSTRACT
Besides the importance of Enterobacter cloacae species complex as a nosocomial pathogen, little is known about the frequency of each species/genotype. Here, we describe a strain of E. hormaechei subsp. hormaechei isolated from a bronchial secretion of a patient, in the Intensive Care Unit at the General Hospital of Cumaná, Venezuela, who died due to complications of his infection. The molecular identification was done by sequencing the 16S rRNA gene and comparing it to sequences from the GenBank. This strain showed resistance to multiple families of antibiotics (MDR), and the genes blaKPC and blaVIM were detected by PCR. This is the first time E. hormaechei has been identified in Venezuela.
A pesar de la importancia de las especies del complejo Enterobacter cloacae como patógeno nosocomial, poco se conoce sobre la frecuencia de cada especie/genotipo. Aquí se describe una cepa de E. hormaechei subsp. hormaechei aislada de una secreción bronquial de un paciente internado en la Unidad de Cuidados Intensivos del Hospital General de Cumaná, Venezuela, quien murió producto de complicaciones de su infección. La identificación molecular fue hecha por secuenciación del gen ARNr 16S y porcomparación con las secuencias del GenBank. Esta cepa mostró resistencia a múltiples familias de antibióticos (MDR) y se detectaron los genes blaKPCyblaVIMpor PCR. Este es el primer reporte de E. hormaechei en Venezuela.
Subject(s)
Humans , Male , Middle Aged , Drug Resistance, Multiple, Bacterial , Enterobacter/classification , Enterobacter/drug effects , Enterobacteriaceae Infections/microbiology , Venezuela , Microbial Sensitivity Tests , Fatal Outcome , Enterobacter/isolation & purificationABSTRACT
Abstract This study assessed the antimicrobial efficacy and surface tension of established irrigating solutions with a new experimental chelating solution in infected dentin tubes. Twenty-five specimens were randomly assigned to each of the irrigating solutions. Twenty specimens were used as negative and positive controls. After 21 days of contamination with E. faecalis, the irrigating solutions MTAD, QMiX and Tetraclean NA were delivered into each infected root canal. The solutions were removed and dentin samples were withdrawn from the root canals with sterile low-speed round burs with increasing ISO diameters. The dentin powder samples obtained with each bur were immediately collected in separate test tubes containing 3 mL of BHI broth. After that, 100 μL from each test tube was cultured on blood agar. The grown colonies were counted and recorded as colony-forming units (CFU). The surface tension of the irrigants was measured using a Cahn DCA-322 Dynamic Contact Angle Analyzer. A Kruskal Wallis nonparametric ANOVA and a Friedman test were used (p<0.05). Tetraclean NA showed lower surface tension and CFU values than MTAD and QMiX. Better antibacterial action and low surface tension were observed for Tetraclean NA, probably due to the improved penetration into the root canal and dentinal tubes
Resumo Este estudo avaliou a eficácia antimicrobiana e tensão superficial de soluções irrigadoras e uma nova solução quelante em tubos de dentina infectada. Vinte e cinco espécimes foram aleatoriamente distribuídos conforme as soluções irrigantes. Decorrifdos 21 dias de contaminação com E. faecalis, a soluções de irrigação MTAD, QMiX e Tetraclean NA foram distribuídas em cada canal radicular infectado. As soluções foram removidas e as amostras de dentina foram retiradas dos canais radiculares com brocas esféricas de baixa velocidade com diâmetros ISO sucessivamente maiores. As amostras do pó de dentina obtidas com cada broca foram imediatamente colocadas em tubos de ensaio separados contendo 3 mL de caldo BHI. A seguir, 100 μL de cada amostra do tubo de teste foi cultivada em agar de sangue. As colônias crescidas foram contadas e registadas como unidades formadoras de colônias (UFC). A tensão superficial das soluções irrigantes foi medida utilizando o método de Wilhelmy. A análise não paramétrica de Kruskal-Wallis e o teste de Friedman foram utilizados (p<0,05). Tetraclean NA apresentou menor tensão de superfície e menores valores de UFC do que MTAD e QMiX. A melhor ação antibacteriana e baixa tensão superficial foram observadas para Tetraclean NA, provavelmente devido à melhor penetração no canal radicular e túbulos dentinários.
Subject(s)
Animals , Cattle , Anti-Infective Agents/pharmacology , Chelating Agents/chemistry , Surface Tension , Surface-Active Agents/chemistry , Enterobacter/drug effects , Root Canal IrrigantsABSTRACT
Besides the importance of Enterobacter cloacae species complex as a nosocomial pathogen, little is known about the frequency of each species/genotype. Here, we describe a strain of E. hormaechei subsp. hormaechei isolated from a bronchial secretion of a patient, in the Intensive Care Unit at the General Hospital of Cumaná, Venezuela, who died due to complications of his infection. The molecular identification was done by sequencing the 16S rRNA gene and comparing it to sequences from the GenBank. This strain showed resistance to multiple families of antibiotics (MDR), and the genes blaKPC and blaVIM were detected by PCR. This is the first time E. hormaechei has been identified in Venezuela.
Subject(s)
Drug Resistance, Multiple, Bacterial , Enterobacter/classification , Enterobacter/drug effects , Enterobacteriaceae Infections/microbiology , Enterobacter/isolation & purification , Fatal Outcome , Humans , Male , Microbial Sensitivity Tests , Middle Aged , VenezuelaABSTRACT
In Enterobacteriaceae, the blaNDM genes have been found in many different genetic contexts, and a wide diversity of plasmid scaffolds bearing those genes has been found. In August 2013, we identified NDM-1-producing Escherichia coli and Enterobacter hormaechei strains from a single rectal swab sample from a patient hospitalized in Rio de Janeiro, Brazil, who had no history of travel abroad. Complete DNA sequencing using the Illumina platform and annotation of the two plasmids harboring the blaNDM-1 gene, one from each strain, showed that they belonged to incompatibility groups IncFIIK and IncX3 and harbored a novel transposon named Tn3000. Similar genetic structures have been identified among other isolates in Brazil but also on plasmids from other continents. Our findings suggest that the blaNDM-1 gene may be transmitted by Tn3000 in different parts of the world.
Subject(s)
DNA Transposable Elements/genetics , Enterobacter/isolation & purification , Escherichia coli/isolation & purification , beta-Lactamases/metabolism , Anti-Bacterial Agents/pharmacology , Aztreonam/pharmacology , Bacterial Proteins/genetics , Base Sequence , Brazil , Conjugation, Genetic , Conserved Sequence , Enterobacter/drug effects , Enterobacter/genetics , Enterobacter/metabolism , Enterobacteriaceae Infections/microbiology , Escherichia coli/drug effects , Escherichia coli/genetics , Escherichia coli/metabolism , Fosfomycin/pharmacology , Humans , India , Microbial Sensitivity Tests , Morocco , Nepal , Plasmids , Rectum/microbiology , beta-Lactamases/geneticsABSTRACT
The most important resistance mechanism against ß-lactam drugs is the production of carbapenemases. In this study, we report the first identification of Klebsiella pneumoniae carbapenemase (KPC)-2 and New Delhi metallo-ß-lactamase (NDM)-1 in Enterobacter hormaechei subps. oharae from Brazil. The detection of carbapenemases was done by phenotypic assays, PCR, and DNA sequencing, whereas the identification was performed by conventional techniques, sequencing of the 16S rDNA gene, and hsp60-genotyping. Molecular typing was performed using pulsed-field gel electrophoresis, and antimicrobial susceptibility was surrogated by the Etest methodology. Using the whole genome sequencing approach, we searched for resistance genes, plasmid incompatibility group genes, and the genetic environment of blaNDM and blaKPC. The plasmid identification was done by restriction digests with the S1 nuclease followed by hybridization using digoxigenin labeled specific probes. The isolate was considered multiresistant, being susceptible to amikacin and polymyxin B. We observed the following resistance genes: blaCTX-M-15, blaACT-7, blaTEM-1, blaOXA-1, aadA1, aadA2, strA, strB, aac(3)-II, qnrB1, and aac(6')-Ib-cr and incompatibility group plasmid genes IncA/C, IncHI2, and IncN. The blaKPC gene was found associated to the transposon Tn4401 isoform b in plasmid with 50 kb (IncN) and blaNDM-1 was flanked by a truncated ISAba125 and bleMBL in plasmid with 160 kb (IncA/C). This study showed the coproduction of two important carbapenemases (KPC-2 and NDM-1) associated with mobile genetic elements of worldwide epidemiological importance (Tn4401 and ISAba125, respectively), reinforcing the idea that urgent measures are necessary to reduce and prevent the spreading of these carbapenemases primarily in the hospital settings.
Subject(s)
Enterobacter/genetics , beta-Lactamases/genetics , Adult , Amikacin/therapeutic use , Anti-Bacterial Agents/therapeutic use , Bacterial Proteins/genetics , Brazil , DNA Transposable Elements/genetics , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial/drug effects , Drug Resistance, Multiple, Bacterial/genetics , Enterobacter/drug effects , Female , Genotype , Humans , Klebsiella Infections/drug therapy , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Plasmids/genetics , Polymyxin B/therapeutic use , beta-Lactams/pharmacologyABSTRACT
The present study proposed the isolation of arsenic resistant bacteria from wastewater. Only three bacterial isolates (MNZ1, MNZ4 and MNZ6) were able to grow in high concentrations of arsenic. The minimum inhibitory concentrations of arsenic against MNZ1, MNZ4 and MNZ6 were 300 mg/L, 300 mg/L and 370 mg/L respectively. The isolated strains showed maximum growth at 37 ºC and at 7.0 pH in control but in arsenite stress Luria Bertani broth the bacterial growth is lower than control. All strains were arsenite oxidizing. All strains were biochemically characterized and ribotyping (16S rRNA) was done for the purpose of identification which confirmed that MNZ1 was homologous to Enterobacter sp. while MNZ4 and MNZ6 showed their maximum homology with Klebsiella pneumoniae. The protein profiling of these strains showed in arsenic stressed and non stressed conditions, so no bands of induced proteins appeared in stressed conditions. The bacterial isolates can be exploited for bioremediation of arsenic containing wastes, since they seem to have the potential to oxidize the arsenite (more toxic) into arsenate (less toxic) form.
Subject(s)
Anti-Bacterial Agents/metabolism , Arsenic/metabolism , Drug Resistance, Bacterial , Enterobacter/drug effects , Klebsiella pneumoniae/drug effects , Wastewater/microbiology , Arsenites/metabolism , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Enterobacter/classification , Enterobacter/growth & development , Enterobacter/isolation & purification , Hydrogen-Ion Concentration , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/growth & development , Klebsiella pneumoniae/isolation & purification , Microbial Sensitivity Tests , Oxidation-Reduction , Proteome/analysis , Ribotyping , /genetics , TemperatureABSTRACT
The present study proposed the isolation of arsenic resistant bacteria from wastewater. Only three bacterial isolates (MNZ1, MNZ4 and MNZ6) were able to grow in high concentrations of arsenic. The minimum inhibitory concentrations of arsenic against MNZ1, MNZ4 and MNZ6 were 300 mg/L, 300 mg/L and 370 mg/L respectively. The isolated strains showed maximum growth at 37 ºC and at 7.0 pH in control but in arsenite stress Luria Bertani broth the bacterial growth is lower than control. All strains were arsenite oxidizing. All strains were biochemically characterized and ribotyping (16S rRNA) was done for the purpose of identification which confirmed that MNZ1 was homologous to Enterobacter sp. while MNZ4 and MNZ6 showed their maximum homology with Klebsiella pneumoniae. The protein profiling of these strains showed in arsenic stressed and non stressed conditions, so no bands of induced proteins appeared in stressed conditions. The bacterial isolates can be exploited for bioremediation of arsenic containing wastes, since they seem to have the potential to oxidize the arsenite (more toxic) into arsenate (less toxic) form.
Subject(s)
Anti-Bacterial Agents/metabolism , Arsenic/metabolism , Drug Resistance, Bacterial , Enterobacter/drug effects , Klebsiella pneumoniae/drug effects , Wastewater/microbiology , Arsenites/metabolism , DNA, Ribosomal/chemistry , Enterobacter/classification , Hydrogen-Ion Concentration , Klebsiella pneumoniae/classification , Microbial Sensitivity Tests , Oxidation-Reduction , Proteome/analysis , RibotypingABSTRACT
There are a growing number of reports of antibiotic resistance (ATBR) in bacteria living in wildlife. This is a cause for concern as ATBR in wildlife represents a potential public health threat. However, little is known about the factors that might determine the presence, abundance and dispersion of ATBR bacteria in wildlife. Here, we used culture and molecular methods to assess ATBR in bacteria in fecal samples from howler monkeys (Alouatta palliata), spider monkeys (Ateles geoffroyi), tapirs (Tapirus bairdii) and felids (jaguars, Panthera onca; pumas, Puma concolor; jaguarundis, Puma yagouaroundi; and ocelots, Leopardus pardalis) living freely in two regions of the Mexican state of Veracruz under different degrees of human influence. Overall, our study shows that ATBR is commonplace in bacteria isolated from wildlife in southeast Mexico. Most of the resistances were towards old and naturally occurring antibiotics, but we also observed resistances of potential clinical significance. We found that proximity to humans positively affected the presence of ATBR and that ATBR was higher in terrestrial than arboreal species. We also found evidence suggesting different terrestrial and aerial routes for the transmission of ATBR between humans and wildlife. The prevalence and potential ATBR transfer mechanisms between humans and wildlife observed in this study highlight the need for further studies to identify the factors that might determine ATBR presence, abundance and distribution.
Subject(s)
Drug Resistance, Multiple, Bacterial , Enterobacter/drug effects , Enterobacter/genetics , Feces/microbiology , Monkey Diseases/microbiology , Alouatta , Animals , Atelinae , Bacterial Typing Techniques , Enterobacter/isolation & purification , Felidae , Mexico , Microbial Sensitivity Tests , PumaABSTRACT
Biofilm formation depends on several factors. The influence of different osmolarities on bacterial biofilm formation was studied. Two strains (Enterobacter sp. and Stenotrophomonas sp.) exhibited the most remarkable alterations. Biofilm formation is an important trait and its use has been associated to the protection of organisms against environmental stresses.