RESUMO
This study demonstrates the effect of fermented botanical product (FBP) on Ralstonia pseudosolanacearum-induced bacterial wilt disease and unravels its action mechanism. Soaking with diluted FBP solutions (0.1%-0.5%) significantly suppressed bacterial wilt in tomato plants, and FBP-treated tomato plants grew well against R. pseudosolanacearum infection. Growth assays showed that FBP had no antibacterial effect but promoted R. pseudosolanacearum growth. In contrast, few or no R. pseudosolanacearum cells were detected in aerial parts of tomato plants grown in FBP-soaked soil. Subsequent infection assays using the chemotaxis-deficient mutant (ΔcheA) or the root-dip inoculation method revealed that FBP does not affect pathogen migration to plant roots during infection. Moreover, FBP-pretreated tomato plants exhibited reduced bacterial wilt in the absence of FBP. These findings suggest that the plant, but not the pathogen, could be affected by FBP, resulting in an induced resistance against R. pseudosolanacearum, leading to a suppressive effect on bacterial wilt.
Assuntos
Fermentação , Fertilizantes , Doenças das Plantas , Ralstonia , Solanum lycopersicum , Solanum lycopersicum/microbiologia , Solanum lycopersicum/crescimento & desenvolvimento , Ralstonia/efeitos dos fármacos , Ralstonia/crescimento & desenvolvimento , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Raízes de Plantas/microbiologiaRESUMO
Bacterial wilt incited by Ralstonia pseudosolanacearum (Rps) race 4 biovar 3 is a serious threat to ginger (Zingiber officinale Rosc.) cultivation throughout the ginger growing tracts and warrants effective remedial measures since most of the strategies failed at field level implementation. After a series of experiments, calcium chloride was found to be effective against Rps both in vitro and in planta and its prophylactic effect has been successfully demonstrated under field conditions. CaCl2 at a concentration of > 2% significantly inhibited Rps under in vitro conditions. Calcium is an important nutritional element imparts a major role in plant disease resistance, and numerous studies have demonstrated the mitigating effect of calcium for disease management. CaCl2 being inhibitory to Rps, the mechanism of inhibition by CaCl2 against Rps was elucidated by a series of in vitro assays including swarming motility and biofilm formation. Direct inhibition was also studied using Scanning Electron Microscopy (SEM). The minimum bactericidal concentration and minimum inhibitory concentration were found to be around 3% while the EC 90 value was found to be 2.25%. The SEM analysis revealed the destruction of cell structure by making perforations on the cell surface. CaCl2 at the targeted concentrations inhibited biofilm formation as well as swarming motility of Rps. These findings suggest that CaCl2 exhibits strong antibacterial activity against Rps and has the potential to be used as an effective bactericide for Rps in managing bacterial wilt in ginger.
Assuntos
Cloreto de Cálcio/farmacologia , Ralstonia/efeitos dos fármacos , Zingiber officinale/microbiologia , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Doenças das Plantas/microbiologia , Ralstonia/ultraestruturaRESUMO
We have demonstrated that chemotaxis to l-malate facilitated motility of Ralstonia pseudosolanacearum MAFF 106611, a causative agent of bacterial wilt, to plant roots. Here, we evaluated the assumption that the disruption of chemotaxis to l-malate leads to inhibition of plant infection by R. pseudosolanacearum MAFF 106611. Chemotactic assays revealed that chemotaxis to l-malate was completely or partially inhibited in the presence of l-, d-, and dl-malate, respectively. Moreover, l-malate served as a carbon and energy source for R. pseudosolanacearum MAFF 106611, while d-malate inhibited the growth of this bacterium. In the sand-soak inoculation virulence assay for tomato plants, the addition of l-, d-, and dl-malate to sand suppressed the plant infection. We concluded that supplementation of l- and dl-malate suppresses tomato plant infection with R. pseudosolanacearum MAFF 106611 by disrupting its chemotaxis to l-malate, while d-malate suppresses it by both the disruption of l-malate chemotaxis and inhibition of growth.
Assuntos
Quimiotaxia/efeitos dos fármacos , Raízes de Plantas/microbiologia , Ralstonia/patogenicidade , Solanum lycopersicum/microbiologia , Malatos/farmacologia , Ralstonia/efeitos dos fármacos , Ralstonia/crescimento & desenvolvimentoRESUMO
Cupriavidus taiwanensis LMG19424, a ß-rhizobial symbiont of Mimosa pudica, harbors phc and tqs quorum sensing (QS), which are the homologous cell-cell communication systems previously identified from the plant pathogen Ralstonia solanacearum and the human pathogen Vibrio cholerae, respectively. However, there has been no experimental evidence reported that these QS systems function in C. taiwanensis LMG19424. We identified (R)-methyl 3-hydroxymyristate (3-OH MAME) and (S)-3-hydroxypentadecan-4-one (C15-AHK) as phc and tqs QS signals, respectively, and characterized these QS systems. The expression of the signal synthase gene phcB and tqsA in E. coli BL21(DE3) resulted in the high production of 3-OH MAME and C15-AHK, respectively. Their structures were elucidated by comparison of EI-MS data and GC/chiral LC retention times with synthetic standards. The deletion of phcB reduced cell motility and increased biofilm formation, and the double deletion of phcB/tqsA caused the accumulation of the metal chelator coproporphyrin III in its mutant culture. Although the deletion of phcB and tqsA slightly reduced its ability to nodulate on aseptically grown seedlings of M. pudica, there was no significant difference in nodule formation between LMG19424 and its QS mutants when commercial soils were used. Taken together, this is the first example of the simultaneous production of 3-OH MAME/C15-AHK as QS signals in a bacterial species, and the importance of the phc/tqs QS systems in the saprophytic stage of C. taiwanensis LMG19424 is suggested.
Assuntos
Cupriavidus/genética , Percepção de Quorum/genética , Rhizobium/genética , Biofilmes , Cromatografia Gasosa , Coproporfirinas/metabolismo , Escherichia coli/metabolismo , Deleção de Genes , Genes Bacterianos/genética , Mimosa/química , Mutação/genética , Ralstonia/efeitos dos fármacos , Ralstonia solanacearum , Espectrometria de Massas por Ionização por Electrospray , Vibrio cholerae/efeitos dos fármacosRESUMO
BACKGROUND: Ralstonia picketti, Ralstonia mannitolilytica, and Ralstonia insidiosa have recently been regarded as emerging pathogens of infectious diseases, in particular as the pathogens responsible for nosocomial infection in immunocompromised patients. R. insidiosa differs from R. picketti and R. mannitolilytica, and its related infections are rarely reported. METHODS: Clinical data from two nosocomial bloodstream infection cases were extracted and analyzed. The causable isolates were identified by the VITEK 2 Compact system, matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and molecular identification methods using PCR with universal and species-specific primers. Antimicrobial susceptibility testing was performed using the broth microdilution method. Both of the isolates were subjected to whole genome sequencing using a HiSeq X10 Sequencer. Antimicrobial resistance genes, virulence factors, and plasmid replicons were identified from assembled genomes. A real-time RT-PCR experiment and a cloning experiment were conducted to explore the related class D ß-lactamase-encoding genes. RESULTS: Both patients recovered under therapy with antibiotics. Isolates were initially misidentified as R. mannitolilytica by the VITEK 2 Compact system rather than R. insidiosa, as identified by both MALDI-TOF MS and 16S rRNA gene sequencing. Both isolates were resistant to aminoglycosides, ß-lactams, and polymyxin B. One isolate harboring blaOXA-570 was resistant to carbapenems. The whole genome sequencing data confirmed species identification based on average nucleotide identity (ANI) and revealed two variants of class D ß-lactamase-encoding gene blaOXA (blaOXA-573 and blaOXA-574). The real-time RT-PCR experiment showed no difference in gene expression between blaOXA-570 and blaOXA-573 in our strains. The cloning experiment showed that variant OXA-573 had no carbapenem hydrolase activity. CONCLUSIONS: We described two cases of nosocomial bloodstream infection caused by R. insidiosa strains. MALDI-TOF MS was cost-effective for rapid species identification. Clinicians should be aware that R. insidiosa can be resistant to commonly used antibiotics, even carbapenems.
Assuntos
Infecção Hospitalar/diagnóstico , Ralstonia/isolamento & purificação , Adulto , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Carbapenêmicos/farmacologia , Carbapenêmicos/uso terapêutico , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana , Humanos , Masculino , Testes de Sensibilidade Microbiana , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/metabolismo , Ralstonia/classificação , Ralstonia/efeitos dos fármacos , Ralstonia/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Sequenciamento Completo do GenomaRESUMO
BACKGROUND: Silver nanoparticles (AgNPs), particularly those entrapped in polymeric nanosystems, have arisen as options for managing plant bacterial diseases. Among the biopolymers useful for the entrapment of AgNPs, chitosan is promising because of its low cost, good biocompatibility, antimicrobial properties and biodegradability. The present study aimed: (i) to greenly-synthesize AgNPs using different concentrations of aqueous extract of tomato leaves followed by entrapment of AgNPs with chitosan (CH-AgNPs); (ii) to characterize the optical, structural and biological properties of the nanosystems produced; (iii) to evaluate the antimicrobial activities of AgNPs and nanomaterials; and (iv) to assess the effectiveness of AgNPs and nanomaterials for controlling tomato bacterial wilt caused by Ralstonia solanacearum. RESULTS: Spherical and oval AgNPs had incipient colloidal instability, although the concentration of the tomato leaf extract influenced both size (< 87 nm) and the polydispersity index. Nanomaterials (< 271 nm in size) were characterized by a highly stable matrix of chitosan containing polydisperse AgNPs. Free AgNPs and CH-AgNPs were stable for up to 30 days, with no significant alteration in physicochemical parameters. The AgNPs and nanomaterials had antibacterial activity and decreased bacterial growth at micromolar concentrations after 48 h. Morphological changes in R. solanacearum cells were observed after treatment with CH-AgNPs. The application of CH-AgNPs at 256 µmol L-1 reduced the incidence of bacterial wilt in a partially resistant tomato genotype but not in the susceptible line. CONCLUSION: Greenly-synthesized chitosan-derived nanomaterials containing AgNPs produced with leaf extracts from their own species appear to comprise a promising and sustainable alternative in an integrated management approach aiming to reduce the yield losses caused by bacterial wilt. © 2019 Society of Chemical Industry.
Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Quitosana/química , Química Verde/métodos , Doenças das Plantas/microbiologia , Extratos Vegetais/química , Prata/farmacologia , Solanum lycopersicum/química , Antibacterianos/química , Portadores de Fármacos/química , Composição de Medicamentos , Solanum lycopersicum/microbiologia , Nanoestruturas/química , Folhas de Planta/química , Ralstonia/efeitos dos fármacos , Ralstonia/crescimento & desenvolvimento , Prata/químicaRESUMO
Ralstonia species, often regarded as an environmental organism of low pathogenicity, can cause significant disease in certain at-risk patient groups, including those with cystic fibrosis. Difficulties with its identification in the clinical laboratory mean that it may be misidentified and therefore under recognised as a cause of disease. A number of outbreaks have been associated with the use of devices for inhaled respiratory therapy, putting those with chronic respiratory conditions at risk. Antimicrobial treatment of infection is challenging and limited due to frequent antimicrobial resistance. This review highlights issues regarding the identification, treatment and prevention of infection due to Ralstonia spp. in children with cystic fibrosis.
Assuntos
Fibrose Cística , Infecções por Bactérias Gram-Negativas , Ralstonia , Antibacterianos/uso terapêutico , Fibrose Cística/microbiologia , Fibrose Cística/terapia , Erros de Diagnóstico/prevenção & controle , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/etiologia , Humanos , Ralstonia/efeitos dos fármacos , Ralstonia/isolamento & purificação , Ralstonia/patogenicidadeRESUMO
Non-aerated compost teas (NCTs) are water extracts of composted organic materials and are used to suppress soil borne and foliar disease in many pathosystems. Greenhouse trials were used to test the effectiveness of NCTs to suppress potato bacterial wilt caused by Ralstonia solanacearum on plants grown in soils inoculated with a virulent isolate of the pathogen (biovar II). NCTs prepared from matured compost sources: agricultural waste (AWCT), vermicompost (VCT) and solid municipal waste (SMWCT) were evaluated at three initial application times (7 days before inoculation, at time of inoculation and 7 days after inoculation) prior to weekly applications, in a randomized complete-block design. AWCT applied initially at the time of inoculation resulted in the greatest disease suppression, with the disease severity index 2.5-fold less than the non-treated plants and the "area under the disease progress curve" (AUDPC) 3.2-fold less. VCT and SMWCT were less suppressive than AWCT regardless of initial application time. Next generation sequencing of the v4 region of 16S rRNA gene and the internal transcribed spacer region (ITS1) revealed that diversity and composition of the bacterial and fungal communities across the NCTs varied significantly. Dominant bacterial phyla such as Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, Verrucomicrobia, Chloroflexi, Planctomycetes, Acidobacteria, and a fungal phylum Ascomycota were detected in all NCTs. AWCT had optimum physico-chemical measurements with higher bacterial Shannon diversity indices (H) and fungal richness (S) than the other treatments. We conclude that bacterial wilt of potatoes grown in controlled conditions can be suppressed by a non-aerated compost tea with a high microbial diversity when applied at planting and weekly thereafter.
Assuntos
Antibacterianos/farmacologia , Bactérias/classificação , Bactérias/isolamento & purificação , Solo/química , Solanum tuberosum/microbiologia , Bactérias/efeitos dos fármacos , Bactérias/genética , Resistência à Doença , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Doenças das Plantas , RNA Ribossômico 16S/genética , Ralstonia/efeitos dos fármacos , Ralstonia/isolamento & purificação , Distribuição Aleatória , Microbiologia do Solo , Solanum tuberosum/crescimento & desenvolvimentoRESUMO
OBJECTIVE: To investigate the drug resistance, ß-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding genes of Ralstonia mannitolilytica, and to explore its structure and pathogenic function. METHODS: The strain was isolated by plate streaking method and identified by automatic bacteria detection system and 16S RNA gene PCR. Microdilution method was applied for drug susceptibility test. ß-lactamases, extended spectrum ß-lactamases (ESBL) and carbapenemases were detected using nitrocefin-disk, Kirby-Bauer disk, and Hodge test, respectively. Five ß-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding gene of the isolate were amplified by PCR for sequencing. Bioinformatic softwares were used to analyze the structure and function of the product of protoporphyrin ferrochelatase-encoding gene. RESULTS: A strain belonging to Ralstonia mannitolilytica was isolated. This isolate was sensitive to cefepime, ciprofloxacin, ofloxacin and tigecycline, but resistant to five penicillins, four cephalosporins and two carbapenems antibiotics. The isolate produced ß-lactamases but did not produce ESBL and carbapenemases. The isolate had five distinct ß-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding gene. The product of protoporphyrin ferrochelatase-encoding gene contained two functional domains of protoporphyrin ferrochelatase belonging to type â ¡ chelatase superfamily that presented the most closely genetic relationship with the protoporphyrin ferrochelatase of Neisseria meningidis. CONCLUSIONS: The isolate of Ralstonia mannitolilytica has a higher resistance to ß-lactam antibiotics and its ß-lactamase-encoding genes are different with the common bacterial ß-lactamase-encoding genes. Protoporphyrin ferrochelatase may act as an important virulence factor of Ralstonia mannitolilytica.
Assuntos
Ferroquelatase , Protoporfirinas , Ralstonia , Antibacterianos/farmacologia , Resistência a Medicamentos/genética , Ferroquelatase/química , Ferroquelatase/genética , Ralstonia/efeitos dos fármacos , Ralstonia/enzimologia , Ralstonia/genética , beta-Lactamases/genéticaRESUMO
Because biofilms have resistance to antibiotics, their control using minimum amounts of chemicals and energy becomes a critical issue particularly for resource-constrained long-term space and deep-sea explorations. This preliminary study investigates how ultrasound promoting penetration of antibiotic-loaded liposomes into alginate-based bacterial biofilms, resulting in enhanced bacterial (Ralstonia insidiosa) killing. Nano-sized liposomes are used as a delivery vehicle for the antibiotic gentamicin. Alginate-based synthetic biofilms, which are widely acknowledged as biofilm phantoms, filled with liposome solution are formed at the bottoms of six-well Petri dishes and exposed to ultrasound (frequency = 2.25 MHz, 10% duty cycle, and spatially and temporally averaged intensity ISAPA = 4.4 W/cm(2)). Gentamicin is released from liposomes after they are lysed using detergent solution (0.05% sodium dodecyl sulfate, 1.0% Triton X-100) and incubated for 20 min. The alginate biofilm is dissolved and diluted, counting of colony-forming units shows about 80% of the bacteria are killed. It has also been shown the liposome-capture density by the alginate film increases linearly with the ultrasound intensity up to ISAPA = 6.2 W/cm(2) reaching approximately threefold that without ultrasound. Measurement by using particle-image velocimetry has demonstrated the acoustic streaming with modification by thermal convection controls the enhancement of the liposome capture rate.
Assuntos
Antibacterianos/administração & dosagem , Biofilmes , Desinfecção/métodos , Gentamicinas/administração & dosagem , Lipossomos , Ondas Ultrassônicas , Purificação da Água/métodos , Alginatos , Carga Bacteriana , Portadores de Fármacos , Fluoresceína-5-Isotiocianato/análise , Ácido Glucurônico , Ácidos Hexurônicos , Temperatura Alta , Hidrogéis , Fluxometria por Laser-Doppler , Ralstonia/efeitos dos fármacosRESUMO
A collection of 161 Ralstonia isolates, including 90 isolates from persons with cystic fibrosis, 27 isolates from other human clinical samples, 8 isolates from the hospital environment, 7 isolates from industrial samples, and 19 environmental isolates, was subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) identification and yielded confident species level identification scores for only 62 (39%) of the isolates, including four that proved misidentified subsequently. Whole-genome sequence analysis of 32 representative isolates for which no confident MALDI-TOF MS species level identification was obtained revealed the presence of seven novel Ralstonia species, including three and four that were isolated from cystic fibrosis or other human clinical samples, respectively, and provided the basis for updating an in-house MALDI-TOF MS database. A reanalysis of all mass spectra with the updated MALDI-TOF MS database increased the percentage of isolates with confident species level identification up to 77%. The antimicrobial susceptibility of 30 isolates mainly representing novel human clinical and environmental Ralstonia species was tested toward 17 antimicrobial agents and demonstrated that the novel Ralstonia species were generally multi-resistant, yet susceptible to trimethoprim/sulfamethoxazole, ciprofloxacin, and tigecycline. An analysis of genomic antimicrobial resistance genes in 32 novel and publicly available genome sequences revealed broadly distributed beta-lactam resistance determinants.IMPORTANCEThe present study demonstrated that a commercial matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification database can be tailored to improve the identification of Ralstonia species. It also revealed the presence of seven novel Ralstonia species, including three and four that were isolated from cystic fibrosis or other human clinical samples, respectively. An analysis of minimum inhibitory concentration values demonstrated that the novel Ralstonia species were generally multi-resistant but susceptible to trimethoprim/sulfamethoxazole, ciprofloxacin, and tigecycline.
Assuntos
Antibacterianos , Testes de Sensibilidade Microbiana , Ralstonia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Humanos , Ralstonia/efeitos dos fármacos , Ralstonia/genética , Ralstonia/isolamento & purificação , Ralstonia/classificação , Antibacterianos/farmacologia , Fibrose Cística/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Farmacorresistência Bacteriana Múltipla/genética , Farmacorresistência Bacteriana , Genoma Bacteriano/genética , Sequenciamento Completo do GenomaRESUMO
Ralstonia mannitolilytica constitutes a rare isolate in clinical specimens and to date very few infections with this Gramnegative bacillus have been reported. The first case of peritonitis in a pediatric patient due to R. mannitolilytica in the setting of peritoneal dialysis is described. It is very important to view this organism as a pathogen rather than contaminant when isolated in children with peritonitis.
Assuntos
Diálise Peritoneal/efeitos adversos , Peritonite/microbiologia , Ralstonia/isolamento & purificação , Antibacterianos/farmacologia , Criança , Feminino , Humanos , Peritonite/etiologia , Ralstonia/efeitos dos fármacosRESUMO
Fixed-bed reactors are usually designed for wastewater biotreatments, where the biofilm establishment and maintenance play the most important roles. Biofilm development strictly relies on different types of bacterial motility: swimming, swarming, and chemotaxis, which can be altered by the microenvironment conditions. The aim of this work is to do an integrated study on the effects of Cu(II), Cd(II), Zn(II) and Cr(VI) on swimming, swarming and chemotaxis of Pseudomonas veronii 2E, Delftia acidovorans AR and Ralstonia taiwanensis M2 to improve biofilm development and maintenance for metal loaded wastewater biotreatment in fixed-bed bioreactors. Swimming, swarming and chemotactic response evaluation experiments were carried out at different metal concentrations. P. veronii 2E motility was not affected by metal presence, being this strain optimal for fixed-bed reactors. D. acidovorans AR swarming was inhibited by Cd and Zn. Although R. taiwanensis M2 showed high resistance to Cu, Cd, Cr and Zn, motility was definitively altered, so further studies on R. taiwanensis M2 resistance mechanisms would be particularly interesting.
Assuntos
Quimiotaxia/efeitos dos fármacos , Delftia/efeitos dos fármacos , Metais Pesados/metabolismo , Pseudomonas/efeitos dos fármacos , Ralstonia/efeitos dos fármacos , Águas Residuárias/microbiologia , Biodegradação Ambiental , Biofilmes/efeitos dos fármacos , Reatores Biológicos , Delftia/fisiologia , Pseudomonas/fisiologia , Ralstonia/fisiologiaRESUMO
Differentiation of the growing nosocomial infectious threats, Ralstonia pickettii and Ralstonia insidiosa, based on nitrate reduction, desferrioxamine susceptibility, arabinose, N-acetyl-glucosamine and phenylacetate assimilation is described. These tests can be used for preliminary identification of Ralstonia pickettii and Ralstonia insidiosa resulting in more accurate identification of these species.
Assuntos
Infecção Hospitalar/diagnóstico , Infecção Hospitalar/microbiologia , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/microbiologia , Ralstonia/classificação , Ralstonia/metabolismo , Acetilglucosamina/metabolismo , Antibacterianos/toxicidade , Arabinose/metabolismo , Técnicas de Tipagem Bacteriana , Desferroxamina/toxicidade , Humanos , Nitratos/metabolismo , Oxirredução , Fenilacetatos/metabolismo , Ralstonia/efeitos dos fármacos , Ralstonia/isolamento & purificaçãoRESUMO
The effect of nickel on the microbial community in the activated sludge of a sequencing batch reactor (SBR) reactor was investigated by continuously dosing nickel from 60 to 240 mg Ni(II) L(-1). The diversity of the microbial community was investigated by polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis of the variable V3 region of the bacterial 16S rDNA. The experimental results showed that the community structure changed significantly after dosing with nickel with a shift in dominant species, the disappearance of some original species and the emergence of some new species. The existence of a nickel resistant gene was also investigated using PCR. The obtained nickel resistance gene had a maximum homology with the plasmid pMOL30 of Ralstonia metallidurans CH34. The quantitative real-time PCR results indicated that the quantity of the nickel resistance gene was related to the nickel concentration loaded to the reactor.
Assuntos
DNA Bacteriano/genética , DNA Ribossômico/genética , Níquel/farmacologia , Ralstonia/efeitos dos fármacos , Ralstonia/genética , Esgotos/microbiologia , Análise por Conglomerados , DNA Bacteriano/química , DNA Ribossômico/química , Eletroforese em Gel de Gradiente Desnaturante , Ralstonia/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Poluentes Químicos da Água/farmacologiaRESUMO
The present study aimed to investigate the potential of simple sugars for use as protection agents in the control of tomato bacterial wilt caused by Ralstonia pseudosolanacearum. Based on the sugar assimilation patterns of the pathogen, four unassimilable sugars (L-arabinose, maltose, D-raffinose, and D-ribose) were selected from 10 representative sugars present in tomato root exudates. These sugars were evaluated for their effects on bacterial wilt using a tomato seedling bioassay. The application of 0.25% L-arabinose significantly reduced disease severity and was, thus, selected as a candidate for further evaluations in a pot experiment under glasshouse conditions. The results obtained showed that the disease suppressive effects of L-arabinose slightly increased at higher concentrations; drench treatments at 0.1, 0.25, and 0.5% reduced disease severity by ca. 48, 70, and 87%, respectively. The drench treatment with 0.5% L-arabinose significantly reduced the pathogen population in the rhizosphere and stem tissues of tomato plants without any antibacterial activity. Real-time reverse-transcription PCR revealed that the expression of salicylic acid-dependent and ethylene-dependent defense genes was significantly enhanced in the stem tissues of L-arabinose-treated tomato plants following the pathogen inoculation. These results suggest that soil drenching with L-arabinose effectively suppresses tomato bacterial wilt by preventing pathogen proliferation in the rhizosphere and stem tissues of tomato plants. This is the first study to report the potential of L-arabinose as a safe, eco-friendly, and cost-effective plant protection agent for the control of tomato bacterial wilt.
Assuntos
Arabinose/farmacologia , Doenças das Plantas/prevenção & controle , Ralstonia/efeitos dos fármacos , Solanum lycopersicum/microbiologia , Doenças das Plantas/microbiologia , Raízes de Plantas/microbiologia , Ralstonia/crescimento & desenvolvimento , Microbiologia do SoloRESUMO
The Lascaux Cave contains a remarkable set of paintings from the Upper Palaeolithic. Shortly after discovery in 1940, the cave was modified for public viewing and, in 2001, was invaded by a Fusarium solani species complex. Benzalkonium chloride was used from 2001 to 2004 to eliminate the fungal outbreak. In this study, we carried out a sampling in most of the cave halls and galleries. Sequence analysis and isolation methods detected that the most abundant genera of bacteria were Ralstonia and Pseudomonas. We suggest that, as a result of years of benzalkonium chloride treatments, the indigenous microbial community has been replaced by microbial populations selected by biocide application.
Assuntos
Compostos de Benzalcônio/farmacologia , Desinfetantes/farmacologia , Fungos/efeitos dos fármacos , Pseudomonas/efeitos dos fármacos , Ralstonia/efeitos dos fármacos , Alcaligenes/efeitos dos fármacos , DNA Bacteriano/efeitos dos fármacos , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , DNA Fúngico/efeitos dos fármacos , DNA Fúngico/genética , DNA Fúngico/isolamento & purificação , Escherichia coli/efeitos dos fármacos , França , Fungos/crescimento & desenvolvimento , Sedimentos Geológicos/microbiologia , Legionella/efeitos dos fármacos , Pinturas , Pseudomonas/crescimento & desenvolvimento , Ralstonia/crescimento & desenvolvimentoRESUMO
In this study, a type of thiazolium-labeled 1,3,4-oxadiazole thioether bridged by diverse alkyl chain lengths was constructed. The antimicrobial activity of the fabricated thioether toward plant pathogenic bacteria and fungi was then screened. Antibacterial evaluation indicated that title compounds possess specific characteristics that enable them to severely attack three phytopathogens, namely, Xanthomonas oryzae pv. oryzae, Ralstonia solanacearum, and Xanthomonas axonopodis pv. citri with minimal EC50 values of 0.10, 3.27, and 3.50 µg/mL, respectively. Three-dimensional quantitative structure-activity relationship models were established to direct the following excogitation for exploring higher active drugs. The in vivo study against plant bacterial diseases further identified the prospective application of title compounds as alternative antibacterial agents. The proteomic technique, scanning electron microscopy patterns, and fluorescence spectrometry were exploited to investigate the antibacterial mechanism. Additionally, some target compounds performed superior inhibitory actions against three tested fungal strains. In view of their simple molecular architecture and highly efficient bioactivity, these substrates could be further explored as promising surrogates for fighting against plant microbial infections.
Assuntos
Antibacterianos/farmacologia , Oxidiazóis/farmacologia , Sulfetos/farmacologia , Antibacterianos/síntese química , Antibacterianos/química , Testes de Sensibilidade Microbiana , Oxidiazóis/síntese química , Oxidiazóis/química , Doenças das Plantas/microbiologia , Ralstonia/efeitos dos fármacos , Sulfetos/síntese química , Sulfetos/química , Xanthomonas/efeitos dos fármacosRESUMO
A new catecholate-containing siderophore, labrenzbactin (1), was isolated from the fermentation broth of a coral-associated bacterium Labrenzia sp. The structure and absolute configuration of 1 was determined by spectroscopic methods and Marfey's analysis. Overall, 1 showed antimicrobial activity against Ralstonia solanacearum SUPP1541 and Micrococcus luteus ATCC9341 with MIC values of 25 and 50 µg ml-1, respectively, and cytotoxicity against P388 murine leukemia cells with an IC50 of 13 µM.
Assuntos
Alphaproteobacteria/química , Antozoários/microbiologia , Antibacterianos/isolamento & purificação , Catecóis/isolamento & purificação , Oxazóis/isolamento & purificação , Sideróforos/isolamento & purificação , Alphaproteobacteria/isolamento & purificação , Animais , Antibacterianos/farmacologia , Antibióticos Antineoplásicos/isolamento & purificação , Antibióticos Antineoplásicos/farmacologia , Catecóis/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Fermentação , Leucemia P388/tratamento farmacológico , Camundongos , Testes de Sensibilidade Microbiana , Micrococcus luteus/efeitos dos fármacos , Estrutura Molecular , Oxazóis/farmacologia , Ralstonia/efeitos dos fármacos , Sideróforos/farmacologiaRESUMO
Members of the species Ralstonia pickettii and R. mannitolilytica, although ubiquitous and lacking major virulence factors, have been associated with nosocomial outbreaks. Tolerance to metals, antibiotics, and disinfectants may represent an advantage for their ubiquity and opportunistic pathogenic potential. In this study, we compared five strains that differed on the origin (hospital effluent, tap water, mineral water) and in the susceptibility to aminoglycosides, regarding their tolerance to metals and disinfection. The growth kinetics and biofilm formation capacity were tested in four R. pickettii strains and one R. mannitolilytica at sub-inhibitory concentrations of aminoglycosides or arsenite. The survival to UV radiation, chlorine, or hydrogen peroxide was also compared in aminoglycoside resistant and susceptible strains. Aminoglycoside-resistant strains presented a higher tolerance to arsenite than the susceptible ones and either aminoglycosides or arsenite was observed to stimulate the biofilm formation. Sub-inhibitory concentrations of the aminoglycoside gentamicin or arsenite significantly decreased the growth rate and yield, but only arsenite caused a significant increase of the lag phase. Hydrogen peroxide presented higher disinfection effectiveness against aminoglycoside susceptible than against resistant strains, an effect that was not observed for UV or chlorine. Although this conclusion needs validation based on a larger number of isolates, including clinical, the results suggest that aminoglycoside resistance may be associated with traits that influence Ralstonia spp. fitness in the environment.