RESUMO
Aim: To evaluate the activity of five antimicrobials against young and mature Stenotrophomonas maltophilia biofilms. Materials & methods: Nineteen clinical strains from hemoculture of hemodialysis patients were tested for biofilm kinetics, MIC and minimum biofilm inhibitory concentration (MBIC) in young and mature biofilms. Results: All strains were moderate biofilm producers. MIC showed total susceptibility to levofloxacin and trimethoprim-sulfamethoxazole and partial resistance to ceftazidime (63.2%) and gentamicin (21%). Young and mature biofilms showed the lowest MBIC/MIC ratio for gentamicin, chloramphenicol and levofloxacin, respectively. The highest MBIC/MIC was for trimethoprim-sulfamethoxazole (young) and ceftazidime (mature). Conclusion: Gentamicin displayed surprising activity against S. maltophilia biofilms. Chloramphenicol was indicated as a good option against young S. maltophilia biofilms, and trimethoprim-sulfamethoxazole showed limited antibiofilm activity.
Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Infecções por Bactérias Gram-Negativas/microbiologia , Stenotrophomonas maltophilia/efeitos dos fármacos , Ceftazidima/farmacologia , Farmacorresistência Bacteriana Múltipla , Humanos , Levofloxacino/farmacologia , Testes de Sensibilidade Microbiana , Minociclina/farmacologia , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Stenotrophomonas maltophilia/fisiologia , Combinação Trimetoprima e Sulfametoxazol/farmacologiaRESUMO
Introduction. Stenotrophomonas maltophilia has emerged as one of the most common multi-drug-resistant pathogens isolated from people with cystic fibrosis (CF). However, its adaptation over time to CF lungs has not been fully established.Hypothesis. Sequential isolates of S. maltophilia from a Brazilian adult patient are clonally related and show a pattern of adaptation by loss of virulence factors.Aim. To investigate antimicrobial susceptibility, clonal relatedness, mutation frequency, quorum sensing (QS) and selected virulence factors in sequential S. maltophilia isolates from a Brazilian adult patient attending a CF referral centre in Buenos Aires, Argentina, between May 2014 and May 2018.Methodology. The antibiotic resistance of 11 S. maltophilia isolates recovered from expectorations of an adult female with CF was determined. Clonal relatedness, mutation frequency, QS variants (RpfC-RpfF), QS autoinducer (DSF) and virulence factors were investigated in eight viable isolates.Results. Seven S. maltophilia isolates were resistant to trimethoprim-sulfamethoxazole and five to levofloxacin. All isolates were susceptible to minocycline. Strong, weak and normomutators were detected, with a tendency to decreased mutation rate over time. XbaI PFGE revealed that seven isolates belong to two related clones. All isolates were RpfC-RpfF1 variants and DSF producers. Only two isolates produced weak biofilms, but none displayed swimming or twitching motility. Four isolates showed proteolytic activity and amplified stmPr1 and stmPr2 genes. Only the first three isolates were siderophore producers. Four isolates showed high resistance to oxidative stress, while the last four showed moderate resistance.Conclusion. The present study shows the long-time persistence of two related S. maltophilia clones in an adult female with CF. During the adaptation of the prevalent clones to the CF lungs over time, we identified a gradual loss of virulence factors that could be associated with the high amounts of DSF produced by the evolved isolates. Further, a decreased mutation rate was observed in the late isolates. The role of all these adaptations over time remains to be elucidated from a clinical perspective, probably focusing on the damage they can cause to CF lungs.
Assuntos
Fibrose Cística/complicações , Infecções por Bactérias Gram-Negativas/microbiologia , Pulmão/microbiologia , Stenotrophomonas maltophilia/genética , Adulto , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Farmacorresistência Bacteriana , Feminino , Genótipo , Infecções por Bactérias Gram-Negativas/etiologia , Humanos , Masculino , Mutação , Fenótipo , Filogenia , Escarro/microbiologia , Stenotrophomonas maltophilia/efeitos dos fármacos , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Stenotrophomonas maltophilia/isolamento & purificação , Adulto JovemRESUMO
The excessive use of agrochemicals negatively impacts the environment, making the development of sustainable technologies for the reduction of contaminants in soil necessary. Hexazinone is the herbicide most used for sugarcane crops and persists in the environment. Moreover, its main route of degradation in the soil is through microorganisms. Therefore, six microorganisms were selected that presented growth in the presence of the herbicide; SCR1 - Microbacterium arborescens; SCR2 - Bacillus pumilus; SCM3 - Stenotrophomonas maltophilia; SCM4 - Bacillus cereus; SCM5A - M. arborescens; and SCM5B - B. safensis. A test was performed to evaluate the ability of each lineage in phosphate solubilization. For the Ca3(PO4)2 solubilization test, the strains that showed the best results were B. pumilus and S. maltophilia. Subsequently, the inoculants were prepared and the concentrations after plating were 2.71 × 109 CFU mL-1 for B. pumilus, 1.02 × 109 CFU mL-1 for S. maltophilia, and 1.14 × 1010 CFU mL-1 for a combination of the two strains. These were satisfactory values for use as inoculants.(AU)
O uso de agroquímicos resulta em impactos ambientais e torna-se necessário o emprego de tecnologias sustentáveis para diminuição de contaminantes no solo. O hexazinona é o herbicida mais utilizado para a cultura da cana-de-açúcar, e apresenta persistência no ambiente. A principal via de degradação no solo é por meio de microrganismos. Com isso, selecionou-se seis microrganismos que apresentaram crescimento na presença do herbicida: SCR1 - Microbacterium arborescens; SCR2 - Bacillus pumilus; SCM3 - Stenotrophomonas maltophilia; SCM4 - Bacillus cereus; SCM5A - M. arborescens; SCM5B - Bacillus safensis. Foi realizado um teste para avaliar a habilidade de cada linhagem na solubilização de fosfatos, e no caso da solubilização de Ca3(PO4)2, as linhagens que apresentaram melhores resultados foram B. pumilus e S. maltophilia. Posteriormente, os inoculantes foram preparados e a concentração após plaqueamento de 2,71x109 UFC mL-1 para B. pumilus, 1,02x109 UFC mL-1 para S. maltophilia e consórcio com as duas linhagens 1,14x1010 UFC mL-1 apresentaram valores satisfatórios para utilização como inoculantes.(AU)
Assuntos
Tratamento do Solo/métodos , Microbiologia do Solo , Poluentes do Solo/antagonistas & inibidores , Bacillus pumilus/crescimento & desenvolvimento , Stenotrophomonas maltophilia/crescimento & desenvolvimentoRESUMO
Stenotrophomonas maltophilia is a multidrug-resistant opportunistic pathogen. S. maltophilia quorum-sensing system is mediated by the diffusible signal factor (DSF), which synthesis depends on rpfF. It has been reported that rpfF disruption in S. maltophilia K279a leads to a loss of DSF synthesis, reduced levels of extracellular protease, swarming motility and virulence in the Galleria mellonella model. The aim of this work was to attain a deeper knowledge of the role of the rpf/DSF signalling system in S. maltophilia biofilm formation, phenotypic traits associated with biofilm development and virulence and antimicrobial susceptibility. To this end, comparative studies were conducted on S. maltophilia K279a and K279arpfF. The results presented here put in evidence the positive role of DSF in bacterial growth, biofilm formation, swimming and twitching motilities, DNAse, lipases and siderophores production as well as resistance to oxidative stress. Interestingly, DSF seems to be essential for the development of the spatially organised structure seen in mature biofilms. Therefore, DSF from S. maltophlia K279a positively regulates biofilm formation and virulence. Furthermore, DSF is necessary for the induction of L1 and L2 ß-lactamase production in K279a. This is the first evidence of the role of the rpf/DSF signalling system in S. maltophilia ß-lactam resistance.
Assuntos
Proteínas de Bactérias/metabolismo , Biofilmes , Stenotrophomonas maltophilia/metabolismo , Fatores de Virulência/biossíntese , beta-Lactamases/metabolismo , Animais , Proteínas de Bactérias/genética , Regulação Bacteriana da Expressão Gênica , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Mariposas/microbiologia , Sideróforos/metabolismo , Stenotrophomonas maltophilia/genética , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Stenotrophomonas maltophilia/patogenicidade , Virulência , Fatores de Virulência/genética , beta-Lactamases/genéticaRESUMO
Resumo Objetivos: Avaliar o grau de contaminação por fungos e bactérias e o modo de conservação destes colírios hipotensores por parte dos pacientes no ambulatório de Glaucoma da Santa Casa de Ribeirão Preto. Métodos: Foram selecionados aleatoriamente cinquenta e cinco pacientes, em seguimento no ambulatório, e após consentimento dos mesmos os colírios eram coletados e enviados via correio para análise por microbiologista e patologista em até 72 horas. Foi analisado 0,5ml aproximadamente das medicações e os pacientes respondiam a um questionário simples sobre o método de conservação e se consideravam estes adequados. Resultados: Dos 55 colírios analisados, cinco (9,01%) estavam com seu conteúdo líquido contaminado. Entre os microrganismos isolados haviam 4 bactérias Gram negativas, sendo 1 (1,8%) por Serratia marcescens, 1 (1,8%) Pseudomonas aeruginosa e 2 (3,6%) Stenotrophomas maltophilia. Um colírio estava contaminado pelo fungo Cândida ssp Todos pacientes do estudo julgam seus métodos de armazenamento e instilação adequados. Os pacientes que tiveram os colírios positivados eram convocados para exame clínico e passavam por novo questionário pelo investigador. Conclusão: O tempo de abertura dos frascos e os métodos de conservação influenciam na contaminação dos medicamentos, todos os colírios com crescimento de microrganismos no presente estudo estavam abertos entre 30 e 90 dias. O fato de que a maioria dos pacientes levam seus colírios em tarefas cotidianas, aumenta a exposição dos frascos e podem ser um fator relevante para determinar a contaminação destas medicações.
Abstract Objetives: To assess the degree of fungal and bacterial contamination of hypotensive eye drops and the way these are preserved by the patients at the Glaucoma outpatient clinic of Santa Casa Hospital in Ribeirão Preto. Methods: Fifty-five patients were randomly assigned to follow-up in the outpatient clinic and, after their consent, an eye drop was collected per patient and later sent by mail for analysis by microbiologist and pathologist in up to 72 hours. Approximately 0.5ml of the medications were analyzed and the patients were asked to answer a simple questionnaire on the method of drug conservation and whether they considered it adequate. Results: Of the 55 analysed eye drops, five (9.01%) had their liquid contents contaminated. Among the microorganisms isolated there were 4 Gram negative bacteria, 1 (1.8%) by Serratia marcenses, 1 (1.8%) Pseudomonas aeruginosa and 2 (3.6%) Stenotrophomas maltophilia. An eye drop was contaminated by the fungus Candida ssp. All the patients in the study judged their methods of storage and instillation appropriate. The patients who had the positive coliria were summoned for clinical examination and passed through a new questionnaire by the investigator. Conclusion: The time and methods of preservation influence the contamination of medicinal products. All the eye drops that presented growth of microorganisms in the present study were open between 30 and 90 days. The fact that most patients take their eye drops on daily tasks increases the exposure of the bottles and can be a relevant fact to determine the contamination of these medications.
Assuntos
Humanos , Masculino , Feminino , Idoso , Soluções Oftálmicas/análise , Soluções Oftálmicas/uso terapêutico , Glaucoma/tratamento farmacológico , Contaminação de Medicamentos , Pseudomonas aeruginosa/isolamento & purificação , Serratia marcescens/crescimento & desenvolvimento , Bactérias/isolamento & purificação , Candida/crescimento & desenvolvimento , Estudos Transversais , Inquéritos e Questionários , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Armazenamento de Medicamentos , Microscopia com Lâmpada de Fenda , Fungos/isolamento & purificaçãoRESUMO
High copper concentration is toxic for living organisms including humans. Biosorption is a bioremediation technique that can remove copper and other pollutants from aqueous medium and soils, consequently cleaning the environment. The aim of this study was, therefore, to investigate the influence of different copper compounds (Cu(II) as CuCl2; Cu(II) as CuSO4; and Cu(I) as CuCl) on copper bioreduction and biosorption using four copper-resistant bacteria isolated from the rhizosphere of two plants (Avena sativa and Plantago lanceolata) in aqueous matrix. Copper resistance profile, bioreduction, and biosorption after 48 h of incubation were evaluated. The isolates displayed high copper resistance. However, isolate A1 did not grow very well in the CuCl2 and isolate T5 was less resistant to copper in aqueous solutions amended with CuCl (Cu(I)). The best copper source for copper bioreduction and biosorption was CuSO4 and the isolates removed as much as ten times more copper than in aqueous solutions amended with the other copper compounds. Moreover, Cu(I) did not succumb to biosorption, although the microbes were resistant to aqueous solutions of CuCl. In summary, Cu(II) from CuSO4 was furthermost susceptible to bioreduction and biosorption for all isolates. This is an indication that copper contamination of the environment from the use of CuSO4 as an agrochemical is amenable to bioremediation.
Assuntos
Sulfato de Cobre/isolamento & purificação , Cobre/isolamento & purificação , Poluentes Ambientais/isolamento & purificação , Acinetobacter/genética , Acinetobacter/crescimento & desenvolvimento , Acinetobacter/isolamento & purificação , Adsorção , Avena/microbiologia , Biodegradação Ambiental , Biomassa , Farmacorresistência Bacteriana , Oxirredução , Raízes de Plantas/microbiologia , Plantago/microbiologia , Pseudomonas putida/genética , Pseudomonas putida/crescimento & desenvolvimento , Pseudomonas putida/isolamento & purificação , RNA Ribossômico 16S/genética , Rizosfera , Soluções , Stenotrophomonas maltophilia/genética , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Stenotrophomonas maltophilia/isolamento & purificação , ÁguaRESUMO
The aim of this study was to compare the in vitro activity of ethanol, EDTA and levofloxacin (Levo), alone or in combination, on biofilms of Stenotrophomonas maltophilia recovered from patients with catheter-related bloodstream infections (CRBSIs) at a university hospital in Argentina. First, 24 and 48 h biofilms were formed in microtitre plates and challenged with 25 or 40â% ethanol for 1 h. Biofilms, of the 14 local isolates and from the reference strain K279a, were eradicated after both treatments as shown by plate counts and the regrowth technique. Second, 24 h biofilms of all isolates were established in silicone catheter segments and challenged with 25 or 40â% ethanol, Levo (2.5 mg ml(-1)), EDTA (30 mg ml(-1)), 25â% ethanol-EDTA or Levo-EDTA for 1, 3 and 24 h. Viable counts of biofilms treated for 1 h with 25 or 40â% ethanol or 25â% ethanol-EDTA were under the limit of detection. Killing of biofilms by Levo or Levo-EDTA was gradual and it was only after 24 h of treatment that no differences could be seen between the effects of these catheter lock solutions (CLSs) and those of ethanol (P>0.05). Levo-EDTA, in combination, did not act synergistically against biofilms. After 24 h of exposure, EDTA did not eradicate biofilms but reduced biofilm survival rates to 1-5â%. The effect of the different CLSs on biomass reduction, estimated by crystal violet staining, was highly dependent on the isolate, and the most effective agents were 25 and 40â% ethanol. Our results suggest that when used as a CLS for short periods, ethanol at low concentrations, alone or in combination with a chelator, can decontaminate the line from S. maltophilia in cases of CRBSI and help, in conjunction with systemic antibiotics, in the retention of precious vascular catheters.
Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Cateteres de Demora/microbiologia , Ácido Edético/farmacologia , Etanol/farmacologia , Levofloxacino , Ofloxacino/farmacologia , Stenotrophomonas maltophilia/efeitos dos fármacos , Argentina , Bacteriemia/microbiologia , Infecções Relacionadas a Cateter/microbiologia , Quelantes/farmacologia , Contagem de Colônia Microbiana , Hospitais Universitários , Humanos , Testes de Sensibilidade Microbiana , Soluções/farmacologia , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Stenotrophomonas maltophilia/isolamento & purificaçãoRESUMO
A Gram-negative bacterium, designated as strain KB2, was isolated from activated sludge and was found to utilize different aromatic substrates as sole carbon and energy source. On the basis of morphological and physiochemical characteristics and 16S rRNA gene sequence analysis, the isolated strain KB2 was identified as Stenotrophomonas maltophilia. Strain KB2 is from among different Stenotrophomonas maltophilia strains the first one described as exhibiting the activities of three types of dioxygenases depending on the structure of the inducer. The cells grown on benzoate and catechol showed mainly catechol 1,2- dioxygenase activity. The activity of 2,3-dioxygenase was detected after phenol induction. Protocatechuate 3,4-dioxygenase was found in crude cell extracts of this strain after incubation with 4-hydroxybenzoic acid, protocatechuic acid and vanillic acid. Because of broad spectrum of dioxygenases' types that Stenotrophomonas maltophilia KB2 can exhibit, this strain appears to be very powerful and useful tool in the biotreatment of wastewaters and in soil decontamination.
Uma bactéria Gram-negativa, denominada KB2, foi isolada de lodo ativado, verificando-se ser capaz de utilizar substratos aromáticos com única fonte de carbono e energia. Com base nas características morfológicas e físico-químicas, e na análise da sequencia do gene 16SrRNA, esta bactéria foi identificada como Stenotrophomonas maltophilia. Entre as diversas cepas de S. maltophilia já descritas, essa cepa é a primeira com atividade de três tipos de dioxigenases, dependendo da estrutura do indutor. As células cultivadas em benzoato e catecol apresentaram atividade de catecol 1,2-dioxigenase principalmente. A atividade de 2,3-dioxigenase foi detectada após indução com fenol. Após incubação com ácidos 4-hidrobenzoico, ácido protocatecuico evanílico, encontrou-se protocatecuato 3,4-dioxigenase no extrato celular. Devido ao amplo espectro de atividade das diferentes dioxigenases de S. maltophilia KB2, esta cepa parece ser uma ferramenta poderosa e útil para o biotratamento de efluentes e descontaminação do solo.
Assuntos
Lodos Ativados , Dioxigenases/análise , Dioxigenases/genética , Hidrocarbonetos Aromáticos/análise , Sequência de Bases/genética , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Stenotrophomonas maltophilia/isolamento & purificação , Biodegradação Ambiental , Métodos , MétodosRESUMO
Stenotrophomonas maltophilia has emerged as an important nosocomial pathogen capable of causing respiratory, bloodstream, and urinary infections. The treatment of nosocomial infections by S. maltophilia is difficult, as this pathogen shows high levels of intrinsic or acquired resistance to different antimicrobial agents, drastically reducing the antibiotic options available for treatment. Intrinsic resistance may be due to reduced outer membrane permeability or to the multidrug efflux pumps. However, specific mechanisms of resistance such as aminoglycoside-modifying enzymes or the heterogeneous production of metallo-beta-lactamase have contributed to the multidrug-resistant phenotype displayed by this pathogen. Moreover, the lack of standardized susceptibility tests and their interpretative criteria hinder the choice of an adequate antibiotic treatment. Recommendations for the treatment of infections by S. maltophilia are based on in vitro studies, certain nonrandomized clinical trials, and anecdotal experience. Trimethoprim-sulfamethoxazole remains the drug of choice, although in vitro studies indicate that ticarcillin-clavulanic acid, minocycline, some of the new fluoroquinolones, and tigecycline may be useful agents. This review describes the main resistance mechanisms, the in vitro susceptibility profile, and treatment options for S. maltophilia infections.
Assuntos
Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana Múltipla , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Animais , Antibacterianos/farmacologia , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Sinergismo Farmacológico , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Stenotrophomonas maltophilia/efeitos dos fármacosRESUMO
The presence of acetic acid bacteria populations on grape surfaces from several Chilean valleys is reported. The bacteria were analysed at both the species and the strain level by molecular methods such as RFLP-PCR 16S rRNA gene, RFLP-PCR ITS 16S-23S rRNA gene regions and Arbitrary Primed (AP) PCR. Our results show that there are limited numbers of species of acetic acid bacteria in the grapes and that there is a need for an enrichment medium before plating to recover the individual colonies. In the Northernmost region analysed, the major species recovered was a non-acetic acid bacteria, Stenotrophomonas maltophila. Following the North-South axis of Chilean valleys, the observed distribution of acetic acid bacteria was zonified: Acetobacter cerevisiae was only present in the North and Gluconobacter oxydans in the South. Both species were recovered together in only one location. The influence of the grape cultivar was negligible. Variability in strains was found to be high (more than 40%) for both Acetobacteraceae species.
Assuntos
Ácido Acético/metabolismo , Acetobacter/classificação , Acetobacter/metabolismo , Filogenia , Polimorfismo de Fragmento de Restrição , Vitis/microbiologia , Acetobacter/crescimento & desenvolvimento , Chile , Gluconobacter oxydans/classificação , Gluconobacter oxydans/crescimento & desenvolvimento , Gluconobacter oxydans/metabolismo , Reação em Cadeia da Polimerase , RNA Ribossômico 16S , RNA Ribossômico 23S , Especificidade da Espécie , Stenotrophomonas maltophilia/classificação , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Stenotrophomonas maltophilia/metabolismoRESUMO
The ability of Azospirillum brasilense Cd to colonize the niche occupied by 3 bacterial strains previously isolated from sorghum rhizosphere was studied by means of the Biolog system. The isolates were identified by different methods as strains belonging to Pseudomonas putida, Stenotrophomonas maltophilia, and Klebsiella terrigena species. Several C sources, also chosen among the constituents of sorghum root exudates, were used to evaluate the metabolic profiles of Azospirillum and the sorghum rhizobacteria. Azospirillum brasilense Cd exploited the same class of C compounds as the sorghum rhizobacteria and overlapped in their niche requirements. Since structure and functioning of a microbial community are largely affected by the flow rate of nutrient supply, the competitive behavior of A. brasilense Cd was studied in a chemostat mixed culture under C-limited conditions using disodium succinate as C source. Only at high growth rates, i.e., when the C source was highly supplied, A. brasilense Cd appeared to be a good competitor and it became the dominant species, whereas at low growth rates, it was outnumbered by the other species. However, the coexistence of all the strains was always maintained, thus suggesting that interactions other than competition or a potential cross-feeding might occur within the mixed culture.