10 results on '"Renata Katsuko Takayama Kobayashi"'
Search Results
2. High-Frequency Detection of fosA3 and blaCTX–M–55 Genes in Escherichia coli From Longitudinal Monitoring in Broiler Chicken Farms
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Maísa Fabiana Menck-Costa, Ana Angelita Sampaio Baptista, Luiz Eduardo de Souza Gazal, Larissa Justino, Matheus Silva Sanches, Marielen de Souza, Erick Kenji Nishio, Beatriz Queiroz dos Santos, Victor Dellevedove Cruz, João Vitor Monteiro Berbert, Bruna Carolina Gonçalves, Galdino Andrade, Eliana Carolina Vespero, Gerson Nakazato, and Renata Katsuko Takayama Kobayashi
- Subjects
blaCTX–M–55 gene ,fosA3 gene ,broiler chicken farms ,multidrug resistance ,fosfomycin ,longitudinal monitoring ,Microbiology ,QR1-502 - Abstract
Considering the worrying emergence of multidrug resistance, including in animal husbandry and especially in food-producing animals, the need to detect antimicrobial resistance strains in poultry environments is relevant, mainly considering a One Health approach. Thus, this study aimed to conduct longitudinal monitoring of antimicrobial resistance in broiler chicken farms, with an emphasis on evaluating the frequency of resistance to fosfomycin and β-lactams. Escherichia coli was isolated from broiler chicken farms (cloacal swabs, meconium, poultry feed, water, poultry litter, and Alphitobius diaperinus) in northern Paraná from 2019 to 2020 during three periods: the first period (1st days of life), the second period (20th to 25th days of life), and third period (40th to 42nd days of life). Antibiogram tests and the detection of phenotypic extended-spectrum β-lactamase (ESBL) were performed, and they were confirmed by seaching for genes from the blaCTX–M group. The other resistance genes searched were mcr-1 and fosA3. Some ESBL blaCTX–M–1 group strains were selected for ESBL identification by sequencing and enterobacterial repetitive intergenic consensus-polymerase chain reaction analysis. To determine the transferability of the blaCTX–M–1– and fosA3-carrying plasmids, strains were subjected to conjugation experiments. A total of 507 E. coli were analyzed: 360 from cloacal swabs, 24 from meconium samples, 3 from poultry feed samples, 18 from water samples, 69 from poultry litter samples, and 33 from A. diaperinus samples. Among the strain isolate, 80% (406/507) were multidrug-resistant (MDR), and 51% (260/507) were ESBL-positive, with the blaCTX–M–1 group being the most frequent. For the fosA3 gene, 68% (344/507) of the strains isolated were positive, deserves to be highlighted E. coli isolated from day-old chickens (OR 6.34, CI 2.34–17.17), when compared with strains isolated from other origins (poultry litter, A. diaperinus, water, and poultry feed). This work alerts us to the high frequency of the fosA3 gene correlated with the CTX-M-1 group (OR 3.57, CI 95% 2.7–4.72, p < 0.05), especially the blaCTX–M–55 gene, in broiler chickens. This profile was observed mainly in day-old chicken, with a high percentage of E. coli that were MDR. The findings emphasize the importance of conducting longitudinal monitoring to detect the primary risk points during poultry production.
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- 2022
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3. Subinhibitory Concentrations of Biogenic Silver Nanoparticles Affect Motility and Biofilm Formation in Pseudomonas aeruginosa
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Erika Kushikawa Saeki, Amanda Yaeko Yamada, Larissa Amianti de Araujo, Laís Anversa, Doroti de Oliveira Garcia, Renan Luiz Barros de Souza, Heloísa Moreira Martins, Renata Katsuko Takayama Kobayashi, and Gerson Nakazato
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antibiofilm ,antivirulence ,virulence factor ,metal nanoparticle ,quorum sensing ,Microbiology ,QR1-502 - Abstract
Biogenic silver nanoparticles (bio-AgNPs) are increasingly recognized as an antibiofilm and antivirulence strategy against P. aeruginosa, a bacterium that causes chronic infections in immunocompromised and cystic fibrosis patients. This study aimed to investigate the effects of subinhibitory concentrations of bio-AgNPs on motility and biofilm formation in P. aeruginosa. Bio-AgNPs were synthesized via reduction of ionic silver catalyzed by cell-free culture filtrate from Fusarium oxysporum. A total of 17 P. aeruginosa isolates and strains were evaluated for swarming, swimming, and twitching motility in the presence and absence (control) of bio-AgNPs, including 10 clinical isolates from patients with and without cystic fibrosis, 5 environmental isolates obtained from the public water supply system, and 2 reference strains (PAO1 and PA14). Isolates were identified by biochemical and molecular methods. Minimum inhibitory concentrations (MICs) were determined by the broth microdilution method. Swarming, swimming, and twitching motility assays were performed in Petri dishes. Biofilm formation capacity was assessed quantitatively by the crystal violet method. MIC values ranged from 15.62 to 62.50 µM. The results showed that subinhibitory concentrations of bio-AgNPs (½ MIC, 7.81–31.25 µM) significantly increased (p < 0.05) swarming, swimming, and twitching motility in 40.0, 40.0, and 46.7% of isolates, respectively. Subinhibitory bio-AgNP treatment enhanced (p < 0.05) biofilm formation capacity in PA14 and a cystic fibrosis isolate (P11). It is concluded that subinhibitory concentrations of bio-AgNPs increased biofilm formation and swarming, swimming, and twitching motility in PA14 and some P. aeruginosa isolates. These virulence factors are directly involved with quorum-sensing systems. Further research should investigate the effects of AgNPs on P. aeruginosa quorum sensing to help elucidate their mechanism of action at subinhibitory concentrations.
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- 2021
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4. Detection of ESBL/AmpC-Producing and Fosfomycin-Resistant Escherichia coli From Different Sources in Poultry Production in Southern Brazil
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Luís Eduardo de Souza Gazal, Leonardo Pinto Medeiros, Miriam Dibo, Erick Kenji Nishio, Vanessa Lumi Koga, Bruna Carolina Gonçalves, Tiela Trapp Grassotti, Taiara Carolaine Leal de Camargo, João Juliano Pinheiro, Eliana Carolina Vespero, Kelly Cristina Tagliari de Brito, Benito Guimarães de Brito, Gerson Nakazato, and Renata Katsuko Takayama Kobayashi
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Avian ,multidrug resistance (MDR) ,Enterobacteriaceae ,fosfomycin ,poultry litter ,public health ,Microbiology ,QR1-502 - Abstract
This study discussed the use of antimicrobials in the commercial chicken production system and the possible factors influencing the presence of Extended-spectrum β-lactamase (ESBL)/AmpC producers strains in the broiler production chain. The aim of this study was to perform longitudinal monitoring of ESBL-producing and fosfomycin-resistant Escherichia coli from poultry farms in southern Brazil (Paraná and Rio Grande do Sul states) and determine the possible critical points that may be reservoirs for these strains. Samples of poultry litter, cloacal swabs, poultry feed, water, and beetles (Alphitobius sp.) were collected during three distinct samplings. Phenotypic and genotypic tests were performed for characterization of antimicrobial resistant strains. A total of 117 strains were isolated and 78 (66%) were positive for ESBL production. The poultry litter presented ESBL positive strains in all three sampled periods, whereas the cloacal swab presented positive strains only from the second period. The poultry litter represents a significant risk factor mainly at the beginning poultry production (odds ratio 6.43, 95% confidence interval 1–41.21, p < 0.05). All beetles presented ESBL positive strains. The predominant gene was blaCTX–M group 2, which occurred in approximately 55% of the ESBL-producing E. coli. The cit gene was found in approximately 13% of the ESBL-producing E. coli as AmpC type determinants. A total of 19 out of 26 fosfomycin-resistant strains showed the fosA3 gene, all of which produced ESBL. The correlation between fosA3 and blaCTX–M group 1 (blaCTX–M55) genes was significant among ESBL-producing E. coli isolated from Paraná (OR 3.66, 95% CI 1.9–9.68) and these genetic determinants can be transmitted by conjugation to broiler chicken microbiota strains. Our data revealed that poultry litter and beetles were critical points during poultry production and the presence of fosfomycin-resistant strains indicate the possibility of risks associated with the use of this antimicrobial during production. Furthermore, the genetic determinants encoding CTX-M and fosA3 enzymes can be transferred to E. coli strains from broiler chicken microbiota, thereby creating a risk to public health.
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- 2021
- Full Text
- View/download PDF
5. Distribution of ExPEC Virulence Factors, blaCTX-M, fosA3, and mcr-1 in Escherichia coli Isolated From Commercialized Chicken Carcasses
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Paula Signolfi Cyoia, Vanessa Lumi Koga, Erick Kenji Nishio, Sébastien Houle, Charles M. Dozois, Kelly Cristina Tagliari de Brito, Benito Guimarães de Brito, Gerson Nakazato, and Renata Katsuko Takayama Kobayashi
- Subjects
ESBL ,multidrug-resistance ,phylogenetic groups ,CTX-M ,fosfomycin ,Microbiology ,QR1-502 - Abstract
Pathogenic Escherichia coli found in humans and poultry carcasses harbor similar virulence and resistance genes. The present study aimed to analyze the distribution of extraintestinal pathogenic E. coli (ExPEC) virulence factors (VF), blaCTX−M groups, fosA3, and mcr-1 genes in E. coli isolated from commercialized chicken carcasses in southern Brazil and to evaluate their pathogenic risk. A total of 409 E. coli strains were isolated and characterized for genes encoding virulence factors described in ExPEC. Results of antimicrobial susceptibility testing confirmed that the strains were resistant to β-lactams, fosfomycin, colistin, and others resistance groups. The highest prevalence of VFs was observed in isolates belonging to the CTX-M groups, especially the CTX-M-2 group, when compared to those in other susceptible strains or strains with different mechanisms of resistance. Furthermore, ESBL strains were found to be 1.40 times more likely to contain three to five ExPEC virulence genes than non-ESBL strains. Our findings revealed the successful conjugation between ESBL-producing E. coli isolated from chicken carcass and the E. coli recipient strain J53, which suggested that genetic determinants encoding CTX-M enzymes may have originated from animals and could be transmitted to humans via food chain. In summary, chicken meat is a potential reservoir of MDR E. coli strains harboring resistance and virulence genes that could pose serious risks to human public health.
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- 2019
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6. Synergistic and additive effect of oregano essential oil and biological silver nanoparticles against multidrug-resistant bacterial strains
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Sara eScandorieiro, Larissa Ciappina de Camargo, Cesar Armando eContreras, Sueli Fumie eYamada-Ogatta, Celso Vataru eNakamura, Admilton Gonçalves de Oliveira Junior, Célia Guadalupe Tardeli Andrade, Nelson Eduardo eDurán-Caballero, Gerson eNakazato, and Renata Katsuko Takayama Kobayashi
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Antibacterial ,Synergism ,Oregano oil ,Multidrug-resistant bacteria ,biological silver nanoparticles ,Microbiology ,QR1-502 - Abstract
Bacterial resistance to conventional antibiotics has become a clinical and public health problem, making therapeutic decisions more challenging. Plant compounds and nanodrugs have been proposed as potential antimicrobial alternatives. Studies have shown that oregano (Origanum vulgare) essential oil (OEO) and silver nanoparticles have potent antibacterial activity, also against multidrug-resistant strains; however, the strong organoleptic characteristics of OEO and the development of resistance to these metal nanoparticles can limit their use. This study evaluated the antibacterial effect of a two-drug combination of biologically synthesized silver nanoparticles (bio-AgNP), produced by Fusarium oxysporum, and OEO against Gram-positive and Gram-negative bacteria, including multidrug-resistant strains. OEO and bio-AgNP showed bactericidal effects against all seventeen strains tested, with minimal inhibitory concentrations (MIC) ranging from 0.298 to 1.193 mg/mL and 62.5 to 250 µM, respectively. Time-kill curves indicated that OEO acted rapidly (within 10 min), while the metallic nanoparticles took 4 h to kill Gram-negative bacteria and 24 h to kill Gram-positive bacteria. The combination of the two compounds resulted in a synergistic or additive effect, reducing their MIC values and reducing the time of action compared to bio-AgNP used alone, i.e., 20 min for Gram-negative bacteria and 7 h for Gram-positive bacteria. Scanning electron microscopy (SEM) revealed similar morphological alterations in Staphylococcus aureus (non-methicillin-resistant S. aureus, non-MRSA) cells exposed to three different treatments (OEO, bio-AgNP and combination of the two), which appeared cell surface blebbing. Individual and combined treatments showed reduction in cell density and decrease in exopolysaccharide matrix compared to untreated bacterial cells. It indicated that this composition have an antimicrobial activity against S. aureus by disrupting cells. Both compounds showed very low hemolytic activity, especially at MIC levels. This study describes for the first time the synergistic and additive interaction between OEO and bio-AgNP produced by F. oxysporum against multidrug-resistant bacteria, such as MRSA, and β-lactamase- and carbapenemase-producing Escherichia coli and Acinetobacter baumannii strains. These results indicated that this combination can be an alternative in the control of infections with few or no treatment options.
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- 2016
- Full Text
- View/download PDF
7. Subinhibitory Concentrations of Biogenic Silver Nanoparticles Affect Motility and Biofilm Formation in Pseudomonas aeruginosa
- Author
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Renata Katsuko Takayama Kobayashi, Laís Anversa, Gerson Nakazato, Amanda Yaeko Yamada, Doroti de Oliveira Garcia, Renan Luiz Barros de Souza, Erika Kushikawa Saeki, Larissa Amianti de Araujo, and Heloísa Moreira Martins
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0301 basic medicine ,Microbiology (medical) ,Silver ,030106 microbiology ,Immunology ,lcsh:QR1-502 ,Virulence ,Motility ,Metal Nanoparticles ,medicine.disease_cause ,Microbiology ,virulence factor ,Virulence factor ,lcsh:Microbiology ,03 medical and health sciences ,Cellular and Infection Microbiology ,Fusarium ,medicine ,Humans ,antivirulence ,Original Research ,metal nanoparticle ,biology ,Pseudomonas aeruginosa ,Chemistry ,antibiofilm ,Broth microdilution ,Biofilm ,quorum sensing ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Anti-Bacterial Agents ,Quorum sensing ,030104 developmental biology ,Infectious Diseases ,Biofilms ,Bacteria - Abstract
Biogenic silver nanoparticles (bio-AgNPs) are increasingly recognized as an antibiofilm and antivirulence strategy against P. aeruginosa, a bacterium that causes chronic infections in immunocompromised and cystic fibrosis patients. This study aimed to investigate the effects of subinhibitory concentrations of bio-AgNPs on motility and biofilm formation in P. aeruginosa. Bio-AgNPs were synthesized via reduction of ionic silver catalyzed by cell-free culture filtrate from Fusarium oxysporum. A total of 17 P. aeruginosa isolates and strains were evaluated for swarming, swimming, and twitching motility in the presence and absence (control) of bio-AgNPs, including 10 clinical isolates from patients with and without cystic fibrosis, 5 environmental isolates obtained from the public water supply system, and 2 reference strains (PAO1 and PA14). Isolates were identified by biochemical and molecular methods. Minimum inhibitory concentrations (MICs) were determined by the broth microdilution method. Swarming, swimming, and twitching motility assays were performed in Petri dishes. Biofilm formation capacity was assessed quantitatively by the crystal violet method. MIC values ranged from 15.62 to 62.50 µM. The results showed that subinhibitory concentrations of bio-AgNPs (½ MIC, 7.81–31.25 µM) significantly increased (p < 0.05) swarming, swimming, and twitching motility in 40.0, 40.0, and 46.7% of isolates, respectively. Subinhibitory bio-AgNP treatment enhanced (p < 0.05) biofilm formation capacity in PA14 and a cystic fibrosis isolate (P11). It is concluded that subinhibitory concentrations of bio-AgNPs increased biofilm formation and swarming, swimming, and twitching motility in PA14 and some P. aeruginosa isolates. These virulence factors are directly involved with quorum-sensing systems. Further research should investigate the effects of AgNPs on P. aeruginosa quorum sensing to help elucidate their mechanism of action at subinhibitory concentrations.
- Published
- 2021
8. Detection of ESBL/AmpC-Producing and Fosfomycin-Resistant Escherichia coli From Different Sources in Poultry Production in Southern Brazil
- Author
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Taiara Carolaine Leal de Camargo, Renata Katsuko Takayama Kobayashi, Vanessa Lumi Koga, Leonardo Pinto Medeiros, Gerson Nakazato, Bruna Carolina Gonçalves, Erick Kenji Nishio, Benito Guimarães de Brito, Kelly Cristina Tagliari de Brito, João Juliano Pinheiro, Miriam Dibo, Tiela Trapp Grassotti, Eliana Carolina Vespero, and Luís Eduardo de Souza Gazal
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Microbiology (medical) ,Veterinary medicine ,Avian ,lcsh:QR1-502 ,Fosfomycin ,medicine.disease_cause ,Microbiology ,lcsh:Microbiology ,03 medical and health sciences ,Enterobacteriaceae ,Genotype ,medicine ,poultry litter ,Escherichia coli ,fosfomycin ,Poultry litter ,030304 developmental biology ,Original Research ,0303 health sciences ,biology ,030306 microbiology ,business.industry ,public health ,Broiler ,Poultry farming ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Antimicrobial ,business ,multidrug resistance (MDR) ,medicine.drug - Abstract
This study discussed the use of antimicrobials in the commercial chicken production system and the possible factors influencing the presence of Extended-spectrum β-lactamase (ESBL)/AmpC producers strains in the broiler production chain. The aim of this study was to perform longitudinal monitoring of ESBL-producing and fosfomycin-resistant Escherichia coli from poultry farms in southern Brazil (Paraná and Rio Grande do Sul states) and determine the possible critical points that may be reservoirs for these strains. Samples of poultry litter, cloacal swabs, poultry feed, water, and beetles (Alphitobius sp.) were collected during three distinct samplings. Phenotypic and genotypic tests were performed for characterization of antimicrobial resistant strains. A total of 117 strains were isolated and 78 (66%) were positive for ESBL production. The poultry litter presented ESBL positive strains in all three sampled periods, whereas the cloacal swab presented positive strains only from the second period. The poultry litter represents a significant risk factor mainly at the beginning poultry production (odds ratio 6.43, 95% confidence interval 1–41.21, p < 0.05). All beetles presented ESBL positive strains. The predominant gene was blaCTX–M group 2, which occurred in approximately 55% of the ESBL-producing E. coli. The cit gene was found in approximately 13% of the ESBL-producing E. coli as AmpC type determinants. A total of 19 out of 26 fosfomycin-resistant strains showed the fosA3 gene, all of which produced ESBL. The correlation between fosA3 and blaCTX–M group 1 (blaCTX–M55) genes was significant among ESBL-producing E. coli isolated from Paraná (OR 3.66, 95% CI 1.9–9.68) and these genetic determinants can be transmitted by conjugation to broiler chicken microbiota strains. Our data revealed that poultry litter and beetles were critical points during poultry production and the presence of fosfomycin-resistant strains indicate the possibility of risks associated with the use of this antimicrobial during production. Furthermore, the genetic determinants encoding CTX-M and fosA3 enzymes can be transferred to E. coli strains from broiler chicken microbiota, thereby creating a risk to public health.
- Published
- 2021
9. Editorial: Nanotechnology for Antimicrobials
- Author
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Renata Katsuko Takayama Kobayashi and Gerson Nakazato
- Subjects
Microbiology (medical) ,medicine.medical_specialty ,therapy ,synthesis ,Chemistry ,disease prevention ,lcsh:QR1-502 ,Antimicrobial ,Microbiology ,lcsh:Microbiology ,Drug delivery ,drug delivery ,medicine ,Disease prevention ,nanoparticles ,microbial pathogens ,Intensive care medicine - Published
- 2020
10. Escherichia coli Bloodstream Infections in Patients at a University Hospital: Virulence Factors and Clinical Characteristics
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Vanessa Lumi Koga, Renata Katsuko Takayama Kobayashi, João Gabriel Material Soncini, Marsileni Pelisson, Marcia Regina Eches Perugini, Ana Paula Daga, Eliana Carolina Vespero, and Caroline Martins de Matos
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0301 basic medicine ,Microbiology (medical) ,medicine.medical_specialty ,CTX- M-15 ,medicine.drug_class ,030106 microbiology ,Immunology ,Cephalosporin ,virulence factors ,lcsh:QR1-502 ,Virulence ,bloodstream infection ,medicine.disease_cause ,Microbiology ,lcsh:Microbiology ,03 medical and health sciences ,phylogenetic group ,Cellular and Infection Microbiology ,Epidemiology ,Genotype ,medicine ,Escherichia coli ,Original Research ,ExPEC ,Extraintestinal Pathogenic Escherichia coli ,biology ,biology.organism_classification ,Pathogenicity island ,030104 developmental biology ,Infectious Diseases ,ESBL ,Bacteria - Abstract
Extraintestinal pathogenic Escherichia coli (ExPEC) isolates are responsible for many bloodstream infections. The aim of this study was to characterize E. coli isolated from the bloodstreams of patients (n = 48) at the University Hospital in Brazil. Epidemiological data were obtained through the analysis of medical records and laboratory tests. By PCR analysis, we investigated the presence of virulence factors (VFs), pathogenicity islands (PAIs), extended-spectrum β-lactamase (ESBL), phylogenetic classifications (A, B1, B2, C, D, E, and F) and molecular genotype by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The mortality analysis showed that 33.3% of the deaths were associated with bacteraemia due to E. coli infections; in addition, an age between 60 and 75 years (p < 0.001; OR = 6.3[2.1–18.9]) and bacteraemia with an abdominal origin (p = 0.02; OR = 5[1.2–20.5]) were risk factors for the severity of the infection. Additionally, the presence of the afa gene was associated with mortality due to E. coli bacteraemia (p = 0.027; OR = 11.4[1.5–85.7]). Immunosuppression (27.1%), intestinal diseases (25.0%) and diabetes (18.8%), were prevalent among patients, and most of the bacteraemia cases were secondary to urinary tract infections (50.0%). The serum resistance gene traT was present in 77.1% of isolates, group capsular 2 (kpsMT II) was present in 45.8% and the K5 capsule was present in 20.8% of isolates. The isolates also showed a high prevalence for the siderophore yersiniabactina (fyuA) (70.8%) and PAI IV536 (77.1%). Phylogenetic analysis showed that group B2 (45.8%) was the most prevalent, and was the phylogroup that had a higher prevalence of VFs and PAIs. However, in this study, a considerable number of isolated bacteria were classified as group B1 (18.8%) and as group E (14.6%). Eight (16.7%) isolates were resistant to third and fourth generation cephalosporin and group CTX-M-1 (CTX-M-15) was the most prevalent ESBL type. The molecular genotyping showed two clonal lineages and several isolates that were not related to each other. This study provides additional information on the epidemiological and molecular characteristics of E. coli bloodstream infections in Brazil.
- Published
- 2019
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